Attorney Docket No.0113409-005WO0 COFFEE AND BEAN-LESS COFFEE COMPOSITIONS HAVING CHLOROGENIC ACIDS AND/OR RHAMNOGALACTURONANS FIELD OF THE INVENTION Aspects of the invention relate generally to compositions and methods for improving organoleptic properties of food and beverages (e.g., coffee and coffee- substitute compositions), and more particularly to methods for the isolation and deliberate processing of plant-derived CGA extracts having utility to improve organoleptic properties of coffee and coffee substitute products. Additional particular aspects relate to polysaccharide-based compositions comprising non-starch polysaccharides (NSPs) and methods using same to improve organoleptic properties of food and beverages. Even more particular aspects relate to polysaccharide-based compositions comprising rhamnogalacturonans (RGs) (e.g., galacturonic acid/rhamnose copolymers with arabinose and xylose side chains, optionally bound to protein) and methods using same to improve organoleptic properties (e.g., roastiness, body, pungency, acidity, sweetness, etc.) of food and beverages (e.g., such as coffee and/or coffee-substitute compositions and beverages, cross-Maillardized coffee and coffee-substitute compositions and beverages including but not limited to extractable coffee and coffee-substitutes and extracts thereof, and including kernels, grounds, beverages, concentrates, flavorings, etc., based thereon, all which are preferably made without coffee beans). BACKGROUND Chlorogenic acids. Nearly all properties of coffee beverages depend upon the constituents of the raw (green) coffee seeds (or beans). The compositions of coffee seeds will vary significantly with growing conditions, soil, altitude and myriad other factors, but even within these variations there are important generalities. One of the most crucial for the unique qualities of coffee beverages is the high levels of chlorogenic acids (CGAs). Chlorogenic acids are a family of ester-linked caffeoylquinic acids and di-caffeoylquinic acids. In coffee, the most prevalent CGAs are 3-, 4- and 5-O- caffeoylquinic acids. In the aggregate, total levels of CGA generally range from 4-7% of the total mass of any given coffee seed. While CGAs are present in the tissues of a wide variety of plant species, very few contain such high levels. 1 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 While the native forms of these CGAs have been linked to the flavor and texture of coffee beverages, it is the mixture of CGAs and their thermal reaction products that are essential to create—or recreate—the distinctive flavor and texture of roasted coffee. Two significant such reaction products are the chlorogenic lactones (CGLs), produced by dehydration of the CGAs, and phenylindanes, the thermal breakdown products of CGLs. Notably, CGLs produce a unique and (to many) pleasant bitter sensation, a unique “coffee-like” bitterness. CGLs are present in varying levels in coffees based on their degree of roast. Phenylindanes, by contrast, produce a harsher bitter taste than the CGLs from which they are derived. Phenylindanes are more prevalent in darker roasted coffee. While this description may appear to suggest phenylindanes should be avoided in all coffee, they contribute to the overall experience of drinking darker roasted coffee—for example, a classic Italian espresso—and are essentially to creating coffee, regardless of the raw materials used to create it. Polysaccharides. When discussing the creation of flavor in roasted coffee, the basic treatises focus on Maillard Reactions. Namely, the complexation of a typically isolated reducing sugar (e.g., fructose) with an isolated amino acid (e.g., leucine). While such discussions and approaches are not incorrect, they are certainly incomplete. Coffee seeds, like most natural products, contain significant fractions of both their [reducing] sugars (e.g., reducing sugars) and amino acids in the forms of polymers, namely polysaccharides and proteins (or polyamides). At first blush, these reagents existing in coffee seeds in polymeric forms may seem a kinetic hindrance. After all, they are generally less mobile and less reactive when attached to tens, hundreds or thousands of similar building blocks. Many such polymers degrade under conditions of coffee roasting, however, liberating mono- and oligosaccharides for flavor creation, though only after sufficient heating. Thus the polysaccharide fraction might be expected to contribute insignificantly to the finished coffee product. The kinetics of these thermal reactions should be considered in the context of the coffee roasting process. The overall roasting process for coffee generally takes 20 minutes or less, depending on the particulars of the bean, the target result, the roaster equipment and other factors. These roast times are not only dictated by Maillard reactions creating volatile flavor compounds, however. The mono and disaccharides present in raw coffee seeds react quite quickly with amino acids to produce myriad volatile compounds. 2 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Yet other precursors, such as chlorogenic acids (CGAs) and trigonelline, require more time for the desired level of conversion to make the best coffee. This presents a potential problem. These essential Maillard-derived compounds (comprising such reactive species as aldehydes, ketones and thiols) may themselves be further reacted, degraded or driven off as the roast continues. However, meaningful quantities of these volatiles remain in coffee roasted for significantly longer times than required to produce these compounds from their the fast-reacting monosaccharide and amino acid reagents. Partway through the roasting process, when much of the initially occurring monomeric species have been consumed, the previously hindered saccharides (e.g., present in polymeric forms, such as those present in polysaccharides) begin to contribute monomeric and/or oligomeric species. By requiring the prior heating steps to make these sugars available for reactions, the inclusion of polysaccharides can be viewed alternatively as incorporating a time release mechanism into key reagents in the creation of essential coffee qualities, such as complex flavor and unique texture. Rather than relying solely on the fast acting reducing sugars initially present in the bean, the presence of these polysaccharides allows the continuation of roast development, via Maillard and caramelization reactions, through longer roasting processes. This time release contribution is not limited to polysaccharides. In an analogous fashion, amino acids or short-chain fractions of previously complete proteins are liberated during the roasting process as the bean structure changes and breaks down. With much of the original monosaccharide content already consumed, these newly reactive amine species require an infusion of new sugar to continue Maillard reactions. The time released polysaccharides, breaking down coincidentally with the polyamides, provide such an infusion of fresh sugars at later stages of coffee roasting. This is clearly illustrated when considering the case of an especially important odorant for roasted coffee, 2-furfuryl thiol (2-FFT). This compound is both volatile and highly reactive with its pendant thiol. It can be inferred that—through this time release mechanism—such a delicate compound can still be present in roasted coffee even after 10+ minutes of roasting, as it originates from arabinogalactans (a type of polysaccharide) rather than monosaccharides such as fructose (Grosch, W., 1999. Key odorants of roasted coffee: Evaluation, release, formation. In: Proceedings of the 19th International Scientific Colloquium on Coffee, Helsinki, Finland, ASIC, Paris, France, pp.17-26). In spite of its reactivity, 2-FFT is generally observed to increase continuously with greater 3 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 roast degree, unlike many odorants that peak then decay with continued heating (i.e., darker roast degrees) past what the coffee industry typically terms a “medium” roast. In addition, these polysaccharides benefit the overall texture of resulting coffee beverages. Intact polysaccharides (e.g., those undergoing essentially no thermal breakdown) are found in finished coffee extractions (e.g., beverages). These are generally high molecular weight, and bear resemblance to food additives like guar gum. However, the texture is also positively impacted thanks to the presence of the thermal breakdown products of certain key polysaccharides, as well as the products of reactions between these breakdown products and other constituents of coffee seeds. Partially degraded polysaccharides are key reagents in the formation of melanoidins. These complex molecules, with somewhat stochastic chemical and physical structures, are essential for much of the color and texture of the resulting coffee and have been implicated in several of the health benefits attributed to coffee consumption. Without the larger, antecedent polysaccharides present in the raw coffee seed, the levels of these crucial constituents would be negatively impacted. In coffee, the polysaccharide content (nearly 50% of dry matter) is largely comprised of 3 main families: cellulose (ca. 3%), galactomannans (≥19%) and arabinogalactans (14-17%). Amongst these, cellulose is perhaps the least involved in the above described reactions. Cellulose, being essentially an insoluble, structural polysaccharide, is involved mostly indirectly. Cellulose reinforces cell walls, which in turn support high pressures inside coffee cells as internal temperatures rise during roasting. These higher pressures in turn influence the reactions that occur. Galactomannans, while generally soluble, are not especially active in these reactions, either. Analyses of roasted coffee beans show that these large polysaccharides remain largely intact through the roasting process (Bradbury, A.G.W., 2001. Chemistry I: non-volatile compounds in: Coffee. Blackwell Science Ltd., pp 1-17). Due to their solubility, however, they can contribute to coffee texture in the finished beverage in a limited fashion (Arya, M., & Rao, L. J. M. (2007). An Impression of Coffee Carbohydrates. Critical Reviews in Food Science and Nutrition, 47(1), 51–67. https://doi.org/10.1080/10408390600550315). The arabinogalactans, in contrast to the prior two species of polysaccharide, are highly active in the roasting process and contribute significantly to the finished product. These polysaccharides comprise a galactan backbone with branches (i.e., side chains) of arabinose and arabinans. During roasting, these arabinose-based branches detach 4 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 from the main chain, and these liberated arabinans are what primarily participate in the reactions described prior. The arabinogalactan in coffee is further differentiated by its native structure in coffee. Rather than existing purely as a complex polysaccharide, it is instead largely present in arabinogalactan proteins (AGPs), even more complex structures wherein arabinogalactans are covalently bound to proteins. It would stand to reason that the specific qualities of roasted coffee would have its origin in the particulars of its most active constituents. As disclosed herein below, it has been surprisingly found, however, in the context of Applicant’s work creating coffee products that contain no products from the seeds of the Coffea genus, that the presence of AGPs is not necessary for the creation of these kinds of flavors and textures. Furthermore, it has been surprisingly found that arabinogalactans are not the only polysaccharides that can be utilized to create these coffee qualities. According to particular aspects, in the place of arabinogalactans, other polysaccharides having unique chemical and physical structures can provide these essential qualities, and have been overlooked in overly simplistic prior art understanding of the underlying mechanisms for the creation of coffee’s unique qualities, and because coffee largely does not contain them. Alleged coffee substitutes, derived from raw materials other than coffee beans/cherries, have historically been pursued for numerous reasons including, for example, coffee bean shortages or limited availability, excessive cost, and caffeine avoidance. Exemplary substitute ingredients include chicory (e.g., in Europe), acorns (e.g., North America), yerba mate (e.g., South America), date seeds (e.g., Middle East), etc. A given substitute will typically have at least some structural and/or compositional similarities to coffee beans, and thus will frequently be treated and processed as if it was coffee bean material in an attempt to produce a coffee-like beverage from it. For example, a coffee substitute raw material may be harvested, cleaned, roasted, ground and extracted as if it were coffee beans, but since none of these ingredients has the same structure and/or composition as green coffee beans, they do not produce, upon such processing, beverages that accurately replicate the organoleptic properties of coffee; that is, traditional coffee substitutes, despite being subjected to traditional coffee bean processing steps and conditions, do not recapitulate or sufficiently approach the coffee 5 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 experience, and the results are at best an alternative, not an organoleptic substitute to the familiar coffee experience. While research in recent years has been directed at identifying key components of green and/or roasted coffee that contribute to its distinctive aroma, taste, texture and color, alternative raw materials may (and typically do) either lack particular key coffee components, contain excessive amounts of particular coffee components, and/or contain different components that may generate undesirable properties upon application of traditional coffee processing steps. For the same reasons, traditional flavor ingredients, alone or in combination with such alternative raw materials (e.g., augmented raw materials) do not sufficiently recapitulate or approach the coffee experience. Additionally, certain compounds found in coffee seeds and coffee beverages may be problematic for organoleptic qualities or for human health. For example, the amino acid asparagine is known to produce the undesirable toxin acrylamide during the coffee roasting process. There is, therefore, a pronounced need for methods that functionally (e.g., chemically and organoleptically) integrate exogenous ingredients/reactants with endogenous reactive components of traditional coffee, or of alternative non-coffee raw materials to provide improved coffee and more organoleptically accurate coffee-substitutes, and which also allow for coffee and coffee-substitute formulations in which desired or undesired compounds may be omitted, removed, degraded, diminished, altered, modulated or increased prior to or during processing. SUMMARY OF EXEMPLARY EMBODIMENTS OF THE INVENTION Particular CGA/RG related embodiments of the disclosure can be described in view of the following clauses: 1. A coffee or bean-less coffee composition, comprising: a substrate carrier material or cross-Maillardized (xMR) substrate carrier material, and/or a traditional coffee ingredient; a chlorogenic acid (CGA) ingredient comprising a coffee or non-coffee CGA- containing plant extract or its thermal reaction product(s); and a rhamnogalacturonan (RG) polysaccharide ingredient, having residues of galacturonic acid, galactose, xylose, arabinose, and rhamnose, derived from a non- coffee seed substrate and containing RG or portions thereof. 6 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 2. The composition of clause 1, wherein the RG is present in the composition in a range of 0.5 wt.% to 60 wt.%, or any subrange thereof, preferably in a range of 0.5 wt.% to 50 wt.%, 0.5 wt.% to 40 wt.%, 0.5 wt.% to 30 wt.%, 0.5 to 20 wt.%, 0.1 wt.% to 10 wt.%, 0.1 wt.% to 5 wt.%, 20 wt.% to 40 wt.%, or 5 wt.% to 20 wt.%, wherein wt.% is in terms of dry mass, and/or wherein the CGA is present at an amount in a range selected from 0.1-40 wt.% or any subrange thereof, preferably in a range of 0.2-8 wt.%, 20-40 wt.%, 4-15 wt.%, 6-20 wt.%, 0.3-6 wt.%, or 0.2-3 wt.%, wherein wt.% is in terms of dry mass. 3. The composition of clause 1, wherein the non-coffee CGA-containing plant is at least one selected from the group consisting of sunflower (e.g., seeds, leaves, stems), artichoke, yerba mate, nightshades (e.g., tomatoes, potatoes, eggplants, tobacco), chicory, prune, Eucommia ulmoides, honeysuckle and combinations thereof, and where the coffee CGA is that of a coffee plant material, and/or wherein the RG comprises an RG from at least one source selected from the group consisting of beet, carrot, potato, strawberry, raspberry, blueberry, blackberry, carob, jackfruit, bell pepper, tomato, pumpkin, ginseng, okra, grapefruit, aronia, acerola cherry, fenugreek seeds, flax seeds, coffee fruit (not seed), and combinations thereof. 4. The composition of clause 1, wherein the substrate carrier material or the cross-Maillardized (xMR) substrate carrier material (e.g., having LWACMP and/or HWACMP cross-Maillard reaction products as defined herein) comprises one or more natural and/or a processed or restructured plant materials selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the mustard family (Brassicaceae), watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, apricot, jackfruit (Artocarpus heterophyllus), cereal and non- cereal grains, and/or coffee. 5. The composition of clause 4, wherein the cross-Maillardized substrate carrier material has been conditioned and/or heated (e.g., roasted) in the presence of fenugreek (e.g., defatted fenugreek seeds). 6. The composition of clause 1, wherein the CGA-containing plant extract comprises at least one extract selected from the group consisting of water and/or alcohol extract (e.g., ethanol, methanol, benzyl alcohol, and combinations thereof), carbon dioxide extract, glycol extract, acetone extract, fat extract, oil (e.g., mono/di/triglycerides) extract, alkane (e.g., hexane, etc.) extract, ethyl acetate extract, methyl ethyl ketone extract, dichloromethane extract, chloroform extract, and combinations thereof. 7 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 7. The composition of clause 3, wherein a plurality of 2-17 (e.g., 2-8) of the RG sources are processed and/or assembled together with at least one other ingredient, wherein if processed the RG source has been processed to at least one form selected from the group consisting of dried, juiced, filtered, sieved, milled, washed, extracted, sorted, hydrolyzed, fermented, roasted, blanched, steamed or otherwise wet heated, chilled, frozen, concentrated, or combinations thereof. 8. The composition of clause 1, wherein the CGA-containing plant extract is raw, having not been heated above a temperature selected from the group consisting of 120°C, 105°C, 100°C, 85°C, 65°C, 40°C, and 20°C, or wherein the CGA-containing plant extract has been heated or roasted to a temperature in a temperature range selected from 230-250°C, 195-250°C, 200-230°C, 180-200°C, 160-200°C, 120-200°C, and 120-160°C. 9. The composition of any one of clauses 1-8, wherein the combination further comprises a starch ingredient, preferably wherein the total amount of starch is present at an amount in a range selected from the range group consisting of 60-90 wt.%, 40-80 wt.%, 35-50 wt.%, 20-45 wt.%, 5-20 wt.%, and 0.1-10 wt.%, wherein wt.% is in terms of dry mass. 10. The composition of clause 9, wherein the starch ingredient is selected from the group consisting of ramon seeds/maya nuts (Brosimum alicastrum), cereal and non- cereal grain, seed, tuber (e.g., potato, arrow root, etc.), pulse (e.g., lentils, peas, beans), kulthi daal (Macrotyloma uniflorum), fruits (ex: plantains), extracts of the above, and combinations thereof. 11. The composition of any one of clauses 1-10, wherein the composition comprises an assemblage (e.g., combination, compounded, extruded, pelletized, etc.) that takes a form selected from dried, roasted, ground, extracted, concentrated, pasteurized, sterilized, or combinations thereof, to provide a finished coffee or bean-less coffee. composition (e.g., green beans, roasted beans, grounds, ready to drink form (RTD), or extract). 12. The composition of clause 11, further comprising one or more additional components selected from the group consisting of caffeine, bioactive agents, flavors, colors, gums, texture modifiers, pH adjusters/regulators, fruit or vegetable powders (non- compounded/pelletized) or their extracts, and combinations thereof. 8 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 13. The composition of clause 11 or 12, in the form of intact pieces of comparable size to coffee beans, ground materials with particle sizes generally < 2 mm or extracts made from these mixtures. 14. The bean-less coffee product of clause 13, wherein the intact pieces are raw (subjected to temperatures below 120ºC) or roasted (subjected to temperatures ≥ 120ºC). 15. The composition of any one of clauses 11-13, wherein the assembled coffee composition comprises Ramon seed, green banana, sunflower seed extract, strawberry fiber, potato fiber, carrot, black aronia berry, carob, and optionally caffeine. 16. A method for creating a coffee or bean-less coffee composition, comprising assembling (e.g., combining compounding, extruding, pelletizing, etc.) a composition according to any one of clauses 1-15. 17. A coffee or beanless coffee composition, comprising a chlorogenic acid (CGA)-containing plant extract or its thermal reaction product(s) assembled (e.g., combined, compounded, extruded, pelletized, etc.) with at least one other ingredient to provide an assembled coffee or bean-less coffee composition. 18. The composition of clause 17, wherein the CGA-containing plant extract or its thermal reaction product(s) is that of a non-coffee plant material. 19. The composition of clause 17, wherein the CGA-containing plant extract or its thermal reaction product(s) is that of a coffee plant material. 20. The composition of clause 18, wherein the non-coffee plant is one or more selected from the group consisting of sunflower (e.g., seeds, leaves, stems), artichoke, yerba mate, nightshades (e.g., tomatoes, potatoes, eggplants, tobacco), chicory, prune, Eucommia ulmoides, honeysuckle. 21. The composition of any one of clauses 18-20, wherein the CGA-containing plant extract comprises a water and/or alcohol extract of the CGA-containing plant material. 22. The composition of any one of clauses 18-20, wherein the s CGA- containing plant extract comprises a carbon dioxide extract, glycol extract, acetone extract, fat extract, oil (e.g., mono/di/triglycerides) extract, alkane (e.g., hexane, etc.) extract, ethyl acetate extract, methyl ethyl ketone extract, dichloromethane extract, or chloroform extract of the CGA-containing plant material. 23. The composition of clause 21, wherein the alcohol is one or more selected from the group consisting of ethanol, methanol, benzyl alcohol, and combinations thereof. 9 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 24. The composition of any one of clauses 17-23, wherein the CGA-containing plant extract is compounded with the at least one other ingredient, and optionally formed and/or optionally ground. 25. The composition of clause 24, wherein the CGA-containing plant extract is compounded with a starch comprising ingredient, preferably wherein the total amount of starch is present at an amount in a range selected from the range group consisting of 60- 90 wt.%, 40-80 wt.%, 35-50 wt.%, 20-45 wt.%, 5-20 wt.%, and 0.1-10 wt.%, wherein wt.% is in terms of dry mass. 26. The composition of clause 25, wherein the starch comprising ingredient is selected from the group consisting of ramon seeds/maya nuts (Brosimum alicastrum), cereal and non-cereal grain, seed, tuber (e.g., potato, arrow root, etc.) , pulse (e.g., lentils, peas, beans), kulthi daal (Macrotyloma uniflorum), fruits (ex: plantains), extracts of the above, and combinations thereof. 27. The composition of clause 24, wherein the CGA-containing plant extract is compounded with fenugreek (e.g., defatted fenugreek seeds). 28. The composition of any one of clauses 17-27, wherein the exogenous CGA-containing plant extract is raw, having not been heated above a temperature selected from the group consisting of 120°C, 105°C, 100°C, 85°C, 65°C, 40°C, and 20°C. 29. The composition of any one of clauses 17-27, wherein the CGA-containing plant extract has been heated or roasted to a temperature in a temperature range selected from 230-250°C, 195-250°C, 200-230°C, 180-200°C, 160-200°C, 120-200°C, and 120-160°C. 30. The composition of any one of clauses 17-29, wherein the composition is blended or otherwise combined with a substrate carrier material or a cross-Maillardized substrate carrier material having cross-Maillard reaction products (e.g., LWACMP and/or HWACMP as defined herein). 31. The composition of clause 30, wherein the substrate carrier material comprises one or more selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the mustard family (Brassicaceae), watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, apricot, jackfruit (Artocarpus heterophyllus), cereal and non-cereal grains, and/or coffee. 10 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 32. The composition of clause 30 or 31, wherein the cross-Maillardized substrate carrier material has been conditioned and/or heated (e.g., roasted) in the presence of fenugreek (e.g., defatted fenugreek seeds). 33. The composition of any one of clauses 30-32, wherein the composition is additionally blended or otherwise combined with fruit powder, vegetable powder, gums, caffeine, colors, flavors, or combinations thereof, to provide a finished ground(s) suitable to produce a beanless coffee or coffee beverage (e.g., at home, in a café, in an industrial production setting, etc.). 34. A method of preparing a beanless coffee or coffee composition, comprising: preparing a chlorogenic acid (CGA) extract by extracting a CGA-containing plant material; and combining the CGA extract with a a substrate carrier material or a cross- Maillardized substrate carrier material having cross-Maillard reaction products (e.g., LWACMP and/or HWACMP as defined herein). 35. The method of clause 34, further comprising compounding the CGA extract with another ingredient, and optionally forming and/or optionally grinding. 36. The method of clause 35, wherein the exogenous CGA-containing plant extract is compounded with a starch comprising ingredient. 37. The method of clause 36, wherein the starch comprising ingredient is selected from the group consisting of ramon seeds/maya nuts (Brosimum alicastrum), cereal and non-cereal grain, seed, tuber (e.g., potato, arrow root, etc.), pulse (e.g., lentils, peas, beans), kulthi daal (Macrotyloma uniflorum), fruits (ex: plantains), extracts of the above, and combinations thereof. 38. The method of any one of clauses 34-37, wherein the substrate carrier material comprises one or more selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the mustard family (Brassicaceae), watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal, apricot, jackfruit, (Artocarpus heterophyllus) and non-cereal grains, and/or coffee. 39. The method of any one of clauses 34-38, further comprising blending or otherwise combining fruit powder, vegetable powder, gums, caffeine, colors, flavors, or combinations thereof, to provide a finished ground(s) suitable to produce beanless coffee or coffee beverages (e.g., at home, in a café, in an industrial production setting, etc.). 11 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 40. The method of any one of clauses 34-39, wherein the cross-Maillardized substrate carrier material has been conditioned and/or heated (e.g., roasted) in the presence of fenugreek (e.g., defatted fenugreek seeds). 41. A coffee or bean-less coffee composition, comprising at least one rhamnogalacturonan (RG) polysaccharide ingredient(s), each derived from non-coffee seed substrates and containing RG or portions thereof, assembled (e.g., combined, compounded, extruded, pelletized, etc.) with at least one other ingredient to provide an assembled coffee or bean-less coffee composition. 42. The composition of clause 41, wherein the RG or portions thereof comprise residues of galacturonic acid, galactose, xylose, arabinose, and rhamnose. 43. The composition of clause 41 or 42, wherein the RGs are present in the assemblage composition in a range of 0.5 wt.% to 60 wt.%, or any subrange thereof, preferably in a range of 0.5 wt.% to 50 wt.%, 0.5 wt.% to 40 wt.%, 0.5 wt.% to 30 wt.%, 0.5 to 20 wt.%, 0.1 wt.% to 10 wt.%, 0.1 wt.% to 5 wt.%, 20 wt.% to 40 wt.%, or 5 wt.% to 20 wt.%, wherein wt.% is in terms of dry mass. 44. The composition of any one of clauses 41-43, wherein the RG comprises an RG from at least one source selected from the group consisting of beet, carrot, potato, strawberry, raspberry, blueberry, blackberry, carob, jackfruit, bell pepper, tomato, pumpkin, ginseng, okra, grapefruit, aronia, acerola cherry, fenugreek seeds, flax seeds, coffee fruit (not seed), and combinations thereof. 45. The composition of clause 44, wherein a plurality of 2-17 of the sources are assembled together with at least one other ingredient, preferably wherein 2-8 of the sources are assembled together with at least one other ingredient. 46. The composition of clause 44 or 45, wherein the at least one RG source is processed, wherein the RG source has been processed to at least one form selected from the group consisting of dried, juiced, filtered, sieved, milled, washed, extracted, sorted, hydrolyzed, fermented, roasted, blanched, steamed or otherwise wet heated, chilled, frozen, concentrated, or combinations thereof. 47. The composition of any one of clauses 41-46, wherein the at least one other ingredient comprises at least one ingredient selected from the group consisting of starch, one or more chlorogenic acid (CGA) species, an RG, a traditional coffee ingredient, and combinations thereof. 48. The composition of clause 47, wherein the starch comprises starch from at least one source selected from the group consisting of cereal or non-cereal grains, 12 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 pulses, tubers, Ramon seeds, products of the Musa genus, and combinations thereof; and/or wherein the CGA comprises a CGA extract from a least one source selected from the group consisting of sunflower, artichoke, yerba mate, nightshades (e.g., tomatoes, potatoes, eggplants, tobacco), chicory, prune, Eucommia ulmoides, honeysuckle, and combinations thereof. 49. The composition of clause 47 or 48, wherein the total amount of starch is present at an amount in a range selected from the range group consisting of 60-90 wt.%, 40-80 wt.%, 35-50 wt.%, 20-45 wt.%, 5-20 wt.%, and 0.1-10 wt.%, wherein wt.% is in terms of dry mass, and wherein the total of all CGAs is present at an amount in a range selected from 0.1-40 wt.% or any subrange thereof, preferably in a range of 0.2-8 wt.%, 20-40 wt.%, 4-15 wt.%, 6-20 wt.%, 0.3-6 wt.%, or 0.2-3 wt.%, wherein wt.% is in terms of dry mass. 50. The composition of any one of clauses 41-49, wherein the assembled composition takes a form selected from dried, roasted, ground, extracted, concentrated, pasteurized, sterilized, or combinations thereof, to provide a finished composition (e.g., green beans, roasted beans, grounds, ready to drink form (RTD), or extract). 51. The composition of clause 50, wherein the finished composition is further combined with a substrate carrier material or cross-Maillardized substrate carrier material having cross-Maillard reaction products (e.g., LWACMP and/or HWACMP as defined herein). 52. The composition of clause 51, wherein the substrate carrier material comprises one or more selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the mustard family (Brassicaceae), watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal, apricot, jackfruit, (Artocarpus heterophyllus) and non-cereal grains, and/or coffee. 53. The composition of clause 50, wherein the finished composition is further combined with one or more additional components selected from the group consisting of a different finished bean-less coffee composition, date seeds, cross-Maillardized (xMR) date seeds, traditional coffee seeds, caffeine, bioactive agents, flavors, colors, gums, texture modifiers, pH adjusters/regulators, fruit or vegetable powders (non- compounded/pelletized) or their extracts, and combinations thereof, to provide an assembled product. 13 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 54. The bean-less coffee product of clause 53, in the form of intact pieces of comparable size to coffee beans, ground materials with particle sizes generally < 2 mm or extracts made from these mixtures. 55. The assembled product of clause 54, wherein the intact pieces are raw (subjected to temperatures below 120ºC) or roasted (subjected to temperatures ≥ 120ºC). 56. The composition of clause 50, wherein the assembled coffee composition comprises Ramon seed, green banana, sunflower seed extract, strawberry fiber, potato fiber, carrot, black aronia berry, carob, and optionally caffeine. 57. The composition of clause 56, wherein the finished bean-less coffee composition is further combined with xMR date seeds and traditional coffee seeds. 58. The composition of claim 57, in the form of intact pieces of comparable size to coffee beans, ground materials with particle sizes generally < 2 mm or extracts made from these mixtures. 59. The composition of clause 58, wherein the intact pieces are raw (subjected to temperatures below 120ºC) or roasted (subjected to temperatures ≥ 120ºC). 60. A method for creating a coffee or bean-less coffee composition, comprising assembling (e.g., combining compounding, extruding, pelletizing, etc.) a mixture containing: at least one rhamnogalacturonan (RG) polysaccharide ingredient(s), each derived from a non-coffee seed substrate and containing RG or portions thereof; and at least one other ingredient to provide an assembled coffee or bean-less coffee composition. 61. The method of clause 60, wherein the RGs are present in the assemblage in a range of 0.5 wt.% to 60 wt.%, or any subrange thereof, preferably in a range of 0.5 wt.% to 50 wt.%, 0.5 wt.% to 40 wt.%, 0.5 wt.% to 30 wt.%, 0.5 to 20 wt.%, 0.1 wt.% to 10 wt.%, 0.1 wt.% to 5 wt.%, 20 wt.% to 40 wt.%, or 5 wt.% to 20 wt.%, wherein wt.% is in terms of dry mass. 62. The method of clause 60 or 61, wherein the RG comprises an RG from at least one source selected from the group consisting of beet, carrot, potato, strawberry, raspberry, blueberry, blackberry, carob, jackfruit, bell pepper, tomato, pumpkin, ginseng, okra, grapefruit, aronia and/or acerola cherry, fenugreek seeds, flax seeds, coffee fruit (not seed) and combinations thereof. 14 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 63. The method of clause 62, wherein a plurality of 2-17 of the sources are assembled together with at least one other ingredient, preferably wherein 2-8 of the sources are assembled together with at least one other ingredient. 64. The method of clause 62 or 63, wherein, prior to assembling, the at least one RG source is subjected to at least one processing step selected from the group consisting of drying, juicing (e.g., separating solids from liquids), filtering, sieving, milling, washing, extracting, sorting, hydrolysis (e.g., through elevated temperatures and pressures, pH modifications, and/or enzymatic processes), fermentation, roasting, blanching, steaming or otherwise wet heating, chilling, freezing, concentrating and combinations thereof. 65. The method of any one of clauses 60-64, wherein the at least one other ingredient comprises at least one ingredient selected from the group consisting of starch, one or more chlorogenic acid (CGA) species, an RG, a traditional coffee ingredient, and combinations thereof. 66. The method of clause 65, wherein the starch comprises starch from at least one source selected from the group consisting of cereal or non-cereal grains, pulses, tubers, Ramon seeds, products of the Musa genus, and combinations thereof; and/or wherein the CGA comprises a CGA extract from a least one source selected from the group consisting of sunflower, artichoke, yerba mate, nightshades (e.g., tomatoes, potatoes, eggplants, tobacco), chicory, prune, Eucommia ulmoides, honeysuckle, and combinations thereof. 67. The method of clause 66, wherein the total amount of starch is present at an amount in a range selected from the range group consisting of 60-90 wt.%, 40-80 wt.%, 35-50 wt.%, 20-45 wt.%, 5-20 wt.%, and 0.1-10 wt.%, wherein wt.% is in terms of dry mass, and wherein the total of all CGAs is present at an amount in a range selected from 0.1-40 wt.% or any subrange thereof, preferably in a range of 0.2-8 wt.%, 20-40 wt.%, 4-15 wt.%, 6-20 wt.%, 0.3-6 wt.%, or 0.2-3 wt.%, wherein wt.% is in terms of dry mass. 68. The method of any one of clauses 60-67, further comprising subjecting the assembled composition to one or more processing steps selected from the group consisting of drying, roasting, grinding, extracting, concentrating, pasteurizing, sterilizing, and combinations thereof, to provide a finished composition (e.g., green beans, roasted beans, grounds, ready to drink form (RTD), or extract). 15 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 69. The method of clause 68, wherein the finished composition is further combined with a substrate carrier material or cross-Maillardized substrate carrier material having cross-Maillard reaction products (e.g., LWACMP and/or HWACMP as defined herein). 70. The method of clause 69, wherein the substrate carrier material comprises one or more selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the mustard family (Brassicaceae), watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal, apricot, jackfruit, (Artocarpus heterophyllus) and non-cereal grains, and/or coffee. 71. The method of clause 68, wherein the finished composition is further combined with one or more additional components selected from the group consisting of a different finished bean-less coffee composition, date seeds, cross-Maillardized (xMR) date seeds, traditional coffee seeds, caffeine, bioactive agents, flavors, colors, gums, texture modifiers, pH adjusters/regulators, fruit, or vegetable powders (non- compounded/pelletized) or their extracts, and combinations thereof, to provide an assembled product. 72. The method of clause 68, wherein the finished composition comprises Ramon seed, green banana, sunflower seed extract, strawberry fiber, potato fiber, carrot, black aronia berry, carob, and optionally caffeine. 73. The method of clause 72, wherein the finished composition is further combined with xMR date seeds and traditional coffee seeds. 74. A coffee or bean-less coffee composition, prepared by the method of any one of clauses 16, 34-40, and 61-74. Particular embodiments of the disclosure involving compositions comprising an xMR ingredient (e.g., xMR date seeds, prepared as described in working examples 1 and 2 of PCT Patent Application PCT/US2021/025565, published as WO 2021/202989, incorporated by reference herein in its entirety), can be described in view of the following clauses of WO 2021/202989: 1. A method of preparing a beverage component, comprising: contacting a substrate carrier material, having an endogenous Maillard-reactive nitrogen constituent and/or an endogenous Maillard-reactive carbohydrate constituent, with an exogenous Maillard reagent comprising an exogenous Maillard-reactive nitrogen constituent and/or 16 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 an exogenous Maillard-reactive carbohydrate constituent to provide a conditioned substrate carrier material; and adjusting the water activity (a
w) of the conditioned substrate carrier material to a value less than that of the conditioning reaction, and reacting, during the adjusting and/or at the adjusted aw value, the exogenous Maillard reagent with the endogenous Maillard-reactive nitrogen constituent and/or with the endogenous Maillard-reactive carbohydrate constituent to provide a low water activity (low aw) cross-Maillardized substrate carrier material having cross-Maillard reaction products (LWACMP) formed by the reaction between the exogenous Maillard reagent, and the endogenous Maillard-reactive constituent(s). 2. The method of clause 1 wherein the conditioned substrate carrier material, prior to adjusting the aw, comprises a cross-Maillardized substrate carrier material having cross-Maillard reaction products (HWACMP). 3. The method of clause 1 or 2, wherein the endogenous Maillard-reactive nitrogen constituent comprises one or more of amino acids, oligopeptides, polypeptides, and/or proteins, and/or wherein the endogenous Maillard-reactive carbohydrate constituent comprises one or more of mono-, di-, oligosaccharide, and/or polysaccharides. 4. The method of any one of clauses 1-3, wherein the exogenous Maillard- reactive nitrogen constituent comprises one or more of amino acids, oligopeptides, polypeptides, and/or proteins, and/or wherein the exogenous Maillard-reactive carbohydrate constituent comprises one or more of mono-, di-, oligosaccharide, and/or polysaccharides. 5. The method of any one of clauses 1-4, wherein the exogenous Maillard- reactive nitrogen constituent comprises one or more amino acids, and/or wherein the exogenous Maillard-reactive carbohydrate constituent comprises one or more mono- or disaccharides. 6. The method of any one of clauses 1-5, wherein the substrate carrier material comprises a natural and/or a processed or restructured plant material having the endogenous Maillard-reactive nitrogen constituent and/or the endogenous Maillard- reactive carbohydrate constituent. 7. The method of clause 6, wherein the plant material comprises one or more selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the mustard family (Brassicaceae), watermelon seeds, 17 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal and non-cereal grains, and/or coffee. 8. The method of any one of clauses 1-7, wherein contacting the substrate carrier material with the exogenous Maillard reagents comprises contacting with an aqueous solution of the exogenous Maillard reagents. 9. The method of any one of clauses 1-8, wherein contacting the substrate carrier material with the exogenous Maillard reagent comprises contacting at least the surface of the substrate carrier material with the exogenous Maillard reagent, and promoting adsorption, absorption, or adherence (e.g., covalently or physically) of the exogenous Maillard reagent, and/or of reaction products thereof, to at least the surface of the conditioned carrier material. 10. The method of any one of clauses 1-9, wherein contacting the substrate carrier material with the exogenous Maillard reagent comprises contacting at one or more conditioning temperature(s), under conditions and for a time period sufficient to provide for infusion of the exogenous Maillard reagent into at least the surface of the substrate carrier material, and/or solubilization and/or depolymerization of the endogenous Maillard-reactive nitrogen constituent and/or the endogenous Maillard-reactive carbohydrate constituent thereof. 11. The method of any one of clauses 1-10, wherein the LWACMP comprises cross-Maillardized reaction products on at least the surface thereof. 12. The method of any one of clauses 1-11, wherein adjusting the aw comprises adjusting to a value less than or equal to a value selected from the group consisting of 0.95, 0.90, 0.85.0.80, 0.75, 0.70, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15 and 0.1, or less than or equal to a value in a range of 0.10 to 0.95, including adjusting to a value less than or equal to any value in any subranges therein (e.g., 0.20 to 0.85, 0.25 to 0.80, 0.25 to 0.75, 0.25 to 0.70, 0.25 to 0.65, 0.25 to 0.60, 0.25 to 0.55), preferably to a value in a range of 0.25 to 0.70. 13. The method of any one of clauses 1-12, wherein adjusting the aw comprises drying the conditioned substrate carrier material at one or more drying temperatures. 14. The method of any one of clauses 1-13, further comprising restructuring one or more of the substrate carrier material, the conditioned substrate carrier material, and/or the LWACMP. 15. The method of any one of clauses 1-14, wherein the restructuring comprises one or more of fragmenting, grinding, milling, micronizing, depolymerizing, 18 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 solubilizing, permeabilizing, compacting and/or compressing the respective substrate carrier material. 16. The method of any one of clauses 1-15, further comprising heating the LWACMP under conditions sufficient to promote further Maillardization thereof, to provide an elevated temperature, cross-Maillardized substrate carrier material having cross- Maillard reaction products (ET-LWACMP). 17. The method of clause 16, wherein the adjusting the water activity (aw) of the conditioned substrate carrier material to provide the LWACMP, and the heating of the LWACMP to provide the ET-LWACMP are stages of one or more continuous or ramped heating process(es). 18. The method of clause 16 or 17, wherein the further Maillardization comprises further cross-Maillardization relative to the LWACMP. 19. The method of any one of clauses 16-18, wherein the heating is at one or more temperatures greater than the temperature used for adjusting the water activity (aw) of the conditioned substrate carrier material, or than the drying temperature. 20. The method of any one of clauses 16-19, wherein the heating comprises one or more of roasting, toasting, baking, grilling, and/or otherwise thermally treating at elevated temperatures. 21. The method of any one of clauses 16-20, further comprising grinding, or otherwise fragmenting, grinding, milling, micronizing, depolymerizing, solubilizing, permeabilizing, compacting, compressing and/or otherwise restructuring the ET- LWACMP. 22. The method of any one of clauses 1-21, wherein the level of at least one compound present in the conditioned substrate carrier material, the LWACMP, the ET- LWACMP, or in extracts thereof is differentially modulated relative to that of the substrate carrier material or that of the exogenous reagent(s) independently subjected to the method, taken alone or in sum. 23. The method of clause 22, wherein the at least one compound comprises 2,5-dimethylpyrazine, 2,3-butanedione, 1,3-bis[(5S)-5-amino-5-carboxypentyl]-4-methyl- 1H-imidazol-3-ium and/or of γ-butyrolactone. 24. The method of any one of clauses 1-23, further comprising extracting the conditioned substrate carrier material, the LWACMP or the ET-LWACMP to provide an extract, and an extracted retentate substrate carrier material. 19 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 25. The method of clause 24, wherein the extracting comprises suffusing or steeping in a suitable solvent (e.g., water, ethanol, glycol, supercritical CO
2, etc.) at a suitable temperature, wherein the extract comprises an infusion, and wherein the extracted retentate substrate carrier material comprises extracted retentate restructured substrate and/or grounds. 26. The method of clause 24 or 25, further comprising addition of one or more additional ingredients to the extract to provide a blended formula. 27. The method of clause 26, wherein the one or more additional ingredients comprises one or more of dry ingredients, liquid ingredients, oil, and/or gum ingredients. 28. The method of any one of clauses 24-27, comprising concentrating the extract or the blended formula, to provide a concentrated extract or concentrated blended formula. 29. The method of any one of clauses 24-28, further comprising subjecting the extract or the blended formula, or the concentrates thereof, to one or more of a sterilization process (e.g. UHT, retort, microwave, ohmic), a pasteurization process (e.g. HTST), a homogenization process, or non-thermal antimicrobial treatments (e.g. HPP, irradiation) etc., optionally followed by packaging or aseptic packaging. 30. The method of any one of clauses 24-29, further comprising drying of the extracted retentate substrate carrier material to provide a dried, extracted retentate substrate carrier material. 31. The method of clause 30, further comprising addition of one or more additional ingredients to the dried, extracted retentate substrate carrier material to provide a formulated retentate substrate carrier material. 32. The method of clause 31, wherein the addition of the one or more additional ingredients, comprises coating or infusing the dried, extracted retentate substrate carrier material. 33. The method of clause 31 or 32, wherein the one or more additional ingredients comprises one or more of dry ingredients, liquid ingredients, oil, gum ingredients, and/or an extract or lyophilized or dried extract of the LWACMP or of the ET- LWACMP. 34. The method of any one of clauses 24-33, further comprising instantizing the extract, the blended formula, or the concentrates thereof, to provide an instantized beverage component, optionally followed by aseptic packaging. 20 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 35. The method of any one of clauses 1-34, wherein the substrate carrier material comprises or is coffee or spent coffee grounds. 36. A beverage component, comprising a component prepared by the method of any one of clauses 1-35. 37. The beverage component of clause 36, wherein the beverage component comprises one or more of: a conditioned substrate carrier material having cross-Maillard reaction products (HWACMP); a low aw cross-Maillardized substrate carrier material (LWACMP) having cross-Maillard reaction products; an elevated temperature, cross- Maillardized substrate carrier material (ET-LWACMP) having cross-Maillard reaction products formed by heating the LWACMP under conditions sufficient to promote further Maillardization thereof; an extract of the HWACMP, the LWACMP, or the ET-LWACMP, or concentrates, blends or formulations thereof; an extracted retentate substrate carrier material having cross-Maillard reaction products; and a concentrated and/or instantized beverage component; and wherein any of these components are optionally packaged in single-use or multi-use pods, capsule, etc. 38. A cross-Maillardized substrate carrier material, or an extract thereof, comprising: a low water activity (low aw) cross-Maillard reaction product (LWACMP) formed, at an aw value less than or equal to 0.95, between an endogenous Maillard- reactive nitrogen constituent and an exogenous Maillard-reactive carbohydrate constituent, and/or between an exogenous Maillard-reactive nitrogen constituent and an endogenous Maillard-reactive carbohydrate constituent; and/or an elevated temperature, low water activity cross-Maillard product (ET-LWACMP). 39. The cross-Maillardized substrate carrier material, or the extract thereof, of clause 38, comprising LWACMP and ET-LWACMP. 40. The cross-Maillardized substrate carrier material, or the extract thereof, of clause 38 or 39, wherein the endogenous Maillard-reactive nitrogen constituent comprises one or more of amino acids, oligopeptides, polypeptides, and/or proteins, and/or wherein the endogenous Maillard-reactive carbohydrate constituent comprises one or more of mono-, di-, oligosaccharide, and/or polysaccharides. 41. The cross-Maillardized substrate carrier material, or the extract thereof, of any one of clauses 38-40, wherein the exogenous Maillard-reactive nitrogen constituent comprises one or more of amino acids, oligopeptides, polypeptides, and/or proteins, and/or wherein the exogenous Maillard-reactive carbohydrate constituent comprises one or more of mono-, di-, oligosaccharide, and/or polysaccharides. 21 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 42. The cross-Maillardized substrate carrier material, or the extract thereof, of any one of clauses 38-41, wherein the exogenous Maillard-reactive nitrogen constituent comprises one or more amino acids, and/or wherein the exogenous Maillard-reactive carbohydrate constituent comprises one or more mono- or disaccharides. 43. The cross-Maillardized substrate carrier material, or the extract thereof, of any one of clauses 38-42, wherein the substrate carrier material comprises a natural and/or a processed or restructured plant material. 44. The cross-Maillardized substrate carrier material, or the extract thereof, of clause 43 wherein the plant material comprises one or more selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the mustard family (Brassicaceae), watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal and non-cereal grains and/or coffee. 45. The cross-Maillardized substrate carrier material, or the extract thereof, of clause 44 wherein the plant material comprises or is coffee or spent coffee grounds. 46. The cross-Maillardized substrate carrier material, or the extract thereof, of any one of clauses 38-45, wherein the cross-Maillardized substrate carrier material comprises one or more of: a kernel or restructured form of the cross-Maillardized substrate carrier material having LWACMP, of the cross-Maillardized substrate carrier material having ET-LWACMP, or of the cross-Maillardized substrate carrier material having LWACMP and ET-LWACMP; an extract (e.g., aqueous) of the kernel or fragmented form of the cross-Maillardized substrate carrier material having LWACMP, of the cross-Maillardized substrate carrier material having ET-LWACMP, or of the cross- Maillardized substrate carrier material having LWACMP and ET-LWACMP; a concentrated and/or instantized extract of the kernel or fragmented form of the cross- Maillardized substrate carrier material having LWACMP, of the cross-Maillardized substrate carrier material having ET-LWACMP, or of the cross-Maillardized substrate carrier material having LWACMP and ET-LWACMP; and an extracted retentate cross- Maillardized substrate carrier material having LWACMP, having ET-LWACMP, or having LWACMP and ET-LWACMP; and wherein any of these components are optionally packaged in single-use or multi-use pods, capsule, etc. 47. The cross-Maillardized substrate carrier material, or the extract thereof, of any one of clauses 38-46, in the form of a beverage or beverage component. 48. The cross-Maillardized substrate carrier material, or the extract thereof, of any one of clauses 38-47, wherein the level of at least one compound present in the 22 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 LWACMP, in the ET-LWACMP, or in extracts thereof is differentially modulated relative to that of a corresponding non-cross-Maillardized substrate carrier material. 49. The cross-Maillardized substrate carrier material, or the extract thereof, of clause 48 wherein the at least one compound comprises 2,5-dimethylpyrazine, 2,3-butanedione, 1,3-bis[(5S)-5-amino-5-carboxypentyl]-4-methyl-1H-imidazol-3-ium and/or of γ-butyrolactone. 50. A cross-Maillard-primed substrate carrier material, comprising a non-liquid combination of: a substrate carrier material having an endogenous Maillard-reactive nitrogen constituent and/or an endogenous Maillard-reactive carbohydrate constituent; and an exogenous Maillard reagent having an exogenous Maillard-reactive nitrogen constituent and/or an exogenous Maillard-reactive carbohydrate constituent, wherein the non-liquid combination is primed (sufficient or capable) to produce a cross-Maillardized substrate carrier material upon adjustment of water activity (a
w), and/or heating, and/or drying thereof; optionally packaged in single-use or multi-use pods, capsule, etc. 51. The cross-Maillard-primed substrate carrier material of clause 50, wherein: the endogenous Maillard-reactive nitrogen constituent comprises one or more of amino acids, oligopeptides, polypeptides, and/or proteins; and/or wherein the endogenous Maillard-reactive carbohydrate constituent comprises one or more of mono-, di-, oligosaccharide, and/or polysaccharides; and/or wherein the exogenous Maillard- reactive nitrogen constituent comprises one or more of amino acids, oligopeptides, polypeptides, and/or proteins; and/or wherein the exogenous Maillard-reactive carbohydrate constituent comprises one or more of mono-, di-, oligosaccharide, and/or polysaccharides. 52. The cross-Maillard-primed substrate carrier material of clause 50 or 51, wherein adjusting the a
w comprises adjusting to a value greater than 0.95, or to a value less than or equal to a value selected from the group consisting of 0.95, 0.90, 0.85.0.80, 0.75, 0.70, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15 and 0.10, or less than or equal to a value in a range of 0.10 to 0.95, including adjusting to a value less than or equal to any value in any subranges therein (e.g., 0.20 to 0.85, 0.25 to 0.80, 0.25 to 0.75, 0.25 to 0.70, 0.25 to 0.65, 0.25 to 0.60, 0.25 to 0.55), preferably to a value in a range of 0.25 to 0.70; wherein drying comprises adjusting the aw to a value less than or equal to a value selected from the group consisting of 0.95, 0.90, 0.85.0.80, 0.75, 0.70, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15 and 0.10, or less than or equal to a value in a range of 0.10 to 0.95, including adjusting to a value less than or equal to any value 23 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 in any subranges therein (e.g., 0.20 to 0.85, 0.25 to 0.80, 0.25 to 0.75, 0.25 to 0.70, 0.25 to 0.65, 0.25 to 0.60, 0.25 to 0.55), preferably to a value in a range of 0.25 to 0.70; and wherein heating comprises heating at, or to, a temperature above ambient temperature. 53. The cross-Maillard-primed substrate carrier material of any one of clauses 50-52, wherein the non-liquid combination comprises a powder or particle form of either the substrate carrier material, the exogenous Maillard reagent, or both. 54. The cross-Maillard-primed substrate carrier material of any one of clauses 50-53, wherein the substrate carrier material and/or the exogenous Maillard reagent are in the form of a bound or unbound aggregate, a direct compression, a dry granulation, wet granulation, extrusion and in each case may optionally comprise one or more further excipients (e.g., binder, distintegrant, lubricant, etc.). 55. The cross-Maillard-primed substrate carrier material of any one of clauses 50-54, wherein the substrate carrier material and the exogenous Maillard reagent are in the form of a compressed or compacted, bound or unbound, kernel, bean, pellet or other form. 56. The cross-Maillard-primed substrate carrier material of any one of clauses 50-55, wherein the substrate carrier material comprises a natural and/or a processed or restructured plant material. 57. The cross-Maillard-primed substrate carrier material of clause 56, wherein the plant material comprises one or more selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the mustard family (Brassicaceae), watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal and non-cereal grains and/or coffee. 58. The cross-Maillard-primed substrate carrier material of clause 57, wherein the plant material comprises or is coffee or spent coffee grounds. 59. A method of making a cross-Maillard-primed substrate carrier material, comprising combining: a substrate carrier material having an endogenous Maillard- reactive nitrogen constituent and/or an endogenous Maillard-reactive carbohydrate constituent; and an exogenous Maillard reagent having an exogenous Maillard-reactive nitrogen constituent and/or and exogenous Maillard-reactive carbohydrate constituent, to provide a non-liquid combination, wherein the non-liquid combination is primed (sufficient or capable) to produce a cross-Maillardized substrate carrier material upon adjustment of water activity (a
w), and/or heating, and/or drying thereof. 24 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 60. The method of clause 59, wherein: the endogenous Maillard-reactive nitrogen constituent comprises one or more of amino acids, oligopeptides, polypeptides, and/or proteins; and/or wherein the endogenous Maillard-reactive carbohydrate constituent comprises one or more of mono-, di-, oligosaccharide, and/or polysaccharides; and/or wherein the exogenous Maillard-reactive nitrogen constituent comprises one or more of amino acids, oligopeptides, polypeptides, and/or proteins; and/or wherein the exogenous Maillard-reactive carbohydrate constituent comprises one or more of mono-, di-, oligosaccharide, and/or polysaccharides. 61. The method of clause 59 or 60, wherein: adjusting the a
w comprises adjusting to a value greater than 0.95, or to a value less than or equal to a value selected from the group consisting of 0.95, 0.90, 0.85.0.80, 0.75, 0.70, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15 and 0.10, or less than or equal to a value in a range of 0.10 to 0.95, including adjusting to a value less than or equal to any value in any subranges therein (e.g., 0.20 to 0.85, 0.25 to 0.80, 0.25 to 0.75, 0.25 to 0.70, 0.25 to 0.65, 0.25 to 0.60, 0.25 to 0.55), preferably to a value in a range of 0.25 to 0.70; wherein drying comprises adjusting the a
w to a value less than or equal to a value selected from the group consisting of 0.95, 0.90, 0.85.0.80, 0.75, 0.70, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15 and 0.10, or less than or equal to a value in a range of 0.10 to 0.95, including adjusting to a value less than or equal to any value in any subranges therein (e.g., 0.20 to 0.85, 0.25 to 0.80, 0.25 to 0.75, 0.25 to 0.70, 0.25 to 0.65, 0.25 to 0.60, 0.25 to 0.55), preferably to a value in a range of 0.25 to 0.70; and wherein heating comprises heating at, or to, a temperature above ambient temperature. 62. The method of any one of clauses 59-61 wherein the non-liquid combination comprises a powder or particle form of either the substrate carrier material, the exogenous Maillard reagent, or both. 63. The method of any one of clauses 59-62, wherein the substrate carrier material and/or the exogenous Maillard reagent are in the form of a bound or unbound aggregate, a direct compression, a dry granulation, wet granulation, or extrusion, and in each case may optionally comprise one or more further excipients (e.g., binder, disintegrant, lubricant, etc.). 64. The method of any one of clauses 59-63, wherein the substrate carrier material and the exogenous Maillard reagent are in the form of a compressed or compacted, bound or unbound, kernel, bean, pellet or other form. 25 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 65. The method of any one of clauses 59-64, wherein the substrate carrier material comprises or is a natural and/or a processed or restructured plant material. 66. The method of any one of clauses 59-65, wherein the plant material comprises or is one or more selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the Brassicaceae family, watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal and non-cereal grains and/or coffee. 67. The method of clause 66, wherein t the plant material comprises or is coffee or spent coffee grounds. 68. A cross-Maillard-primed substrate carrier material, prepared the method of any one of clauses 59-67. 69. A method for imparting flavor and/or aroma to a cross-Maillardized or non- cross-Maillardized carrier material comprising: obtaining a substrate carrier material; and applying a beverage component according to clause 36 or 37, and/or applying a cross- Maillardized substrate carrier material, or an extract thereof, according to any one of clauses 38-49. 70. The method of clause 69, wherein the carrier material comprises or is a natural and/or a processed or restructured plant material. 71. The method of clause 70, wherein the plant material comprises one or more materials selected from the group consisting of date seeds, chicory root, Yerba mate stems and/or leaves, dandelion, seeds from the Brassicaceae family, watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal and non-cereal grains and/or coffee. 72. The method of clause 71, wherein the plant material comprises or is coffee or spent coffee grounds. 73. A flavor and/or aroma enhanced carrier material prepared by the method of any one of clauses 69-72. BRIEF DESCRIPTION OF THE DRAWINGS Those of skill in the art will understand that the drawings, described below, are for illustrative purposes only. The drawings are not intended to limit the scope of the present teachings in any way. 26 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 FIG.1 schematically shows, by way of non-limiting examples of the present invention, a high-level depiction of a first embodiment of a method for production of coffee-substitute beverage products. FIG.2 depicts the levels of 2,5-DMP generated in various steps, and particularly the low levels of 2,5-DMP generated in the preconditioning and drying steps, for Control, CrossMR, and MR samples. Cross reactions between the exogenous reagents and the substrate are observed, as evidenced by the elevated levels of 2,5-DMP generated when substrate and reagents are reacted together. FIG.3 depicts results from experiments conducted across exemplary example compositions, showing that careful selection of substrate and reagent is key to produce the desired final products and that addition of some Maillard reagents can result in decreased yield of desired compounds. FIG.4 depicts the production of 2,3-butanedione in the various example compositions, showing that flavorful aroma compounds resulting from the interaction of exogenous and substrate materials are also generated in greater yield using these inventive compositions. FIG.5 depicts scanning electron microscopy results showing changes in the cellular structure based on the cross-Maillard reactivity; the Control (left) samples show a highly porous structure, whereas CrossMR (right) samples exhibit a more dense and fuller cellular structure. FIG.6 depicts LC/MS results from semi-quantitation of 1,3-bis[(5S)-5-amino-5- carboxypentyl]-4-methyl-1H-imidazol-3-ium in the Control, Cross-MR and MR sample, showing that this compound is exclusively formed in the Cross-MR approach. FIG.7 shows, according to additional aspects of the invention, modulation of particular coffee aroma compounds in a cross-Maillardized raw ("green") coffee beans composition. FIG.8 shows, according to additional aspects of the invention, generation of particular roast aroma compounds by cross-Maillardization of previously roasted, ground and extracted coffee beans. FIG.9 shows, according to additional aspects of the invention, that initial cracking of the date seeds prior to preconditioning enhances the yield of cross-Mailladization products. FIGS.10A and 10B show, according to additional aspects of the invention, that addition of chlorogenic acid to the preconditioning reaction modulates (in this instance 27 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 decreases) the level of 2,5-dimethylpyrazine generated (FIG.10A), and that while cross- Maillaridization lowers the level of γ-butyrolactone relative to non-cross-Maillardized cracked date seeds (control cracked date seeds), addition of chlorogenic acid to the cross-Maillardization preconditioning mixture enhances the yield of γ-butyrolactone in cross-Maillardized date seeds. FIG.11 shows, according to additional aspects of the invention, that fermenting the date seeds prior to preconditioning enhances the yield of cross-Maillardization products. FIG.12 shows, according to non-limiting exemplary aspects of the present invention, a dried, roasted extract of defatted sunflower seed meal used to create a coffee-enhancing soluble solid. FIG.13 shows, according to non-limiting exemplary aspects of the present invention, the levels of CGLs produced from a sunflower seed extract. FIG.14 shows, according to non-limiting exemplary aspects of the present invention, a spectroscopic signature, of chlorogenic acids and/or lactones, of a roasted extract of defatted sunflower seed meal extruded within a ramon seed matrix indicating survival and/or interconversion of those compounds through the roasting and brewing process (especially the peak at 324 nm). FIG.15 shows, according to non-limiting exemplary aspects of the present invention, that extracts of de-hulled, defatted sunflower seed meal contained chlorogenic acids and/or lactone derivatives thereof, inferred from their spectroscopic signatures (at 200 nm and 324 nm) and liquid chromatography. FIG.16 shows, according to non-limiting exemplary aspects of the present invention, a spectroscopic signature, of chlorogenic acids and/or lactones, of a roasted extract of de-hulled, defatted sunflower seed meal extruded within a ramon seed matrix indicating survival and/or interconversion of those compounds through the roasting and brewing process (especially the peak at 324 nm). FIG.17 shows, according to non-limiting exemplary aspects of the present invention, that the levels of many key odorants for coffee, such as a family of pyrazines, are significantly enhanced by compounding kulthi daal flour and the de-hulled, defatted sunflower meal extract prior to roasting. FIG.18 shows a prior art schematic Pectin diagram (taken from Zdunek, A., Pieczywek, P. M., & Cybulska, J. (2021). The primary, secondary, and structures of higher levels of pectin polysaccharides.); 28 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 FIG.19 shows, by way of non-limiting examples of the present invention, a GC- MS spectrum of coffee-like volatile aroma compounds in the immersion extracts of roasted sugar beet pulp nuggets; FIG.20 shows, by way of non-limiting examples of the present invention, a GC- MS spectrum of coffee-like volatile compounds present in the immersion extracts of ED- bean grounds; FIG.21 shows, by way of non-limiting examples of the present invention, a GC- MS spectrum of coffee-like aroma compounds in immersion extracts of roasted carrot nuggets; FIG.22 shows, by way of non-limiting examples of the present invention, a GC- MS spectrum of coffee-like aroma compounds present in immersion extracts of roasted carrot pulp nuggets; FIG.23 shows, by way of non-limiting examples of the present invention, a GC- MS spectrum of coffee-like aroma compounds immersion extracts of roasted potato fiber nuggets; and FIG.24 shows, by way of non-limiting examples of the present invention, a GC- MS spectrum of coffee-like volatile compounds present in the immersion extracts of roasted strawberry fiber nuggets. DETAILED DESCRIPTION OF THE INVENTION Unless otherwise noted (see “DEFINITIONS” below), terms are to be understood according to conventional usage by those of ordinary skill in the relevant art. Chlorogenic acids and derivatives thereof Provided are coffee or plant-based coffee substitute compositions comprising exogenous chlorogenic acid (CGA) extracts derived from non-coffee or coffee plant material (see, e.g., working Examples 36-46 below). Due to the prevalence of CGAs throughout the plant kingdom, it is possible to extract (and to match coffee’s potency, enrich) CGAs from other plant sources. For example, yerba mate contains comparable CGA levels to coffee, but other plant species are interesting for this endeavor. Though at best second tier CGA producers, sunflowers, various nightshades (eggplants, tomatoes, potatoes and tobacco), Eucommia ulmoides, honeysuckle and various other plants produce sufficient levels of CGA to be commercially viable as sources, especially after enrichment. 29 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 While the present discussion focuses mainly on non-coffee sources (and non- coffee coffee generally), coffee is a great source of CGA. Notably, less desirable coffee could be a valuable source of CGA. As a specific example, robusta coffee (Coffea canephora), while having a devoted following, is generally less desirable than arabica. However, the levels of CGAs in robusta coffee tend to be higher than those in arabica. Extracting CGAs from lower quality coffees—and using them to produce higher quality coffee—could be commercially viable. Once in hand, CGAs are useful additions to “raw” forms of coffee, whether derived from Coffea genus plants or not, where the subsequent thermal processing would be expected to drive the conversion of some or all of the CGAs to CGLs and/or phenylindanes. Significant levels of CGA remain in roasted coffees, especially at lighter roast levels, and thus such exogeneous CGA added to an otherwise “finished” product (“roasted” grounds or a prepared beverage) would contribute important qualities to the finished product. One advantage of separating the CGA from its original matrix is that it opens new avenues for conversion of it into desired finished products. When the conversion occurs by roasting an intact bean, the operator (the coffee roaster) has limited control. They can change the heating profile, but they are fundamentally unable to access the CGA directly. With the CGA isolated, a variety of wet and dry process are available to fine tune the output of the conversion to the finished state. One could, for example, heat the isolated CGA to eliminate side reactions and produce a more purified final product. Coffee roasting is nearly monolithic in the approach, whereas isolated CGA could be transformed through heating processes unavailable to intact beans. Or rather than leaving the local composition of co-reactants with the CGA to the vagaries of the seed’s biology, compounded mixtures of CGA and separate constituents which will serve to control or modulate the reaction, or to react with the CGA are possible. This enables the curated production of these constituents of coffee, rather than the imprecision of the traditional methods. Ultimately, crucial facets of coffee are derived from CGA, whether from the acids themselves or their reaction products. As we contend with new growing challenges for coffee arising due to climate change, and pursue new and interesting experiences in coffee, the flexibility afforded by the isolation and deliberate processing of CGAs is a powerful method to improve the coffee we produce (however one chooses to define that). 30 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Particular Examples (36-46) describe powerful methods for the isolation (e.g., extraction) and deliberate processing (e.g., thermal processing and/or compounding) of plant-derived CGAs to provide compositions having utility to improve coffee and coffee substitute products. The disclosed CGA extraction methods and the CGA extract compositions of Examples (36-46) can be used in combination with cross-Maillardization methods and compositions (e.g. Examples 1-35 and as otherwise described herein), to improve the organoleptic properties of coffee and coffee substitute products. These CGA extracts may be combined with these preconditioned cross-Maillardization compositions prior to the roasting process to, e.g. facilitate cross reactions between the CGA comprising extract, may be combined with materials not treated by the specific preconditioning methods described herein for various purposes (e.g. cross reacting, bundling/forming, etc.) before heating and subsequently combining with the preconditioned materials, or may be heated interpedently and subsequently added to the preconditioned materials. Rhamnogalacturonans (RGs) Additionally provided are compositions and methods of preparation of coffee substitute materials having coffee-like flavors and textures by virtue of incorporation of key polysaccharides (see, e.g., working Examples 47-62 below). Surprisingly, these are not arabinogalactans/arabinogalactan proteins (AGs/AGPs) nor glucose homopolymers (e.g., neither starch nor cellulose) that are found to be plentiful in traditional coffee (for AGP) and many coffee substitutes (e.g., barley or date seeds, for starch/cellulose). Rather, these key polysaccharide polymers contain a backbone primarily comprising galacturonic acid with some rhamnose, and side chains comprising arabinose, xylose and galactose. In particular, it has been discovered that a specific class of polysaccharide— rhamnogalacturonans (RGs)—surprisingly contribute myriad valuable attributes to coffee products made without (or, optionally, with) traditional (i.e., Coffea genus seed based) coffee. RGs are not distinct polymers, per se, but rather distinct domains or blocks within larger pectin polymers (see FIG.18). Most plant tissues contain some kind of pectin, the most basic form of which is a simple linear homopolymer of galacturonic acid. These polymers can be quite long and have sections with varied structures over their length. RGs are sections that contain side chains or branches off the main polymer chain, and these side chains contain specific other saccharide species. Notably, these 31 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 side chains contain rhamnose, arabinose and galactose, as well as other saccharide species. Depending on the type of RG in question, the homogalacturonan (HG) backbone may not be present, instead replaced by an alternating copolymer of rhamnose and galactose. Pectins are a highly diverse group of polysaccharides present in the tissues of myriad plant species. Notably, however, prior analyses indicate that pectin comprises especially low fractions of coffee seeds (<1%) (Bradbury, A.G.W. and Halliday, D.J., J. Agric. Food Chem.1990, 38, 2, 389–392; https://doi.org/10.1021/jf00092a010). Furthermore, coffee substitutes were often produced by utilizing other seed species, for example date seeds, barley seeds and the like, which likewise contain insignificantly low levels of pectin. Herein we describe compositions that produce coffee-like qualities while containing—and in large part due to—the presence of these pectin polymers. This is, in part, accomplished by selecting the flesh of these plants, or of portions containing substantial portions of the flesh along with seeds, as plant flesh generally contains significant levels of pectic polymers. According to particular non-limiting exemplary aspects of the present invention, carrots (Daucus carota) are used as a non-coffee-seed ingredient, which contains sufficient quantities of these pectic polysaccharides (comprising significant fraction RGs) to produce coffee-like effects when properly pre-processed and roasted, even without other added ingredients. For example, the flesh of the plant may be used with essentially no changes to its composition (other than the drying, i.e., removal of water). Alternatively, the pulp (the remains after juice extraction) retains significant amounts of these pectic constituents, and can also be used for the creation of coffee qualities. In additional non-limiting exemplary aspects potato (Solanum tuberosum), naturally starch rich, may be used to produce coffee-like qualities. For example, after processing potatoes to remove most of the starch, through physical or biochemical methods) the relevant fraction of non-glucose polysaccharides (including RGs) can be enriched sufficiently to produce coffee-like qualities. In yet additional non-limiting exemplary aspects sugar beets (Beta vulgaris) may be processed and used to enhance coffee-like qualities. Sugar beet pulp, for example, contains significant portions of pectic polysaccharides, including RGs. While soluble in some conditions, these polymers can be further solubilized by partial hydrolysis. For example, subjecting sugar beet pulp to high pressure and high temperature conditions, or to high temperatures in the presence of acid or alkali, may be used to partially degrade 32 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 these polymers and allow high levels of extraction from the pulp. When filtered, this extract provides a potent source of these polysaccharides and make for excellent coffee substitutes or amendments. In further non-limiting exemplary aspects, such ingredients (pectic polysaccharides, including RGs) can be incorporated into coffee compositions that optionally contain traditional coffee (e.g. seeds of Coffea arabica or Coffea canephora) to enhance key coffee qualities such as roast flavors, acidity and texture/body. However, the presence of traditional coffee is not required for these compositions to enhance the coffee-like qualities, nor to create a mimic of sufficient fidelity that a convincingly coffee- like finished product (ex: extractable grounds or a beverage) can be produced using these ingredients and no traditional coffee whatsoever. Representative Key Polysaccharide Sources/Ingredients and their polysaccharides Beets. Beets (Beta vulgaris) refers to both conventional beetroot (“table beets”) as well as sugar beets. Both varietals not only contain significant quantities of reducing sugar, but also RG-containing polysaccharide components that are particularly noteworthy in the context of the present compositions and methods. Sugar beet pulp, for example, is a byproduct obtained from processing sugar beets, comprising approximately 70-80% carbohydrates, with 45-60% classified as non-starch polysaccharides (NSP) and 10-15% as soluble sugars. Pectin is the predominant polysaccharide in sugar beet pulp. The homogalacturonan (HG) domain, primarily consisting of galacturonic acid with minimal side chains, has a low content (1.45% of dry matter). Conversely, the RG-I domain exhibits a relatively high content (26.06% of dry matter). Within the RG-I domain of sugar beet pulp, the side chains contain neutral sugars such as galactose and arabinose. The prevalence of arabinose (19.36% of dry matter) and absence of galactose indicate abundant arabinan side chains. The high content of neutral sugars in sugar beet pulp suggests extensive branching in the RG-I region, as evidenced by the high molar ratios of (Gal + Ar)/Rh = (5.78) (Baryga, A., Ziobro, R., Gumul, D., Rosicka-Kaczmarek, J., & Miśkiewicz, K. (2023). Physicochemical properties and evaluation of antioxidant potential of sugar beet pulp—preliminary analysis for further use (future prospects). Agriculture, 13(5), 1039). Sugar beet pulp extract may also be obtained through subcritical extraction of sugar beet pulp. For example, as shown herein by Applicant, on the basis of dry matter, 33 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 a subcritical extract of sugar beet pulp was found to contain 68% carbohydrates, including 15% free sugars (e.g., fructose, glucose, and arabinose) and 53% oligo/polysaccharides. After sulfuric acid hydrolysis of the oligo/polysaccharides, a significant amount of arabinose (30%), galactose (8%), and other sugars were detected. In sugar beet pulp extract, the HG domain makes up 5.8%, whereas the RG-I domain exhibits a relatively high content of 37.2%, significantly enriched beyond the levels in native sugar beet pulp. These conditions are known generally to hydrolyze pectic polysaccharides into monomeric sugars, oligomers and smaller polymers. In particular embodiments of the present invention disclosed herein, this hydrolysis could be conducted using acidic or alkaline conditions, enzymes, etc. Carrot and carrot fiber. Carrots are celebrated for their nutrient density, featuring a significant dietary fiber content typically ranging from 6-18% by dry weight. Carrot pulp, comprising the solid residues from extracting the juice—rich in free sugars—from carrots, showcases an even higher concentration of dietary fiber, often exceeding 50% by dry weight. The HG domain accounts for 15% of the mass of dietary fiber in carrots, while the RG-I domain has a relatively high content of 45%. Given that dietary fiber constitutes 6-18% for intact carrots and up to 66% in carrot pulp, the HG domain represents 0.9%- 2.7% for intact carrots and up to 9.9% for pulp (dry weight). The RG-I domain ranges from 2.7-8.1% for intact carrots and 28% for pulp (dry weight). Previous literature characterizes carrot pectin as RG-rich, with highly branched arabinan and slightly branched galactan side chains, featuring comparable proportions of these sugars. However, some studies indicate a slightly higher ratio of galactose. Carrot cell walls also contain xylan, xyloglucan, and cellulose (Hotchkiss Jr, A. T., Chau, H. K., Strahan, G. D., Nuñez, A., Harron, A., Simon, S., ... & Yeom, H. W. (2023). Carrot rhamnogalacturonan I structure and composition changed during 2017 in California. Food Hydrocolloids, 137, 108411. Hotchkiss Jr, A. T., Chau, H. K., Strahan, G. D., Nuñez, A., Harron, A., Simon, S., ... & Yeom, H. W. (2023). Carrot rhamnogalacturonan I structure and composition changed during 2017 in California. Food Hydrocolloids, 137, 108411). Potato. Potatoes are generally thought of for their starch content, and with good reason: potatoes can be 60-80% starch on a dry basis. The remaining dry matter is composed of a variety of constituents, including some amount of protein, ash, fiber and the like. A more interesting form of potato for coffee flavor/texture applications is potato fiber. Potato fiber or pulp is the dry residue left over from potato processing, where most starch is extracted. It is a polysaccharide-rich byproduct considered to have low 34 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 commercial value. On a dry matter basis, potato pulp contains 50-60% dietary fiber, 10- 20% starch, and 6% protein. The dietary fiber is rich in RG-I domains, with a total content of 24-28% (dry matter), while the HG content is 5-6% (dry matter) (Klaveren, J. V. (2021). The viability of pectin extraction from potato fibre (Doctoral dissertation; and Trabert, A., Schmid, V., Keller, J., Emin, M. A., & Bunzel, M. (2022). Chemical composition and technofunctional properties of carrot (Daucus carota L.) pomace and potato (Solanum tuberosum L.) pulp as affected by thermomechanical treatment. European Food Research and Technology, 248(10), 2451-2470). Potato fiber pectins are characterized by a higher content of galactans, evidenced by a galactose to arabinose ratio of 3.5:1, which aligns with findings in the literature (Serena, A., & Knudsen, K. B. (2007). Chemical and physicochemical characterisation of co-products from the vegetable food and agro industries. Animal feed science and technology, 139(1-2), 109-124; and Ring, S. G., & Selvendran, R. R. (1978). Purification and methylation analysis of cell wall material from Solanum tuberosum. Phytochemistry, 17(4), 745-752). Strawberry. Strawberries are also a rich source of RG polysaccharides. Like with potato, the content can be enriched by selective removal of some components. Strawberry fiber, in particular, a byproduct of commercial strawberry juice extraction, contains a significant amount of dietary fiber (~55%), with the majority being insoluble (49%). In strawberry fiber, the HG domain represents 0.45% of the dry weight, whereas the RG-I domain exhibits a higher content of 8%. (Hotchkiss, A. T., Chau, H. K., Strahan, G. D., Nuñez, A., Harron, A., Simon, S., ... & Hirsch, J. (2024). Structural characterization of strawberry pomace. Heliyon, 10(9)). Monosaccharide analysis revealed that the strawberry fiber is primarily composed of glucose and xylose. Additionally, free glucose, fructose, xylose, arabinose, galactose, fucose, and galacturonic acid are detected. Bilberry (European wild blueberries) are another source of RG. Bilberry is notable as it contains another RG block type, RG-II. The pomace of bilberry contains approximately 1.8% RG-II by dry weight (Hilz, H., Williams, P., Doco, T., Schols, H. A., & Voragen, A. G. (2006), The pectic polysaccharide rhamnogalacturonan II is present as a dimer in pectic populations of bilberries and black currants in muro and in juice. Carbohydrate polymers, 65(4), 521-528; and see Hilz, H., Bakx, E. J., Schols, H. A., & Voragen, A. G. (2005). Cell wall polysaccharides in black currants and bilberries— characterisation in berries, juice, and press cake. Carbohydrate Polymers, 59(4), 477- 488). RG/HG Ratio. The ratio of dry weight of RG blocks to HG blocks in a given plant 35 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 tissue varies from one plant species to another and within plant samples taken from the same species, or within different tissues from the same plant/plant species. A higher ratio of RG to HG indicates a higher prevalence of, for example arabinose, galactose, etc. that are implicated in the thermal reactions creating desired coffee qualities as disclosed herein. Without placing specific high and low end limits, an example at the high end of this ratio is potato fiber with an approximate RG-I/HG ratio of 12, while an example on the low end is apple fiber with a ratio of 0.35 (e.g., see Niu, H., Dou, Z., Hou, K., Wang, W., Chen, X., Chen, X., ... & Fu, X. (2023), A critical review of RG-I pectin: Sources, extraction methods, structure, and applications, Critical Reviews in Food Science and Nutrition, 1-21; Ognyanov, M., Remoroza, C., Schols, H. A., Georgiev, Y. N., Petkova, N. T., & Krystyjan, M. (2020), Structural, rheological and functional properties of galactose- rich pectic polysaccharide fraction from leek, Carbohydrate Polymers, 229, 115549; and Kaczmarska, A., Pieczywek, P. M., Cybulska, J., & Zdunek, A. (2023), A mini-review on the plant sources and methods for extraction of rhamnogalacturonan I, Food Chemistry, 403, 134378). This ratio could also be determined between RG-II and HG, or between the sum of both RG-I and RG-II and HG, and, for example, that a high RG-I/HG ratio could be present alongside a relatively low RG-II/HG ratio, etc.. Representative plants having RG to HG ratios falling between these exemplary extreme values include Leek; Aronia; Bell pepper; Blackberry; Raspberry; Okra pods; Citrus subcompressa; Cacao pod husks; Grapefruit; Acerola Cherry; Tomato; Apple; Kiwi fruit; Lime; Mango; Pumpkin; Jackfruit; Banana, fenugreek seeds, flax seeds, coffee fruit (not seed) (e.g., the fruit of Coffea genus), etc., and combinations thereof, in addition to other sources previously identified herein. Monosaccharide Ratios. Further ratios are of interest in describing the relative performance of different varieties of RG—for example originating from different plant varietals or species or from different tissues within the same species—are the ratios of the monosaccharides comprising the RGs. These ratios originate in, and give clues to, the grater polymer structures for the RGs, which in turn modulate their utility in the context of coffee flavor. Arabinose, galactose and rhamnose are three particularly important monosaccharides for this discussion. This is because, in the context of RGs, these monosaccharides are always associated with branching. These monosaccharides may form the branch point in the backbone (i.e. rhamnose in RG-I) or comprise major constituents of the sidechains (all 3). Thuse the prevalence of rhamnose describes the 36 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 amount or frequency of branching, which when paired with RG/HG ratios described above, or arabinose and galactose compared to rhamnose, gives clues about the relative sizes of these branches. An important ratio in this context is the ratio of arabinose to galactose. Both can be found in the sidechains of RGs, but their behavior in flavour creation chemistry differs. In part, this is due to their structure: as a C5 sugar, arabinose reacts more quickly than the C6 galactose. Recall in conventional coffee, it is the arabinose side chains credited with the creation of a number of important coffee odorants, not the galactose present in the AGPs nor the galactomannans. Ratios of arabinose to galactose can vary considerably, for example approximately 1.5:1, 2:1, 3:1 or 4:1 or greater have been observed in plants such as sugar beets. Conversely, in plants sources such as strawberry fiber, this ratio has been observed near unity. Likewise, plant sources like potato and carrots can be quite rich in galactose, with some observed approximate ratios of 1:2, 1:3 or 1:4 or smaller. The estimated size of these side chains vary significantly from one source to the next. It is not uncommon to see an approximately 5:1 ratio of arabinose and galactose to rhamnose, or approximately 10:1, 15:1.20:1 or even 25:1 in particularly long side chain RGs. These, in turn, affect the kinetics of these flavor creation reactions as well as the overall balance of saccharides available for reactions. The present invention includes fundamentally different methods for producing desirable coffee and/or coffee-substitute compositions by integrating exogenous reactants (e.g., exogenous reagents comprising particular reactants) into coffee or non- coffee substrate carrier materials (e.g., raw/natural, crude or processed agricultural (e.g., plant-based) products). The functional/organoleptic coffee and/or coffee-like components are created through cross-reactive processes (e.g., Maillard reactions) occurring between the exogenously introduced reagents/reactants and endogenous reactants of the coffee or the non-coffee substrate carrier materials. Exemplary desirable compounds of interest may be placed into 5 exemplary categories, which in each case can be further divided into subsets of related compounds that perform similar functions in the finished beverage, as follows: 1. Flavor/aroma compounds: volatile molecules responsible for the flavor and aroma of coffee. Within the aroma category, important subcategories may include: Thiols: roasted, sulfury 37 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Pyrazines: roasted, earthy Diacetyl: buttery Furanones: caramel Guaiacols: Smoky Terpenes: Flowery Phenyls: Honey, fruity Phenols: Phenolic, ashy Esters: Fruity. 2. Taste compounds: generally non-volatile molecules that interact with taste receptors to provide sweetness, acidity, bitterness, saltiness and umami. 3. Colorants: molecules that provide the desired color for the beverage. Generally these are chosen to result in an overall brown, low turbidity appearance though this is not a stringent requirement. 4. Texture modifiers: compounds that modify the rheology of the liquid to better match the mouthfeel of coffee. 5. Bioactivity effectors: compounds providing beneficial effects, such as caffeine for alertness or polyphenols for their antioxidant quality. Often a family of compounds, rather than specific compounds, is relevant due similarity of the aroma of compounds with similar structure. The combinations of these compounds present in roasted coffee and coffee beverages is what tends to provide the distinctive coffee aroma/flavor. According to aspects of the invention, the disclosed cross-Maillardization methods and coffee-substitute compositions produce some, many, most or all, of these compounds. In the methods and compositions, individual components may be combined to yield the overall profile desired to create the coffee- substitute product. Exemplary embodiments of the invention, therefore, encompass coffee and/or coffee-substitute compositions and methods for making same, based on integrating (e.g., cross-reacting) exogenous reagent(s) into alternate raw materials (coffee, or non-coffee substrate carrier materials) having endogenous chemically reactive groups. The methods solve a long-standing problem in the art of how to optimally integrate, chemically and organoleptically, exogenous ingredients/reagents into such substrate carrier materials to provide modified substrate carrier materials having cross-reaction products (e.g., cross-Maillardized substrate carrier materials). According to aspects of the present invention, direct cross-reaction (e.g., cross-Maillardization) products may either be coffee 38 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 and/or coffee-like components per se, and/or may act as reactive intermediates that lead to indirect formation of other coffee and/or coffee-like components. Additionally, and/or alternatively, the present applicants have found that direct or indirect cross-reaction (e.g., cross-Maillardization) products may function by augmenting, or modulating (increasing or decreasing) the amount of one or more endogenous components (e.g., 2,5- dimethylpyrazine (2,5-DMP); 2,3-butanedione, etc.) that may be present or generated in some amount even during substrate carrier material processing in the absence of any exogenous reagent(s) (e.g., by altering of one or more chemical reaction pathways governing production of such endogenous components). Aside from cross-Maillaridization reactions, other types of cross-reactions may include caramelization and pyrolysis at higher temperatures. Constituents or reaction products may furthermore cross-react with polyphenols and the corresponding chinones. Radical reactions may take place (e.g., as is well known in lipid oxidation), and the reaction products may cross-react as well with other molecules from Maillard reaction cascades. Maillard-reactive constituents may include hydroxyl groups of polysaccharides, and carbonyl and amino groups of the nitrogen source (e.g. amino acids, polypeptides and proteins) as well as other chemical functions know to occur in the side chain of the N-source (e.g. sulfhydryl, amino, carboxyl, amide, and others). They may decompose, preferably upon thermal treatment, resulting in smaller, often more reactive intermediates favoring further cross reactions, referred to as the Maillard reaction cascade. These Maillard-reactive constituents may cross react with components originating from other reactions (e.g. lipid oxidation, polyphenol oxidation, hydrolysis, caramelization, pyrolysis, Fenton reaction, and others). By varying the relative concentrations and types of exogenous reagents relative to different substrate-specific endogenous reactants (e.g., by varying the relative proportion and types exogenous Maillard reactants relative to endogenous Maillard reactants), different proportions of cross-reaction products relative to endogenous or modulated endogenous reaction products (e.g., of cross-Maillard products relative to endogenous or modulated endogenous Maillard products) may be achieved. The disclosed methods, therefore, can not only be broadly applied to many different substrate carrier materials having different endogenous components and chemistries, but may also be fine-tuned based on their substrate-specific chemistries and the desired organoleptic qualities/characters. As described below in working Examples 9, 10, and 12, application of the disclosed cross-Maillardization methods to different substrate carrier materials, can 39 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 be used to either differentially increase or differentially decrease levels of 2,5-dimethylpyrazine, 2,3-butanedione, or 1,3-bis[(5S)-5-amino-5-carboxypentyl]-4- methyl-1H-imidazol-3-ium, respectively, depending on the substrate carrier material. While not being bound by mechanism, the cross-reaction (e.g., cross- Maillardization) methods are surprisingly effective in providing non-coffee compositions (and cross-reacted coffee compositions) that more accurately recapitulate the true coffee experience by reproducing some, many, most, or all of the aroma, taste, appearance, and texture of conventional/traditional coffee. The cross-reacted substrate carrier materials (e.g., cross-Maillardized substrate carrier materials) and/or extracts thereof, can be optionally combined with yet additional ingredients (e.g., dry, wet, gums, flavors, etc.) to provide finished coffee and/or coffee- substitute compositions and precursors (e.g., extractable cross-Maillardized substrate carrier materials (solids, grounds, whole seeds, restructured coffee-like ‘beans’ and the like), and extracts, beverages, concentrates, instantized solid formulations, flavors, etc., based thereon). In preferred embodiments of the cross-reacted (e.g., cross-Maillardized) substrate carrier materials and/or extracts thereof, etc., there are no coffee beans nor coffee-bean derived ingredients, and yet they replicate traditional coffee with greater fidelity than previously achievable. In additional embodiments, the organoleptic qualities of a flawed or low-quality coffee substrate, may be substantially improved by application of the disclosed cross-reaction methods. Such cross-reacted (e.g., cross-Maillardized) and/or regenerated conventional coffee substrate materials, for purposes of the present invention, may also be considered as coffee-substitutes, or cross-reacted coffee substrates (e.g., cross-Maillardized coffee substrates). The cross-reacted (e.g., cross-Maillardized) substrate carrier materials (e.g., coffee-substitute beverage precursors) are versatile, and not limited in the type of coffee-substitute beverage producible therefrom. Embodiments of the invention encompass compositions containing one or more of the cross-reacted (e.g., cross- Maillardized) substrate carrier material-derived compositions suitable for use as a coffee and/or coffee-like flavoring in other food or beverage items, such as ice creams, bakery items, sauces, etc. Embodiments of the cross-reacted (e.g., cross-Maillardized) substrate carrier material-derived compositions encompass blends thereof (e.g., in packaged forms) for use, for example, in flavorings, ice creams, sauces, bakery items, and the like. Embodiments of the invention also encompass cross-reacted (e.g., cross- Maillardized) substrate carrier material-derived compositions in single-use packaging 40 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 (such as, for example, single or multiple use coffee pods, single-serve capsules, and the like) used for on-demand beverage production. Process for preparing cross-reacted coffee and non-coffee substrate carrier materials (e.g., from raw, non-cross-reacted materials) Various generalized ingredients and methods necessary to create the inventive compositions are described herein. Exemplary raw materials (discussed more fully in the next sections) used to produce the building blocks for the above-described cross- reacted (e.g., cross-Maillardized) substrate carrier material-derived compositions include plant or plant-derived materials that can take many forms, such as seeds/kernels/pits (e.g., date seeds, seeds from the mustard family (Brassicaceae), watermelon seeds, pumpkin seeds, Jerusalem artichokes, sesame seeds, cereal and non-cereal grains, and/or coffee (e.g., beans/cherries, grounds), and the like), leaves/stems/stalks/flowers (e.g., yerba mate stems and/or leaves, honeysuckle, and the like), shells (such as, for example, sunflower, and the like), roots (such as, for example, chicory, dandelion, and the like), extracts derived from the above, and other plant materials and derivations from plant materials, and the like. The raw materials may be transformed into the desired cross-reacted products by a multi-step process, as depicted in the exemplary process embodiment of Figure 1, which typically involves one or more of the following exemplary steps: 1. Pre-treatment (substrate processing by, e.g., cleaning, mechanical processing, enzymatic treatments, and the like); 2. Cross-reaction (e.g., cross-Maillardization); including preconditioning; 3. Work up of cross-reaction (e.g., cross-Maillardization) product by, e.g., separation, draining, extraction, concentration, mechanical processing and the like; 4. Optional replication of one or more of steps 1-3, perhaps using alternative reagents, processing conditions, etc., (e.g., if the intermediate result or material (e.g., grounds or extracted grounds) is itself a precursor to a desired final composition); and 5. Final preparation and formulation steps (finishing steps) to form a completed/finished product component. Such steps may include, for example, mechanical processing (e,g., grinding, milling, crushing, compressing, etc., or otherwise restructuring), incorporation of ingredients (e.g., for texture, flavor, etc.), thermal processing, forming, and packaging. 41 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 In practice for step 1, if required, one or more coffee substrates, and/or one or more non-coffee substrates (substrate carrier materials) selected from the exemplary “Raw Materials” section (see below) may be optionally subjected (either separately or together, in the case of more than one substrate) to one or more pre-treatment processing steps (e.g., as described below). These pretreatment step(s) primarily serve to prepare the raw materials for the cross-reaction that occurs next. For example, residual date flesh may be removed from date kernels prior to subjecting date kernels to step 2. In practice, for step 2, the coffee and/or non-coffee substrate carrier material is conditioned with one or more exogenous reagents, (e.g., through cross-Maillardization reaction(s)) to produce and functionally integrate chemical and organoleptic coffee-like components through cross-reactive processes (e.g., Maillard reactions) occurring between the exogenously introduced reagents/reactants and endogenous reactants of the coffee and/or non-coffee substrate carrier material. Surprisingly, using the methods disclosed herein, the cross-reaction products replicate traditional coffee-like tastes, aromas, colors, and textures with greater fidelity than previously achievable. In the methods, substrate carrier materials may initially comprise a significant percentage, e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the dry matter of the total preconditioning reaction mixture. In the methods, mass ratios of added Maillard-reactive carbohydrate constituent:added Maillard reactive nitrogen constituent may be any value(s), e.g., in the range of 1:20 to 20:1, 1:5 to 10, 1:2 to 5:1, or 1:2 to 2:1, or other suitable value. Above listed clauses 1-73, involving compositions comprising an xMR ingredient. describe aspects of the cross-reaction methods in greater detail. In brief, by applying different conditions of water activity (a
w) and temperature, different cross-reactions may be sequentially used to produce and functionally integrate chemical and organoleptic coffee-like components. For example, an initial conditioned substrate carrier material may comprise a high water activity cross-Maillardized substrate carrier material (HWACMP) having cross-Maillard reaction products formed at a a
w value greater than that resulting from subsequent adjustment of the aw of the conditioned substrate carrier material to a value less than that of the conditioning reaction.. Subsequent to pre- conditioning (also referred to herein as “conditioning”), the a
w of the conditioned substrate carrier material may be adjusted to a value less than or equal to e.g., 0.85 (or, e.g., to less than or equal to another exemplary value as recited in clauses 12, 52 and 65) under 42 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 conditions sufficient provide a low water activity (low aw) cross-Maillardized substrate carrier material (LWACMP) having further cross-Maillard reaction products formed by the reaction between the exogenous Maillard reagent, and the endogenous Maillard-reactive constituent(s) (e.g., see above clauses 1-15). The LWACMP may be heated under conditions sufficient (e.g., wherein the heating is at one or more temperatures greater than the temperature used for adjusting the a
w of the conditioned substrate carrier material) to promote further Maillardization thereof, to provide an elevated temperature, cross-Maillardized substrate carrier material having cross-Maillard reaction products (ET- LWACMP) (e.g., see above clauses 16-20 involving compositions comprising an xMR ingredient). As indicated above, other types (other than cross-Maillardization) of cross-reactions may include caramelization and pyrolysis at higher temperatures. Constituents or reaction products may furthermore cross-react with polyphenols and the corresponding chinones. Radical reactions may also take place (e.g., as well known in lipid oxidation), and the reaction products may cross-react as well with other molecules from the Maillard reaction cascade(s). In practice, for step 3, the conditioned substrate carrier material, the LWACMP or the ET-LWACMP may, for example, be ground and/or extracted to provide an extract, and an extracted retentate substrate carrier material (e.g., see above clauses 21-25 involving compositions comprising an xMR ingredient). In practice, for step 4, after working up the initial cross-reactions product(s), the resulting materials may be subjected to additional runs of one or more of the preceding steps 1-3, e.g., using alternative reagents, processing conditions, etc. In practice, for step 5, after the final workup step, the product(s) are assembled in their final form (finished) (e.g., see above clauses 26-34 involving compositions comprising an xMR ingredient). Raw Materials Exemplary substrate carrier materials: Exemplary grain/cereals and pseudo cereals: corn, maize, oat, barley, rye, wheat, millet, sorghum, tiger nut, rice, quinoa, amaranth, buckwheat, and the like, and including the following exemplary cereal grains: Poaceae family, such as Zea mays (corn, resp. maize), Avena sativa (oat), Hordeum vulgare (barley), Secale cereal (rye), Triticum aestivum (wheat), Pennisetum 43 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 glaucum (millet), and Sorghum sp. (sorghum), Cyperus esculentus (tiger nut), or species of the Oryza genus (rice), f.e. Oryza glaberrima, Oryza sativa, and the like; Amaranthaceae family, such as Chenopodium quinoa (quinoa), Amaranthus (amaranth), and the like; and Polygonaceae family, Fagopyrum esculentum (buckwheat), and the like; Exemplary roots: Chicory, artichoke, sunflower, Jerusalem artichoke, dandelion, Chinese artichoke, ginger, and the like. These include the following exemplary roots/part of roots or seeds; Asteraceae family, such as Cichorium intybus (chicory), Cynara scolymus (artichoke), Helianthus annuus (sunflower), Helianthus tuberosus (Jerusalem artichoke), Taraxacum officinale (dandelion), and the like; Lamiacemae family, Stachys affinis (Chinese artichoke), and the like; and Zingiberaceae family, Zingiber officinale (ginger), and the like; Exemplary fruits, seeds, and shells thereof: Sunflower, date, palm, okra, cocoa, pumpkin, hemp, coffee, ramon tree, fig, soy, milkvetch, lupine, pea, peanut, avocado, olive, hazelnut, acorn, cherry, apricot, plum, raspberry, walnuts, hickory, pecan nut, chestnuts, Orange, lemon, grape, sesame, and mustards, and the like, and including the following exemplary fruits, seeds and shells thereof; Persea family, such as Persea americana (avocado), and the like; Asteraceae family, Helianthus annuus (sunflower), and the like; Arecaceae family, Phoenix dactylifera (date), Elaeis sp. (palm), and the like; Malvaceae family, Abelmoschus esculentus (okra), Theobroma cacao (cocoa), and the like; Cucurbitaceae family, Cucurbita pepo (pumpkin), C. maxima, C. argyrosperma, C. moschata, and the like; Cannabaceae family, Cannabis sativa (hemp), humulus (hops), and the like; Rubiaceae family, in specific, seeds and fruits of the Coffea genus, and the like; Moraceae family, Brosimum alicastrum (Ramon seed), Ficus carica (fig), and the like; Fabaceae family, Glycine max (soy), Astralagus boeticus, Lupinus pilosus (blue lupine), Pisum sativum (pea), Arachis hypogaea (peanut), and the like; Oleaceae family, Olea europaea (olive), and the like; Fagaceae family, Corylus sp. (hazelnut), Quercus sp., Lithocarpus sp. (acorn), and the like; 44 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Fagaceae family, Trigonella genus including T. foenum-graecum (fenugreek), and the like; Rosaceae family, in specific Prunus sp. and subsp., Prunus dulcis (almond), Prunus avium (sweet cherry), Prunus cerasus (sour cherry), Prunus subg. Prunus and their cultivars, Prunus armeniaca (apricot), Prunus domestica subsp. Insititia (plum), Rubus subgenus Idaeobatus (raspberry), and the like; Juglandaceae family, Juglans regia (walnuts), Carya sp. and subsp. (hickory and pecan nut), and the like; Betulaceae family, Castanea sp. (chestnuts), and the like; Rutaceae family, Citrus sinensis (orange), Citrus × limon (lemon), and the like; Vitaceae family, Vitis vinifera (grape), and the like; Brassicaceae family, Sinapis alba (yellow mustard), Brassica hirta (white mustard), Brassica nigra (black mustard), Brassica oleracea and rapa (cabbages), and the like; Leaves and stems: Tea, yerba mate, artichoke, and the like. These include the following exemplary leaves and/or stems; Aquifoliaceae family, Ilex paraguariensis (yerba mate), and the like; Theaceae family, Camellia sinensis (tea), and the like; and Asteraceae family, such as Cynara scolymus (artichoke), and the like. And including the Coffea family, such as Coffea arabica, Coffea canephora (Robusta), and the like. Exemplary Exogenous Reagents Sugars: a) Exemplary monosaccharides (and their corresponding salts (e.g., phosphates)), including but not limited to the following ketoses and aldoses, and the like. i. Ketoses 1. Trioses, such as dihydroxyacetone 2. Tetroses, such as erythrulose 3. Pentoses, such as ribulose, xylulose 4. Hexoses, such as fructose, psicose 5. Heptoses, such as sedoheptulose, mannoheptulose ii. Aldoses 45 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 1. Trioses, such as glyceraldehyde 2. Tetroses, such as erythrose, threose 3. Pentoses, such as ribose, arabinose, xylose 4. Hexoses, such as glucose, mannose, galactose 5. Heptoses, such as glucoheptose, mannoheptose, galactoheptose b) Exemplary deoxysaccharides, such as rhamnose, fucose, deoxyribose, and the like. c) Exemplary disaccharides, such as sucrose, maltose, lactose, lactulose, trehalose, cellobiose, isomaltulose, isomalt, and the like. d) Exemplary oligosaccharides, such as fructooligosaccharides, galactooligosaccharides, maltotriose, and raffinose, dextrins, and the like. e) Exemplary polysaccharides, such as dextrins, starch, inulin, cellulose, arabinogalactan, galactomannan, amylose, pectins and depolymerized pectins, glycosides and the like. f) Exemplary degradation products i. Deoxyosones and didesoxyosones, such as 1-desoxyosones and 3- desoxyosones, and the like. ii. Furanones, such as 4-hydroxy-5-methyl-3(2H)-furanone (norfuraneol), 4-hydroxy-2,5-dimethyl-3(2H)-furanone, 2-methyl-4,5-dihydro- 3(2H)-furanone, and the like. iii. Pyranones, such as maltol, 5-hydroxy-5,6-dihydromaltol, and the like. g) Exemplary uronic acids, such as galacturonic acids, glucuronic acids, and the like. h) Exemplary polyols, such as arabitol, glycerol, polyitol, xylitol, sorbitol, and the like. i) Exemplary amino sugars, such as galactosamine, glucosamine, and the like. j) Exemplary sugar syrups, such as aqueous solutions of the named above and their corresponding thermal processed products, such as caramelized sugar syrups, and the like. 46 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 k) Exemplary raw and processed agricultural products, including the products of their fermentations, and including but not limited to the following exemplary products: i. Ingredients, such as hydrolyzed starch (e.g. hydrolyzed corn starch), processed syrup (high fructose corn syrup, glucose syrup), molasses, malt extract, and the like. ii. Fruit juice, such as those derived from apples, plum, cranberries, lime, lemon, orange, grape and/or currant, and the like. iii. Syrups, such as those derived from maple, date, coconut, rice and/or agave, and the like. iv. Honey, invert sugar, and similar products. v. Extracts or hydrolysates of foods high in carbohydrates, such as extracts or hydrolysates of sugar beet, sugar cane, maize, bananas, apples, and the like. vi. Extracts or hydrolysates of grains, such as malt extracts, and the like. vii. Soft drinks, such as lemonades, cola, root beer, ginger ales, and the like. viii. Dairy and dairy products, such as milk, and similar products. ix. Plant-based milk analogues, such as soymilk, oat milk, nut milk, pumpkin seed milk, and the like. x. Pulps derived from food processing of fruits and vegetables, such as Coffea fruits and seeds, apple pulp, orange pulp, and the like, as well as pomace and must. Exemplary Amino acids a) Amino acids and their derivatives, e.g. modified by acetylation etc., may comprise or be derived from one or more of alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, pyrrolysine, proline, glutamine, arginine, serine, threonine, selenocysteine, valine, tryptophan, tyrosine, selenomethionine, hydroxyproline, ornithine, and the like, alone or in any combination or subcombination. 47 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 b) Peptides, such as dipeptides, oligopeptides and/or polypeptides, derived by synthesis, isolation, chemical and/or thermal hydrolysis, enzymatic digestion/polymerization/crosslinking, and the like. c) Protein and protein hydrolysates or the products of their fermentations i. Derived from animal products, such as meat, dairy, eggs and/or connective tissues, and the like. ii. Derived from plant materials, such as soy, pea, pumpkin, rice, oat, chickpeas, almonds, hemp, wheat, and the like. iii. Saccharide-protein conjugates, such as glycoproteins, and the like. iv. Oil-protein conjugates, such as proteolipids. d) Other glycosidically-bound secondary metabolites, and the like. Exemplary Modifying agents and intermediate products a) Reactive precursors and intermediates, such as Amadori and Heyns compounds, and the like. b) Initiators such as aldehydes and ketones (e.g., glyoxal, methylglyoxal, glycolaldehyde, acetol, dihydroxyacetone), and the like. c) Carbonic acids, such as ascorbic acid, lactic acid, pyruvic acid, acetic acid, citric acid, tartaric acid, quinic acid, and the like. For purposes of the present invention, the amount of α-hydroxy carboxylic acid(s), if used in the cross- reactions mixture(s), preferably is less than 10% by weight. d) Additives and agents i. Reducing agents, e.g. sodium hydrosulfide, ascorbic acid, and the like. ii. Antioxidants, e.g., ascorbic acid and [poly]phenols (see item f below), and the like. iii. Catalytic minerals and mineral salts, such as sodium chloride, sodium sulfate, iron chloride, and copper sulfate, and the like. iv. pH-modifiers and buffering agents, such as acids and their corresponding salts (phosphoric acid, lactic acid, acetic acid, and sodium acetate etc.) or bases, such as carbonates and phosphates (ammonia, potassium or sodium phosphates and carbonates, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium bicarbonate), and the like. 48 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 e) Solvents, such as ethanol, hexane, glycol or polyethylene glycol, and the like. f) Phenols and their corresponding esters, such hydroxycinnamic acids, e.g. coumaric, ferulic and/or caffeic acid, and their corresponding esters with e.g., quinic acid, and the like; including, in particular, chlorogenic acid and the corresponding isomers, and/or feruloyl quinic acid derivatives (e.g., as may be sourced from hops), and the like, as well as the conjugates with saccharides thereof, such as glycophenolic compounds, and the like. g) Polyphenols, such as quercetin, epicatechin, lignans, lignin, flavonoids, and the like. h) Alkaloids, such as trigonelline, caffeine and the like. i) Drying agents, such as calcium chloride, potassium carbonate, sodium sulfate, and the like. j) Surfactants, such as phospholipids, saponins, Acacia gum, mono and diglycerols, propylene glycol esters, lactylated esters, polyglycerol esters, sorbitan esters, ethoxylated esters, succinated esters, fruit acid esters, acetylated mono- and diglycerols, phosphated mono- and di-glycerols, sucrose esters, and the like. k) Enzymes for breaking down larger components (such as, for example, hydrolases, lyases, and the like), forming larger components (such as, for example, ligases, polymerases, and the like), modifying (such as, for example, transferases, oxidoreductases, and the like), isomerization (such as, for example, isomerases, and the like), and the like. Substrate Preparation Prior to the reaction step, raw material substrates (i.e., the coffee and/or non- coffee substrate carrier material) can be treated by a variety of processes to prepare the materials for the cross-reaction step 2 (see above general “Process for preparing cross- reacted non-coffee substrate carrier materials”). Depending on the particular substrate carrier material, any combination of one or more of the following processes can be used in any order. In general, these methods are designed to remove undesirable material from the substrate carrier materials, liberate, or render accessible, useful substrate materials from the matrix of the substrate carrier material, or improve the contact or reactivity between exogenous reagents and the substrate carrier materials with which they can react. 49 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Cleaning and Sorting Substrate carrier materials may require removal of foreign matter, undesirable units (such as, for example, poor quality materials), contaminated units of portions thereof, or residual flesh. Sorting based on criteria important for the subsequent reaction can also be carried out. Such criteria non-exclusively include size, color/coloration, density, geometric factors (such as, for example, aspect ratio), and the like. Washing/Extracting Substrate carrier materials may require a solvent-based treatment step to remove certain undesirable components or compounds prior to the cross-reaction (e.g., cross- Maillardization reaction). This can be for a variety of reasons. These components/compounds could produce undesirable reaction products under subsequent reaction conditions (such as, for example, oils that may go rancid), or may themselves be a desirable product to extract before the cross-reaction (e.g., cross-Maillardization reaction) can alter them (such as, for example, caffeine). Other possibilities include avoiding or modulating interference with the cross-reaction (e.g., cross-Maillardization reaction). This could take the form of modulating (increasing or decreasing) or eliminating compounds that suppress or compete with desired reactions (such as, for example, undesirable sugars), or components like skins or structural materials that inhibit penetration of the reagents into the body of the substrate carrier materials that can be removed chemically and/or physically. Thermal Processing Thermal processing may be necessary to properly prepare substrate carrier materials for the cross-reaction (e.g., cross-Maillardization reaction). Examples include the thermal inactivation of undesirable microbial populations or enzymes that would produce undesirable products if left functional. Thermal processing may also be used to alter the structure or composition of the raw material to make it more suitable for subsequent cross-reaction (e.g., cross-Maillardization reaction). This may include, for example, steaming, blanching, roasting, freezing, dehydrating, or lyophilizing, or the like, to disrupt cellular structures thus allowing easier penetration of reagents. Furthermore, it may convert the substrate carrier material to one more favorable for subsequent cross- reaction when the exogenous reagents are added (e.g., exogenous Maillard reagent(s)). 50 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Methods for thermal treatment may include any approach appropriate for the desired result, demands of the process, and details of the samples. This may, for example, include exposure to increased or decreased temperatures relative to ambient. Mechanical Processing The structure of the substrate carrier material(s) may be unsuitable for subsequent processing. Various mechanical treatments could be used to prepare them, such as comminution (e.g., milling, dicing, and the like), peeling, polishing, cracking/crushing, pressing (such as to remove oils or juices), sonication, perforating, and the like. Modified Atmosphere Processing Substrate carrier material(s) may be treated with a vacuum/low pressure environment to remove undesirable compounds or to collect those that should not participate in the cross-reaction. These conditions may also be used to de-gas and/or dehydrate the substrate carrier material or aid in the infusion of reagents to the inner structures of the substrate carrier material. Alternatively, substrate carrier material may be subjected to high pressures. These may, for example, be for purposes of microbial or enzyme inactivation, to modify the structure of the substrate carrier material to enhance subsequent processing, to aid in the infusion of exogenous ingredients for subsequent steps, or to aid extraction of compounds/components not desired in the cross-reaction. Additionally, these environments can be comprised of specific gas mixtures, rather than air. These may be chosen for their biochemical impact, for example to speed ripening (e.g. ethylene, and the like) or to prevent oxidation (such as, for example, from inert N
2, CO
2, and the like). Additionally, these may be gases that are themselves reagents in subsequent steps. Finally, the humidity may be modified to prepare the substrate carrier material. This may include elevated levels to hydrate plant tissues, or reduced humidity to dry and eliminate undesired water (e.g., to adjust the water activity (aw)). Cycling of these various conditions may be desirable and applied. This may include, for example, a vacuum infusion step to displace trapped air, followed by high pressure to enhance the diffusion. Alternatively, cycles of rapid pressurization/depressurization can be used to modify the structure of the substrate carrier material. Immersion 51 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Substrate carrier materials can be exposed to compositions including additional reagents prior to the cross-reaction (e.g., the cross-Maillardization reaction(s)). This may be for purposes of maximizing contact between the materials in the substrate with the exogenous reagents they are to react with (e.g., delivering sugars and/or amino acids to the center of an intact seed). Such exposure could take the form of immersion in liquid solutions or vapor mixtures under a variety of conditions as described herein in the above “Modified Atmosphere Processing” section. Photonic Treatments Continuous or pulsed photonic treatments may be used to reduce microbial levels or to modify the surface, inner structure, or chemistry of the substrate carrier materials and/or added reagents. Enzymatic treatments Endogenous or exogenous enzymes may be used to further modify the substrate carrier material prior to cross-reaction processing (e.g., cross-Maillardization). Enzymes may be used to break down polymers to liberate particular reagents (e.g., by use of amylases or hemicellulases to release a simple sugar), to soften, solubilize or break down the structure of the substrate carrier material (e.g., by use of cellulases, and the like), or to separate skins/membranes (such as, for example, pectinases, and the like). Similarly, peptidases could be used to either liberate useful components for reactions, increase solubility/availability or to break down the structure of the substrate (such as, for example, to increase porosity, ease or facilitate milling, etc.). Lipases are an additional exemplary class of enzyme that may aid in the production of useful precursors or functional ingredients, or in modifying the structure for the subsequent cross-reaction (e.g., cross- Maillardization). Additionally, enzymes that modify particular components of the substrate carrier material without specifically liberating them (e.g., deaminating asparagine to produce aspartic acid and reduce the production of acrylamide) may be used. Sprouting Seeds may be used as substrate carrier material, or may be sprouted and carried to the desired level of plant maturity to enact desired changes within the seed, such as conversion of storage carbohydrates to simple sugars, the attenuation of relevant antinutritional factors, etc. Sprouts may be thermally treated or dried at this point to halt 52 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 or inactivate the biochemical processes and/or to inactivate any microbial species present. Fermentation Prior to cross-reacting (e.g., cross-Maillardization), the substrate carrier material(s) may be modified by a fermentation step. This may comprise fermentation of a relatively crude form of the substrate carrier material prior to washing, such as, for example, a mass of crushed fruit pulp and intact or fractured seeds, or a relatively processed form of the substrate carrier material, such as a steamed grain with high internal moisture content and compromised cell structure. The organisms to perform the fermentation could be native or inoculated onto the substrate. Organisms could be, for example, bacterial or fungal. Such organisms may be genetically modified to enhance their production of key components or to produce compounds not native to the organism. Such fermentation processes may be used, for example, to convert native substrate to a more usable form (e.g., microbial liberation of simple sugars or amino acids, and the like). Such fermentation processes may also be used to generate useful enzymes for subsequent steps, e.g., for developing flavors, or flavor precursors. Process control for such fermentations may be accomplished through the use of conventional bioreactors. Completion of the fermentation step may include an inactivation step, such as, for example, a thermal treatment or antimicrobial ingredient addition. Cross-reaction (e.g., Cross-Maillardization reaction) The cross-reaction (step 2 of the multi-step process depicted in the exemplary process embodiment of Figure 1) is an important step in the creation of the desired final products from the coffee and/or non-coffee substrate carrier materials and the exogenous reagents (e.g., exogenous Maillard reagent(s)). According to particular aspects of the invention, given the nature and complexity of flavor-forming reactions (e.g., of Maillard reactions), the specific compositions, concentrations, and process parameters are useful to control or direct the cross-reaction towards the efficient creation of desired compounds. As discussed above, direct cross-reaction (e.g., cross-Maillardization) products may either be coffee and/or coffee-like components per se, and/or may act as reactive intermediates that lead to formation of other indirect coffee and/or coffee-like components. Additionally, and/or alternatively, the present applicants have found that direct or indirect cross-reaction products (e.g., direct cross-Maillardization products or 53 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 products derived from or including the direct products) may function by augmenting, or modulating (increasing or decreasing) the amount of one or more endogenous components (e.g., 2,5-dimethylpyrazine; 2,5-DMP) that may be present or generated in some amount even during substrate carrier material processing in the absence of any exogenous reagent(s) (e.g., by altering of one or more chemical reaction pathways governing production of such endogenous components). By varying the cross-reaction conditions, and the relative concentrations and types of exogenous reagents relative to different substrate-specific endogenous reactants (e.g., by varying the relative proportion and types exogenous Maillard reactants relative to endogenous Maillard reactants), different proportions of cross-reaction products relative to endogenous or modulated endogenous reaction products (e.g., of cross-Maillard products relative to endogenous or modulated endogenous Maillard products) may be achieved. The disclosed methods, therefore, can not only be broadly applied to many different coffee and/or non-coffee substrate carrier materials having different endogenous components and chemical pathways, but may also be fine-tuned based on their substrate-specific chemistries and the desired organoleptic qualities/characters. As previously mentioned, cross-reactions may also include, but are not limited to, reactions with phenols/chinones, lipid degradation products, and reactions with other (plant) constituents. Overall, cross-Maillard reaction products may further react with molecules resulting from caramelization, pyrolysis, lipid and (poly)phenol oxidation, and the like. During the cross-reaction step, ingredients, for example, from the Raw Materials section (e.g., one or more substrate carrier materials, and particular exogenous reactants) may be combined in any suitable means, blended with solvents (including water) appropriate to the nature of the cross-reaction and thermal processes, and adjustment of water activity may be employed and the cross-reactant products formed in one or more appropriate reaction vessels—which one or more reaction vessels could optionally be a final packaging form—as dictated by the necessary conditions of the cross-reaction. Various factors may be used as controls in the production of the cross-reaction products/compositions. A particular cross-Maillardization product may be intermediate or a final, finished product. Generally provision of final products will involve workup and final assembly steps to produce finished products. In particular aspects, the cross- reaction could produce a finished product if one or more of following exemplary conditions are satisfied: reaction media are loaded into heat-stable, chemically inert packaging prior 54 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 to reacting; the substrate carrier material and exogenous reagents require no further processing after the cross-reaction; and thermal processes sufficient to render a safe product, given the product characteristics and format, are utilized in the reaction. In these cases, the packaging may serve as the reaction vessel. Some examples of types of products (e.g., cross-Maillardized substrate carrier materials, and extracts thereof) that may be produced from such an operation include but are not limited to the following: ready-to-drink (RTD) beverages in a can or bottle, perhaps as a concentrate; single serve pods; ‘grounds’ for subsequent extraction by the user, in a can/jar or similar vessel; intact or restructured seeds/kernels/beans for grinding and extraction by a user, in a can, jar or similar or suitable vessel; liquid or powdered flavors in, for example, glass bottles. Temperature & Time Temperature control may be used to control the production rates of desired cross- reaction products and to limit microbial concerns. Reaction times (e.g., the cross- Maillardization reaction times) and temperatures may be varied to achieve the desired results (e.g., desired chemical and/or organoleptic properties imparted to the substrate carrier materials and/or to extracts thereof). Particular reaction temperatures may favor specific cross-reactions and cross-reaction products and may be selected according to the results desired. Additionally, multiple temperature steps may be used, based on the particulars of a given cross-reaction and substrate carrier material. In general, cross-reactions (e.g., cross-Maillardization) in aqueous media may be conducted at temperatures from, e.g., 0°C to 170°C (e.g., from 55°C to 170°C, from 55°C to 125°C, etc.), with temperatures in excess of 100°C typically requiring above ambient pressure. Reactions in ostensibly dry conditions will typically occur, at least partially, at higher temperatures, e.g., above 170°C. To facilitate the incorporation or use of certain reagents, for example, unstable or highly reactive ones, low temperature steps, including those below 0°C may be incorporated into the cross-reaction methods. Time-varying temperature profiles are desirable in many situations, such as in the roasting of solids or in multi-step reactions. These temperature changes may be timed according to other reaction parameters, such as ingredient additions, pH changes or sufficient progress in a given reaction or cross-reaction (e.g., cross-Maillardization reaction(s)). Cross-reactions (e.g., cross-Maillardization) may also occur during drying (reducing the overall a
w), which may, for example, be conducted at temperatures from 55 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 0°C to 130°C. Drying, for example, may comprise heating of a moist conditioned carrier material (e.g., in an electric oven, roaster, etc.), at temperatures from about 90°C to about 130°C, from about 40°C to about 90°C, from about 50°C to 70°C, etc. Cross-reactions (e.g., cross-Maillardization reactions) may occur during heating (e.g., roasting). In particular aspects, heating (e.g., roasting) of the dried conditioned carrier material may comprise roasting at one, or more temperatures in a range (e.g., ramped range), which may vary with the particular substrate carrier materials (e.g., leaves and roots, seeds, etc.), from about 110°C to about 300°C, from about 140°C to about 160°C; from about 190°C to about 225°C; from about 170°C to about 190°C; about 170°C at the maximum; about 180°C at the maximum; about 190°C at the maximum, etc. In particular aspects, roasting of the dried conditioned carrier material may preferably comprise roasting at one or temperatures in a range from about 180ºC to about 220ºC (e.g., from about 200°C to about 220°C). In particular aspects, roasting may comprise varying (e.g., ramping) the temperature from about 20°C to about 220°C. In particular aspects, the roasting comprises varying (e.g., ramping) the temperature from about 200°C to about 216°C. Heat may be applied or removed in any number of suitable ways based on the form factor of the substrate carrier material(s). Non-exclusively, these include ovens and steam ovens, steaming chambers, kettles and thermal processing vessels, retorts, heat exchangers, ohmic heating devices, screw extruders, immersion cookers, jet cookers, and others as recognized in the art or foreseeable based thereon. Heat may be applied to bulk reaction mixtures or in individual containers each containing a portion of the total cross-reaction mixture. Cooling devices may include, but are not limited to heat exchangers, blast chillers, spiral freezers, etc. Agitation of liquids, solids, or final containers is optionally applied, and is typically useful. The pH of the cross-reaction may be varied for determining the products of the reaction. By setting, or changing, the pH to one or more desired value/range, the products of the reaction may change. Furthermore, the exogenous and endogenous reagents themselves (including precursors and intermediates) may be pH-sensitive and thus may require specific pH values during their introduction and cross-reaction. Preferably, the pH for the cross-reactions (e.g., the cross-Maillardization reactions) is from about pH 5.0 to about 8.5. Particular cross-reactions, however, may 56 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 involve the use of pH levels beyond (below or above) this range. Particular cross- reactions, for example, may provide a desired outcome when performed at pH 6.0 - 8.5, while others may involve preferred ranges from pH 2.5 - 5.0. Higher and lower pH values are possible, and in general, the pH should be returned to suitable ranges for food products prior to release into commerce. pH values can be controlled, for example, through explicit addition of appropriate acids and bases so as to reach a desired pH value. As the cross-reaction can produce compounds that themselves alter the pH over time, control of the pH is a method to enhance the yield, efficiency and organoleptic qualities of the cross-reaction and its products. pH control may, for example, take the form of physical pH buffers, compositions of which were described previously, or active monitoring and control systems with metered dosing of appropriate acids and bases (organic or inorganic). Depending on the cross-reaction and substrate carrier material, a time-dependent pH value that favors different reactions at different times may be used. This change in pH may be coordinated with the progress of certain reactions (for example, production of desired products or consumption of particular reagents), different temperature steps, or the addition of reagents at later stages. Water Activity (a
W) According to particular aspects of the present invention, the water activity (a
w) of a cross-reaction mixture (e.g., of a cross-Maillardization reaction mixture) is useful in controlling the specific cross-reaction products generated and/or modulation of the levels of endogenous components present or produced during the cross-reaction (e.g., modulation of non-cross-reaction products or indirect cross-reaction products). Much like temperature and pH, different ranges of aw may favor the production of different reaction products and/or cross-reaction products, or levels thereof. Control of the a
w is may be accomplished in various ways, for example: 1) Explicit addition or removal of water by, for example, blending, diluting, conditioning, dehydrating, etc. 2) Environmental/atmospheric control during the reaction, such as by humidistatic control in the heating chamber (e.g., steam ovens). 3) Sealing of the heating chamber, thereby limiting or preventing the addition or removal of moisture from the cross-reaction mixture, as in the following exemplary embodiments: 57 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 a) sealing single- (e.g., Nespresso,® K-Cup®) or multi-serve, heat-stable packaging materials after charging with all the ingredients necessary to produce a coffee-like composition; b) an embodiment of a) in which the contents are consumed or dispensed directly from the packaging; c) an embodiment of a) in which the contents of the packaging comprise a coffee-substitute precursor, such as a liquid concentrate that is further diluted to produce a beverage; d) an embodiment of a) in which the contents of the packaging comprise solid materials from which a liquid (likely aqueous) extraction is performed to produce a coffee-like beverage either by manual means (e.g., by gravity/pour-over filtration), semi-automatic (e.g., grounds loaded into a conventional drip machine), or fully automatic (e.g., push button operation such as K-Cup® or Nespresso®- style single-serve vending machines); e) an embodiment of a) in which the packaging materials are recyclable; and f) an embodiment of a) in which the contents of the packaging is compostable/biodegradable. Atmosphere The atmosphere that the substrate carrier material and exogenous reagents are exposed to may be used to influence the cross-reaction products (e.g., influence the cross-Maillardization products of the cross-Maillardized substrate carrier materials and extracts thereof) produced. As previously discussed, the water activity (and thus atmospheric moisture), may be determinants of the final reaction products. Moreover, particular atmospheric components may contribute directly to the reaction. Oxygen, for example, comprises approximately 20% of the native atmosphere and can oxidize labile, flavorful components, thus producing other flavorful components (e.g., desirable and/or undesirable components), especially at elevated temperatures. Additionally, the atmosphere may have a time-varying nature. As the cross- reaction(s) (e.g., cross-Maillardization reactions) take place, volatile components may be created that serve to both modify the composition of the atmosphere as well as alter (e.g., increase) the pressure, which changes may then influence the products of the cross- reaction(s) (e.g., cross-Maillardization reactions). Moreover, increasing (or decreasing) 58 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 pressure may lead to altered product and/or cross-reaction product composition, and is thus an additional control variable in producing the desired compositions comprising cross-reaction products (e.g., cross-Maillardization products). Atmospheric control may be accomplished in ways analogous to humidity control. For example, sealed chambers, whether large reaction vessels or single-serve end-user packaging, may be held fixed to allow native atmospheric changes to take place. The cross-reaction (e.g., cross-Maillardization reactions) in such sealed chambers, for example, could begin with a native atmosphere, or one composed of a particular gas or mix of desired gasses (e.g., comprising an inert gas such as N
2 to prevent oxidation), that will evolve as the cross-reaction proceeds. Alternatively, process (e.g., cross-Maillardization) vessels can be subjected to vacuum conditions, vented, flushing and/or bubbling with preferred gasses, and/or pressurized by addition of a sufficient quantity of one or more desired gases, so as to arrive at the intended atmospheric condition and pressure. These and other atmospheric changes, can be controlled and time-varying to optimize or tailor the cross-reaction(s) (e.g., cross-Maillardization reactions) taking place over time. Reagent Timing These reactions and cross-reactions can be further optimized by delaying the introduction of certain reagents—or replenishment of consumed reagents—by later addition of additional reaction ingredients. These could be, for example, the creation of a precursor from the raw materials before adding the reagent needed to react with the precursor to produce the desired final composition. For example, the reaction could begin with a relatively simple mixture of a substrate material rich in reducing sugar, and one or more exogenous amino acids. After production of Amadori/Heyns products from these starting materials, additional carbohydrate or amino acid sources could be added to create desired products. Alternatively, these additions can be made on a continuous basis—rather than stepwise—to maintain ideal reaction conditions. Furthermore, these additions (continuous or stepwise) could be made based on continuous monitoring of the reaction by suitable measurement or analytical techniques and adjusted to optimize conditions on an ongoing basis. Interfaces 59 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 In some cases, reagents may be insoluble in an acceptable shared solvent. However as close contact is necessary to react two components together, one strategy to enable such reactions is to bring two insoluble phases together and drive the reaction at the interface between these two phases. A liquid biphasic system, for example, of two insoluble solvents produces a planar interface at which the reaction could take place. Alternatively, one insoluble phase could be dispersed into a continuous phase (a colloidal dispersion). Each phase could itself be solid, liquid, or gas (excepting gas-gas dispersions), perhaps stabilized by the addition of emulsifiers or other structuring agents or continuous mixing, bubbling, etc., to prevent undesirable separation. Continuous or dispersed phases could include any of the exemplary ingredients listed in the Raw Materials section, including immobilized catalysts/enzymes on solid carriers, whole or milled substrates, etc. Work Up After completion of the cross-reaction (e.g., cross-Maillardization reaction), in step 3 of the multi-step process depicted in the exemplary process embodiment of Figure 1, the crude product may be treated (e.g., worked-up), and generally is treated, to convert it to a component of a finished or intermediate composition. This may include one or more optional steps, such as separation, concentration, extraction, thermal processing, and the like, which may be performed in any suitable order and combined in any suitable way to provide for the finished or the intermediate component. The products of such workup steps may generally provide the inventive compositions, and in some cases may be essentially finished products (e.g., subjected to optional additional steps described in the next section for completion), or may be used as a component of an additional reaction or step (e.g., used as an intermediate component). Separation Insoluble or immiscible components may be separated by various means, such as decanting, filtration, centrifugation, and the like. Such methods may be further implemented to fractionate products based on size or density. Moreover, vacuum, high pressure, modified atmospheres, and the like may be used to aid in this process. Extraction Solvent or supercritical fluid extractions may be performed to remove undesirable reaction products or to isolate desirable reaction products. Such extractions may include, 60 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 for example, one or more of liquid-liquid extractions, solid phase assisted extractions, chromatographic extractions/separations, and the like. A variety of conditions may be applied, including, e.g., various solvents, pHs, temperatures, contact times, and atmospheres (including pressure/vacuum), and the like. Concentration For liquid fractions, the overall concentration of a component can be modulated (e.g., increased or decreased) if desired. This may be accomplished by using techniques such as reverse and/or forward osmosis, nano-, ultra- or micro-filtration, solvent removal/desolventizing including lyophilization, distillation/evaporation and vacuum distillation, spray drying, extrusion, freeze concentration, and the like. Thermal Processing Exemplary thermal processing methods are described in the “Pre Treatment” section covering relevant processing methods. Mechanical Processing Exemplary mechanical processing methods are described in the “Pre Treatment” section covering relevant processing methods. As mentioned above, optional replication of one or more of steps 1-3 of the exemplary process embodiment of Figure 1 may be employed, perhaps using alternative reagents, processing conditions, etc., (e.g., if the intermediate result is itself a precursor to a desired final composition). Finishing After the workup of step 3 (and optionally step 4) of the exemplary process embodiment of Figure 1 for creation of the inventive compositions, final assembly into a finished product may be necessary or desired as in step 5. This may include combining inventive compositions with any necessary or desired extra ingredients, as well as optional forming, packaging, and/or thermal processing to produce safe products. Exemplary products of this section include various format embodiments in accordance with the present invention, including but not limited to the following: ready-to-drink beverage; grounds in a capsule or other single usage pack or a concentrate for dilution by the end-user; instantized granules or powders; grounds for general usage; constructed beans or other formed solids in both reacted ("roasted") and unreacted 61 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 ("green") forms; and intact coffee-substitute “beans” derived from intact or fragmented processed substrate materials (e.g., to be ground and extracted by an end user). Formulation Individual inventive compositions (e.g., products derived directly or indirectly from the cross-reactions, e.g., cross-Maillardization reactions) may be combined with other inventive compositions or with other ingredients necessary to complete the desired format. Such compositions and/or ingredients include, for example, colorants, flavors, texture and pH modifiers, functional ingredients, nutritional and bioactive ingredients, plant or animal milks in various formats (liquids, dry powders, etc.), and the like. Depending on the format of the desired finished product, solid or liquid forms of the above may be used. The composition(s) may be processed into grounds, such as in a single serve packaging, or single or bulk packed loose grounds or a formed product, and for these purposes, may be further blended with a carrier-type material. For example, upcycled plant materials not previously processed using the disclosed reaction scheme may be used as a carrier matrix for the inventive compositions and other ingredients and optionally with solvents if needed or preferred to produce the desired blend(s). The compositions may be further processed to adopt a particular shape (e.g., see the “Forming/Pelletizing” section herein below), and for such purposes ingredients crucial to or desired for processing may additionally be added. For example, binders, moisture control ingredients and other materials or ingredients that facilitate the forming process, the retention of the given shape or the shelf-life of formed products may be added at this further processing stage. Flavor compounds, either produced by the inventive processes, or added during finishing, may be heat and oxygen sensitive, and may develop harsh or bitter qualities if over processed. Exemplary ingredients that may optionally be added at this stage, therefore, also include phenols and polyphenols, which may be employed, for example, as antioxidants/radical scavengers to limit the production of undesirable oxidized flavors during subsequent thermal processing (e.g., at elevated temperatures) or extended storage. By adding antioxidants at this stage, and subjecting the product compositions to only the heat needed for a safe food product, particular flavors (e.g., typical coffee-like bitterness or astringency) remain closer to the familiar coffee flavors. Concentrating 62 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 The finished composition may be concentrated after formulation, and for such purposes see, e.g., the previous “Concentration” section for exemplary methods and details. Instantizing Individual components, for example flavorful liquid extracts or finished beverages, may be instantized by procedures such as those multi-step procedures known in the art. This may include the separation of volatile flavor components prior to drying, recovery of these compounds, and subsequent reintroduction prior to the drying process, as detailed below, resulting in the finished product. The process of volatile flavor collection may be accomplished by, for example, vacuum-assisted evaporative means, including the recovery of components using cryo traps. The deodorized liquid extract might be concentrated to a suitable total solid (TS, typically around 50%) by processes such as evaporation and freeze concentration, or the like. The concentrated liquid extract can then be combined with the volatile flavor fraction to be dried by processes such as spray drying, freeze drying, or the like. If desired, the previously separated flavor may then be added back to the residue (e.g., by coating, soaking, infusing, etc.). Instantizing may also be accomplished with or without separating volatile flavor components prior to drying, by using, for example, refractance window drying, and/or microwave assisted techniques, etc. Forming/Pelletizing Liquid, slurry, or powder materials may be formed, prior to packaging, into shapes, useful for or desired by the end-user, by processes such as agglomeration, granulation, extrusion, or the like. These include, but are not limited to, spheres, lozenges, coffee bean-like shapes, or other shapes that are easily ground, grated, shaved, or otherwise prepared for subsequent extraction to form a coffee beverage or incorporation into another food or beverage item (e.g., a powdered coffee topping). Such formed items may then be further coated with other ingredients to improve their utility or usable shelf-life. These include, for example, anti-caking agents to prevent sticking, or barrier materials to limit the diffusion of aroma compounds out of the formed product and thus preserve the shelf-life of the flavor and aroma. Such coatings may be functional in the beverage as well as for the above purposes, for example a powdered colorant or flavor, a gum that hydrates when water is added. 63 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Formed items may be subjected to thermal processing, as further detailed in the “Thermal Processing” section. Packaging Products may be filled into packaging appropriate to their format (such as, for example, cans, bottles, jars, bags, boxes, and the like) prior to, after or in the absence of thermal processing. Packaging can be single serve, multi-serve, bulk, industrial, or any other reasonable format. The product entering the packaging need not be “complete” per se when it is added to the container. For example, liquid nitrogen can be added before sealing the pack to both produce an inert headspace or to produce nitrogen bubbles when the pack is opened. Other gases, or alternate phases of compounds that are gaseous at room temperature, e.g., dry ice, may be added (e.g., for purposes such as prolonging shelf- life/excluding oxygen). Thermal Processing Final thermal processing may be conducted to ensure product quality and/or safety. The specifics may depend on the format of the product. As discussed in the “Cross-reaction” section, such thermal processing methods generate flavors and can be used not only to make a safe, lasting product, but also to drive desired changes to produce a final composition in a pack. Liquid products, such as RTD beverages, concentrates, liquid flavors, etc., may be subjected to one or more of a sterilization process (e.g. UHT, retort, microwave, ohmic), a pasteurization process (e.g. HTST), a homogenization process, or non-thermal antimicrobial treatments (e.g. HPP, irradiation) etc., chilling, freezing, and/or other methods not enumerated herein that are useful or sufficient to mitigate microbial risk (if required or desired). These methods may be, or include, in-container heat treatments. Alternatively, filling may occur after heat treatment. Solid or powdered products, such as grounds, single use capsules, restructured or substitute “beans,” and the like, may likewise be heated before or after being placed in their packaging materials if necessary or desired to produce a particular composition. For compositions having a sufficiently low aW (e.g., grounds or formed solids with pre- conditioned moisture levels), heating may not be necessary. However, as discussed previously, this final heating step may nonetheless be utilized to produce final flavors in 64 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 a sealed container, which prevents their egress. Additionally, depending on the nature of ingredients added during formulation, thermal means may be used to remove solvents. Augmented and/or Modified Coffee Substrates or Derivatives Thereof As stated above, the inventive methods are not only applicable to non-coffee substrates, but also provide for improving the organoleptic qualities of a low-quality, flawed, or depleted (e.g., previously extracted or ‘spent’ grounds) coffee material. For example, coffee (e.g., a low quality or flawed coffee) may be used in the methods disclosed herein as the substrate carrier material having an endogenous Maillard-reactive nitrogen constituent and/or an endogenous Maillard-reactive carbohydrate constituent, and may be reacted with an exogenous Maillard reagent comprising an exogenous Maillard-reactive nitrogen constituent and/or and exogenous Maillard-reactive carbohydrate constituent to provide a conditioned coffee substrate carrier material, which may be, for example, dried, roasted, etc., to provide cross- Maillardized beverage components made from coffee. In additional such aspects, traditional, low-quality, or depleted (e.g., previously extracted or ‘spent’ grounds) coffee material, or spent non-coffee material may be rejuvenated/regenerated/reformulated, for example, by addition of exogenous cross- Maillardized beverage components (e.g., concentrated extracts) made from coffee or from non-coffee substrate materials. Such regeneration/reformulation of spent coffee grounds, for example, may be performed as described above in relation to the above- described “Work-up” and “Finishing” steps, wherein e.g., exogenous cross-Maillardized concentrate extracts or flavors, etc. may be added to dried, traditional spent coffee grounds, or spent non-coffee materials, optionally along with other additives to provide for finished regenerated/reformulated coffee grounds, or finished non-coffee materials, which may then be extracted to provide for organoleptically satisfying beverage products. In preferred aspects, these regeneration/reformulation methods provide a solution for recycling traditional spent coffee grounds on a commercial scale. In further methods, spent grounds from non-coffee substrate materials processed by the disclosed methods (cross-Maillardized or not), can likewise be regenerated/rejuvenated. DEFINITIONS. Unless otherwise noted, terms are to be understood according to conventional usage by those of ordinary skill in the relevant art. “Maillard-reactive nitrogen constituent,” as used herein, refers to nitrogen 65 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 constituents (e.g., of one or more of amino acids, peptides, oligopeptides, polypeptides, and/or proteins) that may react to form conjugates thereof with a Maillard-reactive carbohydrate constituent (e.g., sugars (mono-, di-, oligo- or polysaccharides), organic acids, and phenolic compounds. “Maillard-reactive carbohydrate constituent,” as used herein, refers to carbohydrate constituents (e.g., mono-, di-, oligosaccharide, and/or polysaccharides) and/or derivatives thereof covalently bond to other constituents (e.g., organic acids, phenolic acids) that may react with a Maillard-reactive nitrogen constituents to form conjugates thereof (e.g., Amadori and/or Heyns compounds). Maillard-reactive carbohydrate constituents preferably comprise a reducing function (e.g., carbonyl group, reducing sugar), however, non-reducing sugars (e.g., saccharose) may also be converted to reducing components (e.g., glucose and fructose) by hydrolysis or heat treatment. “Exogenous Maillard reagent,” as used herein, refers to an agent that is added or placed to be in contact with the substrate carrier material for purposes of forming one or more cross-Maillardization reaction products with Maillard-reactive moieties/groups that are endogenous to the substrate carrier material. In particular substrate-transactivation aspects, a substrate carrier material may be initially treated with one or more agents (e.g., enzymes, etc.) that may render (activate) or expose otherwise non-Maillard-reactive endogenous moieties/groups as Maillard-reactive endogenous moieties/groups (e.g., transactivation, by exposing and/or releasing them from the substrate material) and in such cases the trans-activated Maillard-reactive endogenous moieties/groups may cross-react with other endogenous Maillard-reactive groups, in which case such trans- activated Maillard-reactive moieties/groups may be considered as exogenous Maillard reagents. “Substrate carrier material,” as used herein, refers to a carrier material (e.g., natural and/or a processed or restructured plant material) having an endogenous Maillard-reactive nitrogen constituent and/or an endogenous Maillard-reactive carbohydrate constituent. Preferably, the substrate carrier material comprises an insoluble natural and/or a processed or restructured plant material having an endogenous Maillard-reactive nitrogen constituent and/or an endogenous Maillard- reactive carbohydrate constituent. “Conditioned substrate carrier material,” as used herein, refers to a substrate carrier material having an endogenous Maillard-reactive nitrogen constituent and/or an 66 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 endogenous Maillard-reactive carbohydrate constituent, which substrate has been contacted with an exogenous Maillard reagent comprising an exogenous Maillard- reactive nitrogen constituent and/or and exogenous Maillard-reactive carbohydrate constituent under conditions sufficient to provide for cross-reaction products, preferably cross-Maillard reaction products, formable by the reaction between the exogenous Maillard reagent, and the endogenous Maillard-reactive constituent(s). Preferably, a conditioned substrate carrier material is one which is cross-reacted and/or cross- Maillard-reacted. “Water activity (a
w),” as used herein, refers to the art-recognized meaning, e.g., the partial vapor pressure of water in a substance divided by the standard state partial vapor pressure of water. In the field of food science, the standard state is most often defined as the partial vapor pressure of pure water at the same temperature. Using this particular definition, pure distilled water has a water activity of exactly one. “Cross-Maillardized substrate carrier material,” as used herein, refers to a substrate carrier material (e.g., having been at least conditioned as described herein) having cross-Maillard reaction products formed by the reaction between the exogenous Maillard reagent(s), and the endogenous Maillard-reactive constituent(s). These reactions take place more readily at elevated temperatures (e.g., >60°C) and low water activity (e.g., <0.8) depending on the availability of the Maillard reactants. The cross- Maillardization products can be volatile or non-volatile, or even of polymeric nature. It is generally known that, at a given temperature, the MR rate increases with decreasing water activity. “Natural plant material,” as used herein, includes but is not limited to those exemplary plant materials listed herein that come from plants, and may include restructured (e.g., fragmenting, grinding, milling, micronizing, depolymerizing (e.g., chemically, enzymatically, etc.), solubilizing, permeabilizing, compacting and/or compressing) plant material. “High water activity cross-Maillard reaction products,” “high aw cross-Maillard products,” or “HWACMP,” as used herein, refer to cross-Maillard reaction products formed with the substrate carrier material under preconditioning water activity (aw) reaction conditions providing a conditioned (pre-conditioned) substrate carrier material as referred to herein. Operationally, high a
w at the conditioning reaction step is selected to be higher than that resulting from adjusting the water activity (a
w) of the conditioned substrate carrier material to a value less than that of the conditioning reaction. “Low water 67 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 activity cross-Maillard products,” “low aw cross-Maillard products,” or “LWACMPs),” as used herein, refer to cross-Maillard reaction products formed with the substrate carrier material, under conditions of a
w less than that of the conditioning (a.k.a.; pre-conditioning) reaction. Preferably, LWACMPs are those cross-Maillard reactions formed with the substrate under conditions of aw less than or equal to, e.g., 0.85 (or, e.g., to less than or equal to another exemplary value as recited in clauses 12, 52 and 65) by reaction between an endogenous Maillard-reactive nitrogen constituent and/or an endogenous Maillard-reactive carbohydrate constituent, and an exogenous Maillard reagent comprising Maillard-reactive nitrogen and/or Maillard-reactive carbohydrate. “Elevated temperature, low water activity cross-Maillard products,” “elevated temperature, low aw cross-Maillard products,” or “ET-LWACMPs),” as used herein, refer to cross-Maillard reaction products formed with the substrate carrier material under conditions of temperature greater than that used for generating the LWACMPs. “Pulp/pomace” as used herein, refers to the remains of a plant product from a juice extraction process, except when otherwise specified. “Fiber” as used herein, generally refers to the dried pulp/pomace of a plant product, optionally with further processing to remove some portions of the plant (ex: seeds) while retaining the bulk of the insoluble portion of the plant flesh. “Bean-less” as used herein, refers to a coffee composition containing no seeds of the Coffea genus, nor products thereof. In particular embodiments bean-less coffee is prepared by cross-Maillardization (xMR) (e.g., xMR date seeds, prepared as described in working examples 1 and 2 of PCT Patent Application PCT/US2021/025565, published as WO 2021/202989). “GalAc” as used herein, refers to galacturonic acid. “Gal” as used herein, refers to galactose. “Ar” as used herein, refers to arabinose. “Xy” as used herein, refers to Xylose. “Rh” as used herein, refers to rhamnose. “HG” as used herein, refers to homogalacturonan, a homopolymer of galacturonic acid, or a polymer block comprising purely galacturonic acid. “RG” as used herein, refers to rhamnogalacturonan, a pectic polysaccharide block comprising galacturonic acid and rhamnose. RGs are observed in two primary formats in plant tissues. These forms are termed RG-I and RG-II (see FIG.18; taken from Zdunek, A., Pieczywek, P. M., & Cybulska, J. (2021), The primary, secondary, and structures of 68 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 higher levels of pectin polysaccharides), Comprehensive Reviews in Food Science and Food Safety, 20(1), 1101-1117., Chicago). In the context of this disclosure, “RG” refers to the portions that comport with the general RG structures, whether those portions are part of larger pectic structures that contain non-RG blocks, or whether those portions have been removed/isolated from larger structures, or if these portions have been further broken down (by, for example but not limited to, alkaline hydrolysis) into smaller portions/structures (oligomers, monomers) that still maintain the composition requirements of RG: RG-I has a backbone comprising an alternating copolymer of galacturonic acid and rhamnose. The rhamnose moieties can themselves be the sites of sidechains or branches from the main chain. These sidechains are generally composed of arabinose, galactose, mixtures thereof, and may be linear or branching themselves. RG-II resembles most other pectins in that the backbone is a homopolymer of galacturonic acid. Side chains in RG-II include apiose, rhamnose, arabinose, fucose, galactose glucuronic acid, aceric acid, as well as galacturonic acid. “Chlorogenic acids” or CGAs, as used herein, refers to the group of polyphenol ester and di-ester compounds formed from the combination of quinic acid with 1 or 2 hydroxycinammic acids (e.g. caffeic acid, ferulic acid, p-coumeric acid). Exemplary compounds include but are not limited to 3-, 4- and 5-O-caffeoylquinic acids, 3-, 4- and 5-feryloylquinic acids, 3,4-, 3,5- and 4,5-dicaffeoylquinic acids. Furthermore, the thermal breakdown products of these mono- and di-esters, including but not limited to 3-, 4- and 5-chlorogenic acid lactones, are also considered CGAs within the scope of this invention. EXAMPLES The following non-limiting examples are provided to further illustrate embodiments of the invention disclosed herein. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent approaches that have been found to function well in the practice of the invention, and thus can be considered to constitute examples of modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments that are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. 69 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Table 1. Summary of Examples
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Example 1 (Bean-less coffee-substitute beverage products were made from cross-Maillardized date seeds) This example describes making several products initially based on cross- Maillardized date kernels: a) cross-Maillardized date seed extract; b) formulated grounds 73 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 from spent grounds; c) formulated spent grounds extract, d) formulated grounds from roasted grounds; and e) an extract from the formulated roasted grounds: a) cross-Maillardized date seed extract: A bean-less, coffee-substitute beverage was made from a cross-Maillardized date seed extract, as follows: In a first reaction, dry date kernels (e.g., 67 g Deglet Nour; cleaned of excess date flesh, stems and calyces) were added to an aqueous Maillard solution (containing 1% glycine (Ajinomoto), 1% arginine (Ajinomoto), 1% fructose (Tate and Lyle), and adjusted to pH 9.7 with KOH), and reacted for 3 hours at 85°C; Liquid and solids of the first reaction were separated via filtration, and the liquid discarded; The solids were dried at or below 55°C to < 12% moisture (a
w <0.4, approx.15 hrs), to provide a dried, conditioned substrate carrier material; The dried kernels were roasted 4 minutes with predefined temperature profile to finished temp of 217°C, then cooled to provide roasted dried kernels (preferably, the date seeds are roasted to temperatures between 180-220 ºC); The cooled roasted date kernels were ground; milled fine (D10 ca.200 μm, D50 ca.500 μm, D90 ca.800 μm), extracted (brewed) in 92 °C water for 4 minutes before gravity filtration to provide a liquid extract fraction (liquid coffee-substitute base extract fraction) and a retentate extracted grounds fraction (spent grounds fractions), followed by cooling of the liquid extract fraction to 4 °C for storage. For final formulation, the date seed liquid coffee-substitute base extract was combined with caffeine, colorants, gums and flavors, filled into cans with nitrogen, and retorted, providing a beverage with notable coffee-like roasted flavors, as determined by sensory analysis (e.g., as in Example 8). b) formulated grounds from the spent grounds fraction of a): The retentate grounds from the production of the coffee beverage in a) were dried by lyophilization. The resulting dry grounds were mixed with caffeine, flavors and colors in powder form and blended thoroughly. c) formulated spent grounds extract: The resulting formulated grounds were then placed in the portafilter of an espresso machine, tamped with 100 N tamping force and extracted for 15 seconds at 93 °C, 9 bar to provide an extract of the formulated spent grounds. 74 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 d) formulated grounds from roasted grounds: Roasted, ground kernels, as described in a), are combined with caffeine, colors and flavors all in powder form, and mixed to provide formulated roasted grounds. e) extract from the formulated roasted grounds of d): The dry mix (formulated roasted grounds from d)) is blended well, then 20 g is placed into a paper filter cone supported above a mug. Hot water (95 °C) is poured slowly over the grounds until 180 g of total water have been added, and the extract collected, providing a beverage with notable coffee-like roasted flavors and aromas, as determined by sensory analysis (e.g., as in Example 8). Example 2 (A bean-less coffee-substitute beverage was made from a cross-Maillardized date seed extract) Raw, cleaned dried dates were combined with fructose, glycine, and aspartic acid at levels of 98.5% / 0.5% / 0.5% / 0.5% in pH 8.5 water and incubated at 85
oC for 3 hours. The dates, separated from the liquid fraction, are then dried and roasted to a finished temperature of 218
oC. The roasted seeds were then ground and extracted (95
oC / 4 minutes, 90% water, 10% kernels). By organoleptic comparison (as determined by sensory analysis as in Example 8), the extract prepared from date kernels that were processed in the same conditions but with no exogenous reagents, was cloudier, more astringent, and contained less roasted coffee-like character. Example 3 (A bean-less coffee-substitute beverage was made from a cross-Maillardized Chicory root extract) Dried chicory root is crushed / ground to yield pieces < 1 cm in diameter, then combined with a mixture of 1% lysine, 1% leucine, 1% phenylalanine, 0.1% cysteine, and 5% glucose (exogenous Maillard reagents). This mixture is blended with equal parts water to form a paste, which is then dried to aw < 0.6 at 75
oC. The resulting cake is then roasted at 150
oC for 30-60 minutes, then ground and extracted (95
oC / 4 minutes). In organoleptic comparison (as determined by sensory analysis as in Example 8) to chicory root alone (processed without the exogenous Maillard reagents), the resulting cross- Maillardized beverage is darker, with a richer and more roasted aroma, including with notes of chocolate. 75 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Example 4 (A bean-less coffee-substitute beverage is made from a cross-Maillardized Yerba mate extract) Yerba mate leaves and stems are soaked in an equal mass of a solution of 0.5% leucine, 0.5% lysine, and 2% glucose. The substrate is drained and dried to aw < 0.4 at 55 ºC, then toasted at 150 ºC in an oven for 10 minutes. The toasted substrate is extracted in 70 ºC water, then cooled to room temperature before washing with a neutral oil. Example 5 (A bean-less coffee-substitute beverage was made from a cross-Maillardized mustard seed extract) Defatted mustard seed powder (97.4%) was mixed with 1% glucose, 1% glycine, 0.5% chlorogenic acid and 0.1% sodium bicarbonate with just enough water to form a paste (roughly 20% of the dry ingredient mass), then dried below aw < 0.4. The dried mixture was then roasted to 200 ºC over a 5 minute temperature ramp, cooled, extracted for 4 minutes using 95 ºC water at a ratio of 90% water/10% seeds, and then filtered. In organoleptic comparison (as determined by sensory analysis as in Example 8) to a beverage prepared using roasted mustard seed powder alone, the resulting beverage contains more roasted and nutty coffee-like aroma with an increased bitterness and a decreased mustard aroma. Example 6 (A bean-less coffee-substitute beverage is made from cross-Maillardized watermelon seeds, pumpkin seeds, Jerusalem artichoke, and/or roasted sesame) An extract (coffee-substitute beverage component) is prepared starting with a substrate comprising watermelon seeds, pumpkin seeds, Jerusalem artichoke, and/or roasted sesame. The plant material and respective extract, in each case, is prepared in accordance with the cross-Maillardization reaction methods, and other examples described herein. Example 7 (Exemplary composition prepared by combining portions (90%:5%:5%) of respective extracts prepared from cross-Maillardized date kernels, chicory root, and yerba mate) 76 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 A mixed extract (coffee-substitute beverage component) is prepared by combining portions (90%:5%:5%) of respective extracts prepared from cross-Maillardized date kernels, chicory root, and yerba mate, each extract prepared in accordance with the cross-Maillardization reaction methods, and other examples described herein. Example 8 (Sensory analysis was conducted on exemplary compositions) The disclosed cross-Maillardization methods (employing substrate preconditioning reactions with exogenous Maillard reagents) and compositions provide important and crucial components normally found in coffee. The participants in the cross- Maillardization conditioning reactions may be: a substrate carrier material comprising or derived from an agricultural product; and exogenous Maillard reagents (e.g., carbohydrates and/or peptides, etc.) that react with the substrate constituents to create, directly and indirectly, the essential compounds for a coffee-substitute beverage. As described herein, these substrates and reagents may or may not be comprised of or derived from coffee. Additional important steps may comprise, inter alia, a moisture conditioning (or water activity modulating) step (e.g., drying, or alternatively moisturizing of the conditioned substrate carrier material), and/or subsequent a heating (e.g., roasting) step. As described above in more detail at pages 14 and 15, exemplary desirable compounds of interest may be placed into 5 exemplary categories, which in each case can be further divided into subsets of related compounds that perform similar functions in the finished beverage. In this example, several exemplary extract compositions were prepared in accordance with the disclosed cross-reaction (e.g., cross-Maillardization reaction) methods, to demonstrate the creation of some of these categories of aroma compounds: Sample Preparation and Sensory Analysis a) Date Kernel extract preparation: Dried raw, cleaned date kernels are combined with fructose, glycine and aspartic acid at levels of 98.5%/0.5%/0.5%/0.5% in pH 8.5 water and processed at 85 ºC for 3 hours. The conditioned date kernels are then dried to aw < 0.4 and roasted to a finished temperature of 218 ºC. The conditioned, dried, roasted kernels are ground, and extracted (95 ºC for 4 minutes, 90% water, 10% kernels). In comparison to extract from date kernels processed in the same manner/conditions but with no exogenous reagents, the resulting 77 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 sample prepared by the cross-Maillardization method is less cloudy and less astringent and contains a more roasted, coffee-like character. b) Chicory Root and Buckwheat extract preparation Small, dried chicory root pieces < 1 cm in diameter (18.4% of the composition) were combined with raw buckwheat (73.5%) lysine (1%), leucine (1%), phenylalanine (1%), cysteine (0.1%) and glucose (5%). This mixture was blended with just enough water to form a paste (roughly 20% the mass of dry ingredients), which was then dried to aw < 0.3 at 75 ºC. The resulting cake was then roasted at 190 ºC for 10 minutes, ground and extracted (95 ºC/4 minutes, 90% water, 10% grounds). In comparison to chicory root alone processed in the same manner/conditions but with no exogenous reagents, the resulting beverage prepared by the cross-Maillardization method was darker, with a richer and more roasted aroma with notes of chocolate. c) Mustard Seed extract preparation Defatted mustard seed powder (97.4%) was mixed with 1% glucose, 1% glycine, 0.5% chlorogenic acid and 0.1% sodium bicarbonate with just enough water to form a paste (roughly 20% of the dry ingredient mass), then dried below a
w < 0.4. The dried mixture was then roasted to 200 ºC over a 5 minute temperature ramp, cooled, and extracted for 4 minutes using 95 ºC water at a ratio of 90% water/10% seeds and filtered. In comparison to a beverage prepared using roasted mustard seed powder alone, processed in the same manner/conditions but with no exogenous reagents, the resulting beverage prepared by the cross-Maillardization method contains more roasted and nutty, coffee-like aroma, and with an increased bitterness and a decreased mustard aroma. d) Watermelon Seed extract preparation Watermelon seeds were toasted at 160 °C for 10 minutes to reduce water activity to < 0.2. The toasted seeds were ground and the derived powder (20%) was mixed with 8% glucose, 0.8% lysine, 0.8% proline and 0.1% cysteine, blended in water (70.3%) with pH adjusted to 8.5. The mixture was then heated at 75 °C for 24 hours. The thickened reaction mixture was spread out and dried to < 0.2 aw. The dried material was then roasted in an electric oven at 190 °C for 10 minutes, and after cooling, the roasted residue was extracted with water (95 °C/4 min, 10% grounds, 90% water). The resulting beverage had a more sulfury, roasted-like aroma and darker color, compared to a beverage derived from watermelon seeds alone, processed in the same manner/conditions but with no exogenous reagents. e) Mixed Substrate extract preparation 78 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 A mixed composition comprising raw, cleaned date kernels and white mustard seeds was combined with fructose, glycine and aspartic acid at levels of 93.5%/5 %/0.5%/0.5%/0.5% in pH 9.7 water and processed at 85°C for 3 hours. The mixture was then dried to aw <0.3 and roasted to a finished temp of 200 °C. The roasted seeds were ground, and extracted (95 °C/4 minutes, 90% water, 10% kernels). In comparison to a mixture of date kernels and mustard seeds alone that were processed in the same conditions but with no exogenous reagents, the resulting sample is less astringent and contains a more caramel and sulfury, coffee-like aroma. In addition to the above-described sensory/organoleptic evidence, the results described in the following chemistry working Examples 9-12 analyzing the above- described compositions, a)-e), provide additional strong evidence for the cross-reactions (e.g., cross-Maillardization) between substrate and exogenous reagents, in the conditioned, a
w adjusted (e.g., dried), heated (e.g., roasted), extracts and residual extracted material. Example 9 (The cross-Maillardization reaction was shown, relative to controls, to differentially affect the levels of 2,5-Dimethylpyrazine (2,5-DMP) production in different stages of the disclosed methods) In this example, the disclosed cross-Maillardization methods (employing substrate preconditioning reactions with exogenous Maillard reagents) were shown, relative to controls, to differentially provide or enhance important components normally found in coffee. Analytical Characterization – Gas Chromatography / Mass Spectrometry. 2,5-Dimethylpyrazine (2,5-DMP) is a volatile compound well known to contribute to roasted coffee flavor. Specifically, 2,5-DMP is known to contribute to the roasty and earthy flavors of coffee. It was selected for further quantification in the present methods as it is indicative specifically of the Maillard Reaction, and not of simple sugar breakdown (i.e., caramelization). Reactivity between an amino acid source and a carbohydrate source is required to produce this compound. Moreover, 2,5-DMP can be produced from nearly any combination of amino acid and carbohydrate, and thus the selection of amino acids and carbohydrates, as well as the substrate, may influence the rate of formation and the final concentration of 2,5-DMP. Accordingly, stages of the above-described methods leading to compositions a)-e) of Example 8, were analyzed, relative to controls, 79 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 for the generation of 2,5-DMP, and the disclosed cross-Maillardization methods were shown, relative to controls, to differentially provide or enhance important components normally found in coffee. For data collection, each sample (crossMR, control, preconditioning solution and blank) was analyzed by means of Headspace SPME GC/MS (Agilent 5975 MSD, Agilent, Santa Clara, USA). The samples were worked up in a triplicate. For analysis, an aliquot of 5 mL of each sample was transferred into a headspace vial. The Vials were sealed and placed into a cooled (4 °C) autosampler (MSP, Gerstel, Muehlheim an der Ruhr, Germany). The samples were extracted using an SPME fiber (57298-U, 50/30 µm DVB/CAR/PDMS, Stableflex, 1 cm, Supelco, Bellefonte, USA) and transferred on the column in ‘splitless’ mode. The chromatography was carried out using a Stabilwax column (60m, 0.32mm ID, 1µm df, RESTEK, Bellefonte, USA) and a temperature gradient, with an initial temperature of 35 °C and an increase of 7.5 °C/min until a total of 250 °C, holding the final temperature for 5 min. Helium was used as carrier gas. As detector, a single quad mass spectrometer was used. The compounds were ionized using EI in positive mode. The identification of the individual compounds was performed using the NIST-17 library. Data analysis was performed using python v3.7, MS Dial v.4.33 (Yokohama City, Japan) and Masshunter v11 (Agilent, Santa Clara, USA). 2,5-DMP was identified using an internal database and the NIST-11 database and was semi-quantified by normalizing the mass spectrometer intensities (in cps) of 2,5- DMP (mass to charge ratio, m/z, = 109.07 [M+H]
+) with the sample weights. Extract composition a) (prepared from cross-Maillardized date kernels, as described in Example 8 above, was analyzed in comparison to controls to determine if cross reactivity between the substrate carrier material (date kernels) and exogenous Maillard reagents took place. The controls for these experiments comprised both kernels alone and the exogenous reagents alone, processed in otherwise identical fashion. More specifically, the kernels alone (“Control”) were preconditioned in pH 8.5 water at the same temperature and time, but lacked any exogenous reagents. The exogenous reagents alone (“MR”) were preconditioned in a pH 8.5 bath at the same temperature and time, but in the absence of kernels. In each case, the sample workup was performed in duplicate. The samples were prepared identically and were each measured after the preconditioning step (heating in aqueous solution, pH 8.5, 3 hours), after drying (65 °C/15 hours), after roasting (IKAWA 80 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Roaster, 210 °C/7 min), after extraction (18 g/100 mL, immersion brew at 95 °C/4 min) as well as the residue (extracted filtrate residue, dried at 65°C/4 hours). Results. In general, it was found that cross-Maillardization reactions resulted in differential generation of low levels of 2,5-DMP in the preconditioning and drying steps (see Figure 2). Cross reaction between exogenous reagents and substrate were observed, as evidenced by the differentially elevated levels of 2,5-DMP generated when substrate and reagents are reacted together, relative to controls. Furthermore, it was found that the level of 2,5-DMP generated during the thermal reaction step (“Roast”) was substantially greater in the sample containing both substrate and exogenous reagents (“CrossMR”), relative to the control samples combined. This is compelling evidence for a cross-Maillard reaction between these endogenous and exogenous groups, since the “CrossMR” level of 2,5-DMP far surpasses the value of the independent reaction of kernel components amongst themselves combined with that of the exogenous materials amongst themselves. Note for purpose of Figure 2, the “MR” values were normalized by taking into account that approximately 11.25 % (as determined by mass analysis) of the exogenous Maillard reagents were present (absorbed by the substrate) in the post- conditioned, separated and dried substrate material. Similar experiments were conducted across all example compositions. The differentially increased 2,5-DMP yield was not universal across all examples (see Figure 3, comparing the roasting stage values among the different substrates). Specifically, and surprisingly, while some combinations of reagents and substrates showed a differential increase in 2,5-DMP yield (i.e., as in examples a), d), and e), relating to date seed, watermelon seed, and mixed substrates, respectively), others showed a decrease in yield (as in b and c). According to particular aspects of the invention, therefore, selection of substrate and reagents may be used in the inventive methods to produce the desired type of products, such as volatile aroma compounds yielding roasted, fruity, etc. notes. As in Figure 2, the normalized values for the “MR” samples (exogenous reagents alone) in these cases were negligible, and thus are not shown in Figure 3). Careful selection of substrate and reagent, therefore, provides flexibility in producing desired final products, and surprisingly, combination of some substrates and exogenous Maillard reagents can result in a decreased yield of one or more particular, potentially desired compounds. Example 10 81 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 (The cross-Maillardization reaction was shown, relative to controls, to differentially affect the levels of diacetyl production in different stages of the disclosed methods) In this example, relating to compositions a)-e) of Example 8, the disclosed cross- Maillardization methods (employing substrate preconditioning reactions with exogenous Maillard reagents) were shown, relative to controls, to differentially provide or enhance important components normally found in coffee. According to additional aspects of the present invention, reactions can occur in the disclosed cross-reactions systems wherein reactants of substrate and exogenous systems interact, but do not ultimately result in compounds formed directly therefrom (i.e., do not result in direct cross-reaction product molecules). For example, the presence of both the substrate and the exogenous reagents may, indirectly (e.g., by affecting the reaction pathway leading to a desirable compound), enhance the generation of desirable compounds even if that desirable compound is not the direct, or even indirect, reaction product of a reaction between endogenous and exogenous reagents. For example, 2,3-butanedione is an art-recognized marker for caramelization reactions as well as the Maillard reactions, and its formation involves mainly carbon atoms of the carbohydrate source. 2,3-Butanedione was identified using an internal database and the NIST-11 database and was semi-quantified by correcting the mass spectrometer intensities (in cps) of 2,3-butanedione (m/z = 87.09 [M+H]
+) with the sample weights (in g). For data collection, each sample (crossMR, control, preconditioning solution and blank) was analyzed by means of Headspace SPME GC/MS (Agilent 5975 MSD, Agilent, Santa Clara, USA). The samples were worked up in triplicate. For analysis, an aliquot of 5 mL of each sample was transferred into a headspace vial. The Vials were sealed and placed into a cooled (4 °C) autosampler (MSP, Gerstel, Muehlheim an der Ruhr, Germany). The samples were extracted using an SPME fiber (57298-U, 50/30 µm DVB/CAR/PDMS, Stableflex, 1 cm, Supelco, Bellefonte, USA) and transferred on the column in ‘splitless’ mode. The chromatography was carried out using a Stabilwax column (60m, 0.32mm ID, 1µm, RESTEK, Bellefonte, USA) and a temperature gradient, with an initial temperature of 35 °C and an increase of 7.5 °C/min until a total of 250 °C, holding the final temperature for 5 min. Helium was used as carrier gas. A single quad mass spectrometer was used for detection. The compounds were ionized using EI in positive mode. The identification of the individual compounds was performed using the NIST-17 library. Data analysis was performed using python v3.7, MS Dial v.4.33 (Yokohama City, Japan) and Masshunter v11 (Agilent, Santa Clara, USA). 82 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 As demonstrated in Figure 4, the presence of exogenous amino acids and substrate/carrier material impact the formation rate and reaction kinetics of 2,3- butanedione in the CrossMR samples, whether they are explicitly part of the reaction pathway (Maillard) or not (caramelization). As in Figures 2 and 3, the normalized values for the “MR” samples (exogenous reagents alone) in these cases were negligible, and thus are not shown in Figure 4). As demonstrated in Figure 4, the presence of exogenous amino acids and substrate/carrier material impact the formation rate and reaction kinetics of 2,3- butanedione in the CrossMR samples, whether they are explicitly part of the reaction pathway (Maillard) or not (caramelization). As in Figures 2 and 3, the normalized values for the “MR” samples (exogenous reagents alone) in these cases were negligible, and thus are not shown in Figure 4). According to particular aspects, therefore, flavorful aroma compounds are differentially produced resulting from the interaction of exogenous and substrate materials using the inventive methods. Example 11 (The cross-Maillardization reaction was shown, relative to controls, to differentially affect the cellular structure of the conditioned substrate carrier material) In this example, the disclosed cross-Maillardization methods (employing substrate preconditioning reactions with exogenous Maillard reagents) were shown, relative to controls, to differentially affect the cellular structure of the conditioned substrate carrier material. Following the procedure for generating the extract composition of example a) of above Example 8 (date kernels), a crossMR sample (date kernels conditioned with exogenous reagents) and a control (date kernels alone) were prepared. Both the conditioned samples and the control were drained and then dried at 65 °C for 15 hours. Dried samples were fractured to expose the inner structures, and the fractured samples analyzed by means of scanning electron microscopy (SEM) using an FEI Quanta FEG- SEM at 2 kV accelerating voltage. The differences between control and combined samples are readily observable using such imaging conditions (see Figures 5A-5D). Kernels preconditioned without exogenous reagents show an open, porous structure (panels A, C), whereas kernels preconditioned with the exogenous reagents show a relatively denser, fuller structure 83 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 (panels B and D). The images in Figure 5 show changes in the cellular structure mediated by the cross-Maillard reaction, wherein the Control (panels A, C) samples show a highly porous structure, whereas CrossMR (B and D) samples exhibit a more dense and fuller cellular structure. This suggests the entry of the reagents into the kernel tissues, consistent with cross-reactivity, particularly given the short lifetimes of some Mallard intermediates. If the exogenous reagents only existed on the outer surface(s) of the substrate, cross reactions would be significantly limited and independent reactions of exogenous reagents and substrate tissues would more likely predominate. As evidenced by the above-described 2,5-DMP data, however, significant cross-Maillardization reaction products are produced by this combination. Example 12 (1,3-bis[(5S)-5-amino-5-carboxypentyl]-4-methyl-1H-imidazol-3-ium (imidazolysine) production was shown, relative to controls, to be differentially regulated by the disclosed cross-Maillardization reaction) In this example, the disclosed cross-Maillardization methods (employing substrate preconditioning reactions with exogenous Maillard reagents) were shown, relative to controls, to differentially regulate production of 1,3-bis[(5S)-5-amino-5-carboxypentyl]-4- methyl-1H-imidazol-3-ium (imidazolysine). Liquid Chromatography / Mass Spectrometry The liquid extract composition of example a) (prepared from date kernels) of above Example 8, was analyzed in comparison to controls to look for cross reactivity. The kernels alone (“Control”) were preconditioned in pH 8.5 water at the same temperature and time, but lacked any exogenous reagents. The exogenous reagents alone (“MR”) were preconditioned in a pH 8.5 bath at the same temperature and time, but in the presence of no kernels. The sample workup was performed in duplicate. The samples were prepared identically and were each measured after the preconditioning step (heating in aqueous solution, pH 8.5, 3 hours), after drying (65 °C/15 hours), roasting (IKAWA Roaster, 210 °C/7 min) and extraction (18 g/100 mL, immersion brew at 95 °C/4 min followed by gravity filtration). The individually prepared extract samples were then prepared for analysis by diluting each sample to a concentration of 1 mg/mL, followed by membrane filtration. The analysis was performed by means of a high-resolution ultra-performance liquid 84 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 chromatography system, coupled to an ion mobility time of flight mass spectrometer for detection, and 2 μL of each sample (biological duplicate (two separate workups), five injections each, technical quintuplicate) were injected for analysis. The measurement was performed in electrospray ionization (ESI) in both, positive and negative mode. The data was then evaluated by statistical tools, such as principal component analysis (PCA) and partial least square analysis (PLSA). More specifically, for data collection each sample (crossMR, control, preconditioning solution and blank) was analyzed by means of UPLC-ToF/MS (Agilent 6500 Q-ToF, Agilent, Santa Clara, USA). Each of the samples was worked up in duplicate and injected five times for profiling analysis. An aliquot of 5 mL of each sample was diluted 1:1000 (v/v, sample/Millipore water), filtered (Minisart Syringe Filter, pore size 0.22 µm, Sartorius, Goettingen, Germany) and transferred into LC vials. The vials were placed into the autosampler of the device and an aliquot of 2 µL was injected. The chromatography was carried out using an RP-18 column (Kinetex 1.7 µm C18100 Å, 100 x 2.1 mm, Phenomenex, Aschaffenburg, Germany) as the stationary phase. The stationary phase was preheated at 50 °C. As the mobile phase, water (A, 0.1 % FA, Millipore-Q) and acetonitrile (B, 0.1 % FA, HPLC grade) was used at a flow rate of 0.3 mL/min. The starting conditions were 100% A. After 1 min, B was increased gradually for 4 min to 100% and kept at 100% B for 30 sec. Eluted chromotography samples were ionized using electro spray ionization, and run separately in positive and negative mode. The compounds were identified using their accurate mass, and by their elemental composition, as well as in comparison with internal libaries of reference compounds. Data analysis was performed using python v.3.7, MS Dial v.4.33 (Yokohama City, Japan) and Masshunter v11 (Agilent, Santa Clara, USA). A compound detectable using these techniques is 1,3-bis[(5S)-5-amino-5- carboxypentyl]-4-methyl-1H-imidazol-3-ium; exact mass 341.10999 m/z from negative ESI). The compound might be expected via the breakdown of both exogenous fructose as well as the endogenous glucose (e.g., in date kernels) to methylglyoxal, and its reaction with lysine (from the substrate) to form the dimer. Imidazolysine is a product of prolonged Maillard reaction, and, as known in the case of coffee, primarily contributes its deep yellow-brown color to the roasted beans and beverage. Figure 6 shows, for the liquid extract stages of the samples a) (date kernels) of Example 8, the semi-quantitation of imidazolysine in the “Control,” “CrossMR” and “MR” extract samples. Imidazolysine is only formed in the “CrossMR” samples (2.2 x 10
6 cps/g), whereas the compound was not found in detectable amounts in the “Control” and 85 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 the “MR” sample (Figure 6). This compound is present in conventional coffee, however the yield is relatively lower than in the exemplified extract composition. For example, the level of imidazolysine in a conventional coffee beverage is shown at the far right. According to particular aspects of the present invention, therefore, at least for particular substrate materials, imidazolysine is exclusively formed by the inventive crossMR approach, which provides for production of compounds not attainable by processing of substrates alone, or of exogenous reagents alone. Moreover, the disclosed crossMR approach provides a method of controlling the production level of such compounds (e.g., by varying the concentration/amount of exogenous reagents, exposure time to same, exposure temperature to same, etc.). Example 13 (A coffee-substitute beverage was made from cross-Maillardized cracked date seeds, and the optional use of added chlorogenic acid to the cross-Maillardization preconditioning mixture was shown to enhance the yield of γ-butyrolactone) Cracked date seeds. Prior to the CrossMR process, dry, intact date seeds were cracked into pieces between 2 and 6 mm in diameter. These pieces were then preconditioned (optionally with Eucommia bark extract as a source of chlorogenic acid), roasted, extracted (as in Example 8, composition a)) and analyzed (using SPME-GC/MS) using the same protocol as described in Example 9. The resulting levels of 2,3-butanedione and 2,5-methylpyrazine are summarized in Figure 9, showing that initial cracking of the date seeds prior to preconditioning enhances the yield of cross-Mailladization products. While 2,3-butanedione is a product of multiple chemical pathways, 2,5-dimethylpyrazine is exclusively produced in these systems from a Maillard process. Thus the dramatic enhancement of 2,5-dimethylpyrazine production can be attributed to a significantly greater degree of cross-Maillard reaction taking place when the seeds are initially cracked in the process. Cracked Date Seeds with the addition of Eucommia Bark extract. Prior to the CrossMR process, dry, intact date seeds were cracked into pieces between 2 and 6 mm in diameter. Half of the pieces were preconditioned by first immersing the seed pieces in a solution of 1% fructose, 1% lysine, 0.5% leucine, 0.5% glycine (2:1 w/w ratio solution:cracked seeds). The other half of the pieces were placed in an identical solution, but with the addition of 2.5% chlorogenic acid (sourced from Eucommia ulmoides). Both 86 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 samples were brought to pH 8.5 and then heated to 55 ºC and stirred at that temp for 2 hours. After 2 hours, materials were drained and dried for 15 hours at 55 ºC. Both samples were then roasted, extracted and analyzed (using SPME-GC/MS) using the same protocol as described in Example 9. The resulting levels of 2,3-butanedione and 2,5-dimethylpyrazine are summarized in Figure10A, showing that addition of chlorogenic acid to the preconditioning reaction modulates (in this instance decreases) the level of 2,5-dimethylpyrazine generated. Figure 10B shows that while cross-Maillardization lowers the level of γ-butyrolactone relative to non-cross-Maillardized cracked date seeds (control cracked date seeds), addition of chlorogenic acid to the cross-Maillardization preconditioning mixture enhances the yield of γ-butyrolactone in cross-Maillardized date seeds. Example 14 (A coffee-substitute beverage was made from cross-Maillardized fermented date seeds) Prior to the CrossMR process, date seeds with approximately 10% residual fruit were immersed in twice their combined mass in water and brought to 38ºC. This mixture was covered and allowed to ferment naturally for 48 hours, during which time the fruit was partially digested. After draining and rinsing the remaining fruit, the fermented seeds were dried to aw < 0.6 and preconditioned (optionally with Eucommia bark extract), roasted, extracted and analyzed (using SPME-GC/MS) using the same protocol as described in Example 9. The resulting levels of 2,3-butanedione and 2,5-methylpyrazine are summarized in Figure 11, showing that fermenting the date seeds prior to preconditioning enhances the yield of cross-Maillardization products. As stated above in relation to Example 13, 2,3-butanedione is a product of multiple chemical pathways, whereas 2,5-dimethylpyrazine is exclusively produced in these systems from a Maillard process. Thus the enhancement of 2,5-dimethylpyrazine production can be attributed to a significantly greater degree of cross-Maillardization taking place after subjecting the seeds to a fermentation process. Example 15 (Spent grounds of cross-Maillardized date seeds were reformulated using a cross- Maillardization product made by concentrating an extract of roasted, cross-Maillardized date seeds) 87 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Spent (previously extracted) grounds of Cross-Maillardized date seeds were dried to a
w < 0.4, and 25 g of these dried grounds were initially combined with 5 g of a dry cross-Maillardization product made by concentrating an extract of roasted, cross-Maillardized date seeds to > 99% solids using a refractance window drying system. This mixture was then combined with 0.5 g of soluble fiber, 0.2 g of a dry flavor, 0.14 g of a dry, soluble color, 0.15 g of caffeine and 0.25 g of roasted, ground chicory root. This combined mixture was then extracted using a drip machine to create a hot beverage with notable coffee-like roasted, caramelized flavors, as determined by sensory analysis (e.g., as in Example 8). Example 16 (A coffee-like beverage is made from regenerated spent (previously extracted) cross-Maillardized date seed grounds, using a cross-Maillardization approach) Previously extracted cross-Maillardized date seed grounds are prepared (dried) by adjusting the aw < 0.60 at 55 °C for 16h. The dried spent grounds are combined with an aqueous solution (1:2, wt/wt grounds:solution) containing 2.5% polyhydroxylated phenolic compounds (e.g., as derived from Eucommia bark rich in chlorogenic acid), 5% wt/wt molasses, 2.5% wt/wt pea protein hydrolysate, 1% wt/wt lysine, 1% wt/wt leucine and 0.25% wt/wt cysteine. The mixture is stirred at 60 °C for 6h, the supernatant discharged, and the a
w of the preconditioned spent date grounds is adjusted to < 0.4 by heating the spent grounds at 140 °C for 1.5h, to provide for cross-Maillardization. A coffee-like beverage is prepared by extracting the cross-Maillardized, reconstituted spent date grounds with hot water (e.g., at 92 °C) over a filter. The extract is confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have a distinct coffee-like, and pleasant caramel-like aroma, with a low bitterness. The extract may be combined with one or more of caffeine, gums and/or flavors. A final formulation may be concentrated using, for example, reverse osmosis or microwave-assisted evaporation techniques, to derive a thick, paste-like coffee-base that can be reconstituted with water to prepare a coffee beverage. Example 17 (A coffee-like extract is made from spent (previously extracted) cross-Maillardized chicory root grounds, using a cross-Maillardization approach) Previously extracted cross-Maillardized chicory root (e.g., grounds) are treated 88 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 with hydrolytic enzymes (e.g., cellulase and trypsin), to release mono/di- and oligosaccharides. The a
w of the enzymatic-treated spent grounds is adjusted to <0.6 at 55 °C/16h, before being extracted with hot water (e.g., 92 °C) multiple times, using elevated pressure (e.g., 9 bars). The extracts are collected, pooled and combined with an aqueous solution (1:1, wt/wt pooled extract:solution) containing 2.5% polyhydroxylated phenolic compounds (e.g., as derived from Eucommia bark rich in chlorogenic acid), 5% wt/wt molasses, 2.5% wt/wt pea protein hydrolysate, 1% wt/wt lysine, 1% wt/wt leucine, 0.25% wt/wt cysteine, and 2% caffeine. The mixture is stirred at 60 °C for 6h, and the a
w of the preconditioned spent chicory ground extract is adjusted to <0.2 by drying the mixture using a microwave-assisted evaporation system. The resulting cross-Maillardized concentrate can be used to reconstitute a coffee-like beverage by adding water, where the reconstituted beverage is confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have distinct coffee- like, pleasant caramel and roasted aromas, with a mild astringent bitterness. Example 18 (A coffee-like roasted seed and grounds is made from reconstituted spent (previously extracted) cross-Maillardized date seeds or from pieces/chunks thereof, using a cross- Maillardization approach) Previously extracted cross-Maillardized spent date seeds (e.g., seeds or pieces thereof) are treated in an aqueous environment with hydrolytic enzymes (e.g., cellulase) to expose and/or release saccharides and amino acids from the date material. The treated date material solution is then heated to 80 °C/10 min to deactivate the enzymes, and eucommia bark extract, caffeine, malt extract and yeast extract are added to 2.5%, 1%, 5% and 1.5% wt/wt, respectively. The mixture is dried by adjusting it to aw < 0.6 at 55 °C/24h, and then heated to 140 °C for 20 mins (e.g., ramp to 140 °C, or continuous at 140 °C for 20 mins) in an electric oven, to provide for cross-Maillardization. The derived, reconstituted cross-Maillardized spent date grounds can be then used to prepare coffee- like beverages. Example 19 (A coffee-like roasted grounds is made from reconstituted spent (previously extracted) cross-Maillardized date seed grounds, and from co-roasted raw mustard seeds, using a cross-Maillardization approach) 89 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Previously extracted cross-Maillardized date seed grounds (spent date seed grounds) are dried to a
w < 0.6 at 55 °C for 16h. Mustard seeds (5% wt/wt), eucommia bark extract (rich in chlorogenic acids) (2% wt/wt), molasses (5% wt/wt), and lysine, leucine, and glycine (each at 1% wt/wt) is added to the dried spent grounds. After homogenization, water is added (1:2, w/w homogenate:water) and the pH adjusted to pH = 8.5. The mixture is then stirred at 55 °C for 2 hours, before excess water is removed by adjusting the mixture to a aw < 0.6 at 55 °C for 16h. The dried mixture is then heated in an electric oven at 140 °C for 30 minutes (to provide for cross-Maillardization), immediately cooled down, and homogenized in a grinder. The derived powder is confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have a similar color and flavor profile compared to conventional roasted and ground coffee. The powder can be packed in bags under inert conditions or packed in capsules or comparable single-/multi-serve containers. Example 20 (A coffee-like roasted grounds is made, using a cross-Maillardization approach, from reconstituted spent (previously extracted) cross-Maillardized date seed grounds and from the aroma distillate of separately roasted raw mustard seeds) Previously extracted cross-Maillardized date seed grounds (spent date seed grounds) are dried to a
w < 0.6 at 55 °C for 16h. To the dried spent grounds, eucommia bark extract (rich in chlorogenic acids) (2% wt/wt), molasses (5% wt/wt), and lysine, 1leucine and glycine (each at 1% wt/wt) is added. After homogenization, water is added (1:2, w/w homogenate:water) and the pH adjusted to pH = 8.5. The pH-adjusted mixture is then stirred at 55 °C for 2 hours, before excess water is removed by adjusting the mixture to a aw < 0.6 at 55 °C by drying for 16h. The dried mixture is then heated in an electric oven at 140 °C for 30 minutes, to provide for cross-Maillardization. Mustard seeds are roasted to a final temperature of 220°C and immediately cooled down. The roasted mustard seeds are ground and the aroma fraction is distilled (e.g., by using a distillation apparatus, and a cold-trap containing nonpolar solvent as trapping solvent) and collected in a cooled aroma trap. The aroma distillate is then combined with the previously roasted, cross-Maillardized reconstituted spent date seed grounds. The aromatized cross-Maillardized spent grounds are filled into single-serve capsules, which are packed and sealed under inert gas. Individual capsules are applied/processed on a coffee capsule system to prepare 90 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 an espresso beverage. The resulting coffee-like beverage is confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have intense, fresh roasted aromas, compared to spent date seed grounds, mimicking the aroma profile of conventional coffee capsules. Example 21 (A ground coffee-like product is made by rejuvenating spent cross-Maillardized date seed grounds using a cross-Maillard-derived rejuvenation product/material) The spent date seed grounds retained from a cross-Maillardized date seed extraction are dried to a
w < 0.6. These dried grounds are sieved to remove particles < 100 µm and > 400 µm in size, then combined (e.g., mixed, combined, coated, etc.) with a dry, cross-Maillardized rejuvenation preparation/material, derived originally from a cross-Maillardized material (e.g., prepared as described herein, from one or more of date seeds, chicory root, yerba mate, mustard seed, etc., as in example 35). The grounds may be further reformulated by addition of one or more dry flavorings, caffeine, soluble colors and/or texture modifying ingredients such as gums, etc. The dried rejuvenated, optionally reformulated spent date seed grounds may be extracted (brewed) to provide a reformulated spent date seed grounds extract fraction, confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have particular roasted coffee-like characters reflecting the particular cross-Maillardized rejuvenation material(s) used. Example 22 (A ground coffee-like product is made by reformulating spent cross-Maillardized date seed grounds using various formulation ingredients) The spent date seed grounds retained from a Cross-Maillardized date seed extraction are dried to a
w < 0.6. These dried grounds, optionally sized selected as in example 30, are then reformulated by combining (e.g., mixed, combined, coated, infused, soaked, etc.) with one or more of: flavorings (e.g., dry powder or liquid), caffeine, soluble colors and/or texture modifying ingredients (e.g., gums, etc.), etc. The dried reformulated, optionally reformulated spent date seed grounds may be extracted (brewed) to provide a reformulated spent date seed grounds extract fraction, confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have particular roasted coffee-like characters reflecting the particular reformulation 91 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 ingredients. Example 23 (A ground coffee-like product is made by combining cross-Maillardization-derived materials with a suitable carrier (e.g., sunflower seed shells)) Carrier grounds are produced by milling toasted (e.g., dark brown color) sunflower seed shells to a particle size suitable for various respective coffee machines (ex: drip, espresso, etc.). These grounds are then soaked in a liquid cross-Maillardization-derived concentrate (e.g., prepared as described herein, from one or more of date seeds, chicory root, yerba mate, mustard seed, etc., as in example 35) for 2 hours at room temperature, then dried to aw < 0.6. The grounds may be further reformulated by addition of one or more of: dry flavorings, caffeine, soluble colors, and/or texture modifying ingredients such as gums, etc. Example 24 (A cross-Maillardized coffee beverage was made from green coffee beans) Whole raw (green) coffee beans were washed in hot water (80 °C) for 1 hour. Afterwards, the aqueous extraction media was discarded and the green coffee beans dried by lyophilization. An aqueous solution containing 5% malt extract (carbohydrate source) and 5% pea protein hydrolysate (amino acid source) was added to the washed green coffee (1:5, w/w, coffee:solution), and the mixture placed under a vacuum (<20 mbar) for 20 minutes at room temperature (to enhance infusion into beans). The liquid was drained and the surface of the infused beans rinsed briefly with water. These coffee beans, infused with the exogenous precursor solution, were then adjusted to a
W < 0.6 by dehydrating at 55 °C. The dried, preconditioned coffee was roasted for 6.5 min to a final temperature of 210 ºC, and the roasted, cross-Maillarized coffee then ground, and a beverage prepared by cold immersion brew (4 °C for 16 hours). The resulting beverage was determined by sensory analysis to be more flavorful and showed improved coffee qualities—in particular it showed higher degrees of roasted, nutty, and burnt aroma qualities—in comparison to results obtained by identical processing of green coffee beans but without infusion with the exogenous precursor solution. The samples (and appropriate controls) were further analyzed by Headspace- SPME-GC/MS using methods analogous to those used in Example 9. The results are summarized in Figure 7, were “crossMR” is cross-Maillardized green coffee bean 92 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 material, “MR” is the similarly processed exogenous Maillard reagents alone, and “Control” is green coffee beans (similarly processed but without exogenous Maillard reagents). These data show the production of 2,3-butanedione was enhanced by over 25% by use of these compositions and methods. Simultaneously, the levels of 2,5-dimethylpyrazine are reduced by nearly 50%. These results highlight the utility of the inventive cross-Maillardization methods to shift/tailor the flavor profile of green coffee to a preferred endpoint—in this case, enhancement of buttery flavors and a reduction of earthy, roasted flavors. Example 25 (A cross-Maillardized coffee beverage is made from green coffee bean chunks) Raw (green) coffee chunks (e.g., broken raw coffee) is infused with warm water for (55 °C) for 8 h, the supernatant discharged, and the infused raw coffee lyophilized. An aqueous solution containing amino acids (1% lysine, 1% glycine, 1% leucine) and a reducing sugar (5% xylose), is added to the freeze-dried raw coffee chunks (2:1, w/w, solution:coffee) and stirred for 4 h at room temperature. The surfaces of the chunks are briefly rinsed with water and the infused, rinsed chunks adjusted to a aw < 0.75, by dehydrating at 55 °C. The dried, preconditioned coffee chunks are roasted to a final temperature of 205 °C for 6 minutes to provide roasted, cross-Maillardized coffee chunks, which are then ground and filled into capsules (e.g., single-serve capsules, such as K- cup, Nespresso, etc.). The capsules are then placed in a suitable machine (e.g., Nespresso “Essenza Mini”) and a coffee beverage (e.g., 110 mL) is prepared. The resulting beverage is confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have an improved aroma profile in comparison to non-cross- Maillardized coffee chunks (e.g., with increased intensities of caramel, chocolate, and roasted aromas. Example 26 (A cross-Maillardized coffee beverage is made from steam-treated green coffee) Raw (green) coffee (e.g., beans and/or chunks) (e.g., low quality green coffee beans and/or chunks) is treated with hot steam (160 °C/14 minutes). The steam is condensed to provide a coffee-enriched wastewater, non-volatile compounds (e.g., chlorogenic acids and saccharides) are extracted into the wastewater from the 93 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 steam-treated coffee, and the extract purified using solid-phase assisted extraction. The steam-treated coffee is then combined with an aqueous solution (1:2, w/w coffee:solution), containing 2% of the purified coffee-enriched wastewater extract (containing chlorogenic acids and other polyhydroxylated phenolic compounds) and 1.5% zein hydrolysates, the mixture stirred for 4h at room temperature, and the infused coffee rinsed with water before being adjusted to a
w < 0.75 by dehydrating at 55 °C. The preconditioned, coffee-enriched coffee beans and/or chunks are roasted to a final temperature of 210 °C. The roasted, cross-Maillardized enriched coffee is then ground, and a hot beverage is prepared by, for example, drip filtration (e.g., at 92 °C). The resulting coffee beverage is confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have a more pleasant flavor profile, with decreased robusta-like coffee aromas, a milder bitterness, and more phenolic, cereal-like and chocolate-like aroma notes compared to untreated coffee (identical processing without steam treatment, infusion, and cross-Marillardization). Example 27 (A cross-Maillardized coffee beverage is made from robusta and arabica coffees) Raw (green) robusta coffee (e.g., beans) is washed (e.g., stirred) using hot water (80 °C, 1:1, wt/wt) for 1h, the aqueous phase separated and the remaining green coffee beans dried by lyophilization [a
w < 0.3]. Additionally, arabica coffee (e.g., beans) is soaked with hot water (80 °C, 1:1, wt/wtt) for 1h, and directly lyophilized (without first separating the aqueous phase). Both lyophilized coffees, the washed robusta, and the soaked arabica are combined (75:25, wt/wt), and an aqueous solution, containing 2% molasses, 2% malt extract, 2.5% glycine and 2.5% mung protein hydrolysate, is added (1:2, wt/wt beans:solution). The mixture is stirred at 55 °C for 6h, and the preconditioned coffees briefly rinsed with water, before adjusting the rinsed coffee to a a
w < 0.75 by dehydrating at 55 °C. The dehydrated preconditioned coffee blend is roasted to a final temperature of 210 °C, the roasted, cross-Maillardized coffee ground, and a hot coffee beverage is prepared from the grounds by e.g., drip filtration (e.g., at 92 °C). The prepared coffee beverage is confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have improved sensory qualities, compared to the robusta coffee alone, with a decreased acrylamide content and a more malt- and caramel-like aroma. 94 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Example 28 (A cross-Maillardized coffee extract/flavoring is made from coffee) Raw (green) coffee (e.g., beans or chunks, preferably of low quality) is washed (e.g., stirred) with hot water (80 °C, 1:1, wt/wt) for 1h, the aqueous phase is separated and the remaining green coffee is dried by lyophilization (e.g., aw < 0.3). To the freeze- dried coffee, an aqueous solution (1:2, wt/wt coffee:solution), containing 5% maltodextrins, and 5% of plant protein hydrolysates (e.g., rice protein and/or pea protein hydrolysate) is added, and the mixture stirred for 8h at room temperature. The stirred mixture, including the supernatant, is dried at 55 °C for 16h to adjust the coffee to a a
w < 0.60, and the surface of the preconditioned beans briefly rinsed with water, dried again at 55 °C for 2h (to aw < 0.60), and then roasted to a final temperature of 210 °C. The roasted, cross-Maillardized coffee is ground and the grounds extracted multiple times with hot water (e.g., immersion brew; at e.g., 92 °C). The extracts are combined, and water is removed (e.g., under reduced pressure or by reverse osmosis). The concentrated extract can be used as a coffee-type flavoring for beverages, confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have increased sensory properties compared to non-cross-Maillardized coffee. Example 29 (A roast and ground cross-Maillardized coffee is made from coffee and sesame) Raw (green) coffee (e.g., beans or chunks, preferably of low quality) is washed (e.g., stirred) with hot water (80 °C, 1:1, wt/wt) for 1h, the aqueous phase is separated and the remaining green coffee is dried by lyophilization (e.g., a
w < 0.3). To the freeze- dried coffee, an aqueous solution (1:2, wt/wt coffee:solution) containing 5% maltodextrins, and 5% of plant protein hydrolysates (e.g., rice protein and/or pea protein hydrolysate) is added, and the mixture stirred for 8h at room temperature. The stirred mixture, including the supernatant, is dried at 55 °C for 16h to adjust the coffee to a a
w < 0.60, and the surface of the preconditioned coffee is briefly rinsed with water, dried again at 55 °C for 2h (to aw < 0.60), and then roasted to a final temperature of 210 °C. Additionally sesame (e.g., seeds) is prepared by roasting it to a final temperature of 220 °C in 3 minutes. The roasted preconditioned coffee and the roasted sesame are mixed (95/5, wt/wt coffee:sesame), homogenized, applied to a grinder setup and finely ground. The ground 95 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 product is immediately filled into bags having a CO2 valve for degassing. The interiors of the bags are placed under vacuum to protect produced the formed flavor from oxidation, and the bags sealed for storage. The roast and ground product can be brewed like conventional coffee, with the cross-Maillardized coffee in combination with sesame confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have a more intense coffee- like flavour and roasted aroma, with a higher overall aroma intensity compared to identical processing without cross-Marillardization. Example 30 (A cross-Maillardized coffee beverage is made from coffee and buckwheat) Raw (green) coffee (e.g., beans or chunks, preferably of low quality) is washed (e.g., stirred) with hot water (80 °C, 1:1, wt/wt) for 1h, the aqueous phase is separated and the remaining green coffee is dried by lyophilization (a
w < 03). The freeze-dried coffee and raw buckwheat are combined (75/25, w/w coffee:buckwheat), homogenized, and an aqueous solution (1:2, w/w coffee-buckwheat:solution) containing containing 5% maltodextrins, and 5% of plant protein hydrolysates (e.g., rice protein and/or pea protein hydrolysate) is added, and the mixture stirred for 8h at room temperature. The stirred mixture, including the supernatant, is dried at 55 °C for 16h to adjust the coffee to a aw < 0.60, the surface of the preconditioned coffee-buckwheat mixture briefly rinsed with water to remove residual sugars/amino acid, dried again at 55 °C for 2h (to aw < 0.60), and then roasted together to a final temperature of 195 °C in a hot air roaster. The roasted, cross- Maillardized coffee-buckwheat mixture is ground and extracted multiple times with hot water (e.g., 92 °C, under pressure), with the aroma being stripped and collected separately (e.g., by means of trapping the volatile aroma compounds via molecular distillation, or by simply collecting the volatiles in the headspace in a cold trap (e.g., cooled with liquid nitrogen, dry ice)). The aroma-free extract is then spray-dried, and combined/coated with the previously separately collected aroma fraction. The derived granular, powdery and dry coffee-buckwheat mixture may, e.g., be used as conventional soluble/instant coffee (3g/200 mL), with the preconditioned, cross-Maillardized coffee- buckwheat confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have a more distinct roast, caramel, nutty and chocolate-like aroma profile compared to identical processing without cross-Maillardization. 96 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Example 31 (A coffee-like beverage is made from regenerated traditional spent (previously extracted) coffee grounds) This example describes regenerating traditional spent (previously extracted) coffee grounds to make several product types: a) regenerated/reformulated coffee grounds are prepared from spent coffee grounds; b) reformulated spent coffee grounds extract is prepared; and c) a finished reformulated spent grounds beverage is produced, as follows: a) Dry retentate (spent) grounds from the production of a coffee beverage are formulated (e.g., mixed, combined, coated, infused, soaked, etc.) with an amount of an exogenous cross-Maillardized flavor or beverage component (e.g., a concentrated extract or lyophilized form thereof, made from coffee or from non-coffee substrate materials by the presently disclosed cross-Maillardization methods), the amount sufficient to coat and/or infuse the retentate grounds to rejuvenate the organoleptic quality potential thereof; b) Dried reformulated spent grounds of a) are extracted (brewed) in. e.g., 92 °C water for 4 minutes before gravity filtration, or alternatively extracted in a portafilter of an espresso machine), in either case to provide a reformulated spent coffee grounds extract fraction) and a retentate extracted reformulated coffee grounds fraction (spent reformulated coffee grounds fraction), followed by cooling (e.g., to 4 °C) of the liquid reformulated coffee grounds extract fraction for storage; and c) For final formulation, the liquid reformulated coffee grounds extract fraction from b) may be combined with one or more of caffeine, colorants, gums and/or flavors, filled into cans with nitrogen (e.g., under nitrogen atmosphere and/or flushed with nitrogen to replace trapped CO
2) and retorted. In further examples, spent grounds from non-coffee substrate materials may likewise be regenerated/rejuvenated by formulating with an amount of an exogenous cross-Maillardized flavor or beverage component. Example 32 (A coffee-like beverage is made from regenerated traditional spent (previously extracted) coffee grounds) 97 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Previously extracted (spent) coffee grounds were treated with an exo-protease (Novozymes Flavourzyme
TM, 0.1%) in an aqueous solution. The enzymes were deactivated at 80 °C for 10 min, and the a
w adjusted by dehydrating to < 0.7 at 55 °C for 16 hours, leaving the enzymatically-treated spent grounds. The dried, treated spent grounds were then combined with an aqueous solution (1:2, w/w grounds:solution) containing 1% caffeine, 2% w/w chlorogenic acid derivatives (e.g., derived from eucommia bark), 1% w/w leucine, 1% w/w lysine, 2.5% w/w pea protein hydrolysate and 5% w/w molasses. The mixture was stirred at 60 °C for 3 hours at pH 8.5, and the water removed by dehydrating at 55 °C for 16 hours to achieve a a
w < 0.6. The dried, preconditioned spent grounds were then heated to 140 °C for 30 min in an electric oven, to provide for cross-Maillardization. The derived regenerated spent coffee grounds were then used to prepare a drip coffee beverage (23 g/320 mL) that was confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have sensory characteristics similar to coffee prepared from non-spent coffee grounds. This rejuvenated composition was further analyzed by Headspace-SPME-GC/MS, using methods analogous to Example 9, and the results summarized in Figure 8. Figure 8 shows that in this example, while the levels of 2,3-butanedione (diacetal) are relatively unchanged, the level of 2,5-dimethylpyrazine was substantially enhanced by cross- Maillardization, in this case in the presence of optionally added chlorogenic acid, of previously roasted, ground and extracted coffee beans. According to particular aspects, use of added chlorogenic acid tends to favor Maillard reactions over carmelization (e.g., more pyrazine, whereas the 2,3-butanedione level is relatively unchanged). Pyrazines in coffee contribute to the earthy, roasted-type aroma characteristic of the roasted product and beverages made from it. These data reveal that the disclosed compositions and cross-Maillardization methods effectively rejuvenate spent (previously roasted, ground and extracted) coffee grounds, such that key aroma compounds like 2,5- dimethylpyrazine can be created in-situ and are available for subsequent extraction using conventional coffee production techniques. Example 33 (Spent coffee grounds were reformulated using a cross-Maillardization product made by concentrating an extract of roasted, cross-Maillardized date seeds) Spent (previously extracted) coffee grounds were dried to a
w < 0.4. The dried spent grounds were reformulated by initially combining 25 g of the dried grounds with 5 98 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 g of a dry, cross-Maillardization product (made by concentrating an extract of roasted, cross-Maillardized date seeds to > 99% solids using a refractance window drying system), and then adding 0.6 g of soluble fiber, 0.15 g of dry flavor, 0.1 g of dry, soluble color, 0.11 g of caffeine, and 0.1 g of roasted, ground chicory root. The resulting mixture was blended and extracted using a drip percolation system. The resulting beverage was determined by sensory analysis (e.g., as in Example 8) to resemble freshly brewed coffee in taste, appearance and texture, with prominent dark roasted notes, dark color, moderate body and the expected levels of caffeine from a fresh brew. Example 34 (A ground coffee-like product is made by rejuvenating spent coffee grounds using liquid cross-Maillardization-derived products) Spent coffee grounds are dried to a
w < 0.6. These dried grounds are then soaked in a liquid cross-Maillardized date seed extract (e.g., as prepared in Example 1 a), or in a liquid concentrate thereof, for 2 hours at room temperature, then dried to a aw < 0.6. The rejuvenated grounds may be further formulated by addition of dry flavoring preparations, caffeine, soluble color compounds and/or texture modifying ingredients such as gums. The dried rejuvenated, optionally reformulated spent grounds may be extracted (brewed) to provide a reformulated spent coffee grounds extract fraction, confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have a roasted coffee-like character. Example 35 (A ground coffee-like product is made by rejuvenating spent coffee grounds using dried cross-Maillardization-derived products/materials) A dried Cross-Maillard rejuvenation material is produced by taking a liquid extract of a cross-Maillardized substrate (e.g., prepared as described herein from one or more of date seeds, chicory root, yerba mate, mustard seed, etc.), or concentrate thereof (e.g., prepared by optionally concentrating using an osmotic or low pressure process), and further dehydrating it using a process such as microwave drying, refractance window, vacuum belt drying, etc., to provide a dry powder. The dry, cross-Maillardization derived powder is then added (e.g., mixed, combined, coated, infused, soaked, etc.) to spent coffee grounds previously dried to a
W < 0.6. These rejuvenated grounds may be further 99 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 formulated by addition of one or more of: dry flavorings, caffeine, soluble colors and/or texture modifying ingredients such as gums, etc. The dried rejuvenated, optionally reformulated spent coffee grounds may be extracted (brewed) to provide a reformulated spent coffee grounds extract fraction, confirmed by sensory analysis (e.g., using methods as described above in Example 8) to have particular roasted coffee-like characters reflecting the particular cross-Maillardized rejuvenation material(s) used. Example 36 (An extract of defatted sunflower seed meal (press cake) was used to create a coffee- enhancing soluble solid) Extraction. Forty (40) g of ethanol and 60 g of water were premixed, then added to 40 g of defatted sunflower seed meal. This mixture was sealed and stored at room temperature for 8 hours. After 8 hours, it was pressed through a 200 μm filter mesh, filtered to 11 μm, centrifuged at 3,500 relative centrifugal force (RCF) for 5 minutes to remove oil and heavy sediment, then reduced in a rotary evaporator until it reached >50% solids. These samples were further dried and toasted in an oven at ~200°C until the liquid was completely evaporated and the remaining solids browned (e.g., about 5 min.) to provide a toasted extract; representative example as shown in Figure 12. The toasted extracts were recovered and added to ground, roasted, preconditioned date seeds (prepared as described below) to yield 3, 5, or 10 wt% in the mixture, which was then extracted in a 1:16 grounds:water ratio using 95°C water and a commercial conical paper filter. A control sample was produced using analogous methods but omitting the sunflower extract and replacing it with the ground, roasted, preconditioned date seeds. The brewed, filtered samples were analyzed for pH, dissolved solids (Brix), and tasted (sensory panel). Preconditioned Date Seeds. Clean, dry, intact date seeds were cracked into pieces between 2 and 6 mm in diameter.1 L of water was added to a beaker, and 10 mM of potassium carbonate was added, along with 60 g of fructose and 50 g of pea protein isolate and 15 g of defatted fenugreek seeds. This was mixed thoroughly with an overheard mixer until homogeneous, then the pH adjusted to 9.3 with potassium hydroxide. While stirring, 500 g of the date seed pieces were added to the mixture. The pH was checked and adjusted back to 9.3. The batch was then heated to 55 °C. Once this 100 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 temperature was reached, it was maintained for 2 hours. The seed pieces were then drained through a sieve and rinsed of surface deposits. Once rinsed, the seed pieces were dried for 15 hours at 55 °C. Dried, preconditioned date seed pieces were roasted in a fluid bed coffee roaster (RFB-S, Neuhaus Neotec) to a final temperature of 205 °C. These roasted, preconditioned date seed pieces were ground to an equivalent particle size of conventional drip coffee grounds. Results. Compared to the control sample, the sample containing defatted sunflower seed meal extract showed increased body, coffee-like bitterness, roastiness, and a preferred acidity. In addition, pH was slightly higher (by 0.2 for 5 wt% extract in grounds) as were dissolved solids (by 0.2 °Bx for 5 wt% extract in grounds). Finally, brews with added defatted sunflower seed meal extract were cloudy, whereas brews without the sunflower extract were clear. Separately, samples of the toasted extract were dissolved in water for chlorogenic acid constituent analysis by liquid chromatography. Liquid chromatography analyses were conducted using a Sciex 5600 TOF in ESI- and paired with a Waters TUV detector set at λ = 324 nm. An Agilent Eclipse Plus C18 (2.1 x 50 mm) was used. Solvent and gradient conditions used are listed in the table below: TABLE 2. Solvent and gradient conditions used in liquid chromatography.
The roasted sample described above was analyzed using this method to determine its level of chlorogenic acid and chlorogenic lactones. As discussed in the introduction, chlorogenic acid provides many essential qualities of traditional coffee. Amongst them, the flavor provided by the acids and the lactones, a bitterness that is unique to coffee, is essential to produce or replicate the full profile of traditional coffee. Figure 13 demonstrates that the roasting process implemented in this example causes a near complete conversion of the 3 isomers of chlorogenic acid to the corresponding CGLs. These lactones, when added to the beverage described earlier in this example, provided the desirable qualities of the distinctive bitterness, the enhanced 101 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 mouthfeel, and the perception of more roasted flavor. Example 37 (An extract of defatted sunflower seed meal (press cake) was extruded within a ramon seed matrix to create a coffee-enhancing component) Extraction. Forty (40) g of ethanol and 60 g of water were premixed, then added to 40 g of defatted sunflower seed meal. This mixture was sealed and stored at room temperature for 8 hours. After 8 hours, it was pressed through a 200 μm filter mesh, filtered to 11 μm, centrifuged at 3,500 RCF for 5 minutes to remove oil and heavy sediment, then reduced in a rotary evaporator until it reached 50% solids. Thirty (30) g of the concentrated extract was combined with 100 g of fine ramon seed flour (D75 ≤ 250 μm; i.e., 75th percentile particle diameter is 250 μm, on a mass or volume basis). This mixture was pressed through a preheated single screw extruder at 95 °C with a 5 mm die. Pieces were cooled to room temperature and cut into 8-10 mm segments. These segments were roasted in a fluid bed roaster (IKAWA Pro) for 9 minutes to a final temperature of 205°C. Cut, roasted samples were ground and brewed with a 1:16 grounds:water ratio using 95°C water as in Example 36. Brewed samples were analyzed for pH, dissolved solids (Brix), and tasted by a sensory panel. The samples were also subjected to CGA chemical analysis by UV-Visible absorption spectroscopy. UV Protocol. UV-VIS data were collected on a Molecular Devices SpectraMax Mini. Scan settings were set from a wavelength range of 200-500 nm and step of one nm. 96 well Greiner UV Star plates were used, and samples were shaken 5 seconds before analysis to ensure homogeneity. Samples were diluted to a concentration range for improved reproducibility in the measurement (OD ≤2.0). Results. As shown in Figure 14, roasted samples contained a spectroscopic signature of chlorogenic acids and/or lactones, indicating survival and/or interconversion of those compounds through the roasting and extraction during the brewing process (especially the peak at 324 nm). Identical roasted samples without the sunflower seed press cake extract, i.e. containing only extruded ramon seed flour, lacked signatures of chlorogenic acids and/or lactones. Compared to an identical preparation made without the defatted sunflower seed meal (press cake) extract, in which it was replaced with extruded ramon seed flour, the sample containing sunflower extract showed increased body, coffee-like bitterness, 102 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 roastiness, and a preferred acidity. As shown in Table 3, the brews with the defatted sunflower seed meal extract showed higher pH and Brix. Table 3. Values for pH and dissolved solids.
Example 38 (An extract of commercially available, de-hulled, defatted sunflower seed meal was extruded within a ramon seed matrix to create a coffee-enhancing component) Extraction. Forty (40) g of ethanol and 60 g of water were premixed, then added to 40 g of de-hulled, defatted sunflower seed meal (often marketed as a sunflower protein). This mixture was sealed and stored at room temperature for 8 hours. After 8 hours, it was pressed through a 200 μm filter mesh, filtered to 11 μm, centrifuged at 3,500 RCF for 5 minutes to remove oil and heavy sediment, then reduced in a rotary evaporator until it reached 50% solids. Thirty (30) g of this concentrated extract were combined with 100 g of fine ramon seed flour (D75 ≤ 250 μm). This mixture was pressed through a preheated single screw extruder at 95 °C with a 5 mm die. Pieces were cooled to room temperature and cut into 8-10 mm segments. These segments were roasted in a fluid bed roaster (IKAWA Pro) for 9 minutes to a final temperature of 205°C. These extruded, cut pieces were roasted, ground, size-sorted to select particles between 200 – 800 µm diameter, and then brewed using a 1:16 grounds:water ratio using 95°C water. A control sample was produced using only ramon seed flour and water without the sunflower seed extract. Brewed samples were analyzed for dissolved solids (Brix) and taste by a sensory panel, and UV-Visible spectroscopy to measure the CGA content. These data were acquired using the same protocol as described in Example 37. Results. As seen in Figure 15, the control sample (extracted from extruded materials comprising ramon seed flour and water only) contains very little signature of CGA. The results from Example 37 are replotted here for reference (defatted sunflower seed meal), along with a standard solution containing 20 μg/mL of 3-O-caffeoylquinic acid. 103 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 When compared in an organoleptic evaluation, the sample containing sunflower extract showed increased body, coffee-like bitterness, roastiness, and preferable acidity in comparison to the control sample with no sunflower extract. Brew pH and dissolved solids increased with the addition of sunflower protein extract, but less than when sunflower seed press cake extract was added. Table 4. Values for pH and dissolved solids.
Example 39 (An extract of commercially available, de-hulled, defatted sunflower seed meal was extruded within a Macrotyloma uniflorum (kulthi daal) matrix to create a coffee- enhancing component) Extraction. Two-hundred forty (240) g of ethanol and 160 g of water were premixed, then added to 40 g of de-hulled, defatted sunflower seed meal. This mixture was covered and heated to 50 °C while stirring. After 4 hours under these conditions, it was cooled and then pressed through a 200 μm filter mesh, filtered to 11 μm, centrifuged at 3,500 RCF for 5 minutes to remove oil and heavy sediment, then reduced in a rotary evaporator until it reached 50% solids. Thirty-eight (38) g of the concentrated extract was first acidified with 1.8g of 50% malic acid solution and then combined with 130 g of fine kulthi daal (Macrotyloma uniflorum) flour. This mixture was pressed through a preheated single screw extruder at 95 °C with a 5 mm die. Pieces were cooled to room temperature and cut into 8-10 mm segments. These segments were roasted in a fluid bed roaster (IKAWA Pro) for 7.5 minutes to a final temperature of 210°C. Cut, roasted samples were ground and brewed with a 1:16 grounds:water ratio using 95°C water. Brewed samples were analyzed for pH, dissolved solids (Brix), and taste by a sensory panel. A control sample of the kulthi daal alone were made by an analogous protocol, omitting the sunflower extract and using only water. The control sample of the sunflower extract alone was made by the same roasting methods of Example 36. Results. The sensory panel reported a reduction in both overall flavor intensity 104 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 (e.g., roastiness) and perceived off-notes (green bean water, peanut shell). Additionally, as shown in Table 5, compared to an identical preparation made without the de-hulled, defatted sunflower seed meal extract, and instead using water to replace the liquid and kulthi daal flour to replace the solids, brews with the de-hulled defatted sunflower seed meal extract showed similar pH and Brix, although the one with de-hulled defatted sunflower meal was slightly lower in both. Table 5. Values for pH and dissolved solids.
This result contrasts with those in Examples 36-38 above, where the addition of CGA extract from defatted sunflower seed meal, or dehulled, defatted sunflower seed meal, caused an increase in amount of dissolved solids in the final brew. The addition of sunflower seed meal extract to the extruded kulthi daal flour did not, suggesting that the constituents (kulthi daal flour and CGA extract) cross reacted during roasting, changing the amount of soluble solids such that the total amount of dissolved solids in the final brew was similar to that found for control samples (i.e. insoluble products were formed from the soluble CGA extract starting material). Additional evidence of cross reaction between the kulthi daal flour and CGA extract is visible in the volatile spectrum of the brew. The volatile profile was collected on an Agilent 8890 GC paired with an Agilent 7250 Q-TOF, and Gerstel Dynamic Headspace (DHS) for headspace analysis. The samples were analyzed using an Agilent DB-WAX column (60m × 0.250mm × 0.25µm). The initial oven temperature was set to 40°C and ramped to 120°C at a rate of 6°C/min and held for 5 min, and then ramped to 230°C at a ramp rate of 8°C/min and held for 2 min. During DHS, the incubation phase was set to a 3 min sample incubation period at 50°C, then a trapping phase at 30.0 mL/min and 25°C. The sample inlet was set to 40°C initially, then ramped to 280°C at a rate of 720°C/min and held for 3 min. As can be seen in Figure 17, the levels of many key odorants for coffee, such as a family of pyrazines, are significantly enhanced by compounding the kulthi daal and sunflower extract prior to roasting. Specifically, as seen in Figure 17, the kulthi daal flour + de-hulled, defatted sunflower seed meal extract composition described in this example provides significant 105 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 enhancements in the production of a variety of pyrazine compounds, namely 2,3- dimethyl, 2,5-dimethyl-, 2,6-dimethyl-, trimethyl-, ethyl-, 2-ethyl-5-methyl-, 2-ethyl-6- methyl- and 3-ethyl-2,5-dimethyl-pyrazines. Such compounds are significant constituents of the roasted flavor profile of coffee. Notably, some of these compounds, such as 2,3- dimethyl- and 2-ethyl-5-methyl-pyrazine are undetectable in the individual ingredients’ volatile spectra. Also enhanced are certain aldehydes, namely 3-furaldehyde and 5- methyl-2-furancarboxaldehyde, notable for its caramel-like aroma, likewise welcome in coffee. The combination also has suppressive effects on the creation of certain compounds. The addition of the de-hulled, defatted sunflower seed meal extract and the kulthi daal flour completely suppresses the creation of propane diol in the compounded form. Example 40 (Commercially available purified chlorogenic acid (CGA) was extruded within a ramon seed matrix to create a coffee-enhancing component) Twenty (20) g of purified CGA from Eucommia ulmoides was dissolved in water and neutralized to pH 7 with potassium hydroxide. This solution was then concentrated to 50% CGA (wt%) using a rotary evaporator. Aliquots of this concentrated CGA solution were combined with fine ramon seed flour (D75 ≤ 250 μm) and water to make 5 wt% and 15 wt% CGA mixtures with a final mass of 100 g dry matter and 20% (wt%) moisture content. These mixtures were pressed through a preheated single screw extruder at 95 °C with a 5 mm die. Pieces were cooled to room temperature and cut into 8-10 mm segments. These segments were roasted in a fluid bed roaster (IKAWA Pro) to a final temperature of 206°C. Roasted samples were ground and brewed with a 1:16 grounds:water ratio using 95°C water. Brewed samples were analyzed for pH, dissolved solids (Brix), and taste by a sensory panel. As shown in Table 6, compared to an identical preparation made without the purified CGA, and instead using water to replace the liquid and ramon seed flour to replace the solids, brews with purified CGA showed similar pH and Brix (see table below). Moreover, for longer roast times, the dissolved solids appeared to be inversely proportional to the amount of CGA. The sensory panel reported an increase in perceived body and a change in the quality of the astringency. 106 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Table 6. Values for pH and dissolved solids at different roast times.
Like the data of Example 39, this result contrasts with those in Examples 36-38-3 above, where the addition of CGA extracted from de-hulled and/or defatted sunflower seed meals caused an increase in amount of dissolved solids in the final brew. The addition of highly purified CGA from to the extruded ramon seed did not, suggesting that the constituents (ramon seed flour and purified CGA) cross reacted during roasting, changing the amount of soluble solids such that the total amount of dissolved solids in the final brew was similar to that found for control samples (i.e. insoluble products were formed from the soluble purified CGA starting material). Example 41 (Coffee bean-less coffee grounds were made using ground, roasted, preconditioned date seeds, combined with defatted sunflower seed meal and ramon seed flour and other ingredients) The roasted grounds (i.e., defatted sunflower seed meal extruded within a ramon seed flour, ground and roasted) from Example 37 above were combined with ground, roasted, preconditioned date seeds (prepared as described in Example 36), dehydrated lemon powder, ground roasted millet malt, caffeine and baking soda to make coffee bean- less coffee grounds. The following brewing formats are intended to be exemplary and non-limiting. Other formats may be substituted. Drip coffee. Fifteen (15) g of these assembled grounds were placed in a paper cone filter that had been pre-rinsed with tap water inside a plastic filter cone holder. Over a series of 5 equal additions, 240 g (total) of 95 °C water was poured over the grounds and the resulting extract collected in a vessel below the filter. This beverage was evaluated by a sensory panel while hot, and found to contain roasted coffee notes, as well as notes of chocolate and distinct, coffee-like bitterness. Additionally, panelists noted a mouthfeel consistent with traditional coffee bean-derived coffee. Dissolved solids of 1.6 Brix were observed, consistent with percolated brewed traditional coffee. 107 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Espresso. Twenty (20) g of the assembled grounds were placed in an espresso machine portafilter and tamped with 30 lbs of pressure. The portafilter was loaded into a LaMarzocco GS3 espresso machine and extracted at 10 bars of pressure. The extraction was halted when 40 g of total extract was yielded. This extract was then evaluated by a sensory panel, which found prominent roast coffee and chocolate notes, espresso-like bitterness, a layer of foam (crema) consistent with traditional espresso, and a rich, syrupy mouthfeel despite yielding slightly low total dissolved solids (7 Brix) in comparison to traditional espresso. Example 42 (An extract of artichokes is obtained to create a coffee-enhancing component) Forty (40) g of ethanol and 60 g of water are premixed, then added to 40 g of artichoke powder. This mixture is sealed and stored at room temperature for 8 hours. After 8 hours, it is pressed through a 200 μm filter mesh, filtered to 11 μm, centrifuged at 3,500 relative centrifugal force (RCF) for 5 minutes to remove any oil and heavy sediment, then reduced in a rotary evaporator until it reaches >50% solids. These samples are further dried and toasted in an oven at ~200°C until the liquid is completely evaporated and the remaining solids browned. The dried, toasted extracts are recovered and added to ground, roasted, preconditioned date seeds (as described in Example 36) to yield 5 or 10 wt% in the mixture, which is then brewed in a 1:16 grounds:water ratio using 95°C water. Brewed samples are analyzed for pH, dissolved solids (Brix), and tasted (sensory panel). Separately, samples of the pure extracts are dissolved in water for chlorogenic acid constituent analysis by liquid chromatography, UV-Visible absorption spectroscopy, and gas chromatography-mass spectrometry. According to particular aspects, extracts of artichoke powder provide a useful source of CGA and other components for use as a coffee-enhancing component as described herein. Example 43 (A commercially available extract of artichokes was extruded within a ramon seed matrix to create a coffee-enhancing component) Sixty-eight (68) g of a commercial artichoke extract was used to prepare a coffee composition. The CGA content of the commercial extract powder was determined using 108 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 the UV-Visible spectroscopic protocol described in Example 37 to be 7%. This extract was prepared by extracting artichokes with water, vacuum evaporating to produce a liquid concentrate, and then spray drying to produce a low moisture powder. The resulting extract was then combined with 177 g of fine ramon seed flour (D75 ≤ 250 μm). This mixture was pressed through a preheated single screw extruder at 95 °C with a 5 mm die. Pieces were cooled to room temperature and cut into 8-10 mm segments. These segments were roasted in a fluid bed roaster (IKAWA Pro) for 8 minutes to a final temperature of 203°C. An analogous control preparation was prepared by omitting the artichoke extract and extruding ramon seed flour alone. Cut, roasted samples were ground and brewed with a 1:16 grounds:water ratio using 95°C water as in Example 36. Brewed samples were analyzed for dissolved solids (Brix) and tasted by a sensory panel. Compared to the control preparation made without the artichoke extract the sensory panel noted coffee-like roastiness, an increase in coffee-like body, as well as flavor notes of herbal, medicinal and green. Dissolved solids were higher in the 1:16 brew than in an equivalent control brew (1.61°Bx vs.1.26°Bx). Example 44 (A commercially available extract of yerba mate was extruded within a ramon seed matrix to create a coffee-enhancing component) Fifteen and a half (15.5) g of a commercial yerba mate extract containing 15 - 35 % CGA was prepared by extracting yerba mate with water, vacuum evaporating to produce a liquid concentrate, and then spray drying to produce a low moisture powder. This extract was combined with 232 g of fine ramon seed flour (D75 ≤ 250 μm). This mixture was pressed through a preheated single screw extruder at 95 °C with a 5 mm die. Pieces were cooled to room temperature and cut into 8-10 mm segments. These segments were roasted in a fluid bed roaster (IKAWA Pro) for 9 minutes to a final temperature of 212°C. An analogous control sample was made by omitting the artichoke extract. Cut, roasted samples were ground and brewed with a 1:16 grounds:water ratio using 95°C water as in Example 36. Brewed samples were analyzed for dissolved solids (Brix) and tasted by a sensory panel. Compared to the control preparation, the sensory panel noted greater coffee-like roastiness, an increase in coffee-like body,astrigency, and strong bitterness in the yerba 109 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 mate extract containing composition. Dissolved solids were similar in the 1:16 brew to the control brew (1.26°Bx vs.1.26°Bx). Example 45 (An extract of tobacco stems is obtained, and extruded within a ramon seed matrix to create a coffee-enhancing component) Forty (40) g of ethanol and 60 g of water are premixed, then added to 40 g of tobacco stems. This mixture is sealed and stored at room temperature for 8 hours. After 8 hours, it is pressed through a 200 μm filter mesh, filtered to 11 μm, centrifuged at 3,500 relative centrifugal force (RCF) for 5 minutes to remove oil and heavy sediment, then reduced in a rotary evaporator until it reaches >50% solids. These samples are further dried and toasted in an oven at ~200°C until the liquid is completely evaporated and the remaining solids browned. The dried, toasted extracts are recovered and added to ground, roasted, preconditioned date seeds (as described in Example 36) to yield 5 or 10 wt% in the mixture, which is then brewed in a 1:16 grounds:water ratio using 95°C water. Brewed samples are analyzed for pH, dissolved solids (Brix), and taste (sensory panel). Separately, samples of the pure extracts are dissolved in water for chlorogenic acid constituent analysis by liquid chromatography, UV-Visible absorption spectroscopy, and gas chromatography-mass spectrometry. According to particular aspects, extracts of tobacco(e.g., stems) provide a useful source of CGA and other components for use as a coffee-enhancing component as described herein. Alternatively, forty (40) g of ethanol and 60 g of water are premixed, then added to 40 g of tobacco stems. This mixture is sealed and stored at room temperature for 8 hours. After 8 hours, it is pressed through a 200 μm filter mesh, filtered to 11 μm, centrifuged at 3,500 RCF for 5 minutes to remove any oil and heavy sediment, then reduced in a rotary evaporator until it reaches 50% solids. Thirty (30) g of the concentrated extract is combined with 100 g of fine ramon seed flour (D75 ≤ 250 μm). This mixture is pressed through a preheated single screw extruder at 95 °C with a 5 mm die. Pieces are cooled to room temperature and cut into 8-10 mm segments. These segments are roasted in a fluid bed roaster (IKAWA Pro) for 9 minutes to a final temperature of 205°C. 110 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Roasted samples are ground and brewed with a 1:16 grounds:water ratio using 95°C water. Brewed samples were analyzed for pH, dissolved solids (Brix), and taste by a sensory panel. The samples are also subjected to CGA chemical analysis by liquid chromatography, UV-Visible absorption spectroscopy, and gas chromatography-mass spectrometry. According to particular aspects, extracts of tobacco (e.g., stems) combined with ramon seed flour and roasted provide a useful source of CGA and other components for use as a coffee-enhancing component as described herein. Example 46 (Exemplary ranges of components, temperatures, and solvent compositions useful to create a coffee-enhancing components according the above Examples 36-45 ) Ranges (listed as wt%): CGA/CGL contents added to “raw” (e.g., green, etc.) preparations ^ 1-100% ^ 5-25% ^ 80-100% ^ 4-8% ^ 20-40% ^ 25-50% ^ 1-10% ^ 40-80% CGA or its thermal reaction products (e.g., CGL) contents present in “finished” preparations (e.g., grounds, roasted bean-less beans, finished beverages, etc.) ^ 0.01-30% ^ 4-6% ^ 4-8% ^ 0.01-1% ^ 0.5-5% ^ 5-15% ^ 10-30% Fraction CGA remaining after thermal conversion (i.e., CGA unconverted to thermal products like CGL) 111 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 ^ 0-100% CGA ^ 80-100% CGA ^ 0-20% CGA ^ 5-40% CGA ^ 20-45% CGA ^ 40-70% CGA ^ 50-85% CGA Ethanol:water blends ^ 50-70% ethanol ^ 30-50% ethanol ^ 30-70% ethanol ^ 20-80% ethanol ^ 20-40% ethanol ^ 0-80% ethanol ^ 0-20% ethanol Maximum temp for “raw” preparations/minimum temp for roasted: ^ 105 °C ^ 100 °C ^ 120 °C ^ 85 °C ^ 65 °C ^ 40 °C ^ 20 °C Final roasting temps ^ 195-250 °C ^ 200-230 °C ^ 180-200 °C ^ 230-250 °C ^ 160-200 °C ^ 120-200 °C ^ 120-160 °C Compounding temperature ranges 112 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 ^ 80-140 °C ^ 100-140 °C ^ 80-100 °C ^ 100-165 °C ^ 160-200 °C ^ 175-225 °C ^ 190-250 °C ^ 0-50 °C ^ 30-80 °C Example 47 (Exemplary polysaccharide types, levels and ratios, along with exemplary reaction conditions, and ranges thereof, relating to exemplary processes used in the working examples were determined.) This example describes exemplary polysaccharide types, levels and ratios, along with representative reaction conditions, and ranges thereof, relating to exemplary processes used in the working examples. Polysaccharide types and levels (e.g., Rhamnogalacturonan). One rhamnogalacturonan type, rhamnogalacturonan I (RG-I), is an important component of a larger pectin family, and has a backbone composed of galacturonic acid and rhamnose residues linked glycosidically. Attached to the rhamnose residues of the RG-I backbone are side chains that can either be linear or branched arabinogalactan polymers or homopolymers of arabinose or galactose. The relative proportions and chain lengths of these side chains can vary depending on the plant source. RG-Is are present in varying amounts in the plant kingdom due to their roles in cell wall structure and function. Consistent with this, and according to particular aspects of the present invention, not all plants containing RG-I polysaccharides contribute equally to coffee-like attributes in the disclosed compositions and processes. According to particular aspects of the disclosed methods and compositions, the rhamnose:galacturonic acid molar ratio may be used as an important indicator used to assess whether RG-containing plants are suitable or optimal/preferred for providing coffee-like organoleptic properties. A higher content of rhamnose relative to galacturonic acid implies more branch points for neutral sugars to attach, and in turn provision of time- 113 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 release and/or enhanced or modified coffee-like organoleptic properties. According to additional aspects of the methods and compositions, the relative side chain lengths between and among RG sources can be used to assess whether particular RG sources are suitable or optimal/preferred for providing time release and/or enhanced or modified coffee-like organoleptic properties—with longer side chain length implying more potentially reactive neutral sugars. According to additional aspects of the disclosed methods and compositions, another important indicator is the ratio of rhamnose + galactose + arabinose to galacturonic acid, which may be used to evaluate the degree of branching in RGs. Generally, a higher ratio of neutral sugars (e.g., rhamnose + galactose + arabinose) to galacturonic acid indicates more side chains in comparison to HG, which could potentially break down and release more fresh sugars during later stages of coffee roasting (e.g., providing a time-release source of reactive sugars). According to particular aspects of the disclosed methods and compositions, therefore, varying (e.g., increasing) the ratio of neutral sugars (e.g., rhamnose + galactose + arabinose) to galacturonic acid (e.g., by selecting particular sources and/or preparations of RG) can be used to enhance or modify coffee-like organoleptic properties. According to additional aspects of the disclosed methods and compositions, varying the relative ratios of the neutral sugars (e.g., varying the ratios of rhamnose, galactose and arabinose to each other) in the context of the overall ratio of neutral sugars to galacturonic acid may be used to enhance or modify coffee-like organoleptic properties. Accordingly, the RG source may be selected or modified, in terms of its RG characteristics (e.g., the relative proportions/distributions of various sugars) in order to provide enhanced or modified time-release effects and/or enhanced or modified coffee- like organoleptic properties. Exemplary levels of RG and RG-derived saccharides ranging from 0.5% to 40% in the final reactant formulation have been tested. For example relatively moderate or low levels of such saccharides (e.g., 20 wt.% strawberry fiber in reactant formula, containing 0.78% dry matter RGs), provide for enhancements in coffee-like qualities compared to the respective control lacking such polysaccharides, including provision of increased body, as well as coffee-like acidity and sweetness. Exemplary total RG levels, expressed as wt.% of dry matter, are as follows: ≥0.25%, ≥0.5%, ≥1%, ≥2%, ≥3%, ≥4%, ≥5%, ≥10%, ≥20%, ≥30%, ≥40%, ≥50%, ≥60%, or a value in ranges of 0.25-65%, of 0.25-50%, of 0.25-40%, of 0.25-30%, of 0.25- 114 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 20%, 0.25-10%, 1-8%, 2-15%, 5-25%, 10-40%, or 35-65%. Preferably, the total RG level, expressed as wt.% of dry matter, is between 0.25% and 65%, or any subrange therein, but may be greater or lesser depending on the RG source/composition. AGP levels. Arabinogalactan proteins (AGPs) are present in a variety of plant tissues, and notably comprising approximately one-third (1/3) of the polysaccharides of traditional coffee seeds. Additionally, the arabinogalactan side chains of RGs can in some cases bond to proteins, forming AGP-like structures that are both bound to proteins and galacturonic acid polymers. Herein, we refer to AGPs as being those protein- polysaccharide complexes lacking connection to a pectic polysaccharide. Exemplary total AGP levels (those not present in the form of complexes with pectic polysaccharides), expressed as wt.% of dry matter (reactant formulation), are as follows: <0.5%, <1%, <4%, <7%. <10%, <14%, <25%, <35%, <50%. Preferably, the total AGP level, expressed as wt.% of dry matter, is in the range of 0.5-14%, 0.5-50%, or any subrange therein, but may be greater or lesser, depending on the desired product characteristics. Polysaccharide modification conditions. Various exemplary conditions may be applied for polysaccharide modification of the RG source materials as follows (and where any of the exemplary conditions may be further enhanced through the use of pectin- degrading enzymes, for example polygalacturonase): Ambient pressure condition set: a pH in the range of 1-3, 4-5, 6-8, or 7-12; a process time (in hours) in the range of 1-2, or 2-4; a process temperature (in °C) in the range of 25-80, 25-70, 25-60, 25-50, 25-40, 25-30, or in any subranges of the preceding ranges, such as 60°C, 70°C or 80°C; in particular embodiments the ambient condition set is pH 1-3, 2-4 h, and 80°C; in particular embodiments the ambient condition set is pH 7-12, 2-4 h, and 80°C. Subcritical extraction condition set: a temperature (in °C) in the range of 105-115, 105-125, or 105-135; a process time (in hours) in the range of 1-2, 1-3, 1-4, or 2-4; a pressure (in bar) of 0.5, 1, 2, or in the range of 0.2 to 2 bar. 115 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Supercritical extraction condition set: a process temperature (in °C) in the range of 400-500, 400-450, or 450- 500; a process time (in hours) in the range of 1-2, 1-3, 1-4, or 2-4; a pressure (in MPa) in the range of 30-50, such as 30, 40, or 50. Extrusion conditions. Particular methods may comprise extrusion, wherein the following material temperatures conditions may apply: Extrusion condition set: a temperature (in °C) in the range of 60-210, 60-180, 60-100, 80-120, 80- 140, 100-160, or 120-210. Roasting conditions. Particular methods may comprise roasting, wherein the following maximum temperatures may apply: Roasting condition set: attaining a temperature (in °C) in the range of 120-250, 180-210, 200-250, or 120-180. Example 48 (Negative control compositions comprising potato starch were found to be weak in strength and bland providing no meaningful coffee-like qualities, and may actually displace constituents that contribute coffee-like qualities.) Methods. A 100 g amount of potato starch was hydrated with water to achieve a moisture content of 20%. This mixture was then pressed through a single screw extruder with a preheated barrel at 85°C and a 6 mm die opening, before being cut into 12-15 mm long segments to create compounded nuggets. These potato starch nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 9 minutes and 30 seconds, reaching a final temperature of 205°C. The roasted nuggets were sensory evaluated using three distinct methods: espresso, immersion, and percolation. For the espresso method, 6 g of the roasted nuggets were milled and blended with 14 g of finely ground, roasted, cross-Maillardized (xMR) date seeds (prepared as described in working examples 1 and 2 of PCT Patent Application PCT/US2021/025565, published as WO 2021/202989). The 20 g mixture was extracted using a Decent DE1PRO espresso machine, and the resulting beverage was tasted to assess its 116 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 organoleptic qualities. A control sample was prepared using 20 g of ground, roasted xMR date seeds. For the immersion method, 10 g of roasted nuggets were milled and then immersed in 170 g of boiling water for 3 minutes. After this period, the sample was tasted using the cupping technique traditionally used for coffee to evaluate its organoleptic qualities (A System to Assess Coffee Value: Understanding the Specialty Coffee Association's Value Assessment (June 2024). https://static1.squarespace.com/static/584f6bbef5e23149e5522201/t/667182ffdde8a50 81afc2d8c/1718715138872/SCA+-+A+System+to+Asssess+Coffee+Value+- +June+2024+(Secured) .pdf). A control sample was prepared using 10 g of preconditioned date seed grounds (xMR date seeds; prepared as described in working examples 1 and 2 of PCT Patent Application PCT/US2021/025565, published as WO 2021/202989). In the percolation method, a control sample was first prepared by placing 15 g of a customized roasted coffee blend into a Hario V60 dripper equipped with filter paper. This blend consisted of 50% coffee substitute (comprising 30% Ramon seed flour, 65% preconditioned date seed (xMR date seeds, as used above), and 5% millet, lemon, guava blend) and 50% traditional coffee. The 15 g blend was placed in a Haro V60 dripper equipped with filter paper, and 240 g of hot water at 95 °C was slowly poured over it. For the inventive composition, 2.5 g (17 wt.%) of the customized roasted coffee blend was replaced with the milled roasted nuggets mentioned above. The resulting brews were then tasted to evaluate their organoleptic properties. Results. Across all three brewing methods, the composition comprising potato starch was found to be weak in strength and bland in taste compared to the xMR and/or traditional coffee controls. Negligible coffee-like attributes were detected in the potato starch composition. Whether alone (immersion), paired with a bean-less coffee (espresso) or traditional coffee (percolation), starch provides no meaningful coffee-like qualities. Furthermore, when roasted, purified starch is used as a portion of a mixture with traditional coffee or bean-less coffee compositions, it makes the resulting mixture less coffee-like by displacing constituents that contribute coffee-like qualities. Example 49 (A bean-less espresso coffee composition was improved through inclusion of a mixture of Ramon seed flour and an extract of sugar beet pulp.) 117 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Methods. Fifty (50) grams of ground sugar beet pulp were mixed with 500 grams of water and placed in an autoclave. The mixture was autoclaved for 240 minutes at 135°C, then filtered through a 200 μm filter mesh followed by a 50 μm filter. It was subsequently concentrated using a rotary evaporator until it reached 50% solids. 100 g of finely milled Ramon seed flour (D75 = 250 μm) was combined with 20 g of the sugar beet extract described above and mixed thoroughly. The moisture content of this mixture was adjusted to 18% total. This mixture was then pressed through a single screw extruder with a preheated barrel (100°C) and a 6 mm die opening, then cut into 12-15 mm length segments to create compounded nuggets. These compounded nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 8 minutes to a final temperature of 205°C. A negative control sample was prepared using the same methodology but omitting the sugar beet pulp extract. Six (6) g of the milled nugget composition was ground and blended with 14 g of finely ground, roasted, xMR date seeds, a known coffee substitute (prepared as described in working examples 1 and 2 of PCT Patent Application PCT/US2021/025565, published as WO 2021/202989). An equivalent control xMR composition was made using the negative control nuggets. These 20 g samples were separately extracted in a Decent DE1PRO espresso machine and tasted to evaluate their organoleptic qualities. Results. In comparison to the control xMR sample, the sensory panel showed a preference for the inventive composition (milled nuggets plus xMR). The inventive composition demonstrated superior coffee-like attributes, particularly in sweetness, roastiness, and body. Another notable observation was the increase in brix of the inventive composition, relative to the control xMR composition, from 4.89 to 6.23. Without being bound by mechanism, this increase is likely a contributing factor to the enhancement in the body, since traditional espresso is generally in the range of 8-12 ºBx and the espresso made from date seeds and Ramon seeds alone (without the processed sugar beet pulp) was significantly less. According to particular aspects, therefore, a beanless espresso coffee composition was improved through inclusion of a mixture of Ramon seed flour and an extract of sugar beet pulp. 118 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Example 50 (Bean-less espresso coffee compositions were improved through inclusion of a mixture of Ramon seed flour and extracts of sugar beet pulp produced using varying pH conditions.) Methods. Three samples were prepared where each sample contained 50 grams of ground sugar beet pulp mixed with 500 grams of water. The pH of each mixture was adjusted to 2.5 or 12 using citric acid or potassium hydroxide, respectively, or left unadjusted. After incubating each sample at 80°C for 2 hours, they were separately filtered initially through a 200 μm mesh filter followed by a 50 μm filter. Following filtration, all samples were concentrated using a rotary evaporator until they reached a solids concentration of 50%. 100 grams of finely milled Ramon seed flour (with a D75 of 250 μm) were mixed thoroughly with 20 grams of sugar beet ambient pressure extract (pH unchanged, adjusted with acid or alkali as described earlier). The moisture content of this mixture was adjusted to 18% total. This mixture was then pressed through a single screw extruder with a preheated barrel (100°C) and a 6 mm die opening, then cut into 12-15 mm length segments to create compounded nuggets. These compounded nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 8 minutes to a final temperature of 205°C. A negative control sample was prepared using the same methodology but omitting the sugar beet pulp extract. Six (6) g of the milled nugget composition was ground and blended with 14 g of finely ground, roasted, xMR date seeds, a known coffee substitute (prepared as described in working examples 1 and 2 of PCT Patent Application PCT/US2021/025565, published as WO 2021/202989). An equivalent control xMR composition was made using the negative control nuggets. These 20 g samples were separately extracted in a Decent DE1PRO espresso machine and tasted to evaluate their organoleptic qualities. Results. Compared to the control xMR sample, the sensory panel exhibited a preference for the novel composition, particularly favoring the alkali-processed extract. The innovative blend demonstrated enhanced coffee-like qualities, notably in sweetness, roastiness, and body. While other extracts also showed slight improvements in coffee- like attributes, particularly in body, these enhancements were not as pronounced as those observed with the alkali-processed extract, particularly in terms of roastiness. 119 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Example 51 (Sugar beet pulp was used to create a coffee-like composition, and further used as a component of a coffee blend with traditional coffee.) Methods. A 100 g portion of finely milled sugar beet pulp flour was adjusted to a moisture content of 25%. This mixture was then processed through a single screw extruder with a preheated barrel set to 100°C and a 6 mm die opening. The extruded material was cut into segments of 12-15 mm in length, forming nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 9 minutes and 30 seconds, reaching a final temperature of 205°C. The roasted nuggets were evaluated using two distinct methods: immersion and percolation. For the immersion method, an extract was made using 10 g of the milled nuggets following the process from Example 48. When tasted hot, the immersion sample showcased a robust coffee-like body with roasted notes reminiscent of coffee, complemented by pleasant sweetness, vegetal notes, and a pronounced acidity. As the sample cooled, it revealed sweet and acidic flavors, a coffee-like upfront bitterness and astringency. In the percolation method, a control sample was first prepared by placing 15 g of a customized roasted coffee blend into a Hario V60 dripper equipped with filter paper. This blend consisted of 50% coffee substitute (comprising 30% Ramon seed flour, 65% preconditioned date seed, and 5% millet, lemon, guava blend) and 50% traditional coffee. The 15 g blend was placed in a Haro V60 dripper equipped with filter paper, and 240 g of hot water at 95°C was slowly poured over it. For the inventive composition, 2.5 g (17 wt.%) of the customized roasted coffee blend was replaced with the milled roasted nuggets mentioned above. The resulting brews were then tasted to evaluate their organoleptic properties. Results. When tasted hot, the percolation sample containing the milled nuggets exhibited a slightly diminished overall coffee flavor compared to the 50:50 control and had increased astringency, yet showed more coffee-like roastiness. Upon cooling, the percolation sample presented a pleasant aroma with notes of sweetness, honey, coffee- like earthiness, roastiness, and caramel, along with a hint of savory. Additionally, the volatile spectrum of the immersion sample of these sugar beet pulp nuggets were analyzed using GC-MS. This analysis comprises 3 steps: trapping headspace volatiles, collecting data in the GC-MS and the final analysis of these results 120 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 to identify the compounds present in the extract. In all cases, a sample of the immersion extract of roasted potato starch nugget (Example 48) is provided for comparison. Highlighted peaks are those that both are relevant for coffee flavor and increase in magnitude using an RG-containing composition instead of the negative control, which contains essentially zero RG. Headspace SPME volatiles trapping A 75 µm Car-DVB-PDMS (Supelco) SPME fiber with manual holder was used throughout the study. Immersion extracts (2 mL) were added to a 20 mL headspace vial and the vial capped. The sample was equilibrated at 50 ºC for 20 minutes with magnetic stirring (400 RPM). The SPME needle was then pierced through the septum and the fiber was exposed to the sample headspace for 15 minutes. After trapping, the fiber was retracted, injected, and exposed in the GC inlet set at 240 ºC in split-less injection mode. The desorption time was 60 seconds followed by a 60 second fiber cleaning at 240 ºC and 50 mL/minute helium purge flow. GCMS analysis Volatiles were analyzed on a 8890 GC coupled with a 7250 Q-TOF detector. The desorbed volatiles were separated on a DB-Wax UI column (60 m x 0.25 mm x 0.25 µm) with helium carrier gas flow of 1.82 mL/min. The oven was programmed to ramp from 40 ºC to 160 ºC at a 6 ºC/min rate, held for 5 minutes at 160 ºC, and finally a 8 ºC/min ramp to 230 ºC with a final hold of 2 minutes. The MS was operated in Scan mode. The mass spectrometer scan parameters were as follows: 35-400 Da mass scan range with a scan speed of 1948 mass units per second and frequency of 5 scans/second. Data extraction and compound identification Data were extracted using Agilent MassHunter Unknowns Analysis software. A retention time window size factor of 100 was used for deconvolution with an absolute area filter of 1000. Compounds were identified based on comparison of their mass spectra with the spectra in the NIST 20 database and a minimum match factor threshold of 70 was set for identification. FIG.19 shows the resulting spectrum including highlighting the aroma compounds important for coffee. The furan compounds contribute various sweet/caramel roast notes, while pyrrole and pyrazine family compounds contribute a variety of earthy roast flavors to the resulting coffee. The 2/3-methylbutanal contributes chocolate-like aroma and the methyl ester of hexanoic acid contributes a fruity (pineapple-like) aroma. According to particular aspects, therefore, sugar beet pulp was used to create a 121 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 coffee-like composition, and further used as a component of a coffee blend with traditional coffee. Example 52 (An extract of sugar beet pulp was used to create a coffee-like composition and as a component of a coffee blend with traditional coffee.) Methods. The 50% solids sugar beet extract of Example 49 was poured into silicon molds shaped like coffee beans (dimensions: 1 cm x 1.6 cm x 1.3 cm) and dried at 55°C in a food dehydrator. The dried sugar beet extract beans, hereafter referred to as “extract-derived beans” or “ED-beans” were then toasted in a belt oven at 425°C for 3 minutes and 30 seconds before being ground into a fine powder. The toasted ED-beans were evaluated using two distinct methods: immersion and percolation. In the immersion method, 10 g of the toasted milled ED-beans were immersed in 170 g of boiling water for 3 minutes. Subsequently, the sample was tasted using the cupping technique to assess its organoleptic properties. When tasted hot, the immersion sample exhibited a robust body with a texture reminiscent of a Moka pot, pronounced coffee-like roastiness and sweetness, strong acidity with a slight sourness, and a lingering bitterness. Upon cooling, the sample presented flavors akin to toasted marshmallow, with a sweet, toasty profile, strong body, and coffee-like astringency. This is especially notable, as the texture of a traditional coffee beverage brewed using a Moka pot is considerably more viscous than a cupping method, due to the higher concentration of the extracted beverage. In the percolation method, a control sample was first prepared by placing 15 g of a customized roasted coffee blend into a Hario V60 dripper equipped with filter paper. This blend consisted of 50% coffee substitute (comprising 30% Ramon seed flour, 65% preconditioned date seed, and 5% millet, lemon, guava blend) and 50% traditional coffee. Then, 240 g of hot water at 95°C was slowly poured over the mixture. For the inventive composition, 2.5 g (17 wt.%) of the customized roasted coffee blend was replaced with the milled ED-beans mentioned above. The resulting brews were then tasted to evaluate their organoleptic properties. Results. Although approximately 17 wt.% of the customized roasted coffee blend was substituted with roasted sugar beet extract beans, the inventive composition retained a similar taste profile to the control sample. Moreover, certain attributes, such as body, 122 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 were enhanced to be more reminiscent of traditional coffee. Specifically, in hot percolation samples, the inventive composition exhibited comparable sweetness and roastiness but was noted to be more astringent than the control. It also featured coffee- like earthy notes and a more pronounced "coffee breath." Upon cooling, the percolated sample demonstrated improved body compared to the control, with a clean acidity profile reminiscent of malic acid rather than citric acid. The volatile spectrum of the immersion sample of ED bean grounds were also analyzed using GC-MS, using the same instrument and protocol as described in Example 51. The resulting spectrum is depicted in FIG.20. The furan and pyrrole contribute roast- like flavors, and 2/3-methylbutanal chocolate flavors, as described previously. Acetoin, 2,3-pentanedione and 2.3-butanedione contribute buttery aroma, while benzaldehyde contributes an almond-like aroma. Example 53 (A coffee composition was improved through the inclusion of a carrot composition with Ramon seeds.) Methods. Eighty (80) g of finely milled Ramon seed flour (D75 = 250 μm) was combined was 20 g of dehydrated carrot powder and mixed thoroughly. The moisture content of this mixture was adjusted to 18% total. This mixture was then pressed through a single screw extruder with a preheated barrel (100°C) and a 6 mm die opening, then cut into 12-15 mm length segments to create compounded nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 8 minutes to a final temperature of 205°C. A control sample was prepared using the same methodology but omitting the dehydrated carrot powder. Six (6) g samples of the inventive composition were blended with 14 g of finely ground, roasted, preconditioned date seeds (roasted xMR date seeds, as above). An analogous control sample was prepared using the control nuggets. These 20 g samples were separately extracted in a Decent DE1PRO espresso machine and tasted to evaluate their organoleptic qualities. Results. In contrast to the control sample, the inventive composition showcased superior coffee aroma and coffee taste quality, featuring heightened sweetness, acidity, body, strength, and mouthfeel. During sensory evaluation, a pleasant balance between sweetness and acidity stood out as noteworthy. Participants also remarked on a syrupy mouthfeel that closely mirrored the qualities of espresso. The brix value in the inventive 123 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 composition increased from 4.89 to 6.43 compared to the control sample. Furthermore, the pH shifted from 5.36 to 4.85 in the inventive composition, especially noteworthy as carrots are not known for being acidic. Without being bound by mechanism, this likely indicates that the roast converted constituents of the carrot into flavorful acids, present at levels appreciated by trained coffee tasters, in an analogous fashion to traditional coffee. Example 54 (A compounded mixture of Ramon seed flour and dehydrated carrot pulp powder was combined with roasted preconditioned date seeds to create a coffee composition.) Methods. Eighty (80) g of finely milled Ramon seed flour (D75 = 250 μm) was combined was 20 g of dehydrated carrot pulp powder and mixed thoroughly. The moisture content of this mixture was adjusted to 18% total. This mixture was then pressed through a single screw extruder with a preheated barrel (100 °C) and a 6 mm die opening, then cut into 12-15 mm length segments to create compounded nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 8 minutes to a final temperature of 205°C. A control sample was prepared using the same methodology, but omitting the dehydrated carrot pulp powder. Six (6) g samples of the inventive composition were blended with 14 g of finely ground, roasted, preconditioned date seeds (roasted xMR date seeds, as above). A analogous control composition was made using the control nuggets. These 20 g samples were separately extracted in a Decent DE1PRO espresso machine and tasted to evaluate their organoleptic qualities. Results. In contrast to the control sample, the innovative composition demonstrated an enhanced coffee aroma and coffee flavor profile, specifically showing a substantial increase in brightness and upfront acidity, along with improved body and strength. In the inventive composition, the brix value experienced a rise from 4.89 to 5.63, bringing this composition closer to traditional espresso despite containing no coffee beans. Additionally, there was a shift in the pH from 5.36 to 4.97 in the inventive composition, especially noteworthy as carrots are not known for being acidic. When interpreted in the context of the prior example, even the ”spent” remains of carrots— lacking much of the small sugars extracted during the juicing process—retain important, generally polymeric, constituents for the creation of improved coffee flavors and textures. 124 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 Example 55 (Dehydrated carrot powder was used to create a coffee-like composition and as a component of a coffee blend with traditional coffee.) Methods. A 100 g of dehydrated carrot powder was adjusted to a moisture content of 25%. This mixture was then processed through a single screw extruder with a preheated barrel set to 100°C and a 6 mm die opening. The extruded material was cut into segments of 12-15 mm in length, forming nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 4 minutes and 30 seconds, reaching a final temperature of 205°C. The roasted nuggets were evaluated using two distinct methods: immersion and percolation. Using the immersion method, an extract was prepared with 10 grams of milled carrot nuggets, following the procedure outlined in Example 48. When tasted hot, the immersion sample exhibited a mid-intensity coffee-like acidity, a robust body, pleasant caramel notes, and pronounced bitterness with a roasty flavor. As the sample cooled, its flavor profile improved, displaying coffee-like sweetness, a strong body, lingering bitterness, and notes reminiscent of dark, caramelized fruit sugars and dried fruit. In the percolation method, a control sample was first prepared with 15 g of a customized roasted coffee blend (same as in Example 52), following the procedure outline in Example 52. For the inventive composition, 2.5 g (17 wt.%) of the customized roasted coffee blend was replaced with the milled roasted carrot nuggets mentioned above. The resulting brews were then tasted to evaluate their organoleptic properties. Results. Although approximately 17 wt.% of the coffee blend was substituted with roasted carrot powder nuggets, the innovative composition maintained a taste profile similar to the control sample in terms of coffee-like attributes, with an even stronger body and enhanced sweetness. The hot percolation sample exhibited similar roastiness to the control but was less bitter, had a slightly better body, and tasted like roasted sweet root. Upon cooling, the percolated sample demonstrated stronger bitterness compared to the control, along with some dark fruit flavor and an improved body. The volatile aroma compounds of the immersion sample of these roasted nuggets were analyzed using GC-MS, using the same protocol as described in Example 51. The resulting volatile spectrum is depicted in FIG. 21, with the coffee-like compounds highlighted. As mentioned previously, compounds from the furan family contribute roasted flavors, 2/3-methybutanal contributes chocolate flavor and 2,3-pentanedione 125 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 contributes buttery notes. Ionene contributes a floral (rose-like) aroma. Example 56 (Dehydrated carrot pulp was used to create a coffee-like composition and as a component of a coffee blend with traditional coffee.) Methods. A 100 g of dehydrated carrot pulp powder was adjusted to a moisture content of 25%. This mixture was then processed through a single screw extruder with a preheated barrel set to 100°C and a 6 mm die opening. The extruded material was cut into segments of 12-15 mm in length, forming nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 4 minutes and 30 seconds, reaching a final temperature of 205°C. The roasted nuggets were evaluated using two distinct methods: immersion and percolation. Utilizing the immersion technique, an extract was prepared with 10 grams of milled carrot pulp nuggets, adhering to the procedure described in Example 48. Upon tasting the heated immersion sample, it exhibited sensory characteristics of coffee-like roastiness, a pleasant acidity, sweetness similar to that of brown sugar, and a flavor reminiscent of unripened persimmon, accompanied by a pleasant coffee breath. As the sample cooled, the sweetness remained, but there was an increase in body and bitterness. In the percolation method, a control sample was first prepared with 15 g of a customized roasted coffee blend (same as in Example 49), following the procedure outline in Example 52. For the inventive composition, 2.5 g (17 wt.%) of the customized roasted coffee blend was replaced with the milled roasted carrot pulp nuggets mentioned above. The resulting brews were then tasted to evaluate their organoleptic properties. Results. Despite substituting approximately 17 wt.% of the coffee blend with roasted carrot pulp powder nuggets, the novel composition maintained a taste profile akin to the control blend. The hot percolation sample exhibited similar roastiness to the control but with intensified sweetness and bitterness. Upon cooling, the percolated sample retained its initial characteristics while developing notes of dark cherry and blackberry, accompanied by improved body. In comparison to carrot nuggets, carrot pulp nuggets generally exhibited greater bitterness and reduced sweetness. The volatile aroma compounds present in the immersion extract of roasted carrot 126 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 pulp nuggets were analyzed by GC-MS, using the same protocol as described in Example 51. These results are depicted in FIG.22. Many compounds (furans, 2/3-methyl butanal, 2,3-pentanedione, methyl hexanoate, ionene) contribute the same notes discussed in prior examples.2-methyl propanal contributes a pungent floral aroma. Example 57 (A compounded mixture of Ramon seed flour and potato fiber powder was combined with roasted preconditioned date seeds to create a coffee composition.) Eighty (80) g of finely milled Ramon seed flour (D75 = 250 μm) was combined was 20 g of dehydrated potato fiber powder and mixed thoroughly. The moisture content of this mixture was adjusted to 18% total. This mixture was then pressed through a single screw extruder with a preheated barrel (100°C) and a 6 mm die opening, then cut into 12-15 mm length segments to create compounded nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 8 minutes to a final temperature of 205°C. A control sample was prepared using the same methodology but omitting the dehydrated potato fiber powder. Six (6) g samples the inventive composition were blended with 14 g of finely ground, roasted, preconditioned date seeds. An analogous control sample was made using the control nuggets. These 20 g samples were separately extracted in a Decent DE1PRO espresso machine and tasted to evaluate their organoleptic qualities. In contrast to the control sample, the innovative composition revealed an improved coffee aroma and coffee flavor profile, particularly highlighting heightened body and aroma pungency. Example 58 (Potato fiber was used to create a coffee-like composition and as a component of a coffee blend with traditional coffee.) Methods. In this example, A 100 g of dehydrated potato fiber powder was adjusted to a moisture content of 25%. This mixture was then processed through a single screw extruder with a preheated barrel set to 100°C and a 6 mm die opening. The extruded material was cut into segments of 12-15 mm in length, forming nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 7 minutes and 30 seconds, reaching a final temperature of 205°C. The roasted nuggets were evaluated using two distinct methods: immersion and 127 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 percolation. Using the immersion method, an extract was prepared with 10 g of milled potato fiber nuggets, following the procedure outlined in Example 48. Upon tasting the heated immersion sample, it displayed sensory characteristics with a substantial body, earthy and coffee-like roastiness, accompanied by a hint of smokiness. As the sample cooled, the roastiness persisted, and a more pronounced acidity and sweetness emerged, along with nutty notes reminiscent of peanuts. In the percolation method, a control sample was first prepared with 15 g of a customized roasted coffee blend (same as in Example 52), following the procedure outline in Example 52. For the inventive composition, 2.5 g (17 wt.%) of the customized roasted coffee blend was replaced with the milled roasted potato fiber nuggets mentioned above. The resulting brews were then tasted to evaluate their organoleptic properties. Results. Despite substituting approximately 17 wt.% of the coffee blend with roasted potato pulp powder nuggets, the new composition maintained a taste profile similar to the control, though with heightened aspects such as roastiness, nuttiness, and bitterness. Specifically, the hot percolation sample exhibited a complex and layered flavor profile, prominently showcasing roastiness, good acidity, and a distinctive coffee varietal essence that is unmistakably coffee-like. Upon cooling, the percolated sample retained its initial attributes while developing nuances of vanilla, marshmallow, nuttiness reminiscent of peanuts, along with well-balanced acidity and a pleasant upfront bitterness that transitions smoothly. The volatile aroma compounds of these roasted potato fiber nuggets were analyzed using GC-MS, using the same protocol as described in Example 51. The resulting spectrum is depicted in FIG.23. Unlike the arabinose-rich examples (carrot and beet), which were rich in furan family compounds, these nuggets create a plethora of pyrazine, pyridine and pyrrole family compounds along with some furan compounds.2- heptanone contributes a fruity (banana-like) aroma, while dimethyl disulfide is a pungent aroma compound associated with roasted coffee. A precursor to dimethyl disulfide is 2- FFT, a crucial odorant for roasted coffee described earlier. Example 59 (A compounded mixture of Ramon seed flour and strawberry fiber powder was combined with roasted preconditioned date seeds to create a coffee composition.) Methods. Eighty (80) g of finely milled Ramon seed flour (D75 = 250 μm) was 128 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 combined was 20 g of dehydrated strawberry fiber powder and mixed thoroughly. The moisture content of this mixture was adjusted to 18% total. This mixture was then pressed through a single screw extruder with a preheated barrel (100°C) and a 6 mm die opening, then cut into 12-15 mm length segments to create compounded nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 8 minutes to a final temperature of 205°C. A control sample was prepared using the same methodology but omitting the dehydrated strawberry fiber powder. Six (6) g samples the inventive composition were blended with 14 g of finely ground, roasted, preconditioned date seeds (roasted xMR date seeds, as above). An analogous control sample was made using the control nuggets. These 20 g samples were separately extracted in a Decent DE1PRO espresso machine and tasted to evaluate their organoleptic qualities. Results. In contrast to the control sample, the innovative composition displayed an intensified coffee aroma and flavor profile, characterized by a fuller body, heightened aroma intensity, and a distinctive lingering sweet bitterness. Moreover, the inventive blend reveals a unique strawberry jam-like flavor. Example 60 (Strawberry fiber was used to create a coffee-like composition and as a component of a coffee blend with traditional coffee.) Methods. A 100 g of dehydrated strawberry fiber powder was adjusted to a moisture content of 25%. This mixture was then processed through a single screw extruder with a preheated barrel set to 100°C and a 6 mm die opening. The extruded material was cut into segments of 12-15 mm in length, forming nuggets. These nuggets were roasted in a fluid bed roaster (IKAWA Pro) for 4 minutes and 30 seconds, reaching a final temperature of 195°C. The roasted nuggets were evaluated using two distinct methods: immersion and percolation. Results. Utilizing the immersion method, an extract was prepared with 10 grams of roasted, milled strawberry fiber nuggets, following the procedure described in Example 48. Upon tasting the hot immersion sample, it exhibited sensory characteristics with a pronounced acidity reminiscent of malic acid, a tea-like astringency, and a sweetness akin to coffee. As the sample cooled, the sweetness endured, and more prominent 129 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 fruitiness and roastiness emerged, while the acidity became more assertive. In the percolation method, a control sample was first prepared with 15 g of a customized roasted coffee blend (same as in Example 52), following the procedure outline in Example 52. For the inventive composition, 2.5 g (17 wt.%) of the customized roasted coffee blend was replaced with the milled roasted strawberry fiber nuggets mentioned above. The resulting brews were then tasted to evaluate their organoleptic properties. Despite substituting approximately 17 wt.% of the coffee blend with roasted strawberry fiber powder nugget grounds, the new composition maintained a taste profile akin to the control, albeit with heightened coffee-like aspects such as fruitiness, sweetness, and acidity. Specifically, the hot percolation sample exhibited a fruitier taste compared to the control, with a lingering bitterness and strong acidity, while maintaining the same level of roastiness. Upon cooling, the percolated sample retained its initial attributes. The volatile spectrum in the immersion extract of these nuggets was also analyzed by GC-MS, using the same protocol as described in Example 51. The resulting spectrum is depicted in FIG.24. The furan family compounds, 2-methyl propanal and benzaldehyde contribute the same notes as described prior. Additionally, fruity aromas are contributed by ethyl formate (raspberries) and methyl acetate, while pentanedione contributes creamy/buttery aroma. Example 61 (A coffee composition was made by compounding Ramon seeds, green banana flour, a sunflower seed extract containing CGA, strawberry fiber, carrot powder, potato fiber, black aronia berry powder, carob pod powder.) Methods. A flour blend weighing 100 g, composed of 4 g of carrot powder, 23.9 g of green banana flour, 6.4 g of strawberry fiber, 1.6 g of aronia powder, 4 g of potato fiber powder, 7.6 g of unroasted carob powder, 40.4 g of milled Ramon seed flour, and 12.1 g of SFE powder, was adjusted to achieve a moisture content of 25%. This mixture underwent compounding by a single screw extruder with a preheated barrel set at 100°C and a 6 mm die opening. The extruded material was then cut into segments measuring 12-15 mm in length, forming nuggets. These nuggets were subsequently roasted in a fluid bed roaster (IKAWA Pro) for 9 minutes and 30 seconds, reaching a final temperature of 205°C. The roasted nuggets were assessed using a percolation method. A control sample 130 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 was prepared using traditional coffee in the same percolation method. Fifteen (15) g of milled nuggets or coffee ground was placed in a Hario V60 dripper fitted with filter paper, and 240 g of hot water at 95°C were slowly poured over it. The resulting brews were evaluated for their sensory characteristics. Results. The innovative composition exhibited many similarities to the control, particularly in terms of acidity and body. However, it showed slightly lower roastiness while demonstrating increased fruitiness and sweetness compared to the control. Example 62 (A coffee blend was made with a mixture of date seeds, RG-rich plant materials and traditional coffee seeds.) Methods. Coffee substitute nuggets were prepared using the same protocol described in Example 58.3.75 g of these roasted grounds were blended with 3.75 g of ground, roasted, xMR date seeds and 7.5 g of ground roasted traditional coffee seeds. Samples for organoleptic evaluation were prepared using the method described in Example 58. A control sample was prepared using 15 g of the traditional roasted coffee grounds. Results. In comparison to the control sample, the inventive composition maintained crucial quality attributes of the traditional coffee—such as roasted flavors, texture and usage qualities such as extraction time and bloom—while adding in extra layers of flavor. In particular, the one-note sharp citric note of the control sample was enhanced by incorporating notes of berries and producing a more balanced overall cup. The various methods and techniques described above provide a number of ways to carry out the invention. Of course, it is to be understood that not necessarily all objectives or advantages described can be achieved in accordance with any particular embodiment described herein. Thus, for example, those skilled in the art will recognize that the methods can be performed in a manner that achieves or optimizes one advantage or group of advantages as taught herein without necessarily achieving other objectives or advantages as taught or suggested herein. A variety of alternatives are mentioned herein. It is to be understood that some preferred embodiments specifically include one, another, or several features, while others specifically exclude one, another, 131 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 or several features, while still others mitigate a particular feature by inclusion of one, another, or several advantageous features. Furthermore, the skilled artisan will recognize the applicability of various features from different embodiments. Similarly, the various elements, features and steps discussed above, as well as other known equivalents for each such element, feature or step, can be employed in various combinations by one of ordinary skill in this art to perform methods in accordance with the principles described herein. Among the various elements, features, and steps some will be specifically included and others specifically excluded in diverse embodiments. Although the application has been disclosed in the context of certain embodiments and examples, it will be understood by those skilled in the art that the embodiments of the invention extend beyond the specifically disclosed embodiments to other alternative embodiments and/or uses and modifications and equivalents thereof. In some embodiments, the numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth, used to describe and claim certain embodiments of the application are to be understood as being modified in some instances by the term “about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the application are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. In some embodiments, the terms “a” and “an” and “the” and similar references used in the context of describing a particular embodiment of the application (especially in the context of certain of the following claims) can be construed to cover both the singular and the plural. The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (for example, “such as”) provided with respect to certain embodiments herein is intended 132 4869-6285-1820v.10113409-005WO0
Attorney Docket No.0113409-005WO0 merely to better illuminate the application and does not pose a limitation on the scope of the application otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the application. Preferred embodiments of this application are described herein. Variations on those preferred embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. It is contemplated that skilled artisans can employ such variations as appropriate, and the application can be practiced otherwise than specifically described herein. Accordingly, many embodiments of this application include all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the application unless otherwise indicated herein or otherwise clearly contradicted by context. All patents, patent applications, publications of patent applications, and other material, such as articles, books, specifications, publications, documents, things, and/or the like, referenced herein are hereby incorporated herein by this reference in their entirety for all purposes, excepting any prosecution file history associated with same, any of same that is inconsistent with or in conflict with the present document, or any of same that may have a limiting affect as to the broadest scope of the claims now or later associated with the present document. By way of example, should there be any inconsistency or conflict between the description, definition, and/or the use of a term associated with any of the incorporated material and that associated with the present document, the description, definition, and/or the use of the term in the present document shall prevail. The embodiments of the application disclosed herein are illustrative of the principles of the embodiments of the invention. Other modifications that can be employed can be within the scope of the application. Thus, by way of example, but not of limitation, alternative configurations of the embodiments of the application can be utilized in accordance with the teachings herein. Accordingly, embodiments of the present application are not limited to that precisely as shown and described. 133 4869-6285-1820v.10113409-005WO0
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