CN1205690A - Natural carotenoid concentrate from plant material and method for its preparation - Google Patents

Abstract

Chromosomes and/or chloroplasts are isolated from plant material. The wet clumps are digested in an aqueous medium with pectin- and/or protein-degrading enzymes to remove insoluble substances. The resulting colloidal dispersion is acidified to obtain a carotenoid-rich precipitate. The precipitate is heated with aqueous ethanol or isopropanol at an alkaline pH to remove the major components of the degraded proteins, lipids, and other minor substances. The carotenoid-rich precipitate is mixed with an antioxidant, and may be dried if necessary, to obtain a carotenoid concentrate free of toxic residual solvents.

Description

Natural carotenoid concentrate from plant material and method for its preparation

field of invention

The present invention relates to a carotenoid concentrate obtained by processing chromoplasts and chloroplasts isolated from plant material and a process for its preparation.

Background of the invention

The carotenoids mentioned here are a class of unsaturated hydrocarbons and their lipid-added complexes, that is, isopentenyl esters, and unsaturated hydrocarbons contain isoprene units or their derivatives substituted by various functional groups .

It is well known that a healthy diet includes the consumption of moderate amounts of high-quality vegetables and fruits. Compared with different components of these foods, carotenoids are essential components for maintaining the body's health. Carotenoids such as lycopene are powerful antioxidants that trap harmful free radicals that are constantly being produced in the body. In addition, some carotenoids also function as provitamins.

In addition to the function of provitamins, the main task of carotenoids is to deactivate the large number of free radicals formed in the human body due to an increasing number of environmental hazards such as smog, increased UV radiation, air pollution, etc. However, even the best diet does not provide a sufficient and continuous supply of carotenoids, according to data from various scientists, the daily required supply of carotenoids for adults is 6-15 mg, therefore carotenoid compositions are provided as dietary supplements needs have become more urgent.

Epidemiological surveys and other experimental data have confirmed that the morbidity and mortality caused by various cancers are low in areas and populations with high consumption of vegetables and fruits. The data suggest that this diet provides an adequate supply of carotenoids, suggesting that carotenoids may have a preventive role in maintaining health.

Since it is rather difficult and sometimes impossible to provide a continuous supply of carotenoids through the diet, there is a need to supplement the diet with carotenoid compositions prepared from various sources and in this way ensure the required supply of carotenoids.

It is worth emphasizing that continuous carotenoid intake is a prerequisite for optimal results. Free radicals are constantly produced in the organism and therefore they must also be constantly detoxified. This can only be done with sufficient levels of antioxidants in the body, carotenoids are not the only compounds with antioxidant properties, tocopherols are also antioxidants, but carotenoids are integral, and together with tocopherols they can even act synergistically active.

In conclusion, carotenoids should be constantly present in the human body and at appropriate levels in blood plasma to preventively protect the organism from abnormal processes initiated by free radicals. Since this effect is preventive, the dietary schedule should be designed accordingly, ie the supplementation of carotenoids in the diet should be regularized.

Taking the above aspects into consideration, as a continuous food additive, a carotenoid composition as a medical agent or possibly a drug (as a vitamin preparation) should meet the following requirements:

a) The range of carotenoids should be similar to that in human plasma, according to literature data, the range is as follows:

α-carotene 0.12-0.05μmol/l,

β-carotene 0.40-0.80μmol/l,

Lutein (lutein + zeaxanthin) 0.28-0.35μmol/l,

Lycopene 0.49-0.74 μmol/l;

b) The raw materials used to prepare the composition will not cause any toxic damage, that is, will not cause any nutritional health problems;

c) Since the composition will be used continuously as a therapeutic agent, its production process must be designed to ensure that the final product is free of toxic contaminants such as residual solvents.

d) The composition must contain a sufficiently high content of the active ingredient so as to avoid the use of large quantities of medicaments or food additives [correspondingly, the concentration of carotenoids indicated in a) should be increased by at least 10 to 100 times. Therefore, our goal is to prepare a product with a concentration of 50,000-100,000m/kg from carrots with a carotene content of 100-200mg/kg].

Based on the bioavailability of carotenoids, an adult must consume about 1 to 2 kg of carrots and tomatoes per day in order to meet the daily requirement of 10-15 mg/kg of carotenoids. This naturally leads to undesirable dietary side effects such as excess carbohydrate consumption, poor palatability and unbalanced complex diets. Additionally, in addition to the aforementioned aspects, obtaining satisfactory carotenoid levels from an adequate supply of vegetables and fruits is limited by region and even more often by financial resources.

In view of the foregoing, there are clear advantages in producing and using a suitable carotenoid composition.

If one considers that although fresh, uncooked carrots contain 150-200 mg/kg of carotene, prepared (processed or cooked) foods contain considerably lower amounts of carotene: 7.5-8.1 mg α- and β- Carotene/100g [M. S． Micozzi et al., J Nat. Cancer． Inst. 82, 282-285 (1990); and Heinonen et al., J. Agric． Food Chem． 37,655-659 (1989)], the advantage of carotene composition is more prominent. This new product can avoid the large loss of carotene, and has higher absorption and bioavailability.

current technology

The characteristics of the existing products and their production methods are significantly different from the products and methods of the present invention.

The water-soluble or water-dispersible compositions disclosed in U.S. Patent No. 3,206,316 are most likely prepared from synthetic carotenoids, and the chlorinated solvents used in the process are not used in the process of the present invention.

WO 86/04059 mentions a process in which the juice obtained by mechanically mashing and pressing carrots is treated with pectin degrading enzymes and concentrated by ultrafiltration. The residue is utilized as a wet concentrate or in dry form, the color bodies are not further purified, so that the concentration in the final product is 500-600 mg carotene/100 g product, whereas in the product according to the invention the carotene content is 10 higher -20 times, namely 2500-5000-10,000mg carotene/100g product.

WO 92/18471 mentions the extraction of carotenoids from natural sources, especially carrots, resulting in virtually the same products as described in WO 86/04059. In the chromosomes (obtained as final product) precipitated with calcium chloride as auxiliary material, the carotene concentration was equal to 12.44 mg/g dry matter, whereas the process according to the invention allowed a ten-fold increase in concentration.

US Patent 2.412,707 mentions an oily emulsion obtained by cooking (heating) wet raw materials in edible (cooking) oil. No matter its product or method all can not be compared with product and method of the present invention.

U.S. Patent No. 2,412,707 discloses a process in which plant chromophores are separated from an aqueous suspension and precipitated by agglutination. Chromosomes were not purified, nor processed, and no attempt was made to increase the carotene concentration.

Ind. Eng． Chem． , 46, 2279 (1954) mentioned a complex extraction method in which carotenoids were selectively separated from raw materials using various solvents. Undesirable solvents such as hexane, heptane, acetone, etc. are used in this method. In contrast to our purification method, in this method the active ingredient is not filtered out of the chromosomes, but additional accompanying components are removed.

There is also a method in which carotenoids are isolated from palm oil, here carotenoids are dissolved in palm oil. Palm oil is heated (cooked) under alkaline conditions, followed by extraction of carotenoids with hexane.

Hungarian patent application P 93 03605 mentions a method in which the chromoplast fraction is flocculated in the press liquid of mechanically processed (mashed, pressed) carrots, and then most of the protein is digested with protease at pH 3.5-4. At 5 o'clock, a precipitate containing carotenoids was formed and dried.

Summary of the invention

In the course of experiments it was found that compositions with favorable properties can be prepared by refining, ie concentrating, carotenoids in chromoplasts and chloroplasts isolated from plant material.

The method is based on the observation that carotenoids in chromoplasts are associated with lipoproteins or adducted with various lipid components to form particles in the colloidal range. If lipophilic solvents are used to remove these lipids, or parts thereof, thereby disrupting the lipoprotein/lipid system, the carotenoid concentration in the residue can be adjusted without the use of undesirable or highly toxic solvents (ethane or chlorinated hydrocarbons). Several times higher, residual solvents of this type, even in trace amounts, are highly toxic due to their long-term consumption along with carotenoids (due to their similar lipophilicity to carotenoids), resulting in constant migration to target cells and tissues, where it accumulates and cannot be removed, causing great harm.

Unlike the method of U.S. Patent No. 3,206,316, the method of the present invention does not use chlorinated solvents, nor does it use aprotic nonpolar solvents such as hexane.

Detailed description of the invention

The present invention relates to a product having chromoplast/lipoprotein complexes enriched to various carotenoid concentrations, having the following properties:

a) Compared with the raw material, the concentration of carotenoids is increased by 400-500 times, and the concentration of carotenoids is increased by 2-15 times compared with the crude precipitate of colored bodies.

b) Increase concentration by cleaving the protein/lipid components of lipoprotein complexes and removing salts of polypeptides, amino acids and fatty acids by solubilization of carotenoid side chains.

c) Cleavage of the complex by taking into account the solubility of the degradation products, which are orders of magnitude more soluble in water/alcohol (ethanol, refined alcohol, isopropanol) mixtures than in ethanol, such that toxic solvents remain in the final product Not higher than the LD ₅₀ of ethanol.

d) The carotenoid range of the product obtained by processing chromoplasts isolated from suitably selected raw materials such as carrots or tomatoes meets the general requirements in an optimal manner, eg similar to the carotenoid range in human blood plasma.

The invention also relates to a process for the preparation of a product having the properties described above, which comprises isolating by known methods the color bodies containing carotenoids, preferably from carrots and tomatoes, by sedimentation or centrifugation, followed by enzymatic treatment Chromosomes partially cleave lipoproteins. After removal of accompanying substances, the resulting liquid phase is acidified, thereby precipitating a partially purified crude carotenoid extract, to which an alkaline solution is added and refluxed several times with fresh aqueous alcohol to remove the cleavage complex released during the cleavage complex. Basic salts of peptides, amino acids, fatty acids, etc. The carotenoid concentrate suspension is slightly acidified by adding sodium dihydrogen phosphate and citric acid, the product is settled or centrifuged and mixed with antioxidants, after which, if necessary, it is dried.

The completion of enzymatic cleavage preferably adds 3-5 times of dry weight of water to homogenate the chromoplasts and/or chloroplasts obtained from the plant material. The pH of the homogenate is adjusted to about 8.5 with sodium hydroxide, and then Proteolytic enzymes (preferably also have some lipase activity), such as pancreatin, enzymatically digest the mixture at 37-38° C. for 2-3 hours, while continuously adjusting the pH to the initial level by repeatedly adding sodium hydroxide solution. After enzymatic hydrolysis, the mixture was centrifuged at 1200 g to separate a brown colloidal precipitate. The liquid phase or "supernatant" or "concentrate" is then adjusted to pH 3.5-4.0, preferably with an acid recognized in the food industry. The material is preferably centrifuged at 3000g.

Further refining (purification, concentration) of the carotenoid fraction is as follows: the precipitate obtained from the acidified mixture after centrifugation and enzymolysis is suspended in a 60:40 to 80:20, preferably 70:30 v/v ethanol-water mixture, and 35 g of hydrogen are added Sodium oxide (calculated on the basis of 1 kg of dry matter), sodium hydroxide is preferably added as a 10-35% (w/w) solution in water or the above mixture, and the mixture is refluxed for 2 hours. The insoluble precipitate was centrifuged, and the same amount of sodium hydroxide solution was added to repeat the process.

Preferably, 8 times the volume of a 70:30 ethanol-water mixture is used for the first alkali treatment, and 5 times the volume of an 80:20 ethanol-water mixture is used for the second alkali treatment.

After the second 2-hour reflux centrifugation was repeated, the precipitate was resuspended in 4 times the volume of solvent mixture (without adding alkali), and the third reflux was performed. Half an hour before the end of reflux, 30 g of sodium dihydrogen phosphate (NaH ₂ PO ₄ ) and 200 g citric acid (for 1 kg dry matter). This solution was prepared by dissolving the compound in the same ethanol-water solution [ie (80:20)] as used above.

After centrifugation, usually 45-50% of the original dry matter is obtained, and the rest is washed away as sodium salts of peptides, amino acids, fatty acids, etc. The loss of carotenoids is very small, and its loss depends largely on the separation process grasping skills.

After the above operations, the carotene content of the colored body part is 2 times or even higher than that of the original material.

By this method, fresh carrots with a carotene content of 150 mg/kg are processed to obtain a dry matter containing 7.5-8.5 percent carotene after the color body precipitation operation is completed.

Carotene concentration can be further increased to more than 10-15% by repeated alkali treatment, and some methods can also obtain higher concentration, but there may be some economical problems due to higher solvent, energy and manpower requirements, even in these separation processes loss of carotenoids.

After repeated alkali treatment, according to the dry matter obtained after separation, 6 times the amount of 80:20 v/v ethanol-water solution was added, and the mixture was refluxed for 2 hours under stirring.

Then add an appropriate amount of sodium dihydrogen phosphate and citric acid to the stirred mixture to adjust the pH to 7.0-7.2, which is about the neutral range.

The centrifuged material is mixed (kneaded) with tocopherol and ascorbyl palmitate, dried at a temperature not exceeding 60° C. in ethanol-soaked form, but wet form can also be used.

The sieved dry powder is vacuum packed.

Chloroplasts obtained from photosynthetic plants can be similarly treated. However, in this case, chlorophyll degradation products such as pheophorbide and pheophorbide should be removed with great care if the product is intended for human consumption.

The obvious innovation of the method of the present invention is:

- transfer of chromoplasts and chloroplast cellular components to the liquid phase (pressing liquid, decanting liquid) by mechanical processing partially carried out in an alkaline medium.

- chromophores in the liquid phase or in suspension are denatured by pH adjustment or heat or a combination of both, causing their precipitation from the liquid phase.

- Separation of precipitated flocs (decantation, separation), removal of water-soluble components, relative to the raw material, the concentration of carotenoids in the separated flocs has increased significantly.

- Chromoplasts and chloroplasts contain colored substances, including carotenoids, which are lipoprotein complexes with a very high lipid content that bind carotenoids in a colloidal solution (these carotenoids are practically insoluble in water, and are extremely poorly soluble even in organic solvents), and transport these pigments within the organism. Part of the lipids of the complex is retained in the product and increases the bioavailability of carotenoids.

The new thinking is not that carotenoids should be removed from accompanying substances, but that accompanying substances should be removed from carotenoids as much as possible. This is achieved by cleavage of lipoprotein complexes in an alkaline medium, transferring most of the lipids in their sodium salt form to an aqueous ethanol solution and removing them.

By selecting appropriate raw materials, preparing crude carotenoid concentrates from them, and then mixing them together, it is possible to obtain a product with a range of carotenoids that meets the needs of the human body and is similar to that in human plasma.

These advantages can be achieved by avoiding the use of toxic solvents, which are commonly used to dissolve carotenoids. The result is that the by-products of the process and the waste generated therein are environmentally friendly, biodegradable and can in most cases be used as food or feed additives.

The following examples illustrate the above-mentioned products and their preparation methods in detail, but do not limit the scope of the present invention. % values represent w/w if not stated otherwise.

The analysis of carotenoids was carried out according to the following method:

a) The analysis of β-carotene adopts improved Booth spectrophotometry [M. A． van der Meer et a1. (Wageningen), Z. Lebensmittel Untersuch. und Forschung． 185. 461-467 (1987)].

b) The range of carotenoids was determined by high performance liquid chromatography (HPLC) on a Waters600 multi-solvent transfer system (Waters. Milford, USA), using reversed-phase LiChrosorb RP-18 (10 μm, 250 × 4.6 mm) and Spherisorb ODS (5μm, 250×4.6mm) column, acetonitrile-methanol-tetrahydrofuran (75:15:10v/v) mixture as eluent, photodiode sensor (Photo Diode Array Detector 990 (DAD), Waters, Milford, USA) As a detector in the wavelength range 190-800nm. Purified standard substances were used as internal standards.

Example

Example 1

A concentrate containing 5% carotenoids was prepared from carrot (Daucus carota).

1000kg of defoliated carrots (carotid body with a carotenoid content of 180-200mg/kg) were processed according to the process shown in Figure 1 using methods known in the art. The rinsed carrot body is milled and squeezed, and the residue cake is milled again, homogenized with water and squeezed repeatedly, and then repeatedly milled, and the squeezed fiber cake is extracted in the third step with a sodium hydroxide solution of pH 8.5, The residue and the extract (decanting the concentrate) were separated by a tilt pen centrifuge. Combine the squeezed liquid and the decanted concentrated liquid, and adjust the pH of the combined liquid to 4.0 with 10% citric acid. The colored bodies are aggregated in the form of flocs and placed to settle.

220 liters of precipitate that can pass through the pump, after decanting and separation, adjust the pH to 8.5 with 10% sodium hydroxide solution, add 10 liters of water suspension containing 100 g of trypsin to digest the protein during vigorous stirring at 37 ° C, and Constantly correct for falling pH with 10% sodium hydroxide solution. The treatment lasted 3 hours and the mixture (pH 8.5) was centrifuged to remove insoluble particles not belonging to the colloidal solution. Separated by a tilting pen centrifuge, the resulting pellet is mixed with the edible portion before drying.

The concentrate, also containing the carotenoids, was decanted, acidified to pH 4.0 with 10% citric acid, and the precipitated flocs were separated.

Separated sludge contained 16-18% dry matter, which contained 2.5% carotenoids (calculated on dry matter), had a wet weight of 45-50 kg, and 90 liters of 80:20 (v/v) ethanol- The water mixture was mixed, then 3 liters of 10% sodium hydroxide solution were added, and the mixture was refluxed for 2 hours with vigorous stirring.

The hot mixture was separated, and the separated precipitate (wet weight 28-30 kg) was added to 50 liters of aqueous ethanol (80:20) and 2 liters of 10% sodium hydroxide solution to repeat alkaline treatment, and then the mixture was refluxed for 2 hours. Repeat this separation process to obtain 16-20kg of wet precipitate.

The wet precipitation was first refluxed with 40 liters of aqueous ethanol used above for 1 hour, and during this process, a 3:1 aqueous solution of 10% citric acid and sodium dihydrogen phosphate was added to make the pH approximately neutral (7.0-7. 2). This requires about 300g of citric acid and 100g of monobasic sodium phosphate.

After separation, 14-16 kg of precipitate was obtained, equivalent to 3.2 kg of dry matter, which contained 0.19 kg of carotenoids.

150-300 g of antioxidant mixture (alpha-tocopheryl acetate, alpha-tocopherol and ascorbyl palmitate mixed in 2:2:1 w/w) was added to the pellet. The mixture is thoroughly mixed (kneaded) and dried under vacuum at a temperature not exceeding 50°C.

Dried Final Product Specifications:

Solid matter 3.4kg

Carotenoid content 5.5%

Loss on drying Max. 5% (105°C, 3 hours)

color reddish brown

Carotenoid Range 60-65% Beta-Carotene

                                                                                                   

5-10% other carotenoids

Stable for at least 1 year under vacuum or in protective gas.

Example 2

Preparation of concentrates containing 7-8% carotenoids

The operation process described in Example 1 is applied, but carrots containing 220-250 mg/kg of carotenoids are used as raw materials, and bacterial (Bacillus subtilis) protease is used for protein digestion.

Final product: 3 kg of reddish-brown dry powder or granules, with a carotenoid content of 7.5-8.0%, having a similar range of carotenoids to the product in Example 1.

Example 3

Prepare a concentrate containing approximately 10% carotenoids

The raw material in Example 2 was used for treatment. On the basis of the two-step alkali treatment, the third treatment is performed using the same amount of solvent and alkali as the second alkali treatment. Then, according to Example 1, the wet precipitate was refluxed with 40 liters of 80:20 ethanol-water mixture for 1 hour, and then the resulting mixture was neutralized according to Example 1, the precipitate was separated, the antioxidant mixture was mixed, and vacuum-dried.

Final product: 1.6kg reddish-brown dry product

Drying loss max 6%

Minimum carotenoid content 10.5% (0.17kg)

The range of carotenoids is the same as in the previous embodiment.

Example 4

Preparation of lycopene-rich concentrates from carrots and tomatoes

Chromosomes containing carotenoids are separated from carrots and tomatoes separately, and the collected material is processed (refined).

500 kg of carrots (carotoids with a carotenoid content of 220 mg/kg) were cleaned and defoliated. Obtain juice according to the flow process shown in Figure 1. The flocculated color bodies obtained at pH 4.0 (10% citric acid solution) were allowed to settle and separated by decantation. The dilute sludge with about 8% dry matter was concentrated in the separator to give a precipitate with a dry matter content of 16-20%. It can be directly used in alkaline treatment without protein degradation. For every 20-25 kg of wet precipitate, add 3 times the amount (about 75 kg) of ethanol-water mixture (80:20) and 1.5 liters of 10% sodium hydroxide solution, the mixture is refluxed for 2 hours, separate the hot mixture, and then repeat the process of the precipitate . After two alkali treatments, the precipitate obtained by separation or centrifugation was rinsed with ethanol and neutralized according to the method of Example 1. The precipitate separated at reflux for two hours was the first component of the final product.

1000kg tomatoes are processed separately. The juice, filtered by known methods, contained 7% dry matter and was concentrated to give a concentrate containing 20% dry matter. The concentrate was treated 3 times with ethanol. First use 2 times volume of ethanol with a purity of 96% (about 500 liters) to reflux for two hours, then separate the ethanol phase, and repeat the treatment twice with aqueous ethanol (80:20). In the third treatment, 3 liters of 10% sodium hydroxide solution were added to the stirred reflux mixture. The residue obtained from the third treatment, consisting mainly of pectin, polysaccharides and fibers, is added to the edible composition before drying. Collect the ethanol extract, adjust the pH value to 7.0-7.5 with citric acid/sodium dihydrogen phosphate mixture according to the method described in Example 1, and distill the mixture. The ethanol was recovered to obtain 10 liters of concentrate, which was mixed as the second component with the wet precipitate obtained from the treatment of carrots, mixed with 5% antioxidant, and vacuum-dried.

Final product: 3.6kg reddish-brown dry matter

Loss on drying: 5% max

Carotenoid content: minimum 5.5%

Carotenoid range: Lycopene 1.8%

                                                2.0%

  α-Carotene 1.2%

  Lutein + Others 0.5%

This product is characterized by its high content of lycopene.

Example 5

Preparation of products with high lycopene content

Carrots are processed according to the method described in Example 4.

3000 kg of tomatoes were treated, and the pectin of the tomato juice was digested with pectinase at pH 4.0 according to methods known in the art, and the pectinase was isolated from the Aspergillus culture. The diluted juice after pectin digestion is vacuum concentrated to a dry matter content ranging from 7% to 30% to obtain 540 kg of concentrate, which is then treated with ethanol. The first treatment was with 3 volumes of 96% pure ethanol, and the next two were with an 80:20 ethanol-water mixture. Each ethanol treatment was refluxed for 2 hours. During the third ethanol treatment, 8 liters of 10% sodium hydroxide solution were added to the stirring reflux solution. During the ethanol treatment, the ethanol solution and the insoluble fraction separate. (After the third ethanol treatment, the ethanol is removed from the obtained lycopene-free substance, dried and added to the edible composition).

According to the method described in Example 1, add citric acid/sodium dihydrogen phosphate solution mixture to the collected ethanol solution to adjust the pH to 7.0-7.5, concentrate the neutral mixture to 20 liters, and reclaim ethanol. The concentrate and the wet sediment in the carrot treatment method were mixed homogeneously, then mixed with 5% antioxidant (calculated on dry matter), and dried in vacuum.

Final product: 6.2kg reddish-brown dry matter

Loss on drying 5% max

Carotenoid content minimum 5.5%

Carotenoid range: Lycopene 3.0%

                                                                                                                 

        α-Carotene 0.6%

Other carotenoids 0.7%

This product is characterized by a similar content of lycopene: carotene

Example 6

Concentration of carotenoids from carrots and broccoli h

Process 1000kg of carrots and 1000kg of young cauliflower according to the flow process shown in Figure 1 to prepare juice.

Heat the juice to 56°C, adjust the pH to 4.0 with 10% citric acid solution, and then cool the solution to 20-25°C to further settle the precipitated chromoplasts and chloroplast flocs.

The precipitate was concentrated in a separator to obtain a wet precipitate with a dry matter content of 20%. For every 22 kg of dry matter (approximately 90 kg of wet precipitate), 180 liters of aqueous ethanol (80:20) was added under vigorous stirring, and 6 liters of 10% sodium hydroxide solution was added to adjust the pH to 9.5, and the mixture was refluxed for 2 hours. Separate the hot mixture. This treatment was repeated 3 times. After each treatment, the volumes of sodium hydroxide solution and solvent were reduced proportionally to the dry matter content. Experimental dosage (based on dry matter): 8-10 times the volume of aqueous ethanol (80:20) and 2.25-3.5% sodium hydroxide.

The residue of the fourth alkaline treatment was suspended in 45 liters of aqueous ethanol (70:30), refluxed and neutralized according to Example 1. After separation, 4.1 kg of precipitate was obtained, mixed with 5% antioxidant mixture, and dried under vacuum at a temperature not exceeding 50°C. The product obtained after drying contains a considerable amount of dihydroxycarotenoids (lutein, zeaxanthin).

Final product: Green-brown dry matter

Drying loss max 5%

Carotenoid content minimum 6.5%

Carotenoid range: β-carotene 3.2%

                                                                                                           

    Lutein + Zeaxanthin 1.0%

Lycopene 0.4%

Other carotenoids 0.4%

Example 7

Preparation of carotenoid concentrates with a carotenoid range similar to that found in plasma

The products in Examples 4, 5 and 6 were mixed with antioxidants in a ratio of 1:2:1 in the form soaked in ethanol, stirred evenly, and the mixture was vacuum-dried.

Final product: Green-brown dry matter

Loss on drying 5% max

Carotenoid content minimum 6.2%

Carotenoid range: Lycopene 32.9%

                                                                                                                                      

                                                                                                                                                                                                                                21.7 %

  Lutein + Zeaxanthin 11.6%

Other carotenoids 3.3%

The content of lycopene and dihydroxycarotenoids (lutein and zeaxanthin) in the product is exactly the same as the required plasma level. The product not only ensures a satisfactory supply of α- and β-carotene, but also contains lycopene and dihydroxycarotenoids with strong antioxidant properties, which further highlights the advantages of the product.

Example 8

Preparation of carotenoid concentrates with a carotenoid range similar to that found in plasma

In the manner of Example 7, the dry matter obtained in Examples 4, 5 and 6 was mixed in a mass ratio of 1:2:1, and the range of carotenoids in the product was similar to that obtained in Example 7.

Example 9

Treat Dunaliella or Blakeslea spp.

Obtain Dunaliella or Blakeslea spp. from natural sources or fermentation cultures. Biomass, concentrated by sedimentation or centrifugation to a dry matter content of 8-10%. The dry matter content of carotenoids in the concentrate is 2%. The mass was heated to 100°C by 3 bar saturated steam and this temperature was maintained for 10 minutes with continuous steaming until the mass walls of the mass separated.

According to the amount of 5% of the dry matter dissolved in 20 times the volume of isopropanol-water (70:30 v/v) mixture, sodium hydroxide was added to adjust the pH of the wall separation to 11, and the mixture was refluxed for 2 hours under stirring, and separated The mixture was heated, and the precipitate and liquid phases were separated.

Add 2 volumes of isopropanol-water (80:20) mixture and 0.08 volumes of 10% sodium hydroxide solution to the wet precipitate, reflux the mixture for 2 hours, and repeat the above separation process.

This treatment was repeated two more times.

After the fourth treatment, add 2 volumes of isopropanol-water mixture (80:20) to the wet precipitate, reflux for 1 hour, neutralize the hot sludge, filter or centrifuge as described in Example 1.

The filter residue cake or centrifuge precipitate obtained through neutralization and separation was mixed with an antioxidant as described in Example 1, and finally vacuum-dried.

The dry reddish-brown product has the following properties:

Dry matter content 94-96%

Crude protein (N×6.25) 18-22%

Crude Fiber 5-6%

Sulfated Ash 20-25%

Non-protein minor ingredients 8-10%

Lipid*) 32-46%

The content of carotenoids is 20-25%

*) Lipid-like substances are dissolved in a mixture of chloroform and methanol (unsaponifiable lipid fraction).

The range of carotenoids in the product depends on environmental factors such as the initial culture or fermentation conditions, but the content of carotenoids in the product is not less than ten times the dry matter content of carotenoids in the initial plasmodium isolate, that is, at least enriched ten times.

Claims (9)

1. from chromoplastid and/or chloroplast(id), separate the natural carotenoid concentrates that obtains, it is characterized in that:

A) it contains 1.5-25% (w/w) natural carotenoid and other composition that obtains from handled plant and the antioxidant that can choose interpolation wantonly;

B) it does not contain Toxic matter, especially poisonous residual solvent; And

C) its carotenoid scope can be regulated arbitrarily as required.

2. the enriched material of claim 1 is characterized in that, contains in its carotenoid content:

The 10-30% alpha-carotene

The 22-65% β-Hu Luobusu

The 6-55% Lyeopene and

5-22% dihydroxycarotene (xenthophylls, zeaxanthin) and other carotenoid; And

1.8-3.4% (w/w) alpha-tocopherol acetate,

1.8-3.4% (w/w) alpha-tocopherol and

0.9-1.7% (w/w) Quicifal

(calculating according to dry-matter) is as antioxidant.

3. preparation does not contain the method for the natural carotenoid concentrates of poisonous residual solvent, it is characterized in that:

I) chloroplast(id) of separating from vegetable material and/or chromoplastid part are handled with the enzyme with pectin and/or protein degradation characteristic (also lipase activity can be arranged) in water-bearing media, after digestion is finished, insoluble substance and colloid disperse phase are separated;

But acid is used in ii) colloid disperse phase optionally heating or cooling then, and preferably with the acid of foodstuffs industry approval, acidifying mixture precipitates carotenoid, by isolating precipitation in the liquid phase;

The solid precipitation that iii) contains carotenoid heats in aqueous alcohol under alkaline pH, thereby other submember of filtering degraded product and plant origin after the neutralization, goes out carotenoid by settlement separate; With

Iv) contain precipitation and specified quantitative alpha-tocopherol and/or the VITAMIN E ACETATE and/or the Quicifal mixing of carotenoid; If desired, drying composite; With

If desired, several dryed products with inhomogeneity carotene scope mix with specified proportion; Or

If desired, several dryed products with inhomogeneity carotene scope mix, and add alpha-tocopherol and/or the VITAMIN E ACETATE and/or the Quicifal mixing of specified quantitative then, if desired, and drying composite.

4. the method for claim 3, it is characterized in that, step I) chromoplastid and/or chloroplast(id) partly use the proteolytic enzyme or the animal protease (zymine) in bacterium (as Bacillus subtillis), fungi (as aspergillus oryzae), plant (papoid) source to handle in, handling pH is 4.5 to 11, treatment temp is 28 ℃ to 60 ℃, and the treatment time is 30 to 180 minutes.

5. claim 3 or 4 each methods is characterized in that, at step I i) in add C _2-4Aliphatic carboxylic acid, single-or dihydroxy-two-or tricarboxylic acid, the salt that the oxyhydroxide of phosphoric acid or they and basic metal or alkaline-earth metal and/or basic metal and/or alkaline-earth metal forms, in 8.5 scopes, regulate the iso-electric point of pH value at pH3.0, thereby carotenoid is precipitated out to chromoplastid and/or chloroplast(id).

6. the method for claim 3 is characterized in that, at step I i) in regulate after the pH value, but the mixture optionally heating to be not higher than+60 ℃ or choose wantonly be cooled to be not less than+5 ℃.

7. the method for claim 3 is characterized in that, step I ii) in aqueous pure processing can choose wantonly and repeat several times.

8. the method for claim 3 is characterized in that, step I ii) in used water/ethanol or water/isopropanol mixture water content be 15-40% (v/v), boiling point is lower than 90 ℃.

9. the method for claim 3 is characterized in that, step I v) gained the mixture frost drying or be lower than 60 ℃ of following vacuum-dryings or with wet form utilization.

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