WO2008108001A1 - Galactolipid - Google Patents

Abstract

Disclosed are: a novel monogalactosyldiacylglycerol or digalactosyldiacylglycerol compound or a derivative thereof; a process for production of the compound or the derivative; and a food containing the compound.

Description

 Galactolipid

Technical field

 The present invention relates to galatatolipid. More specifically, monogalactosyldiacylglycerol (MG D G) and digalactosyldiacylglycerol

 (D GD G) relates to the compound. book

Background art

 Ipomoea batatas L. (Satsumaimo) is a tuberous perennial plant grown in many countries whose edible roots are eaten raw or after being cooked in various ways.

 The upper part is herbaceous and is consumed as fresh vegetables in many parts of the world. Sweet potato leaves are an excellent source of biologically active anthocyanin and polyphenol components. 15 different anthocyanin compounds and 6 different polyphenol compounds were identified and quantified. Recent studies on the nutritive composition and physiological functions of sweet potato leaves when used in vegetables, tea, strawberries, bread, and confections have shown that they provide a beneficial food supply for beneficial polyphenolic compounds. Suggested that it could be a source. In addition, they are rich in vitamin B, iron, calcium, sub- and protein.

In particular, there are reports in the patent literature describing the possibility of producing anticancer drugs or ingredients for food and beverages for cancer prevention from various parts of sweet potato (ie leaves, vines, stems and rhizomes). (Japanese Laid-Open Patent Publication No. 7-2 5 8 100). This publication describes that this activity is caused by a glycolipid fraction that can suppress the proliferation of cancer cells and promote differentiation. How this fraction is It has not been confirmed whether it contains any glycolipids.

 With regard to glycolipid structure determination, an analytical approach based on mass spectrometry is used to determine the structure of the fatty acid acyl group of the glyce mouth glycolipid. In detail, GG Orgambide et al. (Lipids, 28th, 1993, p. 975), after methanolysis of darice cellolipid mixtures, followed by GC / MS analysis of fatty acid methyl ester derivatives, Fatty acids were identified by comparing retention times and mass spectra. Another analytical strategy is to compare the collision-dissociation (CID) spectrum pattern of the carpoxy ion (RC00—) generated by negative ion fast atom bombardment mass spectrometry (FAB / MS) of glyce mouth lipids with that of the standard product. Thus, the structure of fatty acids was determined (from NJ Jansen et al., Lipids, 21st, 1986, 580, and NJ Jansen et al., Lipids, 22, 1987, 480). In any case, it was not possible to find out from which component of the fatty acid methyl ester derivative or free carboxy ionic force S, the glyce mouth lipid mixture and from where in their glycerol backbone.

 The structure of monogalactosyldiacylglycerol (MG DG) and digalatatosyldiacylglycerol (DGDG) derived from wheat flour contained in glycate oral lipid was generated by FAB ionization. Sodium was added to galatatolipid. (YH Kim et al., J. Mass Spectrom., 32nd, 1987, p. 968, and YH Kim et al., Estimated by the positive ion CID spectrum of the molecule ([M + Na] +) Microchemical Journal, 68, 2001, p. 143). In this case, it was possible to determine the position of the double bond in the chain, but since this experiment was performed on a pure compound, this represents a reliable technique for mixture analysis. Do not mean.

To determine the binding position of the acyl chain on the glycephthal skeleton of MG DG and DGDG compound classes, HPLC-coupled electrospray ionization quadrupole ion tofutanam mass fraction ίΠ "(electrospray ionization— quadrupole ion tra Although tandem mass spectrometry coupled with HPLC (HPLC-ESI / QITMS / MS) force S was adopted, this approach was not useful in elucidating the position of double bonds in the acyl chain (G. Guella et al. , Rapid Commun. Mass Spectrom., 17th, 2003, p. 1982, and W. Wang et al., Rapid Commun. Mass Spect rom., P. 13, 1999, p. 1189).

 Galactosylglycerols such as MG D G and D G D G are also noted for their bioactivity. For example, it is known to have anti-inflammatory properties (from A. Bruno et al., Eur. J. phar macol., 524, 2005, p. 159) and to inhibit carcinogenesis. Disclosure of the invention

 The present inventors have discovered several novel galactolipids, monogalactosyl diacylglycerol (MG D G) and digalactosyl diacyl glycerol (D G D G) compounds from fractions of the low polar fraction of sweet potato leaves.

 The present invention provides a monogalactosyldiacylglycerol compound or a derivative thereof having the following structural formula (I):

ここで、

here,

Is a palmitate palmitate and R ₂ is a linolenic acid acyl chain;

Is an α-linolenic acid acyl chain, and R ₂ is 9, 12-nonadecadienoic acid Is it a chain?

R ₁ is a 9, 12-nonadecamate asil chain and R ₂ is an α-linolenic acid asil chain;

Is R an arachidate chain and R ₂ is an oleate chain;

Ri is an arachidic acid acyl chain and R ₂ is a linoleic acid acyl chain;

Is an 11-eicosenoic acid acyl chain and R ₂ is a linoleic acid acyl chain;

1 ^ is an 11-eicosenoic acid acyl chain and R ₂ is an α-linolenic acid acyl chain;

Is an 11-eicosenoic acid acyl chain and R ₂ is a 9, 12-nonadecadienic acid chain;

Is an 11-eicosenoic acid chain and R ₂ is an 11-eicosenoic acid chain; or

Shaye is a heneicosanoic acid acyl chain, and R ₂ is an α-linolenic acid acyl chain.

 The present invention also provides a digalactosyldiacylglycerol compound or derivative thereof having the following structural formula (I I):

ここで、

here,

■ Is R a palmitate palmitate and R ₂ is a 9, 12-nonadecadienoic acid chain;

Is a 9, 12-nonadecadienoic acid acyl chain and R ₂ is an α-linolenic acid acyl chain;

Is 11-eicosenoic acid acyl chain and R ₂ is palmitic acid acyl chain;

1 ^ is an 11-eicosenoic acid acyl chain and R ₂ is an α-linolenic acid acyl chain;

Is an 11-eicosenoic acid acyl chain and R ₂ is a 9, 12-nonadecadienic acid acyl chain; or

1 ^ is the 11-eicosenoic acid chain, and R ₂ is the 11-eicosenoic acid chain.

 The present invention also provides a food containing at least one selected from the above-mentioned monogalactosyldiacylglycose oral compounds or derivatives thereof. Also provided is a food containing at least one selected from the group consisting of the above-mentioned digalatatosyldiacylglycerol compounds or derivatives thereof.

 In one embodiment, the food further contains at least one selected from the group consisting of wheat leaf and kale.

 In a further embodiment, the food further contains at least one selected from the group consisting of indigestible dextrin, oligosaccharide, and lactic acid bacteria. The present invention also provides a method for producing the monogalactosyldiacylglycerol compound or derivative thereof, the method comprising:

 (1) Process of subjecting sweet potato leaves to methanol extraction,

 (2) a step of separating the low polarity fraction by solid phase extraction; and

(3) Separation process using high performance liquid chromatography including.

 'The present invention also provides a method for producing the above-mentioned digalatatosyldiacylglycerol compound or a derivative thereof,

 (1) Process of subjecting sweet potato leaves to methanol extraction,

 (2) · a step of separating the low polarity fraction by solid phase extraction; and

 (3) Separation process using high performance liquid chromatography

including. Brief Description of Drawings

 Figure 1 shows that [M + Na] + ion is m / z 829.9]) ^ 000 compound (1 1)

It is a graph which shows a C-ESI / MS / MS spectrum. (A) is the overall spectrum, (b) is the high molecular weight region, and. (C) is the low molecular weight region. Figure 2 shows that [M + Na] + ions are m / z 991.9 0 & 0. It is a graph which shows a C-ESI / MS / MS spectrum of compound (24). (a) is the overall spectrum, (b) is the high molecular weight region, and (c) is the low molecular weight region. Figure 3 is a graph showing the LC-ESI / MS / MS spectrum of the MGDG compound (7) whose [M + Na] + ion is m / z 813.9. (a) is the overall spectrum, and (b) is the high molecular weight region. BEST MODE FOR CARRYING OUT THE INVENTION

We separated the polar fraction of the methanol extract of sweet potato leaves using solid-phase extraction, and liquid chromatography coupled electrosprayed quadrupole time-of-flight tandem mass spectrometry (liquid chromatography coupled) An analytical method based on tandem mass spectrometry (HPLC-ESI / QToFMS / MS) was developed to elucidate the structure of glycolipids contained in sweet potato leaves. The inventors of the present invention have been able to determine the composition and bonding position of the fatty acid acyl group in the glycolipid and to elucidate the position of the double bond in the acyl chain by the above analysis method. The composition of the sugar composition was determined using 1D and 2D NMR (nuclear magnetic resonance). In addition, the determination of the position of the unsaturated double bond of the fatty acid bound to the two carbons of the glycerol skeleton was confirmed, and in some cases, gas chromatography mass spectrometry (GC / MS) of the derivatized fatty acid. ) To determine the three-dimensional structure.

 With this strategy, 16 new opio 10 known galactolipids could be isolated and characterized. To the best of our knowledge, this is the first report of galatatolipid composition in sweet potato leaves.

 Galatatolipid is a kind of glyce mouth lipid. Galatatolipids include monogalactosyl diacyl glycerol (MG D G) and digalactosyl diacyl glycerol (D G D G). MG DG has a structure in which two fatty acid acyl groups are bound to the sn-1 and sn-2 positions of the glycerol skeleton, and one galactose is ether-linked to the sn-3 position of glycerol mouth. Have. D G D G has a structure in which two fatty acid acyl groups are bonded to the sn-1 and sn-2 positions of the glycerol skeleton, and two galactoses are bonded to the glycerol sn 3 position. Hereinafter, in the present specification, the term “galactolipid” refers to both MG D G and D GD G.

 The present invention provides novel galagtolipids as described below.

ここで、 More specifically, the present invention provides a monogalactosyl diacylglycose oral compound or derivative thereof having the following structural formula (I):

here,

Is a palmitate palmitate and R ₂ is an α-linolenic acid acyl chain;

! ^ Is -linolenic acid acyl chain and R ₂ is 9, 12-nonadecadienoic acid chain;

Is a 9, 12-nonadecadienoic acid acyl chain and R ₂ is an α-linolenic acid acyl chain;

R i is an arachidic acid acyl chain and R ₂ is an oleic acid acyl chain;

Is R an arachidate acyl chain and R ₂ is a linoleic acid acyl chain;

Is an 11-eicosenoic acid acyl chain, i.e., R ₂ is a linoleic acid acyl chain;

Is 11-eicosenoic acid acyl chain and R ₂ is α-linolenic acid acyl chain;

Is R ₁ an 11-eicosenoic acid chain and R ₂ is a 9, 12-nonadecaic acid chain;

1 ^ is an 11-eicosenoic acid chain and R ₂ is an 11-eicosenoic acid chain; or

Is a heneicosanoic acid acyl chain, and R ₂ is an a-linolenic acid acyl chain. The present invention also provides a digalactosyldiacylglycerol compound having the following structural formula (II) or a derivative thereof:

ここで、

here,

Is R ₁ a palmitate palmitate and R ₂ is a 9, 12-nonadecamate chain;

Is a 9,12-nonadecadienoic acid acyl chain and R ₂ is an α-linolenic acid acyl chain;

Is 11-eicosenoic acid acyl chain and R ₂ is palmitic acid acyl chain;

Is 11-eicosenoic acid acyl chain and R ₂ is α-linolenic acid asinole chain;

Is an 11-eicosenoic acid acyl chain and R ₂ is a 9, 12-nonadecadienic acid acyl chain; or

R ₁ is an 11-eicosenoic acid chain and R ₂ is an 11-eicosenoic acid chain.

Examples of the fatty acid of the acyl chain in the galactolipid of the present invention include the following: palmitic acid (C16: 0); stearic acid (C18: 0); oleic acid (C18: ln-9); linoleic acid (C18: 2 6-6); α-linolenic acid (C18: 3 n-3); Nona Decadienoic acid (C19: 2), especially with double bonds at 9 and 12 positions; arachidic acid (C20: 0); 11-eicosenoic acid (C20: l n-9); and heneicosanoic acid (C2l: 0) ) In the case of unsaturated fatty acids, the double bond can be cis or trans. Linoleic acid and α-linolenic acid usually have a cis-type double bond. In nonadecadienoic acid or 11'-eicosenoic acid, the double bond can be cis or trans.

 The galatatolipid of the present invention can be produced from sweet potato leaves. The variety of sweet potato is not particularly limited. For example, potato shoots of varieties such as Joy White, Koganesengan, Shiku Yutaka, Sweet Mustache, Ayamurasaki, and Suio can be used.

 The sweet potato leaves used in the present invention are preferably shoot leaves appearing on the ground when sweet potatoes are cultivated. The leaves of shoots grown from the ground are preferably grown to 10 cm or more, more preferably 30 cm or more, more preferably 60 cm or more. Further, when the length from the position where the stem of the sweet potato goes out from the ground to the tip is measured, the length is preferably within 300 cm, more preferably within 200 cm, further Preferably, shoot leaves that are within 150 cm are used. If it exceeds 300 cm, the tip of the shoot will touch the ground, making it more susceptible to harmful insects and the like, and a sufficient amount of leaves may not be obtained.

 In particular, in the present invention, it is preferable to use shoots in a green state, that is, young leaves of sweet potato shoots. As used herein, “young shoots of sweet potato shoots” refers to leaves within 60 cm from the tip of the sweet potato shoots. The sweet potato leaves as described above may be subjected to processing described later, preferably after washing adhering mud or the like with water.

 (1) Heat treatment

Heat treatment is used to stabilize the quality by deactivating enzymes in the sweet potato leaves, and The purpose is to prevent the fascination of the clover leaves. Examples of the heat treatment include branching treatment (for example, blanching), dry heat treatment, microwave treatment, infrared ray or far-infrared treatment, steam treatment, and the like. Of these heat treatments, blanching treatment is preferably used. Furthermore, for the convenience of the treatment process, each treatment may be performed after cutting the sweet potato leaves into a major axis of about 10 to 30 cm as necessary.

 When performing the blanching treatment, the planting treatment may be carried out by a method commonly used by those skilled in the art so that the color of the sweet potato leaves, that is, the color of chlorophyll, which is the color of the green plant, does not fade. An example of such a blanching process is boiling. The optimum conditions for blanching treatment vary greatly depending on the plant used. In some cases, the blanching process may damage flavor and nutrients and deactivate the physiological activity of useful ingredients. Therefore, in the present invention, the pH is preferably 5.4 or more, more preferably 5.6 to 8.4, more preferably 5.6 to 8.0, and most preferably 5.6 to 7.6. Blanching with hot water. By such branching treatment, it is possible to efficiently obtain sweet potato leaves having high activity such as antioxidant activity and high content of components such as polyphenol.

When blanching treatment is performed, salt is added in an amount of 0.1 to 5 mass per hot water in order to improve the flavor. It is preferable to add at a ratio of / ₀ , preferably 0.2 to 3% by mass. By adding sodium chloride in this way, it is possible to obtain sweet potato leaves that are greener and have a better flavor.

When performing dry heat treatment, microwave treatment, infrared or far-infrared treatment, and water vapor treatment, perform these pH adjustment treatments such as spraying a solution adjusted for pH on sweet potato leaves. It is preferable to carry out the treatment. The pH adjustment process is performed by a method commonly used by those skilled in the art. For example, when adjusting to basic conditions, sodium hydroxide, baking soda, calcium carbonate (egg shell calcium , Scallop shell calcium, coral calcium, etc.), and calcium oxide obtained by firing these calcium carbonates can be used for treatment. Further, alkaline ionized water or the like may be used. On the other hand, for adjustment under acidic conditions, a solution of an organic acid such as acetic acid, citrate, ascorbic acid, tartaric acid, malic acid or fumaric acid may be used. The amount of these pH adjusting agents may be adjusted appropriately according to the adjusting agent used.

 The heating temperature in the heat treatment is higher than 80 ° C., preferably 90 ° C. or higher. The heating time is 5 minutes or less, preferably 3 minutes or less, and most preferably 10 seconds to 3 minutes.

 It is preferable to immediately cool the potato leaves after the heat treatment in order to maintain the green opium flavor. Cooling can be done by methods commonly used by those skilled in the art, such as immersing the potato leaves after heat treatment in cooling water or quenching them with cold air. For example, when cooling by immersing in cooling water, water of 30 ° C. or lower, preferably water of 2 ° C. or lower is used. The lower the cooling temperature, the brighter the sweet potato leaves are and the more beautiful they look. The cooling time is an arbitrary time according to the processing amount of the sweet potato leaves, but it is preferable to carry out the cooling until the sweet potato leaves themselves reach a temperature equal to the cooling temperature.

 (2) Drying treatment and powdery soot treatment

 Drying is preferably used because the quality of the sweet potato leaves before processing can be maintained. Drying is performed using any drying method commonly used by those skilled in the art. Drying is performed using a dryer according to the drying method, for example, a hot air dryer, a high-pressure steam dryer, an electromagnetic wave dryer, a direct-fired heater, a rotary draft dryer, a freeze dryer, or a vacuum concentrator. Done. Of these, hot air dryers, direct-fired dryers, and rotary draft dryers are preferable from the viewpoint of cost and drying efficiency.

Drying is preferably performed so that the moisture content in the dried product is 5% by mass or less. The drying treatment is performed at a temperature of about 60 to 1550 ° C. under normal pressure, so that a sweet and colorful dried powder of sweet potato leaves can be obtained. 6 0 under reduced pressure. If it is performed at a temperature below C, preferably above the temperature at which sweet potato leaves freeze and below 60 ° C., drying can be performed while reducing the loss of nutrient components. ''

 When drying the sweet potato leaves as they are, it is preferable to dry them in two stages. The two-stage drying can be performed using, for example, a hot air dryer. In the two-stage drying, first, primary drying is performed at a temperature of 60 to 80 ° C. until the water content becomes 25 mass% or less. Next, secondary drying is performed at a temperature higher than the primary drying until the moisture content of the primary dried potato leaves is 5% by mass or less. At this time, if the drying temperature of the primary drying is less than 60 ° C, the drying speed becomes slow, and if the drying temperature of the secondary drying exceeds 100 ° C, it may cause scorching. Therefore, the temperature of secondary drying is preferably 70 to 90 ° C, and more preferably around 80 ° C to obtain a sweet potato leaf powder having a high polyphenol content and a bright color. Can do.

 The temperature difference between primary drying and secondary drying is preferably about 5 to 15 ° C, and more preferably about 10 ° C. For example, in the case of secondary drying at 90 ° C., the temperature of the subsequent drying is preferably 75 to 85 ° C., more preferably about 80 ° C.

 By performing such a two-step drying process, the drying time is shortened and the green sweet flavor of sweet potato leaves is maintained. By setting the temperature difference within a certain range as described above, water management of the green leaves in the drying process is facilitated and drying is performed efficiently.

The potato leaves dried in this manner may be further powdered to obtain a dry powder. In order to obtain dried powder of sweet potato leaves, from the viewpoint of production cost and drying efficiency, hot air dryer, direct-fired heater, and rotary type A ventilation dryer is preferably used. The dried powder of sweet potato leaves may be further pulverized to further reduce the particle size. Since sweet potato leaves have different parts from the stem, leaf and petiole, it is preferable to go through a coarse pulverization step or a fine pulverization step from the viewpoint of increasing the pulverization efficiency.

 Coarse rice cake 0: The process of cutting the dried green potato leaves with any machine or apparatus known to those skilled in the art such as cutter, slicer, dicer and the like. As for the size of the cut green leaves, the major axis is preferably 2 O mm or less, more preferably 0.1 to L O mm.

The fine pulverization step is a step of finely pulverizing the coarsely pulverized potato leaves to obtain fine pulverized powder. In the pulverization step, 90% by mass is finely pulverized so that it passes through the 200 mesh section. The fine pulverization is performed by using a machine or an apparatus normally used by those skilled in the art, such as a crusher, a mill, a blender, and a stone mill. When heat sterilization is performed, it is performed before the fine powder mashing process. By carrying out this heat sterilization treatment, the coarsely ground sweet potato leaves can be uniformly heated, the flavor of the green leaves can be improved, and efficient sterilization can be performed. This heat treatment is performed at 110 ° C. or higher, and a high-pressure sterilizer, a heat sterilizer, a pressure steam sterilizer, or the like can be used. '' For example, in the case of heat treatment by pressure steam sterilization, coarsely ground sweet potato leaves are, for example, saturated with steam at 110 to 200 ° C under pressure of 0.5 to 10 kg / cm ² , Heat sterilization treatment for 2 to 10 seconds. If necessary, the moisture contained during heating with saturated water steam is further dried.

 Thus, the texture is improved by finely kneading, and preferably, the texture is further improved by sequentially performing the steps of coarse pulverization, heat treatment, and fine pulverization. It becomes easy to mix.

The dried potato leaves may be prepared by coarsely mashing the potato leaves before the drying treatment into fine pieces or pastes, and then performing the above drying treatment. Ie powdering The treatment may be performed using dried potato leaves, or may be performed using undried (eg, fresh leaves, boiled potato leaves, etc.) potato leaves.

 You may give a drying process to said squeezed juice obtained by giving the below-mentioned pressing process etc. to a sweet potato leaf using said dryer. In the case of drying using sweet potato juice, a vacuum concentrator is preferable in terms of production cost and drying efficiency. By performing the drying process in this way, a powder of juice (hereinafter sometimes referred to as “extract powder”) is obtained.

 In addition, when the extract of sweet potato leaves is used as an extract powder, it may be pulverized using a spray dryer such as a spray dryer. When a spray dryer is used, excipients such as dextrin, cyclodextrin, starch, and maltose are added as necessary to increase the recovery rate. Preferably dextrin is used. For example, in order to facilitate powdering by addition of dextrin, the mass ratio of squeezed to dextrin is preferably 1:10 to 5: 1.

 The drying to obtain the extract powder is preferably performed so that the water content in the extract powder is 5% by mass or less. ·

 (3) Press processing

 The pressing process is performed using, for example, a pressing machine. As a result, the juice of sweet potato leaves is obtained. When the obtained juice is used as it is without being subjected to the drying treatment as described above, it is preferable to perform heat sterilization at 80 ° C. to 130 ° C.

Therefore, hereinafter, the term “sweet potato leaf” includes not only fresh leaves but also potato leaves that have been processed as described in (1) to (3) above. The galactolipid of the present invention is obtained by transferring methanol extract of the above sweet potato leaves, methanol, water, Z-acetonitrile, as described in the following examples, for example. It can be separated and recovered by subjecting it to a solid phase extraction method used as a phase. The following examples detail the procedure for separating galactolipids of the present invention, but the method for obtaining galactolipids of the present invention is not limited thereto.

 There is also provided a method for producing the galactolipid or derivative thereof of the present invention, which comprises (1) a step of subjecting sweet potato leaves to methanol extraction, (2) a step of separating a low polarity fraction by solid phase extraction, and (3) Includes a step of separation using high performance liquid chromatography.

 The extraction process is performed by adding methanol (including water-containing methanol) to sweet potato leaves (raw leaves, dry powder, extract powder, etc.).

 The low polarity fraction can be obtained by, for example, extracting the above extract from a C-18 sep-pak cartridge (Waters

Corporation, Milford, Massachusetts, USA). The cartridge can be preconditioned with methanol, water, and 0.2% formic acid, washed with 0.2% formic acid and 0.2% formic acid / methanol (50:50, v / v), and then the low polarity fraction. Can be eluted with methanol. The eluted low polar fraction can be separated by mobile phase (elution method A) of methanol Z water / acetonitrile (90.5: 7: 2.5), for example. In addition, for separation of low polar fractions, the mobile phase of methanolic water Z-acetonitrile (82: 5: 15: 2.5) (elution method)

B) can also be used. Thereby individual galactolipids can be obtained. The galactolipids collected from the sweet potato leaves can be used as needed by synthetic adsorbents (Diaion HP 20, Sephabies SP 8 25, Amberlite XAD4, MC Ige 1 CHP 20 P, etc.), dextran. It may be purified by a method usually used by those skilled in the art using a resin (such as CEFADEX LH-20).

The galatatolipid of the present invention may be in the form of a derivative. Any derivative that can be found in the naturally occurring form, for example, derivatives in which the alcoholic 0H of the galactose residue is substituted with various substituents, and food or pharmaceutically acceptable Any derivative that is capable can be included.

 The galactolipid or derivative thereof of the present invention can be used for pharmaceuticals, foods and beverages, and cosmetics based on its biological activity. Examples of the biological activity include an antitumor action, an anti-inflammatory action, and an antioxidant action. The galagtolipid of the present invention can be used for the production of functional foods and supplements having anticholesterol action, blood flow improvement action and the like, and cosmetics having skin quality improvement effect, beauty effect and moisturizing effect. It can be used with ingredients such as food ingredients, pharmaceutical ingredients, excipients, extenders, binders, thickeners, emulsifiers, colorants, fragrances, or seasonings.

The galatatolipid or derivative thereof of the present invention can be incorporated into foods as described below. For example, royal jelly, propolis, vitamins (A, C, D, E, K, folic acid, pantothenic acid, piotin, derivatives thereof), minerals (iron, magnesium, calcium, zinc, selenium, etc.), α-lipoic acid , Lecithin, polyphenols (flavonoids, derivatives of these), carotenoids (lycopene, fastaxanthin, zeaxanthin, rutin, etc.), xanthine derivatives (caffeine, etc.), fatty acids, proteins (collagen, elastin, etc.), mucos Polysaccharides (such as hyaluronic acid), amino sugars (eg, gnorecosamine, acetylidanorecosamine, galactosamine, acetylenoregalactosamine, neuroamic acid, acetylneuraminic acid, hexosamine, salts thereof), oligo Sugar (isomaltori Sugars, cyclic oligosaccharides, etc.), phospholipids and their derivatives (phosphatidylcholine, sphingomyelin, ceramides, etc.), sulfur-containing compounds (alyain, sepaen, taurine, dartathione, methylsulfonylmethane, etc.), sugar alcohols, lignans (Sesamin, etc.), animal and plant extracts containing these, root vegetables (turmeric, ginger, etc.), green leaves of grasses such as wheat leaves, green leaves of cruciferous plants such as kale, indigestible dextrin (For example, it is made from starch such as corn and straw and marketed. Lactic acid bacteria (especially lactic acid bacteria with high intestinal reach efficiency, for example, spore-forming lactic acid bacteria, lactic acid bacteria prepared by coating enteric substances, etc., specific strains with high intestinal reach Lactic acid bacteria, Bifidobacterium longam, Lactobacillus casei, Lactobacillus acidophilus, Rata topaticilus' Helveticas, Rata tobacillus delpureiki, Streptococcus thermobirarus, etc.) obtain.

 The food in powder form can be dispersed in water or the like and used as a green juice form. In particular, it can be mixed with green leaves such as wheat leaves, kale, tomorrow leaves, and mulberry leaves, which are conventional green juice ingredients, preferably wheat powder or kale powder. Furthermore, the food can also be in the form of beverages such as plant fermented juice, vegetable juice (eg carrot juice), plant extract, fruit juice and the like. Indigestible dextrins, oligosaccharides, and materials such as lactic acid bacteria can also be suitably incorporated into foods.

 Foods are processed into forms such as granules, tablets, capsules (hard capsules, soft capsules, etc.), pills, powders, liquids, tea bags, and baskets, depending on the application. Such processed foods can be eaten as they are, or they can be dissolved in water, hot water or milk. If powder or the like is provided in the form of a tea bag, the extract obtained by soaking in hot water can be used as a beverage. Example

 1. Experimental method

 1.1 Material

Methanol for HPLC (MeOH) was purchased from: [. T. Baker (Baker Mallinckrodt, Phillipsburg, NJ, USA). Acetonitrinore (ACN) and formic acid were obtained from Carlo Erba (Milan, Italy). HPLC water (18 mQ) was prepared using a Millipore Milli-Q water purifier (Millipore Corp., Bedford, MA, USA). 1. 2. Sample preparation

 A sample (3 g) of air-dried sweet potato leaves was extracted with 60 mM MeOH at room temperature for 8 days. The extract was filtered to give a yield of 313.13 mg. To separate low polarity components from polyphenols and anthocyanins, the methanol extract was passed through a C-18 sep-pak cartridge (Waters Corporation, Milford, Massachusetts, USA). The cartridge was preconditioned with methanol, water, and 0.2% formic acid. After washing with 0.2% formic acid and 0.2% formic acid / methanol (50:50, v / v), the low polar components were eluted with methanol and used for chromatographic analysis.

1. 3. HPLC-UV analysis

 Methanol fractions from C-18 sep-pak cartridge separations were analyzed by HPLC using an Alliance HPLC equipped with a dual lambda absorbance detector (Waters Corporation, Milford, Massachusetts, USA). Individual galactolipids were added to a Hypersil BDS C18 column (250 mm x 4.6 mm, 5 ^ m) with a mobile phase (elution method A) of methanol / water Z-acetonitrile (90.5: 7: 2.5) at a flow rate of 1 ml / min ( (Glass, Bellefonte, PA, USA). Detection was performed at 205 ηm.

 A mobile phase of methanolic water Z-acetonitrile (82.5: 15: 2.5) (elution method B) was also used to improve the separation of the more polar galactolipids.

1. 4. Soot Analysis

And ¹³ C NMR spectra were recorded on a Bruker DRX-600 MHz spectrometer at T = 300K. The NMR samples were prepared by dissolving the sample _{CD 3 0D (Sigma Aldri ch,} 99.96% D) in. To match the spectral standards, the solvent signal was used as the internal standard and corrected 1 !, δ = 3.34 ppm; ¹³ C = 49. Oppm). 1.5. Liquid chromatography / electrospray mass spectrometry (LC-ESI / MS and LC-ESI / MS / MS)

 Methanol fractions from C-18 sep-pak cartridge separations were analyzed by on-line LC-ESI / MS. For this analysis, a Waters Q-Tof Micro hybrid quadrupole accelerating time-of-flight mass spectrometer (Waters Corporation, Milford, Massachusetts, USA) connected to the Alliance HPLC and facing the orthogonal Z spray source was used. Using. Individual galatatolipids were separated using the chromatographic conditions described above. In order to verify the contribution to ionization using an acidic solvent, both chromatographic methods in which 0.2% formic acid was added to the eluent were also performed. The eluate is injected directly into the electrospray ion source, and MS1 and MS / MS spectra are obtained in positive ion mode, using software provided by the manufacturer (Waters Mass Lynx 4.0 software). Elucidated using. The desolvation and source temperatures were 180 ° C and 100 ° C, respectively. The operating conditions optimized for the detection of galactolipids were as follows: Kybilly voltage 3400V, sample cone voltage 40V, extraction voltage 1.5V, impact cell voltage 7V. MS / MS spectra were obtained in measurement scan mode using impact energy in the range of 30-45V. MS / MS spectra were recorded in the 50-1500 m / z region. 1.6 Preparation of fatty acid methyl esters

The galactolipid separated by HPLC was exposed to 2.5 ml of a methanol solution containing 2% sulfuric acid at 80 ° C. for 4 hours and subjected to a transesterification reaction. After completion of the reaction, the reaction tube was cooled. Then, 2.0 ml of water and 1.0 ml of hexane were added to each reaction tube, sealed, and mixed by vortexing for 10 minutes. The reaction tube to promote centrifuged to phase separate for 1 minute at 4000 r _P m, and; ^ transferred hexane phases in GC measurement vials. To the remaining aqueous phase was added 1.0 ml of hexane and extraction was performed as described above. This process Repeated twice.

1.7 Gas chromatography mass spectrometry (GC / MS)

 GC Chromatography GC System 6890 Series for fatty acid methyl ester analysis compatible with a single-branch ram (30m x 0.25mm ID, 0.25 / zm film thickness) DB-1 (J & W Scientific, Folsom, CA, USA) GCTTM orthogonal acceleration time-of-flight (oa-T0F) GC mass spectrometer (Waters Corporation, Milford, Massachusetts, USA) equipped with (Agilent Technologies, Palo Alto, CA, USA) The analysis was performed using the following analytical conditions: the injection was in splitless mode, the injector temperature was 280 ° C, the ion source temperature was 200 ° C, and the GC / MS interface temperature was 300. ° C. The initial oven temperature is 40 ° C (5 minutes), programmed to increase to 60 ° C at 4 ° C / minute, then 15 to 150 ° C at Z minutes, and 280 ° at 3 ° C / minute C was heated to 300 ° C in 20 ° CZ minutes. The total execution time was 60 minutes. Helium was a carrier gas and l ml / min was a column flow.

 Mass spectrometry was performed at 70 eV ionization energy and a 50 ^ 500 mass range was analyzed with a scan time of 0.5 seconds.

2 Results

2. 1. UV spectrophotometry HPLC analysis

To optimize the chromatographic conditions for NMR and GC / MS analysis and to isolate pure compounds, HPLC-UV pre-analysis was performed on the methanol fraction from the C-18 sep-pak cartridge separation. . Although the above elution method A separated low polarity compounds well, it was not suitable for analyzing more polar compounds. For this reason, another elution method B with a different ratio of organic solvent to water was set up. A total of 26 compounds were separated and recovered using these elution methods. In detail, 16 compounds were separated and recovered by elution method A and 10 by elution method B (Tables 1 and 2). In Table 1 and Table 2 below, compounds having a superscript a are separated and collected by elution method A, and compounds having a superscript b are separated and collected by elution method B.

2. 2. MR analysis

Analysis of the ¹ H NMR data measured on the isolated pure compound confirmed the presence of glyce mouth lipids. This indicates that the proton signal H-2 of the D-glycerol unit has a characteristic δ 5.29 absorption when the 0-acyl group is bound to the C-2 position of the D-glycerol unit. (GL Sassak i et al., J. Nat. Prod., 62nd, 1999, p. 844). By further examining the absorption bands characteristic of sugar, we identified that these compounds are galatatolipids and that there are two molecular groups. These molecular groups were classified by the difference in the number of galactopyranoses 0-bonded to the di-0-acylglycose skeleton.

For the first group of molecules, the combined analysis of NMR and a selective one-dimensional T0CSY (total correlation spectroscopy) vector identified the presence of one unit of β-D-galactoviranosyl. In fact, the j3 -glycosidation to glycerol shows that the anomeric carbon (C1, position) has a low magnetic field shift (δ 105) and the proton (HI, position) has a high magnetic field shift (δ 4 26), it was confirmed that the coupling constant of ³ JHI- -H2- was very large (J> 7Hz). In addition, the coupling constant of ³ J _H2 , _ _H3 is large (J> 9 Hz), and the coupling constant of the H4 and protons in the sugar part is small (δ 3.86, J <3Hz), the constituent sugar was found to be galactose.

For the second group of molecules, the HSQC (Heteronuclear Single Qua ntum Correlation: By examining the spectrum, it was found that the sugar part consists of disaccharides. A selective one-dimensional T0CSY spectrum confirmed the presence of 1 unit of hy-D-galactopyranosyl and 1 unit of J3-D-galactopyranosyl. M, but [delta] ^{3. 9} 1 Anoma of (J rather ³ Hz) and H ^3, but [delta] ^{3. 53} of (J> ⁹ H z) 'with pro tons signal δ 4. ²⁸ (J = 7.3Hz) From the signal, it was confirmed that the D-glycerol moiety and 3-galactopyranose sugar were bound. H2 "is _{^{S3.82 (3 J Hl i = 3.6Hz}} , 3 J H = 9.5Hz) and H4" Anomashi Gunaru of δ 3.94 (J <3Hz) δ4.90 with pro tons signal (J = 3.6 Hz) Thus, the presence of α-galactopyranose sugar was confirmed.

 Finally, the HMBC (Heteronuclear Multiple Bond Correlation) spectrum places the α-galactose CI 〃 located on the outer side with the H-6 'proton pair of j3-galactopyranose. A significant correlation was found between them, revealing a (1 "→ 6 ')-0-glycoside bond.

 These results indicate that there are two groups of galactolipids (monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG)) and saturation of their fatty acids depending on the number of sugar units in the sugar moiety. It was found that the chain length was different.

2. 3. LC-ESI / MS and LC-ESI / MS / MS analysis

 2. 3. 1. LC-ESI / MS analysis

To further elucidate the structure of galactolipids, methanol fractions separated on C-18 sep-pak cartridges were subjected to LC-ESI / MS analysis using different mobile phases. Specifically, two elution systems with different organic phase rates were used to improve the separation of galactolipids with different polarities (ie, the organic solvent content was changed). In addition, the analysis can be performed with or without 0.2% formic acid in the mobile phase to separate galactolipids and affect their behavior when electrosprayed. I checked the sound. In fact, the behavior of MS fractions of methanol fractions rich in galatatolipid content differed according to the acid strength of formic acid used as solvent.

Under elution conditions with acidic solvents, the mass spectrum is preferentially lost from the galactosyl moiety in addition to [M + Na] + ions, so [M + H- 1 62] + and [ M + H-180] ⁺ 'ions were generated. In addition, these molecular species collide and disintegrate in the L C-ESI / MS apparatus, so that the fatty acid acyl group disappears from the diacylglycerol structure as free fatty acid, and other fragment ions are removed. occured. This is generally observed as [R _x C0 + 74] + ion fragments.

In these experimental conditions, [R _x C0] + ions were also present. Here, X = 1 or 2, which is generated by releasing from the sn-1 position of the glycerol skeleton when X = 1 and from the sn-2 position when X = 2.

 Conversely, in non-acidic elution conditions, spontaneous fragmentation is unlikely to occur in the instrument, and the mass spectrum shows only [M + Na] + ions, [M + H-162] + There were few fragments of M + H-180] +.

 From these results, the fatty acid composition of each galactolipid could be determined. C18: 0 palmitic acid (or hexadecanoic acid); C18: 0 stearic acid (or octadecanoic acid); C18: l octadecenoic acid; C18: 2 octadecadienoic acid; C18: 3 octadecatrienoic acid; C19 : 2 Nonadecadienoic acid; C20: 0 arachidic acid (or eicosanoic acid); C20: l eicosenoic acid; and C21: 0 one or two fatty acids in heneicosanoic acid may be present as fatty acids in galactolipids I understood, but could not determine the three-dimensional structure. 2. 3. 2. LC-ESI / MS / MS analysis

In LC-ESI / MS / MS experiments, [M + Na-00 ₂ produced by [M + Na] ⁺ precursor ions By calculating the relative intensities of the H] + and [M + Na-R ₂ C0 ₂ H] + fragments, the three-dimensional structure of the acyl chain was determined and the galactolipids present in the mixture were identified.

 The acid strength of the elution solvent also affected the fragmentation pattern of the MS / MS spectrum. In fact, the LC-ESI / MS / MS data measured for MG DG compounds under acidic conditions show that fragmentation with [M + H-162] + ion formation occurs in addition to the fragment ions generated by cleavage of the galactose moiety. It showed that it was detected.

 Since the DGDG compound has two galactosyl units, its MS / MS spectrum was derived from the [M + H-162] + MG DG ion derivative by the disappearance of the second sugar (ie, the outer sugar) Product fragment ions were also detectable.

On the other hand, in addition to the fragments mentioned so far, all types of galactolipids (MG DG and DGDG) are obtained by cleavage of glycosidic bonds (this is mainly promoted under neutral conditions). The loss of neutral fatty acids resulted in [M + Na-R ^ O ^] + and [M + Na-R ₂ C0 ₂ H] + fragments. This finding suggests that this is an electronic analytical method that simply provides information on fatty acid composition and the arrangement of their bonds.

 The determination of the degree of unsaturation of fatty acids and the position of double bonds in the galactolipid chain was accomplished by analyzing the high molecular weight region of the MS / MS spectrum.

2.4. Fatty acid GC / MS analysis

In order to confirm what fatty acids are bound to the two carbons of the glycephthal skeleton, pure galactolipids separated by HPLC are subjected to transesterification, and the resulting fatty acid methyl esters are converted to GC / Analyzed by MS. The fatty acid composition of pure galactolipids is consistent with that estimated from the MS / MS spectrum, and in some cases also determines the exact structural form of polyunsaturated fatty acid double bonds. I was able to. GC / MS analysis of fatty acid methyl esters confirmed the presence of the following fatty acids: palmitic acid (C1 ⁶ : 0), stearic acid (C18:

0), oleic acid (C18: ln-9 cis), linoleic acid (C18: 2 n-6 cis), α-linolenic acid (CI ⁸ : ³ η-3 cis), nonadecadienoic acid (CI ⁹ : ² ), Arachidic acid (C20: 0), 11-eicosenoic acid (C20: l n-9 cis), and heneicosanoic acid (C21: 0).

(Example 1)

 The MS / MS spectrum is an MGDG and DGDG upper series with a large number of C (i.e. at least one of the two carbons of the glycerol skeleton is C20: 0, C20: l, and C21: 0 (With some fatty acids attached) produced a more complex spectrum, especially in the high molecular weight region, and a variety of strong fragment ions in addition to the fragments already described.

 In the LC-ESI / MS chromatogram, a compound that showed a peak with a mass spectrum of m / z 829.9 with a retention time of 71.59 minutes by elution method B (compound 1 in Table 1)

1) existed. It corresponded to a molecule in which sodium was added to MGDG where eicosenoic acid (C20: l) and linolenic acid (C18: 3) were bound to the two carbons of the glycerol skeleton.

Figure 1 shows the LC-ESI / MS / MS spectrum of compound (11). (A) is the overall spectrum, (b) is the high molecular weight region, and (c) is the low molecular weight region. In Figure 1 (a), two fragment ion peaks at m / z 519.6 and 551.5 were prominently detected. These were thought to be the result of the disappearance of the two fatty acids, icosenoic acid (C20: 1) and linolenic acid (C18: 3), respectively, thereby determining the fatty acid composition. The binding position of each fatty acid acyl chain in the MGDG molecule was determined by the relative intensity of [M + Na-R _x C0 ₂ H] + ions. Specifically, eicosenoic acid (20: 1) at sn-1 and linolene at sn-2 It was confirmed that the fatty acid chain of acid (18: 3) was present.

 According to literature values, the low molecular weight region shows fragment ions associated with the sugar moiety. The explanation is given below with reference to Fig. 1 (c). Cleavage of the glycosidic bond yields fragment ion peaks at m / z 185.1 and m / z 203.1, which are the hydroxyl groups removed from the sugar ring (m / z 185.1) (M / z 203. 1), and it was determined that the ions were added by sodium.

 The fragment ion peak at m / z 243.2 is the cleavage of two neutral fatty acids simultaneously from the galactosylglycerol skeleton.

In addition, in this spectral region, peaks at m / z 261.3, 293.2, and 335.4 are also seen, which have already been explained [RiCO] +, [R ₂ C0] +, And [R ₂ C 0 + 74] + fragment ions.

 The sodium-bound ions of m / z 301.3 and 315.3 were formed by the disappearance of one fatty acid and the elimination of a part of the acyl chain of the other fatty acid. This detachment occurred in the part of the other fatty acid in the chain of the acyl chain up to two or three positions away from the carbon.

 Information on the position of the double bond in the fatty acid acyl chain was obtained by analysis of the high molecular weight region of the MS / MS spectrum, as described below.

In typical fragmentation of saturated fatty acid acyl chains, only fragment ions resulting from the disappearance of C _n H _2n from the parent ion could be detected. Furthermore, in the case where the unsaturated fatty acid Ashiru strands are present, C _n H disappearance of _2n _ ₂ and C _n H _{2n + 2} have also been reported (YH Kim et al., Supra) ₀

Therefore, the high molecular weight region of the MS / MS spectrum of [M + Na] + ions at m / z 829.9 was analyzed in detail (Fig. 1 (b)). According to this analysis, a peak at m / z 811.9 due to [M + Na-18] + ions was observed. These latter two peaks determined that the first double bond was located 15 carbons away from the carbon carbon of the sn-2 fatty acid acyl chain. Instead, two different fragmentation paths could be assumed, explaining that the origin of the fragment ion is in the range from m / z 771.8 to m / z 5δ1.5. Of these, the first pathway involves the fragmentation of two fatty acid acyl chains exclusively, but it is not energetically dominant and the resulting fragment ions are slightly less intense.

In contrast, if two fatty acid acyl groups are fragmented and detected at the same time, more fragment ion peaks can be explained. For example, starting with the ion at m / z 811.9, the fragment ion at m / z 755.8 is C ₃ H _{4 of the} sn-2 fatty acid acyl group and CH ₄ of the sn-1 fatty acid acyl group. It could be formed by either the disappearance of each, or the disappearance of _two C ₂ H _{4 in} both acyl chains.

 Therefore, the MS / MS spectrum in the molecular weight range of 771.8.—551.5 was analyzed. As a result, it is confirmed that the double bond of the sn-2 acyl group is located at the 9th and 12th carbons, and that the double bond of the sn-1 acyl group is located at the 1st carbon. As a result, the former was confirmed to be 9, 12, 15-octadecatrynoic acid, and the latter was found to be due to 11-eicosenoic acid.

+イオンの断片化経路の分析に よる C_nH_2n、 C_nH_2n_₂、 および C_nH_2n+2の消失の算定もまた有用であった。 To establish the position of the double bond in the fatty acid chain that is attached to the C2 glycerol position, m / z 519.

+ C _n H _2n Analysis fragmentation pathway of ions, calculation of C _n H _2n _ _2, and C _n H _{2n + 2} of the disappearance was also useful.

 Apparently, a fragment ion peak at m / z 649.7 due to [M + Na-180] + ions also occurred in the high molecular weight region of the MS / MS spectrum.

(Example 2)

The same LC-ESI / MS chromatogram as in Example 1 above showed a peak at m / z 9 91.9 by mass spectrum. Compound with retention time 53.08 min by elution method B (2 4 ) Existed. This peak corresponds to the ion of a molecule in which DGDG is attached with sodium. This DGDG has 2 units of gallium at the sn-3 position of the glycerol skeleton. It presents the same fatty acid pattern as the MGDG molecule of Example 1 except that kutosyl is bound.

 Figure 2 shows the LC-ESI / MS / MS spectrum of compound (24). (A) is the overall spectrum, (b) is the high molecular weight region, and (c) is the low molecular weight region. The MS / MS spectrum (FIG. 2) showed a fragmentation pattern very similar to the MGDG molecule of compound 11 of Example 1 above.

As shown in Figure 2 (a), there are two fragment ion peaks at m / z 681.7 and 713.7 associated with [M + Na- RCCyfl + ion and [M + Na-R ₂ C0 ₂ H] + ion. Remarkably detected. These were beneficial in terms of the conformation and composition of the two fatty acids in the molecule being analyzed. It was concluded that this DGDG presents a C20: l-acyl chain at the C1 glycerol position and a C18: 3-acyl chain at the C2 position.

 In this case, the fragmentation of the two fatty acid acyl chains was only involved in one chain. Beginning with the [M + Na] + ion at m / z 991.9, fragmentation at the sn-2 acyl chain eventually resulted in the related propyl derivative / z 769.8; Figure 2 (b)).

 The same is true for fragmentation at the sn-1 chain. In this case, a stronger pattern could also be detected due to the incomplete fragmentation of the sn-2 chain first followed by the involvement of the sn-1 chain.

In the sn-2 chain, the fragment ion peak of m / z 945.9, which starts from [M + Na-18] + ion at m / z 973.9 and is caused by the disappearance of C ₂ H ₄ , is prominently detected. A fragment ion peak at m / z 905.9, which was caused by the disappearance of C ₅ H ₈ , was detected. In addition, the disappearance of C ₈ H _{12 and 2} produced ion peaks at m / z 865.9 and 853.8. This profile also explained the involvement of the presence of one, two, and three double bonds in the fragment sequences analyzed. The resulting fragmentation within the mass range of the fragment sequence considered Ion was analyzed in more detail. This analysis estimated that one carbon was missing from C _n H _2n and C _n H _2n _ ₂ (n = l units were missing), and their respective fragment ion peaks had a molecular weight of 1 Corresponds to 4 or 1 2 and is an indicator of the position of the double bond. In this way, it is determined that the exact position of the double bond in the sn-2 acyl chain is at the 9th, 12th, and 15th carbons, and the fatty acid linked to the C2 glycerol position is 9, 12, 15-octadecatrienic acid was confirmed.

 Similarly, in the sn-1 fatty acid chain, it was found that one double bond was located 11 carbons away from the carbonyl carbon. From this, it was concluded that 11-eicosenoic acid was present at the C1 glycerol position.

There were two fragment ions in the MS / MS spectrum, m / z 829.9 and 676.7. These ions were generated by the disappearance of two consecutive molecular weights 162, each corresponding to a galactose moiety. This confirmed that this sugar was a homologous compound having a higher molecular weight than the MG DG compound of Example 1. In the same manner as in Example 1, two neutral fatty acids are cleaved simultaneously to generate a sodium-bound ion of m / z 405.5, while the other fatty acid chain is lost. And elimination of a portion of the acyl chain of the other fatty acid resulted in sodium-bound ions at m / z 463.5 and 477.5. Instead of detecting [M + Na- Ι ^ 0 ₂ Η] + ion opium [M + Na-R ₂ C0 ₂ H] + ions in m / z 533.7 and 551.7, It was explained that one of the fatty acid acyl chains was lost from the DGDGG molecule lacking the outer galactose.

In addition, for MS / MS spectra, [R ^ 0] +, [R ₂ C0 + 74] + and [R ^ O +74] + for m / z 293.3, 335.5, and 367.1 Fragment ions were also shown.

(Example 3) Galatatolipids belonging to a lower series with a small number of c (ie, fatty acids of C16: 0, 18: 0, 18: 1, 18: 2, 18: 3, 19: 2 on the two carbons of the glycerol skeleton) For MGDG and DGDG with structures that are linked, the MS / MS spectrum did not appear to be as useful as the information from the higher series. Nevertheless, by comparing with the mass spectrum of galactolipids belonging to the upper series, it was possible to carry out precise structure determination for these compounds.

In these spectra, [M + Na-R _x C0 ₂ H] + fragment ions are most prominently detected, and [M + Na-162] +, [M + Na- 180] +, and [R _x C0 + 7 4] + fragment was also detected. Few other fragment ions were generated.

Take as an example a compound (7) that exhibits a [M + Na] + ion at m / z 813.9 in the mass spectrum and that has a retention time of 67.47 min in elution method B in the LC-ESI / MS mouthmatogram. . Figure 3 shows the MS / MS spectrum of compound (7). (A) is the overall spectrum, and (b) is the high molecular weight region. As shown in Fig. 3 (a), the MS / MS spectrum showed two strong ions at m / z 519.5 and 535.5. These correspond to [M + Na- „1]. + Fragment ion and [M + Na-R ₂ C0 ₂ H] + fragment ion. This mass difference is due to nonadecadienoic acid (C1 9: 2) and linolenic acid (C18: 3) In addition, due to the difference in strength between these two fragment ions, the nonadecadienol chain (C19: 2) force binds to the sn-1 position and the other acyl chain However, it became clear that it was bonded to the sn-2 position.

The high molecular weight region of the spectrum (Fig. 3 (b)) shows the two generated fragment ions of m / z 651.8 and 633.8 with [M + Na-162] ⁺ opium [M + Na-180] + respectively. The presence of In addition, in the low molecular weight region of the spectrum, [R ₂ C0] + fragment ions and [R ₂ C0 + 74] + Fragment of m / z 26 1.3 and 35.4 The fragment ion peaks already described at m / z 185.1 and 203.2 were seen with the dopant ions.

 As described in Example 1 or 2, the position of the double bond in the chain was determined by analyzing the fragmentation pattern of the high molecular weight region of the spectrum.

 In the MS / MS spectrum of galactolipids belonging to this subseries, fatty acid chain chain fragmentation was only involved in one chain. This often results in a common fragment ion for both acyl chains, as reported by Kim et al. (Supra), and always begins with the disappearance of the alkyl end (¾).

In the sn-1 chain, C _n H _2n disappeared continuously, resulting in fragment ions of m / z 755 · 8 and 727.7. On the other hand, the fragment ion of m / z 715.9 was generated, which marked the ω-double position in the second carbon. Position of the second double bond, according to the ion of C _n H _2n and C _n H _2n _ ₂ of m / z 701. related to lost 9, 68 7.8, and 675.8, 9 th Proven to be carbon. These results demonstrated that 9,12-nonadecadienoic acid is in the sn-1 glycerol position.

In the fragmentation pattern for the sn-2 acyl chain, the disappearance of CH ₄ first results in a fragment ion of m / z 797.8, and the disappearance of C _n H _2n - ₂ results in m / z 757. 8 fragment ions were generated. C _n H _2n — ₂ is n = 3 and suggests that there is a double bond at the 15th carbon of the chain.

Subsequently, C _n H _{2n and} C _n H _2n _ ₂ disappeared in the sn-2 chain, and CH ₄ disappeared in the sn-1 chain, m / z 743.8, 727. 7 Explained by the detection of fragment ion peaks at 715. 9, 687.8, and 685.8. This confirms that there are two additional double bonds in the 9th opi 1 2nd carbon and that there is 9, 12, 15-octadecatrynoic acid at the sn-2 glycerol position. did.

Separation by HPLC to verify that the position of the double bond is accurate The sample collected by extraction with methanol was subjected to GC / MS analysis. The retention time and mass spectrum were compared with the standard fatty acid methyl ester standard product, and agreed with the above results. As a result, it was confirmed that 9, 12-nonadecadienoic acid and monolinolenic acid were present in this compound. '

These two fatty acids were also observed in the fatty acid composition analysis of another MGDG compound. The compound was eluted on LC-ESI / MS chromatogram by elution method B with a retention time of 73.35 minutes, and exhibited an [M + Na] + ion at m / z 813.55 in the MS / MS spectrum. (Compound 6 in Table 1). In order to explain this result, we considered that the bond positions of the acyl chains are different. Relative intensities of MS / MS spectra m / z 535.5 and 519.5 [M + Na-R ^ O ^] + fragment ion and [M + Na-R ₂ C0 ₂ H] + in this compound It was confirmed that linolenic acid was located at the sn-1 glycerol position and nonadecadienoic acid was located at the sn-2 glycerol position. MS / MS spectral studies determined the positions of the double bonds of both acyl chains and confirmed that they were α-linolenic acid and 9,12-nonadenosic acid.

(Example 4)

Similarly, by using two elution methods (立体 and 二 重) to improve the separation of galactolipids with different polarities, depending on the fatty acid composition, conformation, and position of the double bond of the compound eluted. All galatatolipids present in the mixture were determined. The results are shown in Tables 1 and 2 below. Therefore, in addition to the compounds described in Examples 1 to 3 (compounds 11, 24, 7 and 6), compounds 1, 8 to 10, 1 2 to 14, 17, 22, 23 in Table 1 and Table 2 , 25 and 26, a novel galacto fat was obtained. Tables 1 and 2 below show the various compounds of the identified galactolipids. Table 1 shows MGDG having the structural formula (I), and Table 2 shows DGDG having the structural formula (II). In Tables 1 and 2, and R ₂ represent a fatty acid acyl group bonded to the sn-1 position and a fatty acid acyl group bonded to the sn-2 position, _{respectively.} The notation of fatty acid acyl groups follows the notation used when discussing biosynthesis. The position of the double bond is described with a number from the side opposite to the carbonyl group. For example, the linolenic acid has cis double bonds at the 9th, 12th, and 15th positions. : 3 n-6 cis ".

table 1

Retention time (min) MM + Na ⁺ Ri R ₂

1 69.18 ^a 752.54 ° 775.53 C16: 0 C18: 3 n-3 cis

2 64.35 ^a 778.56 801.55 C18: 2 n-6 cis C18: 2 n- -6 cis

3 47.53 ^a 776.54 799.53 C18: 2 n-6 cis C18.3 n-3 cis

4 55.77 ^a 780.57 803.56 C18: 3 n—3 cis C18: 0

5 36.05 ^a 774.53 797.52 C18: 3 n—3 cis C18: 3 n- 3 cis

6 73.35 ^b 790.56 ° 813.55 C18: 3 n-3 cis C19: 2

7 67.47 ^b 790.56 ° 813.55 C19: 2 C18: 3 n-3 cis

8 17.29 ^a 812.64 ° 835.63 C20: 0 C18: 1 n-9 cis

9 16.63 ^a 810.62 ° 833.61 C20.0 C18: 2 n-6 cis

10 14.26 ^a 808.62 ° 831.59 C20: 1 n-9 transformer C18: 2 n-6 cis

11 806.59 ° 829.58 C20: 1 n-9 卜 Lance C18: 3 n-3 cis

12 15.45 ^b 822.62 ° 845.61 C20: 1 n— 9 卜 lance C19: 2

13 15.97 ^b 838.65 ° 861.64 C20: 1 n—9 transformer C20: 1 n-9 卜 lance

14 15.32 ^b 822.62 ° 845.61 C21: 0 C18: 3 n-3 cis a: Elution method A

b: According to elution method B

c: New galactolipid

Table 2

Retention time (min) MM + Na ⁺ Ri R ₂

15 71.87 ³ 916.61 939.60 C16: 0 C18: 2 n-6 cis

16 53.90 ³ 914.59 937.58 C16: 0 C18: 3 n-3 cis

17 16.33 ³ 930.63 ° 953.62 C16: 0 C19: 2

18 97.78 ^a 942.63 965.62 C18.0 C18: 3 n-3 cis

19 52.64 ^a 940.61 963.60 C18: 2 n-6 cis C18: 2 n-6 cis

20 37.16 ^a 938.59 961.58 C18: 2 n— 6 cis C18: 3 n-3 cis

21 27.53 ^a 936.58 959.57 C18: 3 n—3 cis C18: 3 n-3 cis

22 53.73 ^b 952.61 ° 975.60 C19: 2 C18: 3 n-3 cis

23 15.19 ^a 946.66 ° 969.65 C20: 1 n-9 transformer 016: 0

24 53.08 ^b 968.64 ^c 991,63 C20: 1 n—9 transformer C18: 3 n-3 cis

25 13.10 984.67 ° 1007.66 C20: 1 n-9 transformer C19: 2

26 12.62 ^b 1000.70 ° 1023.69 C20: 1 n-9 Transformer C20: 1 n—9 卜 Lance a: According to elution method A

b: According to elution method B

c: New galactolipid

(Example 5)

 Planted seeds of sweet potatoes and cultivated until the stem length (the length of the part protruding from the ground) reached about 150 cm. Next, a 60 cm portion (young shoot leaves) was cut from the tip of the potato shoots and washed twice with water to obtain 1 kg of potato leaves. '

 The obtained sweet potato leaves were cut to about 5 mm and immersed in 2 L hot water (90 ° C) adjusted to pH 8.0 for 1 minute. Next, it was cooled with water at 25 ° C, and the cooled sweet potato leaves were centrifuged for 30 seconds to be dehydrated to some extent. After dehydration, dry with warm air (primary drying) for 2 hours at 70 ° C in a dryer until the water content reaches about 20% by mass, and further at 80 ° C so that the final water content becomes 3% by mass. Hot air drying (secondary drying) was performed for 4 hours. Next, steam sterilization was performed using saturated steam at 150 ° C for 3 seconds. Next, the water contained in the sweet potato leaves is dried by warm air drying, and then finely ground using a hammer mill so that 90% by mass passes through the 200 mesh section, and the dry powder of sweet potato leaves (80 g) is added. Obtained.

 Using the above powder, foods (tablets; 2.00 mg / tablet) can be produced in the following amounts. ·

<Ingredients> Blending amount (mass%) Sweet potato leaf dry powder 5 Barley young leaf powder 11 Asconorebic acid 10 Crystalline senorelose 14 Sucrose esterole 4 Reduced maltose 30 Silicon dioxide 1 Tre / Rose 25 (Example 6)

 Using the dry powder of Example 5 above, a food (granule) is produced with the following blending amount.

 <Raw material> Blending amount (% by weight) Sweet potato leaf dry powder 2 Kale dry powder 3 3 Tea catechin 1 0 Crystalline cellulose 2 0

(Example 7)

 Using the dry powder and indigestible dextrin of Example 5 above, a food can be produced. (Example 8)

 A food can be produced using the dry powder and oligosaccharide of Example 5 above.

(Example 9)

 A food can be produced using the dry powder and lactic acid bacteria of Example 5 above. Industrial applicability

Based on its biological activity, the galatatolipid of the present invention can be used in pharmaceuticals, foods and beverages, and cosmetics. Examples of the biological activity include an antitumor action, an anti-inflammatory action, and an antioxidant action. Galatatolipids of the present invention are functional foods, supplements, and skin quality improving effects, cosmetic effects, and moisturizing effects that have anti-cholesterol effects, blood flow improving effects, etc. It can be used for the production of cosmetics with fruits.

Claims

The scope of the claims 1. a monogalactosyldiacylglycerol compound having the following structural formula:

Ri is a palmitate palmitate and R ₂ is an α-linolenic acid acyl chain; A force c-linolenic acid acyl chain and R ₂ is a 9, 12-nonadecadienoic acid acyl chain; Is a 9,12-nonadecadienoic acid acyl chain and R ₂ is an α-linolenic acid acyl chain; R is an arachidic acid acyl chain and R ₂ is an oleic acid acyl chain; R is an arachidic acid acyl chain and R ₂ is a linoleic acid acyl chain; Is an 11-eicosenoic acid acyl chain and R ₂ is a linoleic acid acyl chain; Is a 1 卜 eicosenoic acid acyl chain and R ₂ is an α-linolenic acid acyl chain; Is an 11-eicosenoic acid acyl chain and R ₂ is 9, 12-nonadecadien Whether it is an acid chain; Is an 11-eicosenoic acid chain and R ₂ is an 11-eicosenoic acid chain; or 1 ^ is heneicosanoic acid acyl chain, and shaku ₂ is 0; -linolenic acid acyl chain, compound  Or its derivatives. 2. A food comprising at least one selected from the monogalatatosyldiacylglycose oral compound according to claim 1 or a derivative thereof. 3. The food according to claim 2, further comprising at least one selected from the group consisting of wheat leaf and kale. 4. The food according to claim 2 or 3, further comprising at least one selected from the group consisting of indigestible dextrin, oligosaccharide, and lactic acid bacteria. 5. A process for producing the monogalactosyldiacylglycerol compound or derivative thereof according to claim 1, comprising the steps of:  (1) Process of subjecting sweet potato leaves to methanol extraction,  (2) a step of separating the low polarity fraction by solid phase extraction; and  (3) Separation process using high performance liquid chromatography Including the method. 6. A digalactosyldiacylglycose oral compound having the following structural formula:

here, Is a palmitate palmitate and R ₂ is a 9,12-nonadecadienoic acid chain; 1 ^ is a 9, 12-nonadecadienoic acid acyl chain, and ₂ is a hy-linolenic acid acyl chain; Whether R is an 11-eicosenoic acid acyl chain and R ₂ is a palmitic acid acyl chain; Is an 11-eicosenoic acid acyl chain and R ₂ is an α-linolenic acid acyl chain; Is an 11-eicosenoic acid acyl chain and R ₂ is a 9, 12-nonadecadienic acid acyl chain; or 1 ^ is a 1-eicosenoic acid chain, and R ₂ is an 11-eicosenoic acid chain,  Or its derivatives. 7. A digalactosyldiacylglycerol compound according to claim 6 or A food comprising at least one selected from its derivatives. 8. The food product according to claim 7, further comprising at least one selected from the group consisting of barley wakaba opi kale. 9. The food according to claim 7 or 8, further comprising at least one selected from the group consisting of indigestible dextrin, oligosaccharide, and lactic acid bacteria. 10. A method for producing a digalatatosyldiacylglycerol compound or derivative thereof according to claim 6, comprising the steps of:  (1) A step of subjecting the sweet potato leaves to methanol extraction,  (2) separating the low polar fraction by solid phase extraction; and  (3) Separation process using high performance liquid chromatography Including the method.