JP2002531109A - Methods and compositions for producing berry derived products - Google Patents

Abstract

(57) Abstract A method is described for separating a mixture of anthocyanins, bioflavonoids and phenols from edible berries using an adsorbent resin that is regenerated for reuse. Mixtures with consumable carriers are particularly useful in food products and as dietary supplements or nutritional products.

Description

DETAILED DESCRIPTION OF THE INVENTION

CROSS REFERENCE TO RELATED APPLICATIONS This application is filed from US application Ser. No. 09 / 317,310, filed Mar. 24, 1999;
Claim priority to US Provisional Application No. 60 / 120,178, filed on the 16th. No U.S. government sponsored research or development statement

BACKGROUND OF THE INVENTION (1) Summary of the Invention The present invention edible berry derived composition, as more information preparation method cherry, and or food additive as vegetable / nutritional dietary supplements that berry derived composition How to use. In particular, the present invention relates to natural berry compositions containing anthocyanins, bioflavonoids and phenols, in particular cherries, for use as dietary supplements or food additives.

(2) Description of the Related Art Many plant-derived compounds inhibit prostaglandin synthesis due to their ability to serve as cellular antioxidants by maintaining low levels of reactive oxygen intermediates. Foods can also be provided with pharmacological or "nutritional" traits, possibly as anti-inflammatory agents or as inhibitors of enzymes involved in cell growth. These activities are important in ameliorating chronic diseases including cancer, arthritis, and cardiovascular disease (Kinsella et al., Food Tech. 85-89 (1993)). Thus, for natural products, the dietary supplement / food industry and nutrition companies are able to increase food stability as efficiently as synthetic antioxidants but also provide significant health benefits to consumers. Have the opportunity to take advantage of

[0004] Dyes such as anthocyanins are regarded as indicators of quality in tart cherries. Most importantly, recent work has shown that anthocyanins such as cyanidin-3-glucoside have strong antioxidant activity (Tsuda, T. et al., J. Am.
Agric. Food Chem. 42: 2407-2410 (1994)). Addition of antioxidants is one of the popular ways to extend the shelf life of foods thought to be related to lipid peroxides. Natural antioxidants can play an important role in cancer prevention. Dietary antioxidants are effective against peroxidative damage in biological systems (Halliwell, B. and JMC Gutteridge
Free radicals in biology and medicine, Oxford University Press, New York 416-494 (1989); Osawa, T. et al., Role of dietary antioxidants in protection against oxidative disorders, anti-mutagenic and anti-cancer mechanisms; Kuroda, Y .; Shankel, DM, Waters
, MD, Eds .; Plenum Publishing, New York 139-453 (1990)).

Early studies have shown that MONTMORENCY cherries contain anthocyanin cyanidin-3-gentiobioside and cyanidin-3-rutinoside (Li, KC, et al., J. Am. Chem. Soc. 78: 979). -980 (1956)). Cyanidin-3-glucosylrutinoside was also found in six of the seven sour cherry varieties (Harbor
ne, JB, et al., Phytochemistry 3: 453-463 (1964)). Dekazos (Dekazos, ED, J. Foo
d Sci. 35: 237-241 (1970)) converts anthocyanin dyes in montmorancy cherries to cyanidin-3-sophoroside, cyanidin-3-rutinoside and cyanidin.
Reported as peonidine-3-rutinoside, paeonidine and cyanidin along with -3-glucoside. However, cyanidin-3-glucosyl rutinoside as well as cyanidin-3-glucoside, cyanidin-3-sophoroside and cyanidin-3-rutinoside were identified as the main pigments in sour cherry. Using HPLC retention values, Chandra et al. (Chandra, A., et al., J. Agric. Food Chem. 40: 967-969 (1992)) suggested that cyanidin-3-sophoroside and montmorancy cherry grown in Michigan. Cyanidin-3-glucoside was reported to be the major and minor anthocyanins, respectively. Similarly, cyanidin-3-xylosylrutinoside was detected as a minor dye in montmorancy cherry (Shrikhande, AJ and FJ Francis).
, J. Food Sci. 38: 649-651 (1973)).

In the prior art, the production of pure anthocyanins (compounds 1-3 of FIG. 1) from BALATON and Montmorency Cherry secretions involves first converting the dye to AMBERLITE
Performed by absorption on an XAD-2 (Sigma Chemicals) column. The column was washed with water until the eluent had a pH of about 7.0. Dye absorbed with other phenols eluted with MeOH. The resulting crude anthocyanins were fractionated and purified by C-18 MPLC and HPLC, respectively, to give pure anthocyanins for spectroscopic studies. Purification of 500 mg of crude montmorancy anthocyanin by AMBERLITE XAD-2 yielded 60 mg of pure anthocyanins 1-3, compared to 391.43 mg from Balaton. This study showed that the crude anthocyanins from Montmorency obtained from XAD-2 contained a high percentage of other organic compounds. AMBERLITE XAD-2 could not reuse the resin. No attempt was made to use a crude mixture of phenols and anthocyanins for any purpose. Garb No. 5,266,685 for utt, the No. 5,817,354 for No. 5,665,783 and Mo Zaffar for Katzakian et al, describes their use for various absorbent resin and unrelated products. These patents merely illustrate the general state of the art in the use of absorbent resins.

[0007] U.S. Patent No. 5,503,867 for Pleva, describes the use of whole ground cherry and oat bran in ground beef. The amount of cherry used is 10-15% by weight, and oat bran is considered to be added to supplement the secretion of the cherry. In any event, cherries clearly add flavor to the taste of the meat and its generally unacceptable product.

[0008] Recent studies on the stabilization of low-fat ground beef by cherry tissue indicate that this plant source not only inhibits lipid peroxide, but also produces heterocyclic aromatic amines and cholesterol oxides when frying. (Gomaa et al., IFT Abstract No. 68E-7 (1996)). The hypothesis used to explain these observations was that polyphenols such as flavonoids, anthocyanins and anthocyanidins, which are commonly found in vacuoles of higher plants such as cherries, are responsible for this antioxidant effect. There is a need for a natural edible berry-derived composition, especially for use as a dietary supplement / nutrition or food additive.

SUMMARY OF THE INVENTION The present invention relates to a method for producing a mixture comprising edible berry-derived anthocyanins, bioflavonoids and phenols as a composition, comprising the following steps: (a) Berry-derived Supplying an aqueous solution containing anthocyanins, bioflavonoids and phenols of (a); (b) transferring the anthocyanins, bioflavonoids and phenols from the aqueous solution onto a resin surface; (c) eluent of the resin surface Eluting the anthocyanins, bioflavonoids and phenols from the resin surface; and (d) separating the eluent from the anthocyanins, bioflavonoids and phenols.

[0010] The present invention further relates to a method for producing edible berry-derived anthocyanins, bioflavonoids and phenols as a composition, comprising the following steps: (a) Fresh or flash frozen and thawed (B) breaking the berry and separating the pulp from the secretion thereof; (c) dissolving the anthocyanin, bioflavonoid and phenols from the pulp in an aqueous solution (D) transferring the anthocyanins, bioflavonoids and phenols from the aqueous solution containing the anthocyanins, bioflavonoids and phenols separated from the flesh onto adsorbent resin particles; e) washing the resin particles with a lower alkanol, Removing the anthocyanins, bioflavonoids and phenols; (f) separating the alkanols from the anthocyanins, bioflavonoids and phenols; and (g) separating the second batch of berries with the separated alkanols. Repeating steps (a) to (e) with the resin particles from which the anthocyanins, bioflavonoids and phenols have been removed.

The present invention further relates to a consumer composition comprising a mixture of: (a) a dry mixture of anthocyanins, bioflavonoids and phenols isolated from edible berries; and (b) a food grade carrier. However, the mass ratio of (a) to (b) is between about 0.1: 100 and 100: 0.1.

Finally, the invention relates to a method of feeding a mammal comprising providing the mammal with a consumable composition comprising a mixture of: (a) anthocyanins, bioflavonoids isolated from edible berries and A dry mixture of phenols; and (b) a food grade carrier. However, the mass ratio of (a) to (b) is between about 0.1: 100 and 100: 0.1. Preferably, the composition comprises, at least in part, the dried pulp of the berry.

The term “anthocyanin” refers to a compound that imparts color to a carrier. The term "bioflavonoid" refers to isoflavonoids and flavonoid compounds contained in cherries. The term "phenols" refers to compounds having a phenyl group and having one or more hydroxyl groups derived from cherry. Some of the edible berries are cranberry, raspberry, strawberry, blueberry, blackberry, elderberry, red grape, gooseberry, barbados cherry (acerola cherry) and chalk cherry.

An object is therefore an object of the present invention is to provide a in food or natural sources edible berry compositions which can be used as a dietary supplement or nutritional products. It is a further object of the present invention to provide a method for separating this composition on a commercial scale. Finally, it is an object of the present invention to provide a natural source composition that can be prepared economically and is easy to use. These and other objects will become more apparent with reference to the following description and drawings.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The cherries used in the present invention may be sweet or sour, but the latter has high levels of malic acid, which, in addition to other organic acids, contributes to the sour taste of tart cherries. It is preferable because it contains. The method of the present invention separates sugar-containing malic acid and other organic acids that can be used to impart sourness and flavor to foods. Most preferred are Balaton and Montmorency Cherry. The isolated mixture of anthocyanins, bioflavonoids and phenols can be tableted and used as a natural nutrition / dietary supplement. Generally, this tablet is
A daily dose of about 1 to 200 mg, preferably 60 to 100 mg daily, of anthocyanins and bioflavonoids is provided. 100 cherries give 60 to 100 mg of anthocyanins. Phenols (FIG. 4) are given in amounts of 0.1 to 50 mg daily. 100 cherries are 1 ~
This gives 50 mg of phenols. The amounts of anthocyanins, bioflavonoids and phenols can be prepared by isolating the individual compounds and blending them together. The use of a natural mixture of anthocyanins, bioflavonoids and phenols isolated by a resin is preferred.

This resin has a surface on which anthocyanins, bioflavonoids and phenols are adsorbed. A preferred class of adsorption resins is AMBERL, such as AMBERLITE XAD-4 and XAD-16, commercially available from Rohm & Haas Co., Philadelphia, PA.
A polymer crosslinked resin composed of styrene and divinylbenzene, such as the ITE series resin. Other polymeric crosslinked styrene and divinylbenzene adsorption resins suitable for use in the present invention are XFS-4257 from Dow Chemical Company, Midland, Michigan.
, XFS-4022, XUS-40323 and XUS-40322.

It is preferred to use a commercially available, required government approved, styrene-divinyl-benzene (SDVB) cross-linked copolymer resin (eg, AMBERLITE XAD-16). Thus, in a preferred embodiment, AMBERLITE XAD-16, commercially available from Rohm & Haas Company and described in U.S. Patent No. 4,297,220, is used as the resin. This resin is a nonionic hydrophobic crosslinked polyester divinylbenzene adsorbent resin. AMBERLITE XAD-16 has a macroporous structure with a continuous polymer phase and a continuous pore phase. In a particularly preferred embodiment, the resins used in the present invention have a particle size in the range of 100-200 μm (100-200 microns).

Other adsorbents, such as the AMBERLITE XAD adsorbent series of adsorbents, including hydrophobic macroporous resin beads with particle sizes in the range of 100-200 μm (100-200 microns) are also useful in the method of the present invention. it is conceivable that. In addition, various variations of AMBERLITES, such as the AMBERCHROM CG series adsorbents used in particle sizes in the range of 100-200 μm (100-200 microns) are also suitable for use in the present invention. AMBERLITE XAD-16 is used many times (10
It is preferable because it can be reused. However, for food, the government-approved resins of the present invention are considered important and / or desirable.

The adsorbed anthocyanins, bioflavonoids and phenols may be moved using any solvent. Lower alkanols containing one to four carbon atoms are preferred, and ethanol (ethyl alcohol) is most preferred as it is approved for food use. Normally, ethanol is azeotroped with water; however, absolute ethanol can be used. Water containing malic acid and sugars in the cherry passes through the column. This can be collected and used as a flavor in foods.

The anthocyanins, bioflavonoids and phenols are preferably separated from balaton and montmorency cherry. The composition of the cherry was February 1997
No. 08 / 799,788, filed on Dec. 12, and partly in US Application No. 60 / 111,945, filed Dec. 11, 1998. As described in these applications, Montmorency (Prunus cerasus) varieties account for over 95% of Michigan and US tart cherry cultivation. However, a new tart cherry cultivar, Balaton tart cherry (P. cerasus), has been planted in several Michigan orchards in place of Montmorency. This cherry is
It has a high anthocyanin content and is considered a better variant. Montmorency and Balaton tart cherries have been reported for anthocyanin content (Wang et al., 1997; Chandry et al., 1993). However, detailed studies of other phenolic compounds in Balaton tart cherries have not been previously performed. Earlier studies showed that Montmorency Cherry was based on cyanidin-3-gentiobioside and cyanidin-
3-Lutinoside (Li, KC et al., J. Am. Chem. Soc. 78: 979-980 (195
6)). Cyanidin-3-glucosyl rutinoside has also been found in six of the seven tart cherry variants (Harbone, JB et al., Phytochemistry 3: 453-463 (1964));
ekazos (Dekazos, ED, J. Food Sci. 35: 237-241 (1970)) is an anthocyanin dye of Montmorency Cherry, which is used for cyanidin-3-sophoroside and cyanidin-3-.
Reported as peonidine-3-rutinoside, paeonidine and cyanidin along with rutinoside and cyanidin-3-glucoside. However, cyanidin-3-glucoside,
Cyanidin-3-sophoroside and cyanidin-3-rutinoside, as well as cyanidin-3-glucosylsiltinoside, have been identified as major pigments in sour cherry. Using HPLC retention values, Chandra et al. (Chandra, A. et al., J. Agric. Food Chem.
40: 967-969 (1992)) and reported that cyanidin-3-sophoroside and cyanidin-3-glucoside are the major and minor anthocyanins of Montmorency Cherry grown in Michigan, respectively. Similarly, cyanidin-3-xylosylrutinoside was detected as a minor dye in Montmorency Cherry (Shri
khande, AJ and FJ Francis, J. Food Sci. 38: 649-651 (1973).

The term “carrier” or “filler” is used to mean a component that is added to increase the volume of a composition purified from cherry. Preferably dried cherry pulp. These include any edible starch material, proteins such as non-fat dry milk. This group includes flour, sugar, soybean meal, maltodextrin and various spices such as salts, peppers, spices and herbs. Fillers are used in an amount of from about 10 ^-6 to about 10 ⁶ parts by weight of the mixture.

The composition is introduced into the food in an amount of about 0.1 to 10 mg / gm of the active ingredient of the food. The amounts are preferably selected so as not to affect the taste of the food and to achieve the most beneficial results. The food may be high (wet) or low moisture (dry), as is well known to those skilled in the art. When used as a dietary supplement, tablets contain 0.01-1 gram of active ingredient.

Methods for extraction and isolation of phytochemicals (Chandra, A. et al., J. Agric. Food Chem. 41: 1
062 (1992); Wang, H. et al., J. Agric. Food Chem. 45: 0556-2560 (1997)), and rapid screening for antioxidant activity has been developed (Arora, A. and GM Strasburg, J. et al. Am
er. Oil Chem. Soc. 74: 1031-1040 (1997)). Using these methods, antioxidant compounds from Balaton and Montmorency Cherry are identified and characterized. Secreted cherry tissue is continuously extracted with hexane, ethyl acetate and methanol. Both the methanol and ethyl acetate fractions showed strong antioxidant activity in the screening assay. The ethyl acetate fraction was further purified by silica gel suction liquid chromatography to give four subfractions; the subfraction that showed the strongest antioxidant activity was separated by preparative reverse phase HPLC into seven more fractions. Figures 2 and 3 show bioflavonoids isolated from Balaton cherry. Thus, there are numerous analogs or homologs of tart cherry.

Two new phenolic compounds have been identified: I) 1- (3′-4′-dihydroxycinnamyl) -2,3-dihydroxycyclopentane, and II) 1- (3′-4′-). Dihydroxycinnamyl) -2,5-dihydroxycyclopentane. Other compounds isolated from the ethyl acetate extract of cherry fruit and characterized by spectroscopy include: 1- (3′-methoxy, 4 ′
-Hydroxycinnamyl) quinic acid, 2-hydroxy-3- (2'-hydroxyphenyl)
Propanoic acid, methyl 2-hydroxy-3- (2′-hydroxyphenyl) propanoate, D (+)-malic acid, β-sitosterol and β-sitosterol glucoside.
FIG. 4 shows some of the phenols isolated. In addition, the anthocyanin component obtained from the secretion fraction has been identified and fully characterized (Ch
andra, A. et al., J. Agric. Food Chem. 41: 1062 (1992); Wang, H. et al., J. Agric. Food Chem.
45: 2556-2560 (1997)); the results indicate that these compounds contain potent antioxidant activity.

Examples 1 and 2 Individual quick frozen (IQF) cherries (with nuclei removed) were thawed and blended in an industrial WARING blender, as shown in FIG. 10,000rp mixture
The secretions were decanted by centrifugation at m. The residual pulp was further squeezed with cheesecloth to remove any additional secretions. The pulp was lyophilized at 15 ° C. The secretion was treated on AMBERLITE XAD-16 HP resin to obtain cherry acidity, anthocyanins, bioflavonoids and phenols. 1 kg of XAD-16 resin was washed with ethanol (1-2 L) and then with water (6 L). The XAD16 resin was allowed to stand in water for 1 hour, and then loaded into a glass column (10 × 90 cm long) with a cotton plug. After washing the packed column with water (2 L), the secretion liquid was loaded for separation. Purified for every 800 mL of secretion. The secretion liquid was added on the surface of the column, and was stabilized so as not to flow. It was eluted with water and the first IL was discarded. The next 2 L washes were collected because they contained malic acid and sugar from the cherry and sour cherry secretions. The column was further washed with 4 L of water for Balaton and 5 L of Montmorency Cherry secretion. Once the cherry secretion was collected, the residue from the water wash was discarded. Then, the column is diluted with ethanol (1.3 to 1.5 L).
) Eluting and collecting a red solution containing anthocyanins, bioflavonoids and phenols (700-800 mL). After draining the column and washing with 10 L of water, the process was repeated many times (more than 100 times).

The red alcoholic solution was evaporated under reduced pressure (2.7 Pa (20 mtorr)) to remove ethanol, the aqueous solution was stabilized with 50 ppm ascorbic acid,
And freeze-dried. The red powder was collected and stored. Results of Example 1: mass of Balaton cherry IQF cherry 15.74 kg mass of dried pulp 605 g volume of secretion fluid 12.16 L mass of anthocyanins, bioflavonoids and phenols (red powder) 31.35 g sour by-products (malic acid and saccharides) 35L Result of Example 2: Mass of Montmorency Cherry IQF Cherry 30.45kg Mass of dried pulp 895g Volume of secretion fluid 24.03L Mass of anthocyanins, bioflavonoids and phenols (red powder) 47g Sour by-products (malic acid and sugars) 75 L The red powders of Examples 1 and 2 are preferably mixed with the dried pulp as a carrier into tablets of 1-1000 mg including the carrier (1 adult daily).

Various food grade acids can be added to the separated anthocyanins, bioflavonoids and phenols to prevent degradation. Preferably, they do not add flavor. Ascorbic acid (vitamin C) is preferred. The acid can be added either before or after drying the cherry compound. For small-scale processing, lyophilization is used to remove water. For large scale production, drying in an air circulation oven is preferred.

Embodiment 3 As shown in FIG. 6, the open container 10 includes an inlet line 11 and an outlet line 12 having valves 13 and 14, respectively. Resin beads 15 are placed in open container 1
It is supplied within 0. Water is introduced into the container 10 and removed via the outlet line 12 and discarded. As in Example 1, cherry secretion (without pulp or nucleus) is introduced into container 10 and allowed to stand for 25 minutes. The temperature of water and secretion is about 20 ~
30 ° C. Cherry secretion residue containing malic acid and saccharide is discharged through outlet line 1
Remove from 2 and save as food seasoning. Then, the resin 15 in the melter is washed again with water from the inlet line 11, removed, and drained from the outlet line 12. Then, 95% ethanol is introduced from the inlet line 11 to anthocyanin on the resin particles,
Extract bioflavonoids and phenols. The ethanol containing anthocyanins, bioflavonoids and phenols is taken out of the container 10. Ethanol is removed from anthocyanins, bioflavonoids and phenols and dried by flash drying under nitrogen. The resulting powder is preferably mixed with dried cherry pulp or other carriers as in Example 1. The resin particles are washed with water, and the resin and ethanol are reused many times.

Example 4 A crude ethyl acetate extract from cherries (containing anthocyanins, bioflavonoids and phenols) was tested in aqueous solution under various conditions by fluorescence analysis for antioxidant activity. First, the fluorescence analysis will be described. Fluorescence analysis of antioxidant activity (general) : To screen a very large number of compounds or extracts for antioxidant activity, a model that reasonably well represents the structure and functional properties of the composition alone or in food It requires the use of a system (or systems). Also, the tests must be sensitive, fast and inexpensive. To evaluate the antioxidant efficiency, a fluorescence-based assay was used (Arora, A. and G. et al.
M. Straburg, J. Am. Chem. Soc. (1996)). Large unilamellar vesicles were prepared consisting of 1-stearoyl-2-linoleoyl-sn-glycero-3-phosphocholine that closely resemble the biological membrane, one of the major sites of peroxidation. Incorporating a fluorescent probe, 1,6-diphenylhexatrienepropionic acid into the membrane, such that the polar head group anchors the probe near the aqueous interface, while the hydrophobic portion is located parallel to the fatty acid chain. did. The probe reacts with free radicals generated during peroxidation, resulting in a decrease in fluorescence intensity over time. The reaction is initiated using a peroxide inhibitor (such as ferrous ion or the free radical generator AAPH (such as azobis- [2-amidinopropane hydrochloride]), in the presence of the antioxidant composition to be tested. In the absence of it, the kinetics of the decrease in fluorescence is determined: at present, assays for certain concentrations of compounds take only 21 minutes on a simple fluorometer and consume only a few micrograms of fat. It can be done easily.

(MacDonald, R, C. et al., Biochem. Biophys. Acta 1061: 297-30)
38 (1991)) outlined and followed the procedure for 1-stearoyl-2-linoleoyl-sn
Large monolayer vesicles (LUVs) were prepared from -glycero-3-phosphocholine. In short, the lipid was dissolved in chloroform and dried on a rotary evaporator to form a thin film. The dried membrane is resuspended in an aqueous suspension and the Liposofast piposomes (piposom
e) Extruded through a 100 nm pore size polycarbonate filter using an extruder (Avestin, Inc., Ottawa, Canada). Vesicle size uniformity (80-100n
m) and the nature of the monolayer were confirmed by freeze-destructive scanning electron microscopy. During preparation, a fluorescent probe, diphenylhexatriene-propionic acid (DPH-PA) was incorporated into the vesicle at a molar ratio of 1: 350 (probe: lipid). DP for fluorescence experiments
LUVs containing H-PA were purified to a final concentration of 100 μM in 100 mM NaCl, 50 mM Tris-
Suspend in HEPES buffer at pH 7.0. Excitation of the fluorescent probe at 384 nm, 423n
Light emission was detected at m. Lipid oxidation is inhibited in LUVs by the addition of ferrous ions or the free radical generator AAPH; its decrease in DPH-PA fluorescence intensity as a result of reaction with free radicals generated during 21 minutes. Progress was monitored. A plot of the decrease in fluorescence intensity as a function of time was used to determine the kinetics of lipid oxidation. The results show that a mixture of the crude anthocyanin extract with ethylene acetate is effective in inhibiting oxidation.

[0031] Solvent extraction of anthocyanins, bioflavonoids and phenols can be used; however, this is not preferred when the product is used as a food because it is expensive. With the preferred adsorbent resin, this step is not necessary. The components can also be separated and remixed by chromatography; however, this requires high pressure liquid chromatography and is therefore too expensive for the purposes of the present invention.

Example 5 Wang et al., J. Nat. Products 62: 294-296 (1999); Wang et al., J. of Ag. And Food Chemis.
try, 47: 840-844 (1999) and Wang et al., J. of Nat. Products, 62: 86-88 (1999), using cyclooxygenases I and II (COX-I and COX-
II) was used to test the anti-inflammatory activity of the composition. As a result, the composition showed a strong anti-inflammatory activity, especially strong inhibition of COX-I and COX-II.

Examples 6, 7 and 8 Processing of Raspberry, Blueberry and Blackberry : Raspberry (339 g), Blueberry (350 g) and Blackberry (670 g) were thawed and separately 500 mL in an industrial Waring blender Blended with water. The mixture was centrifuged at 10,000 rpm for 20 minutes and the secretion was decanted. The residue (pulp) was further squeezed with cheesecloth to remove any additional secretions. The secretions after centrifugation were approximately 775 mL (raspberry), 700 mL (blueberry), and 1100 mL (blackberry).

The pulp was lyophilized at 15 ° C. Secretions were processed on XAD-16 resin to separate anthocyanins and phenols from sugars and acids. The XAD-16 resin (1 kg) was washed with ethanol (1-2 L) and then with water (6 L). After the resin was allowed to stand in water for 1 hour, a secretion liquid for separation was loaded. Purified for every 800 mL of secretion. The secretion liquid was added on the surface of the column, and was stabilized so as not to flow. Elute with water and discard the first 1 L. The next 2 L washes were collected because they contained sugar and acid. The column was washed with an additional 3 L of water for all secretions. After removal of sugar and acid, the column was ethanoled (1.3 L for raspberry and blueberry, 1.0 L for blackberry).
L) Each wash was performed and a red solution containing anthocyanins and phenols was collected (700 mL). After the column was drained and washed with 10 L of water, this process was repeated.

Then, the red alcoholic solution was evaporated under reduced pressure to remove ethanol, the aqueous solution was stabilized with 50 ppm of ascorbic acid, and lyophilized at 10 ° C. The red powder was collected and stored at 20.degree. Table 1 shows the results.

[0036]

[Table 1]

The foregoing description is merely illustrative of the present invention, and the present invention is limited only by the claims.

[Brief description of the drawings]

FIG. 1. Anthocyanins (pigments) isolated from Balaton and Montmorency Cherry
Wherein aglycone cyanidin has a hydroxyl group at the 3-position.

FIG. 2 shows major bioflavonoids isolated from cherries as described in provisional application number 60 / 111,945, filed Dec. 11, 1998.

FIG. 3 shows major bioflavonoids isolated from cherries as described in provisional application number 60 / 111,945, filed Dec. 11, 1998.

FIG. 4 shows phenols isolated from tart cherry.

FIG. 5 shows the steps of the method of the invention described in Examples 1 and 2.

FIG. 6 is a schematic diagram showing the use of an open container 10 to hold resin beads that transfer anthocyanins and phenols from cherry secretions.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 10 Open container 11 Inlet line 12 Outlet line 13 Valve 14 Valve 15 Resin

────────────────────────────────────────────────── ─── Continued on the front page (31) Priority claim number 09 / 317,310 (32) Priority date May 24, 1999 (May 24, 1999) (33) Priority claim country United States (US) ( 31) Priority claim number 09 / 383,324 (32) Priority date August 26, 1999 (August 26, 1999) (33) Priority claim country United States (US) (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ) , BY, KG, KZ, MD, RU , TJ, TM), AE, AL, AM, AT, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GD , GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, US, UZ, VN , YU, ZA, ZW

Claims (21)

[Claims] 1. A method for producing a mixture containing edible berry-derived anthocyanins, bioflavonoids and phenols as a composition, comprising the following steps: (a) the berry-derived anthocyanins and bioflavonoids And (b) transferring the anthocyanins, bioflavonoids and phenols from the aqueous solution onto a resin surface; (c) eluting the resin surface with an eluent, Transferring the anthocyanins, bioflavonoids and phenols from the resin surface; and (d) separating the eluent from the anthocyanins, bioflavonoids and phenols. 2. A method for producing a mixture of anthocyanins, bioflavonoids and phenols from edible berries as a composition, comprising the steps of: (a) fresh or flash frozen and thawed cherries; Supplying a first batch; (b) breaking the berry and separating the pulp from its secretion; and (c) extracting the anthocyanin, bioflavonoid and phenols from the pulp into an aqueous solution. (D) transferring the anthocyanins, bioflavonoids and phenols from the aqueous solution containing the anthocyanins, bioflavonoids and phenols separated from the flesh onto adsorbent resin particles; (e) the resin Washing the particles with a lower alkanol, from the resin particles, the anthocyanin, (F) separating the alkanol from the anthocyanin, bioflavonoid and phenol; and (g) for a second batch of berries, the separated alkanol and the anthocyanin. Repeating steps (a) to (e) with the resin particles from which bioflavonoids and phenols have been removed. 3. The method according to claim 2, wherein said alkanol is ethanol. 4. The cherry according to claim 2, wherein the cherries are individually quick frozen.
The method described in. 5. The method according to claim 2, wherein the resin particles are in the form of a column. 6. The berry, wherein the berry is cranberry, raspberry, strawberry,
The method according to claim 2 or 3, wherein the method is selected from the group consisting of blueberries, blackberries, elderberries, red grape, gooseberries, barbados cherry (acerola cherry) and chalk cherries. 7. The method of claim 1, wherein the anthocyanins, bioflavonoids and phenols are mixed with the pulp from the berry and then dried. 8. The method of claim 1, wherein the anthocyanins, bioflavonoids and phenols are dried and then mixed with the dried pulp of the berry. 9. The method according to claim 2, wherein the anthocyanins, bioflavonoids and phenols are mixed with the pulp from the berries and then dried. 10. The method according to claim 2, wherein the anthocyanins, bioflavonoids and phenols are dried and then mixed with the dried berry pulp. 11. The method of claim 7, further comprising forming said mixture of said anthocyanins, bioflavonoids and phenols and said pulp into tablets. 12. Further, the anthocyanins, bioflavonoids and phenols are mixed with the pulp from the berries and then dried, and the mixture of the anthocyanins, bioflavonoids and phenols and the pulp is formed into tablets. The method according to claim 2 or 3, wherein the method is performed. 13. The method of claim 11, wherein the anthocyanin, bioflavonoid and phenols are dried and then mixed with the dried pulp of the berry, and the mixture of the anthocyanin, bioflavonoid and phenols and the pulp is formed into a tablet. Item 4. The method according to Item 2 or 3. 14. A mass ratio of (a) to (b) comprising: (a) a dry mixture of anthocyanins, bioflavonoids and phenols isolated from edible berries; and (b) a food grade carrier. Wherein the consumption composition is about 0.1: 100 to 100: 0.1: 15. A mass ratio of (a) to (b) comprising: (a) a dry mixture of anthocyanins, bioflavonoids and phenols isolated from edible berries; and (b) a food grade carrier. Is from about 0.1: 100 to 100: 0.1, and wherein the anthocyanins, bioflavonoids and phenols are prepared by the method of any of claims 1, 2 or 3. 16. The carrier of claim 1, wherein the carrier is dried pulp of the berry.
5. The composition according to 4. 17. A method of feeding a mammal comprising: a mixture of (a) a dry mixture of anthocyanins, bioflavonoids and phenols isolated from edible berries; and (b) a food grade carrier. Providing a consumable composition wherein the weight ratio of (a) to (b) is from about 0.1: 100 to 100: 0.1. 18. The method according to claim 1, wherein the carrier is dried pulp of the berry.
7. The method according to 7. 19. The method of claim 17, wherein said mammal is a human. 20. The method according to claim 1, wherein the mixture of anthocyanins, bioflavonoids and phenols is prepared by the method according to claim 1, 2, or 3.
7. The method according to 7. 21. The method of claim 17, wherein said mammal is an animal.