A deep processing method for ginkgo endosperm

Abstract

The invention discloses a deep processing method of ginkgo fruit endosperm, wherein the ginkgo fruit endosperm is subjected to supercritical CO2 extraction to obtain ginkgo fruit crude oil and endosperm after the crude oil is extracted, the ginkgo fruit crude oil is subjected to vacuum film evaporator, refrigerated centrifugation, vacuum filtration, and molecular distillation to obtain ginkgo essential oil; the endosperm after the crude oil is extracted is subjected to dynamic continuous countercurrent extraction, centrifugal separation, and macroporous adsorption resin enrichment to obtain a medium-polar active component extract, the endosperm after the crude oil is extracted is subjected to dynamic continuous countercurrent extraction to also produce a first solid residue, the first solid residue is subjected to microwave-assisted extraction and ultrafiltration to obtain a filtrate, a second solid residue and a retentate, the filtrate is subjected to reduced pressure vacuum and spray drying to obtain a polar active component extract; the second solid residue and the retentate are mixed and fermented to obtain ginkgo fruit wine and vinasse, and the vinasse is dried and crushed as a biological feed. The invention makes full use of ginkgo fruit endosperm resources according to the active ingredients, nutrients, and specific ingredients contained in ginkgo nuts to meet the needs of different groups of people.

Description

Technical Field

[0001] The invention relates to the technical field of deep processing of ginkgo nut, and in particular to a deep processing method of ginkgo nut endosperm. Background Art

[0002] Ginkgo fruit, commonly known as Ginkgo Semen, is the dried mature seed of Ginkgo biloba L. and is one of the famous nuts.

[0003] Ginkgo endosperm is the part of the ginkgo kernel left after the mesocarp, endocarp and embryo are removed from the ginkgo. The kernel (endosperm) of ginkgo is rich in nutrients, containing 11.27% protein, 73.2% starch, 3.93% fat, 3.17% total sugar, 1.2% crude fiber, 3.4% ash, the content of nucleosides and base components ranges from 772.8 to 1799.6 μg / g, adenosine, guanosine, uridine, adenine, guanine nucleoside components are widely present in ginkgo seeds, and it also contains vitamin A0.245 mg, vitamin C21.65 mg, vitamin E2.33 mg and other ingredients per 100 grams, as well as the content of macroelements K, P, Ca and Mg, and the content of trace elements Fe, Cu, Mn, Zn, Se, Al and B is relatively rich. In addition, it also contains active compounds flavonoids and terpene lactones (Zhou Guisheng. Research on the Chemistry of Ginkgo Seed Resources. Master's Thesis of Nanjing University of Chinese Medicine, 2013.; Zhou Guisheng, Yao Xin, Tang Yuping, et al. Research on the Chemical Components of Ginkgo Kernel. Chinese Pharmaceutical Journal, 2012, 47(17): 1362-1366.; Chen Jinming, Zhuang Pengyu, Zhao Liwei, et al. Research on the Chemical Components of Ginkgo Seeds. Journal of Liaoning University of Traditional Chinese Medicine, 2015, (8): 46-48.).

[0004] 17 chemical components were identified in supercritical extraction of ginkgo oil, of which unsaturated fatty acids accounted for 85.4%; unsaturated fatty acids were mainly 16- and 18-carbon. (Chen Xijuan, Wang Chengzhang, Chen Hongxia, Ye Jianzhong. Study on the extraction process and chemical composition of ginkgo oil. Journal of Northwest Botany, 2010, 30(4):838-843.). Fatty acids are one of the active ingredients of ginkgo, and linoleic acid and conjugated linoleic acid play an important role in lowering blood pressure, lowering blood lipids, and preventing and treating cardiovascular diseases (Ruan Zheng, Wu Moucheng, Hu Xiaobo, et al. Research progress of polyunsaturated fatty acids. China Oils and Fats, 2003, 28(2):55-59.). From the perspective of healthy edible oils, consuming unsaturated fatty oils such as oleic acid can delay aging and reduce the total cholesterol content in the blood (Zhou Guisheng. Research on the chemistry of ginkgo seed resources. Master's thesis of Nanjing University of Chinese Medicine, 2013.)

[0005] Ginkgo biloba has the same active ingredients as ginkgo leaves, namely ginkgolides and flavonoids (Chen Jinming, Zhuang Pengyu, Zhao Liwei, et al. Study on the chemical composition of ginkgo seeds. Journal of Liaoning University of Traditional Chinese Medicine, 2015(8):46-49). It has been reported that the content of ginkgolide in ginkgo seeds is higher than that in leaves (Luo Man, Bao Jiake, Xiong Huilin, Song Xiaoning, Zhang Zhiyong, Zhang Zhiyu. Study on the fingerprint of terpene lactone components in ginkgo kernels. Chinese Traditional Patent Medicine, 2011, 33(9):1465-1468; Zhang Qunqun, Zhang Xuelan, Li Huifen, et al. Comparison of the contents of four terpene lactone components in different parts and different processed products of ginkgo. Journal of Liaoning University of Traditional Chinese Medicine, 2015(10):45-47). Ginkgolide is a recognized platelet activating factor (PAF) receptor antagonist, and its pharmacological effect of antagonizing PAF has been fully confirmed. Ginkgolides are now clinically used to treat coronary heart disease, cerebral thrombosis, cerebral ischemia, brain dysfunction, sequelae of brain injury, nervous system diseases and asthma. Pure ginkgolides have been used in foreign countries for the treatment of stroke, organ transplant rejection, hemodialysis and shock (Hosford D, Braquet P. Antagonists of Platelet activating factor: chemistry, pharmacology and clinical applications. Prog Med Chem, 1990, 27: 325.).

[0006] Using defatted ginkgo nuts as raw materials and adopting the salting-out and salting-out methods, albumin (GAP, 41.99%), globulin (GGP, 48.68%) accounting for more than 90% of the total protein in the kernel, and salt-soluble protein (GSP, 3.98%) were prepared from ginkgo nuts for the first time. The active protein in the kernel accounts for 42% of the total protein. Ginkgo albumin (GAP) has anti-aging effects when administered by oral gavage or intraperitoneal injection. GAP can delay the aging process of naturally aged mice and galactose subacute aging model mice to a certain extent. Its mechanism of action may be related to improving the immune function and antioxidant capacity of mice (Deng Qianchun, Wang Lan, Wu Jia, et al. Study on the anti-aging activity of ginkgo albumin. Chinese Pharmacological Bulletin, 2006, 22(3):352-7.). The seed kernel contains anti-aging substances and can inhibit the spread of cancer cells; the seed kernel has antitussive, antiasthmatic, antibacterial, and anti-fatigue effects (Zhang Weiming, Wu Guorong, Zhao Botao. Study on the health care function of ginkgo seed kernel. Journal of Nanjing Normal University (Natural Science Edition), 1998, 21(3):72-74.).

[0007] Ginkgo polysaccharide isolated from ginkgo biloba has a significant scavenging effect on DPPH free radicals, superoxide free radicals and hydroxyl free radicals; in vivo experiments show that ginkgo polysaccharide can significantly increase the content of SOD, GSH-Px and CAT in serum and liver, and reduce the content of MDA in serum and liver. Therefore, the ginkgo polysaccharide has a significant antioxidant function; ginkgo polysaccharide has a lipid-lowering function and has a certain preventive effect on the occurrence of hyperlipidemia; ginkgo polysaccharide can significantly reduce the atherosclerosis index (AI), indicating that it has a certain preventive effect on coronary heart disease and atherosclerosis. (Yang Qiang, Li Xinhua, Wang Lin, et al. Study on the physicochemical properties and antioxidant activity of ginkgo polysaccharide. Modern Food Science and Technology, 2013, 29 (10): 2395-2400.; Yang Qiang. Study on the extraction, separation and functional characteristics of ginkgo polysaccharide. Doctoral dissertation of Shenyang Agricultural University, 2013.).

[0008] In summary, ginkgo nuts are rich in nutrition, sweet, fragrant and delicious, and can not only nourish the body, but also prevent and treat a variety of diseases. Regular consumption of ginkgo nuts can nourish the yin, nourish the skin, resist aging, dilate capillaries, promote blood circulation, make people's skin and face rosy, refreshed, and prolong life (Zhang Weiming, Wu Guorong, Zhao Botao. Research on the health care function of ginkgo kernels. Journal of Nanjing Normal University (Natural Science Edition), 1998, 1(3): 72-75.; Fang Jiamao. Research on the pharmacological effects of ginkgo cell extracts. Modern Food Science and Technology, 2009, 25(12) 1396-1403.).

[0009] In the long-term consumption, production and practice, it is known that "gingko is poisonous", and children are more sensitive to ginkgo. It is generally believed that the main substance that causes ginkgo allergy is ginkgolic acid (ginkgolic acid substances, also known as alkylphenol compounds). There are also reports that MAP (4-0-Methylpyridoxine) is the main toxic substance. The sensitization mechanism may be inhibition of enzyme activity, cytotoxicity, allergic reaction, etc. (Yang Jianting, Wu Cai'e* Research Progress on Ginkgo Allergenic Components and Their Sensitization Mechanisms. Food Science and Technology, 2009, 34(6): 282-286.). The content and proportion of ginkgolic acid in different parts of ginkgo fruit vary greatly, with the highest content of 64.415μg·g in the germ. -1 , followed by the endocarp content of 16.030 μg·g -1 , the content of the middle seed coat is 5.277μg·g -1 , the content of endosperm is 0.103μg·g -1 (Tian Xiaoqing, Wang Xichang, Wu Wenhui, et al. Determination of Ginkgo Biloba Acid Content in Dafozhi by High-Pressure Liquid Chromatography. Journal of Nutrition, 2010, 32(6): 608-611.); Sample ginkgo acid / (μg·g -1 ), total ginkgolic acid / (μg·g -1) are: embryo 3475.92, 5539.31, endocarp 169.93, 322.32, mesocarp 138.61, 245.90, endosperm 4.73, 9.27 (Li Zhuanmei, Zhang Xuelan, Li Huifen, et al. Quantitative comparison of ginkgolic acid and total ginkgolic acid in different parts and processed products of ginkgo. Chinese Patent Medicine, 2015, 37(1): 164-168.). The content of embryo is hundreds of times higher than that of endosperm. Therefore, it is safer to use endosperm as raw material for ginkgo processed products.

[0010] At present, the processing of ginkgo nuts is still in a relatively primitive stage. Most of them are fast food, and the products are relatively extensive and have a poor taste. There is no response to different groups of people, different types, and different needs. In particular, the ginkgo endosperm resources have not been rationally and fully utilized according to the active ingredients, nutrients, and specific ingredients contained in ginkgo nuts, and deep-processed into high-tech products. Summary of the invention

[0011] The technical problem to be solved by the present invention is to provide a deep processing method for ginkgo endosperm, which overcomes the above-mentioned problems existing in the prior art, makes reasonable and full use of ginkgo endosperm resources according to the active ingredients, nutrients and specific ingredients contained in ginkgo nuts, extracts and separates the ginkgo endosperm from non-polar to polar gradient, and deeply processes it into products with high technical content to meet the needs of different groups of people.

[0012] In order to solve the above problems, the present invention adopts the following technical solutions:

[0013] The present invention provides a deep processing method of ginkgo endosperm, comprising the following steps:

[0014] S1, putting the ginkgo nuts into a vacuum dryer for drying, so that the moisture content of the ginkgo nuts is less than 5%;

[0015] S2, putting the ginkgo nuts with a moisture content of less than 5% obtained in step S1 into a crusher for crushing to 4-40 mesh, and collecting the crushed ginkgo nuts for later use; wherein the crusher has a speed of 2500-3500 r / min;

[0016] S3, adding an entrainer to the ginkgo nut granules obtained in step S2 to perform supercritical CO2 extraction to obtain ginkgo nut crude oil and endosperm after the crude oil is extracted;

[0017] S4, adding the crude ginkgo nut oil obtained in step S3 to a primary falling film vacuum thin film evaporator for vacuum concentration, adding the concentrated crude ginkgo nut oil to a secondary scraper vacuum thin film evaporator for vacuum deodorization, and collecting the ginkgo nut crude oil from the discharge port of the scraper vacuum thin film evaporator;

[0018] S5, subjecting the ginkgo biloba oil obtained in step S4 to refrigerated centrifugation, collecting the supernatant of the refrigerated centrifugation for vacuum filtration, and further subjecting the filtrate to molecular distillation to obtain ginkgo essential oil;

[0019] S6, adding the endosperm after crude oil extraction obtained in step S3 to the extraction solution by a dynamic continuous countercurrent extraction method, and then centrifuging to obtain a first extract and a first solid residue; wherein the volume ratio of the endosperm after crude oil extraction to the extraction solution is 1:4-10, and the centrifugal separation speed is 12000-25000 r / min;

[0020] S7, the first extract obtained in step S6 is subjected to reduced pressure and vacuum recovery at a temperature of 50-70° C. and a vacuum degree of 70-90 Pa to obtain a second extract;

[0021] S8, adsorbing the second extract obtained in step S7 through a macroporous adsorption resin, washing the macroporous adsorption resin with water, collecting the water washing liquid, eluting with 50% to 80 vol% ethanol, and collecting the first alcohol eluate;

[0022] S9, the first alcohol eluate obtained in step S8 is subjected to reduced pressure and vacuum recovery at a temperature of 50-70° C. and a vacuum degree of 70-90 Pa to recover the ethanol solvent, the remaining liquid after the ethanol solvent is recovered is applied to a polyamide column, adsorbed at a flow rate of 1-2 BV / hr, washed with water until the effluent turns light yellow, and about 1.2 BV of impurities are removed, and 2-3 BV of water is continuously eluted, the eluate is collected, and the eluate is concentrated under reduced pressure and vacuum to 1 / 10 of the total volume of the eluate to obtain a first concentrated solution; wherein the volume ratio of the endosperm to the polyamide column is 8:1;

[0023] S10, extracting the first concentrated solution obtained in step S9 with ethyl acetate twice, combining the upper layer solutions, and rotary evaporating to obtain a light yellow-white extract, and then vacuum drying at 40-80° C. for 6-10 hours, crushing, and passing through a 100-mesh sieve to obtain a ginkgo total terpene lactone extract; wherein the volume ratio of ethyl acetate to the first concentrated solution is 1:1;

[0024] S11, eluting the polyamide column obtained in step S9 with 50% to 80 vol% ethanol, collecting the alcohol eluate when the HCI-Mg reaction is positive, and combining the second alcohol eluate when the HCI-Mg reaction is weakly positive;

[0025] S12, subjecting the second alcohol eluate obtained in step S11 to reduced pressure vacuum at a temperature of 50-70° C. and a vacuum degree of 70-90 Pa, and spray drying to obtain a total flavonoid extract; wherein the spray drying conditions are: an inlet air temperature of 160-180° C. and an outlet air temperature of 70-100° C.;

[0026] S13, mixing the total terpene lactone extract of Ginkgo biloba obtained in step S10 and the total flavonoid extract obtained in step S12, stirring evenly to obtain a medium-polarity active ingredient extract;

[0027] S14, subjecting the first solid residue obtained in step S6 to microwave-assisted extraction for 1 to 5 times, and centrifuging to obtain a third extract and a second solid residue; wherein the microwave-assisted extraction uses 0.1 mol / L sodium chloride solution as a solvent, and the volume ratio of the solvent to the first solid residue is 1:15 to 50.

[0028] S15, ultrafiltration of the third extract obtained in step S14, and collection of the filtrate and the retentate;

[0029] S16, the filtrate obtained in step S15 is subjected to reduced pressure vacuum at a temperature of 50-70°C and a vacuum degree of 70-90 Pa, and spray-dried to obtain a brown-yellow dry powder polar active ingredient extract; wherein the spray-drying conditions are: an inlet air temperature of 160-180°C and an outlet air temperature of 70-100°C;

[0030] S17, mixing the second solid residue obtained in step S14 and the intercepted material obtained in step S15, adding distiller's yeast and white sugar, mixing evenly, fermenting at 18 to 22° C. for 12 to 18 days, and centrifuging to obtain ginkgo wine and distiller's grains;

[0031] S18, vacuum-drying the wine lees obtained in step S17, and crushing the lees through an 80-mesh sieve to obtain a biological feed.

[0032] As a preferred technical solution, the crushing speed in step S2 is 2900 rpm, the crushing effect is optimal, and when the particle size of the ginkgo nut particles is 10 mesh, the ginkgo nut germ exposure effect is best.

[0033] As a preferred technical solution, the supercritical CO2 extraction conditions in step S3 are: pressure 25-35MPa, extraction temperature 25-45°C, and extraction time 1.5-3h. Among them, the yield of ginkgo fruit crude oil is best when the pressure is 32MPa, the temperature is 35°C, and the extraction time is 2.5h.

[0034] As a preferred technical solution, the entrainer in step S3 is one of methanol, ethanol, acetone and ethyl acetate. Adding a certain proportion of the entrainer can effectively promote the supercritical CO2 extraction to selectively separate and enrich certain functional substances.

[0035] As a preferred technical solution, the conditions for molecular distillation in step S5 are: vacuum degree 10 -3 ~1Pa, evaporation surface speed 500~1500r / min, evaporation surface temperature 60~100℃, condensation surface temperature 0~10℃. Among them, the vacuum degree is 10 -2When the evaporation surface speed is 1000r / min, the evaporation surface temperature is 80℃, and the condensation surface temperature is 2℃, the yield of ginkgo essential oil is the highest.

[0036] As a preferred technical solution, when the volume ratio of the endosperm after crude oil extraction to the extraction solution in step S6 is 1:8, the extraction effect of the medium-polar components in the ginkgo endosperm is optimal.

[0037] As a preferred technical solution, when the centrifugal separation speed in step S6 is 15000 r / min, the extraction effect of the medium-polar components in the ginkgo endosperm is optimal.

[0038] As a preferred technical solution, the extraction solution in step S6 is selected from ethanol, acetone, ethyl acetate, and n-butanol. When the extraction solution is 60 vol% ethanol, the extraction effect of the polar components in the ginkgo endosperm is the best.

[0039] As a preferred technical solution, the process parameters of the dynamic continuous countercurrent extraction method in step S6 are: temperature 60-100°C, extraction time 1-3h, screw speed 0.2-2r / min. Among them, when the temperature is 85°C, the extraction time is 1.5h, and the screw speed is 1.2r / min, the extraction effect of the polar components in the ginkgo endosperm is the best.

[0040] As a preferred technical solution, the macroporous adsorption resin in step S8 is selected from one of weak polar macroporous adsorption resin, medium polar macroporous adsorption resin and non-polar macroporous adsorption resin. When the macroporous adsorption resin is selected from weak polar macroporous adsorption resin, the second extract has the best effect of adsorbing or desorbing total terpene lactones and flavonoids of Ginkgo biloba.

[0041] As a preferred technical solution, when the volume ratio of endosperm to macroporous adsorption resin during the adsorption process in step S8 is 15:1, the volume ratio of water to macroporous adsorption resin during the water washing process in step S8 is 1.5-5:1, and the volume ratio of ethanol elution amount to resin during the ethanol elution process in step S8 is 4-7:1.

[0042] Among them, when the volume ratio of water to macroporous adsorption resin in the water washing process is 3, the water washing process is optimal.

[0043] Among them, when the volume ratio of ethanol elution amount to resin in the ethanol elution process is 6, the ethanol elution effect is best.

[0044] Among them, the optimal volume ratio of the endosperm to the macroporous adsorption resin is 15:1. If the amount of the macroporous adsorption resin is too small, some active ingredients in the second extract will fall off. If the amount of the macroporous adsorption resin is too large, it will cause the amount of water eluent and alcohol eluent to increase, resulting in a waste of resources.

[0045] As a preferred technical solution, the optimal volume of endosperm and polyamide column in step S9 is 8:1. If the amount of the polyamide column is too small, some active ingredients in the first alcohol eluate will fall off. If the amount of the polyamide column is too large, it will cause waste of polyamide and water eluate.

[0046] As a preferred technical solution, the amount of water washing liquid used in step S9 is 2 to 3 BV. If the amount of water washing liquid used is less than 2 BV, the total terpene lactones in Ginkgo biloba cannot be completely eluted. If the amount of water washing liquid used is greater than 3 BV, it will cause a waste of water washing liquid.

[0047] As a preferred technical solution, when the microwave-assisted extraction in step S14 is performed three times, the extraction effect of the first solid residue is the best.

[0048] As a preferred technical solution, when the volume ratio of the solvent to the first solid residue in step S14 is 1:25, the extraction effect of the first solid residue is optimal.

[0049] As a preferred technical solution, the microwave-assisted extraction conditions in step S14 are: extraction temperature 50-80°C, each extraction time 8-20 minutes. Among them, the microwave-assisted extraction conditions: extraction temperature 70°C, each extraction time 10 minutes, the first solid residue extraction effect is the best.

[0050] As a preferred technical solution, in step S15, the molecular weight of the ultrafiltration membrane is 2-5 kD, and the extraction pressure is 0.06-0.25 MPa. Among them, when the molecular weight of the ultrafiltration membrane is 4 kD and the extraction pressure is 0.14 MPa, the filtration effect of the active ingredient in the third extract is the best.

[0051] As a preferred technical solution, when the fermentation time in step S17 is 14 days, the fermentation effect is optimal.

[0052] The invention uses supercritical CO2 extraction and molecular distillation to process the ginkgo fruit endosperm into highly functional ginkgo essential oil, and other vegetable oils are added to produce a series of blended composite oils. The ginkgo essential oil can also be further made into microcapsules of microparticle carrier type. Microencapsulation improves bioavailability, increases the stability of unstable active substances such as polyunsaturated fatty acids, and is convenient for making various dosage forms, and is convenient for storage and carrying. Ginkgo essential oil contains fat-soluble active ingredients, unsaturated fatty acids account for more than 92% of total fatty acids, is rich in polyunsaturated fatty acids, vitamins E, A, D and carotenoids such as lutein, beta-carotene and alpha-carotene; it can delay aging, reduce the content of total cholesterol in the blood, and has multiple biological functions such as anti-oxidation and anti-cancer, and is a nutrient beneficial to the human body.

[0053] The present invention extracts the endosperm after crude oil extraction by a dynamic continuous countercurrent extraction technology that reduces the amount of alcohol solvent, reduces energy consumption, and has a high extraction rate of effective components, and extracts and separates the active components of polar components, such as ginkgo terpenoid lactones and flavonoids, through a combination of centrifugal separation technology, and enriches them with macroporous adsorption resin to obtain a product with greatly increased content of terpenoid lactones and flavonoids, namely ginkgo crystal, which promotes the circulation of the brain and body limbs. It has an excellent tonic effect on increasing the blood flow to the brain and on the nervous system. It has a good effect on many problems including insufficient blood flow to the brain and intellectual decline in elderly patients, and has a good effect on many possible symptoms of aging, such as anxiety and depression, memory damage, difficulty in concentrating, decreased alertness, decreased intelligence, dizziness, headache tinnitus (ringing in the ears), macular degeneration of the retina (the most common cause of blindness in adults), inner ear disturbance (which can cause partial deafness), poor terminal circulation, and impotence caused by poor blood flow to the penis.

[0054] The present invention uses microwave-assisted extraction-salting method to extract the first solid residue, i.e., the endosperm after the extraction of the polar components, and uses water as a solvent to extract the nutrients rich in amino acids, proteins, polysaccharides, nucleoside components, water-soluble vitamins, polar trace elements, etc., and applies membrane separation technology to remove unnecessary substances and impurities, selectively separate active ingredients, and increase the content of polar active ingredients, which is beneficial to the subsequent product molding process to obtain a high-active ingredient product. It has the effects of improving immune function and antioxidant capacity, beautifying and anti-aging, and has a certain preventive effect on hyperlipidemia, coronary heart disease and atherosclerosis. The microwave-assisted extraction-salting method-membrane separation combined technology has the characteristics of rapidity, energy saving, high efficiency, and green environmental protection.

[0055] The second solid residue produced by the microwave-assisted extraction-salting method of the present invention and the retentate produced after membrane separation, i.e., the endosperm after the extraction of the intermediate polar and polar components, is mainly gelatinized starch. Ginkgo fruit wine can be brewed by adding koji, and fortified functional wine can be prepared by adding ginseng extract.

[0056] According to the present invention, after the wine lees obtained from winemaking are vacuum dried, crushed and sieved, vitamins, animal protein powder and the like can be added according to feed requirements to prepare biological feed.

[0057] In order to meet the needs of different groups of people, different types and different needs and strengthen certain functions, one or more nutritional supplements such as grape seeds, blueberries, sheep placenta, collagen powder, angelica extract, licorice extract, motherwort extract, trans-resveratrol, knotweed extract, L-carnitine, multivitamins, mineral extracts, lily of the valley extract, soybean extract, ginseng extract, etc. can be added to the ginkgo essential oil, medium polar components, polar components, ginkgo fruit wine and distiller's grains obtained by the present invention; coenzyme Q10, tea polyphenols, sea cucumber polypeptide, soybean isoflavone extract, astaxanthin, saw palmetto extract, wolfberry extract, aloe extract, cordyceps polysaccharide, bitter melon extract, etc. can be added to the ginkgo essential oil, medium polar components, polar components, ginkgo fruit wine and distiller's grains obtained by the present invention; Extracts, American ginseng extract, American ginseng saponins, proanthocyanidins, hawthorn flavonoids, sea buckthorn extract, flaxseed extract, ganoderma lucidum extract, shiitake mushroom extract, shiitake polysaccharide, curcumin, saffron extract, ginsenosides, ginseng extract, octacosanol, marigold extract, cranberry extract, α-lipoic acid, lycopene, valerian extract, maca extract, maca enamine, EPA (eicosapentaenoic acid,) DHA (eicosahexaenoic acid), N-acetyl-L-cysteine ​​(free radical scavenger), red yeast rice extract, Nordic deep-sea fish oil, glucosamine, chondroitin sulfate, bromelain, lecithin, garlic oil, royal jelly, etc. It can be processed into soft capsules, hard capsules, pills, tablets, granules, oral liquid preparations, wine preparations, etc.

[0058] The invention uses gradient extraction and separation from non-polar to polar, which is more conducive to the extraction and separation of active ingredients. The ginkgo endosperm resources are reasonably and fully utilized according to the active ingredients, nutrients and specific ingredients contained in the ginkgo nuts. One or more nutritional supplements can be added to the ginkgo essential oil, medium-polar component extract, polar component extract, ginkgo fruit wine and lees obtained by the invention to meet the needs of different groups of people and enhance product functions. DETAILED DESCRIPTION

[0059] Example 1

[0060] Take 1 ton of ginkgo nuts, put them into a stainless steel plate, and put them into a 60°C vacuum dryer for drying until the moisture content of the ginkgo nuts is less than 5%.

[0061] Add the ginkgo nuts obtained above to the crusher, adjust the crusher speed to 2500 / min, control the crushing particle size to 10-20 mesh, and put the crushed ginkgo nuts into the barrel of the supercritical extractor. The pressure of the extraction kettle is 32Mpa, the temperature of the extraction kettle is 28°C, the pressure of the separation kettle is 6Mpa, the temperature of the separation kettle is 55°C, the entrainer is methanol, and the extraction time is 2.5 hours. After the extraction, collect the ginkgo crude oil and the endosperm after the crude oil is extracted from the separation kettle, wherein the yield of ginkgo crude oil is 6.83%. Add the above ginkgo crude oil to the first-stage falling film vacuum thin film evaporator to recover the solvent, and then add it to the second-stage scraper vacuum thin film evaporator for vacuum deodorization, and collect the ginkgo oil from the discharge port. After frozen centrifugation and vacuum filtration, the filtrate is molecularly distilled, and the molecular distillation working parameters are: vacuum degree 10 -1 Pa, evaporation surface speed 1500r / min, evaporation surface temperature 60℃, condensation surface temperature 0℃, and refined ginkgo essential oil with a yield of 4.94%. The molecularly distilled ginkgo essential oil is put into a stainless steel oil tank and sent to the bottling process. Ginkgo essential oil has multiple biological functions such as delaying aging, reducing the content of total cholesterol in the blood, anti-oxidation, and anti-cancer.

[0062] Example 2

[0063] Take 2 tons of ginkgo nuts, put them into a stainless steel plate, and put them into a 60°C vacuum dryer for drying until the water content of the ginkgo nuts is less than 5%.

[0064] The ginkgo nuts obtained above are put into a crusher, the crusher speed is adjusted to 2900 / min, the crushing particle size is controlled at 7-9 mesh, and the crushed ginkgo nuts are loaded into the barrel of the supercritical extractor. The pressure of the extraction kettle is 30Mpa, the temperature of the extraction kettle is 35℃, the pressure of the separation kettle is 6Mpa, the temperature of the separation kettle is 50℃, the entrainer is ethyl acetate, and the extraction time is 2 hours. After the extraction, the ginkgo crude oil and the endosperm after the crude oil is extracted are collected from the separation kettle, and the yield of the ginkgo crude oil is 7.41%. The ginkgo crude oil is added to a first-stage falling film vacuum thin film evaporator to recover the solvent, and then added to a second-stage scraper vacuum thin film evaporator for vacuum deodorization, and the ginkgo initial oil is collected from the discharge port. After freeze centrifugation and vacuum filtration, the filtrate is molecularly distilled, and the molecular distillation working parameters are: vacuum degree 10 -2 Pa, evaporation surface speed 1500r / min, evaporation surface temperature 80°C, condensation surface temperature 2°C, and refined ginkgo essential oil with a yield of 5.36%. The molecularly distilled ginkgo essential oil is put into a stainless steel oil tank and sent to the bottling process.

[0065] Example 3

[0066] Take 1000 g of the ginkgo essential oil prepared in Example 1, add 50 g of PEG4000, mix with 50 g of vitamin E, stir evenly, and put into soft capsules.

[0067] Example 4

[0068] Take 1000g of the ginkgo essential oil prepared in Example 2, add 0.38g of gum arabic powder to a homogenizer, grind evenly, add 1000ml of water at high speed and evenly, to form colostrum, and dilute it with 30000ml of 3% gum arabic solution to form an emulsion. Stir while heating, wait until the temperature rises to 45°C, slowly add 3% gelatin solution preheated to 45°C, the glue solution is at 43°C to 45°C, continue stirring, and adjust pH4.1 to 4.3 with acetic acid. Under the microscope, it can be observed that the emulsion droplets are covered with a condensed film layer. Add 150000ml of water with a slightly lower temperature, continue stirring, and continue stirring when the temperature drops to below 30°C and place an ice bath. Add 1000ml of 3% formaldehyde solution and stir to solidify. Adjust the pH to 7 to 7.5 with 5% sodium hydroxide solution to reduce the mesh of the mesh structure of the gel, and stir for another 30 minutes. Add 4000 ml of 10% raw starch suspension to fully disperse the starch and form an isolation layer between microcapsules. Stir at 10°C for another hour. Separate and wash by suction filtration to remove as much water as possible, granulate with a No. 2 sieve, and dry at 60°C to obtain ginkgo essential oil microcapsules, which are then put into capsules.

[0069] Example 5

[0070] 1000 g of the ginkgo essential oil prepared in Example 1 was added to 9000 g of corn endosperm crude oil, mixed, stirred evenly, and canned to obtain a ginkgo plant blended composite oil and a ginkgo corn endosperm oil product.

[0071] Example 6

[0072] The endosperm obtained from Example 1 after extracting crude oil was extracted by dynamic continuous countercurrent extraction technology, with an ethanol concentration of 60%, a solid-liquid ratio of 1:6, a temperature of 85°C, an extraction time of 1.0 hour, a spiral speed of 2.0 r / min, centrifugal separation of the extract, a speed of 15000 r / min, collecting the first extract and the first solid residue, the extract vacuum degree was controlled within the range of 70-90 kPa, the temperature was 50°C-70°C, and the ethanol solvent was recovered under reduced pressure, and the extract was placed on an AB-8 macroporous adsorption resin adsorption column (endosperm: resin = 15:1), washed with water, the washing amount was 2.5 times the volume of the resin column, the washing was collected, ethanol 70% ethanol was used for elution, the alcohol elution amount was 5 times the volume of the resin column, and the ethanol eluate was collected. The ethanol eluate vacuum degree was controlled within the range of 70-90 kPa, and the temperature was 50°C-70°C, and the ethanol solvent was recovered under reduced pressure. The solution after ethanol recovery was put on a polyamide column (for column chromatography, 30-60 mesh; endosperm: polyamide = 8:1), and after adsorption at a flow rate of 1-2 BV / hr, it was washed with water until the effluent turned light yellow, and about 1.2 BV of impurities were removed by water washing, and elution with water was continued to collect 2.5 BV of eluate (flow rate 1-2 BV / hr), and the eluate was concentrated to 1 / 10 of the volume under reduced pressure; the concentrate was extracted twice with an equal volume of ethyl acetate, filtered through a sand core funnel, and the filtrate was rotary evaporated at 70°C to remove ethyl acetate to obtain a light yellow-white extract, which was vacuum dried at 60°C for 8 hours and crushed through a 100-mesh sieve to obtain a ginkgo total terpene lactone extract.

[0073] The polyamide column after elution with water is eluted with 70% ethanol. When the HCl-Mg powder reaction is positive, the alcohol eluate is collected. When the HCl-Mg powder reaction is weakly positive, the alcohol elution is terminated and the first alcohol eluate is combined. The vacuum degree of the first alcohol eluate is controlled within the range of 70-90 kPa and the temperature is 50°C-70°C. The ethanol solvent is recovered under reduced pressure to obtain the first concentrated solution. The first concentrated solution is spray-dried with an inlet temperature of 160°C-180°C and an outlet temperature of 80°C-90°C to obtain a yellow-brown dry powder as a total flavonoid extract. The total terpene lactone extract of Ginkgo biloba and the total flavonoid extract (extraction rate 91.39%) are combined and stirred evenly to obtain Ginkgo biloba crystals, which have the effect of increasing cerebral blood flow and having an excellent tonic effect on the nervous system. Effectiveness for a wide range of problems including poor blood flow to the brain and mental decline in elderly patients, it has great effects on many possible symptoms of aging such as: anxiety and depression, memory impairment, difficulty concentrating, decreased alertness, decreased intelligence, vertigo, headaches, tinnitus (ringing in the ears), macular degeneration (the most common cause of blindness in adults), inner ear disturbances (which can lead to partial deafness), poor peripheral circulation, and impotence due to poor blood flow to the penis.

[0074] Example 7

[0075] Take the ginkgo crystal obtained in Example 6, add coenzyme Q10, tea polyphenols, and red yeast rice extract, mix well, and bottle to obtain a healthy product for protecting cardiovascular and cerebrovascular vessels.

[0076] Example 8

[0077] Take the solid part after liquid-solid separation left by the dynamic continuous countercurrent extraction of Example 6, that is, the first solid residue, and perform microwave-assisted extraction, using NaCl concentration 0.1mol·L-1 water as the solvent, liquid-solid ratio of 1:25, extraction temperature of 60°C, extraction times 2 times, each extraction time of 8 minutes. Centrifuge the extract at a speed of 15000r / min to achieve liquid-solid separation, and obtain the third extract and the second solid residue. The third extract is separated by ultrafiltration 4kD membrane, pressure 0.20Mpa; water is continuously added and diluted during the ultrafiltration process to ensure the stability of the membrane flux, and the filtrate and retentate are collected. The vacuum degree of the filtrate is controlled within the range of 70-90kPa and the temperature is 70℃-80℃, and part of the solvent is recovered under reduced pressure to obtain a concentrated solution. The concentrated solution is spray dried, the inlet temperature is 160℃-180℃, and the outlet temperature is 80℃-90℃, and the brown-yellow dry powder is obtained as the polar active ingredient extract, that is, the product of ginkgo nutrient powder (extraction rate 90.28%). It has the effects of improving immune function and antioxidant capacity, nourishing the skin and anti-aging, and has a certain preventive effect on hyperlipidemia, coronary heart disease and atherosclerosis.

[0078] Example 9

[0079] Take 1000g of the ginkgo nutrient powder obtained in Example 8, add 100g of maca extract, 2.7g of maca enamine, 0.9g of glucosinolate, 10g of ascorbyl palmitate, 50g of ginseng extract, 90g of epimedium extract, 120g of dodder extract, 100g of vitamin C, 3g of B1, 6g of B6, mix, stir evenly, and bottle. A product that enhances physical endurance, resists fatigue, and supports healthy sexual desire is obtained.

[0080] Example 10

[0081] Take the second solid residue and intercepted material after the liquid-solid separation left by microwave extraction in Example 8, add distiller's yeast, stir evenly, control the fermentation temperature at 18°C-22°C, add sugar, brew for about 14 days, separate the supernatant, put it into a pre-cleaned fermentation barrel, seal and age it. Fill it up without leaving any gaps to prevent contamination by bacteria in the air. After 20 days, perform a second separation, and store it for 3-5 months after separation to obtain the finished ginkgo wine.

[0082] Embodiment 11

[0083] The wine trough after brewing ginkgo wine in Example 10 was vacuum dried, crushed, and sieved to control the mesh size to 20-60, and 2% bone meal, 0.5% sodium bicarbonate, and 3% vitamin and trace element additive premixes accounting for the feed were added, mixed evenly, and prepared into biological feed.

[0084] Example 12

[0085] The ginkgo fruit wine obtained in Example 10 was added with American ginseng extract to strengthen the function of ginkgo, thereby obtaining ginkgo and American ginseng wine.

[0086] Example 13

[0087] Take the ginkgo crystals obtained in Example 6, pass through a 100-mesh sieve, add microcrystalline cellulose and hydroxypropyl cellulose and mix well, make a soft material with 8% starch slurry, pass through an 18-mesh sieve, dry at 60°C, pass through a 20-mesh sieve, add magnesium stearate and mix well, and press into tablets to obtain ginkgo crystals.

[0088] The above-mentioned ginkgo wafer formula is as follows:

[0089] Components content Ginkgo Crystal 40g starch 78g Microcrystalline Cellulose 18g Hydroxypropyl Cellulose 9g Magnesium Stearate 0.8g

[0090] This recipe yields a total of 1,000 ginkgo wafers.

[0091] Embodiment 14

[0092] Analysis of fatty acid components of functional substances in ginkgo essential oil by GC-MS

[0093] (Wang Yingqiang. Analysis of fatty acid composition in linseed oil. Journal of Pharmaceutical Analysis, 1998, 18(3): 176.; Li Yingxia, Zhou Shan, Zhong Ling, Wu Yan, Ge Fahuan. Response surface methodology for optimization of supercritical CO2 fluid extraction of ginkgo oil and its component analysis. Chinese Medicinal Materials, 2011, 34(2): 298-301.)

[0094] Methyl esterification: Take 0.5g of crude ginkgo oil sample / refined ginkgo essential oil in a 25mL stoppered round-bottom flask, add 2mL of 0.5mol·L-1 potassium hydroxide methanol solution, shake well, place in a 60℃ water bath for saponification for 30min (the oil droplets disappear completely), add 2mL of 14% boron trifluoride methanol solution after cooling, shake well, esterify in a 60℃ water bath for 6min, cool, add 2mL each of n-hexane and saturated sodium chloride aqueous solution, shake well, separate and separate clearly, and take the supernatant as the sample for gas chromatography / mass spectrometry analysis.

[0095] Instrument: PE Clarus 680 / SQ8T GCMS,

[0096] Chromatographic column: PE Elite-5 30m 0.32mm 0.25μm,

[0097] Column temperature: Initial temperature 60°C, maintain for 5 min, increase to 240°C at a rate of 3°C / min, maintain for 25 min,

[0098] Inlet temperature: 220°C,

[0099] Detector temperature: 280°C,

[0100] MS interface temperature: 280°C,

[0101] Ion source temperature: 230°C,

[0102] Carrier gas: high purity helium,

[0103] Carrier gas flow rate: 20psi constant pressure mode,

[0104] Split ratio: 100:1,

[0105] Injection volume: 0.1 μL.

[0106] Calculate the percentage of polyunsaturated fatty acids and unsaturated fatty acids in the total fatty acids in ginkgo essential oil.

[0107] Polyunsaturated fatty acids account for % of total fatty acids Unsaturated fatty acids as a percentage of total fatty acids Example 1 53.49 93.17 Example 2 54.08 93.72

[0108] Embodiment 15

[0109] Determination of the content of total flavonoids in Ginkgo biloba crystals

[0110] Determination method: Chinese Pharmacopoeia Committee. Pharmacopoeia of the People's Republic of China: 2015 Edition Volume 1. Beijing: China Medical Science and Technology Press, 2015: 417.

[0111] Test instrument: Agilent 1100 Infinity liquid chromatograph (Agilent 1100 quaternary pump, DAD detector, Agilent 1100 autosampler, Agilent Chemstation software, Agilent Technologies, Inc., USA);

[0112] Agilent Eclipse Plus C18 chromatographic column (column length: 150 mm, column inner diameter: 4.6 mm, particle size: 5 μm).

[0113] Reference substance:

[0114] 1. Quercetin reference substance (Chengdu Ruifensi Biotechnology Co., Ltd., purity> 98%),

[0115] 2. Kaempferol reference substance (Chengdu Ruifensi Biotechnology Co., Ltd., purity> 98%),

[0116] 3. Isorhamnetin reference substance (National Institute for the Control of Pharmaceutical and Biological Products).

[0117] The mobile phase was methanol-0.4% phosphoric acid solution (50:50); the detection wavelength was 360 nm.

[0118] Preparation of reference solution: Accurately weigh an appropriate amount of quercetin reference substance and add methanol to make a solution containing 30 mg per 1 mL.

[0119] Preparation of test solution: Take 40 mg of Ginkgo biloba crystals, weigh accurately, add 25 ml of a mixed solution of methanol-25% hydrochloric acid solution (4:1), heat under reflux in a water bath for 30 minutes, quickly cool to room temperature, transfer to a 50 ml volumetric flask, dilute to the scale with methanol, shake well, filter, and obtain the filtrate.

[0120] Determination steps: Accurately pipette 10μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine. Take the peak area of ​​quercetin reference as reference, calculate the content of quercetin, kaempferol and isorhamnetin according to the corresponding correction factors in the table below, and use the relative retention time of the chromatographic peak of the component to be tested and the chromatographic peak of quercetin to determine the peak position of quercetin, kaempferol and isorhamnetin. The relative retention time should be within ±5% of the specified value (if the relative retention time deviates by more than 5%, it should be confirmed by the corresponding substituted reference substance). See the table below for relative retention time and correction factor (F):

[0121] Component to be measured (peak) Relative retention time Correction factor (F) Quercetin 1.0000 1.0000 Kaempferol 1.7700 1.0020 Isorhamnetin 2.0000 1.0890

[0122] Total flavonol glycoside content = (quercetin content + kaempferol content + isorhamnetin content) X2.51

[0123] The content of total flavonol glycosides in Ginkgo biloba crystal was measured.

[0124]

[0125]

[0126] Example 16 Determination of the content of total terpenoid lactones in Ginkgo biloba crystals

[0127] Determination method: Chinese Pharmacopoeia Committee. Pharmacopoeia of the People's Republic of China: 2015 Edition Volume 1. Beijing: China Medical Science and Technology Press, 2015: 416-417.)

[0128] Measurement instrument: Agilent 1100 Infinity liquid chromatograph (Agilent 1100 quaternary pump, DAD detector, Agilent 1100 autosampler, Agilent Chemstation software, Agilent Technologies, Inc., USA);

[0129] Agilent Eclipse Plus C18 column (column length 150 mm, column inner diameter 4.6 mm, particle size 5 μm);

[0130] Reference substance:

[0131] 1. Ginkgolide A reference substance (National Institute for the Control of Pharmaceutical and Biological Products),

[0132] 2. Ginkgolide B reference substance (National Institute for the Control of Pharmaceutical and Biological Products),

[0133] 3. Ginkgolide C reference substance (National Institute for the Control of Pharmaceutical and Biological Products),

[0134] 4. Bilobalide reference substance (National Institute for the Control of Pharmaceutical and Biological Products).

[0135] The mobile phase was propanol-tetrahydrofuran-water (1:15:84); the detection was performed using an evaporative light scattering detector.

[0136] Preparation of control extract solution: Take an appropriate amount of Ginkgo biloba total lactone control extract, accurately weigh it, and add methanol to make a solution containing 2.5 mg per 1 mL.

[0137] Preparation of test solution: Take 0.15g of Ginkgo crystal, accurately weigh, add 10ml of water, warm in a water bath to dissolve, add 2 drops of 2% hydrochloric acid solution, shake and extract with ethyl acetate 4 times (15ml, 10ml, 10ml, 10ml), combine the extracts, wash with 20ml of 5% sodium acetate solution, take the sodium acetate solution, wash with 10ml of ethyl acetate, combine the ethyl acetate extract and the washing solution, wash with water twice, 20ml each time, take the water solution, wash with 10ml of ethyl acetate, combine the ethyl acetate solution, recover the solvent to dryness, dissolve the residual liquid with methanol and transfer it to a 5ml volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate to obtain.

[0138] Determination steps: Accurately pipette 5μl and 10μl of the control extract solution and 5-10μl of the test solution, respectively, inject into the liquid chromatograph for determination, and use the external standard two-point method logarithmic equation to calculate the contents of bilobalide, ginkgolide A, ginkgolide B and ginkgolide C, respectively.

[0139] Total terpenoid lactones in Ginkgo biloba crystals / % Example 6 3.13

[0140] Embodiment 17

[0141] Determination of total protein content in ginkgo nutrient powder

[0142] Test instrument: Kjeldahl nitrogen analyzer K-355, BUCHI, Switzerland

[0143] Test reagents:

[0144] 1. Potassium sulfate, analytical grade, Nanjing Chemical Reagent Co., Ltd.;

[0145] 2. Copper sulfate, analytical grade, Nanjing Chemical Reagent Co., Ltd.;

[0146] 3. Sodium hydroxide, analytical grade, Nanjing Chemical Reagent Co., Ltd.;

[0147] 4. Sulfuric acid, analytical grade, Sinopharm Chemical Reagent Co., Ltd.;

[0148] 5. Acid methyl red-bromocresol green mixed indicator;

[0149] 6. 0.1 mol / L hydrochloric acid standard solution, Sinopharm Chemical Reagent Co., Ltd.;

[0150] The total protein content of ginkgo powder was determined by Kjeldahl method. 1g of ginkgo nutrient powder was accurately weighed and transferred into a Kjeldahl flask for 3 parallel tests. 10g of potassium sulfate, 0.5g of copper sulfate, 20mL of sulfuric acid and several zeolites or glass beads were added and mixed thoroughly to ensure that the sample was completely soaked with sulfuric acid. Heat it on an electric stove in a fume hood. Shake it from time to time and carbonize it until the foam disappears, then increase the heat until the digestion liquid becomes transparent blue-green, and continue heating for 1-2h.

[0151] Cool the digestion solution, add 50-100mL of distilled water, shake well, completely dissolve the sulfate, cool, and add 2 zeolites. Place a collection bottle containing 50mL of 4% boric acid solution and 5 drops of methyl red-bromocresol green mixed indicator under the condenser, so that the lower end of the condenser is immersed below the liquid surface. Carefully inject 80mL of 40% sodium hydroxide solution along the wall of the flask, immediately connect it to the distillation device, heat and distill, allow the steam to enter the collection bottle, until the volume of the distillate is about 150mL, and lower the collection bottle. Make the lower end of the condenser leave the liquid surface, continue distillation for 1min, rinse the lower end of the condenser with distilled water, collect the washing liquid into the absorption bottle, and stop distillation. Titrate with 0.1mol / L hydrochloric acid standard solution until the color of the solution just changes from blue-green to light purple as the end point, and record the volume of the consumed hydrochloric acid standard solution. Weigh 1g of sucrose instead of the sample and perform a blank determination. The consumption of 0.1mol / L hydrochloric acid standard solution shall not exceed 0.2mL.

[0152] Calculate the total protein content in ginkgo nutritious powder according to the following formula:

[0153] Crude protein (%) = (V1-V0) × T × 0.0140 × f / m

[0154] Where, V1: volume of hydrochloric acid standard solution consumed when titrating the sample, mL;

[0155] V0: the volume of the hydrochloric acid standard solution consumed in the blank titration, mL;

[0156] T: molar concentration of hydrochloric acid standard solution, mol / L;

[0157] m: mass of the sample, g;

[0158] f: protein coefficient 6.25;

[0159] 0.0140: Milligram equivalent of nitrogen.

[0160] Ginkgo nutritional powder total protein content / % Example 8 48.13

[0161] Embodiment 18

[0162] Determination of total polysaccharide content in Ginkgo biloba

[0163] Testing instrument: UV-2500PC ultraviolet spectrophotometer (Shimadzu Corporation, Japan);

[0164] Test reagents: concentrated sulfuric acid, phenol, aluminum sheet, sodium bicarbonate, anhydrous glucose, all of analytical grade (Sinopharm Chemical Reagent Co., Ltd.).

[0165] Preparation of glucose standard curve: Take dry glucose (105℃, 2-3h), accurately weigh it, dissolve it in deionized water, and prepare a 250μg / mL standard glucose solution.

[0166] Take 0.1mL, 0.25mL, 0.4mL, 0.5mL, 0.7mL and 0.8mL of glucose standard solution in different thumb tubes respectively, add deionized water to make up 2.0mL, add 1.0mL of 4% phenol solution, quickly add 7mL of concentrated H2SO4, place in a 40℃ water bath to react for 30min, and place in a cold water bath for 5min to prepare the test solutions.

[0167] Take 2.0 mL of deionized water as blank solution and prepare it according to the above method. Measure the absorbance of the standard test solution and the blank test solution at 490 nm by ultraviolet spectrophotometry and determine the standard curve.

[0168] Determination of sample solution: Take an appropriate amount of ginkgo nutrient powder, weigh it carefully, dissolve it in deionized water, and prepare

[0169] Prepare the sample solution according to the preparation method of glucose standard test solution.

[0170] The total sugar content of the sample to be tested was calculated according to the glucose standard curve.

[0171] Total polysaccharide content of Ginkgo nutrient powder / % Example 8 39.14

[0172] The above specific examples are used to illustrate the present invention, which is only used to help understand the present invention and is not intended to limit the present invention. For those skilled in the art, according to the concept of the present invention, some simple deductions, modifications or substitutions can be made.

Claims

1. A deep processing method for ginkgo endosperm, characterized in that: The following steps are involved: S1, putting the ginkgo nuts into a vacuum dryer for drying, so that the moisture content of the ginkgo nuts is less than 5%; S2, putting the ginkgo nuts with a moisture content of less than 5% obtained in step S1 into a crusher for crushing to 4-40 mesh, and collecting the crushed ginkgo nuts for later use; wherein the crusher has a speed of 2500-3500 r / min; S3, adding an entrainer to the ginkgo nut granules obtained in step S2 to perform supercritical CO2 extraction to obtain ginkgo nut crude oil and endosperm after the crude oil is extracted; S4, adding the crude ginkgo nut oil obtained in step S3 to a primary falling film vacuum thin film evaporator for vacuum concentration, adding the concentrated crude ginkgo nut oil to a secondary scraper vacuum thin film evaporator for vacuum deodorization, and collecting the ginkgo nut crude oil from the discharge port of the scraper vacuum thin film evaporator; S5, subjecting the ginkgo biloba oil obtained in step S4 to refrigerated centrifugation, collecting the supernatant of the refrigerated centrifugation for vacuum filtration, and further subjecting the filtrate to molecular distillation to obtain ginkgo essential oil; S6, adding the endosperm after crude oil extraction obtained in step S3 to the extraction solution by a dynamic continuous countercurrent extraction method, and then centrifuging to obtain a first extract and a first solid residue; wherein the volume ratio of the endosperm after crude oil extraction to the extraction solution is 1:4-10, and the centrifugal separation speed is 12000-25000 r / min; S7, the first extract obtained in step S6 is subjected to reduced pressure and vacuum recovery at a temperature of 50-70° C. and a vacuum degree of 70-90 Pa to obtain a second extract; S8, adsorbing the second extract obtained in step S7 through a macroporous adsorption resin, washing the macroporous adsorption resin with water, collecting the water washing liquid, eluting with 50% to 80 vol% ethanol, and collecting the first alcohol eluate; S9, the first alcohol eluate obtained in step S8 is subjected to reduced pressure and vacuum recovery at a temperature of 50-70° C. and a vacuum degree of 70-90 Pa to recover the ethanol solvent, the remaining liquid after the ethanol solvent is recovered is applied to a polyamide column, adsorbed at a flow rate of 1-2 BV / hr, washed with water until the effluent turns light yellow, and about 1.2 BV of impurities are removed, and 2-3 BV of water is continuously eluted, the eluate is collected, and the eluate is concentrated under reduced pressure and vacuum to 1 / 10 of the total volume of the eluate to obtain a first concentrated solution; wherein the volume ratio of the endosperm to the polyamide column is 8:1; S10, extracting the first concentrated solution obtained in step S9 with ethyl acetate twice, combining the upper layer solutions, and rotary evaporating to obtain a light yellow-white extract, and then vacuum drying at 40-80° C. for 6-10 hours, crushing, and passing through a 100-mesh sieve to obtain a ginkgo total terpene lactone extract; wherein the volume ratio of ethyl acetate to the first concentrated solution is 1:1; S11, eluting the polyamide column obtained in step S9 with 50% to 80 vol% ethanol, collecting the alcohol eluate when the HCI-Mg reaction is positive, and combining the second alcohol eluate when the HCI-Mg reaction is weakly positive; S12, subjecting the second alcohol eluate obtained in step S11 to reduced pressure vacuum at a temperature of 50-70° C. and a vacuum degree of 70-90 Pa, and spray drying to obtain a total flavonoid extract; wherein the spray drying conditions are: an inlet air temperature of 160-180° C. and an outlet air temperature of 70-100° C.; S13, mixing the total terpene lactone extract of Ginkgo biloba obtained in step S10 and the total flavonoid extract obtained in step S12, stirring evenly, to obtain a medium-polarity active ingredient extract; S14, subjecting the first solid residue obtained in step S6 to microwave-assisted extraction for 1 to 5 times, and centrifuging to obtain a third extract and a second solid residue; wherein the microwave-assisted extraction uses a 0.1 mol / L sodium chloride solution as a solvent, and the volume ratio of the solvent to the first solid residue is 1:15 to 50; S15, ultrafiltration of the third extract obtained in step S14, and collection of the filtrate and the retentate; S16, the filtrate obtained in step S15 is subjected to reduced pressure vacuum at a temperature of 50-70°C and a vacuum degree of 70-90 Pa, and spray-dried to obtain a brown-yellow dry powder polar active ingredient extract; wherein the spray-drying conditions are: an inlet air temperature of 160-180°C and an outlet air temperature of 70-100°C; S17, mixing the second solid residue obtained in step S14 and the intercepted material obtained in step S15, adding distiller's yeast and white sugar, mixing evenly, fermenting at 18 to 22° C. for 12 to 18 days, and centrifuging to obtain ginkgo wine and distiller's grains; S18, vacuum-drying the wine lees obtained in step S17, and crushing the lees through an 80-mesh sieve to obtain a biological feed.

2. The deep processing method of ginkgo biloba endosperm according to claim 1, characterized in that: The supercritical CO2 extraction conditions in step S3 are: pressure 25-35 MPa, extraction temperature 25-45° C., and extraction time 1.5-3 h.

3. The deep processing method of a ginkgo endosperm as claimed in claim 1, characterized in that: In step S3, the entrainer is one of methanol, ethanol, acetone and ethyl acetate.

4. The deep processing method of ginkgo biloba endosperm according to claim 1, characterized in that: The conditions for molecular distillation in step S5 are: vacuum degree 10 -3 ~1Pa, evaporation surface speed 500~1500r / min, evaporation surface temperature 60~100℃, condensation surface temperature 0~10℃.

5. The deep processing method of ginkgo endosperm according to claim 1, characterized in that: In step S6, the extraction solution is selected from one of ethanol, acetone, ethyl acetate and n-butanol.

6. The deep processing method of ginkgo endosperm according to claim 1, characterized in that: The process parameters of the dynamic continuous countercurrent extraction method in step S6 are: temperature 60-100° C., extraction time 1-3 hours, and screw speed 0.2-2 r / min.

7. The deep processing method of ginkgo endosperm according to claim 1, characterized in that: In the step S8, the macroporous adsorption resin is selected from one of weak polar macroporous adsorption resin, medium polar macroporous adsorption resin and non-polar macroporous adsorption resin.

8. The deep processing method of ginkgo endosperm according to claim 1, characterized in that: The volume ratio of endosperm to macroporous adsorption resin during the adsorption process in step S8 is 15:1, the volume ratio of water to macroporous adsorption resin during the water washing process in step S8 is 1.5-5:1, and the volume ratio of ethanol elution amount to resin during the ethanol elution process in step S8 is 4-7:

1.

9. The deep processing method of ginkgo biloba endosperm according to claim 1, characterized in that: The microwave-assisted extraction conditions in step S14 are: extraction temperature 50-80° C., and each extraction time 8-20 minutes.

10. The deep processing method of ginkgo endosperm according to claim 1, characterized in that: In step S15, the molecular weight of the ultrafiltration membrane is 2-5 kD, and the extraction pressure is 0.06-0.25 MPa.

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