CN116637130A - Preparation method of ginkgo leaf extract - Google Patents

Preparation method of ginkgo leaf extract Download PDF

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CN116637130A
CN116637130A CN202310619853.2A CN202310619853A CN116637130A CN 116637130 A CN116637130 A CN 116637130A CN 202310619853 A CN202310619853 A CN 202310619853A CN 116637130 A CN116637130 A CN 116637130A
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leaf extract
ginkgo leaf
concentrated solution
ethanol water
concentrating
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陈小明
罗丽媛
吕敬崑
陈思伽
魏允武
吕宏再
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Hunan University of Science and Engineering
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Abstract

The invention discloses a preparation method of ginkgo leaf extract. The method comprises the following steps of: 1) Sun drying folium Ginkgo, grinding into powder, mixing with rhizoma Menispermi, adding fermenting agent, and fermenting for 3-10 days to obtain fermented product; 2) Putting the fermented product into an extraction tank, and carrying out hot reflux extraction by using an ethanol water solution; 3) Dissolving the concentrated solution with water, loading into macroporous adsorbent resin column for adsorption, performing gradient elution, collecting 40-60% ethanol water eluate, and concentrating to obtain concentrated solution; 4) Adding ethanol water solution into the concentrated solution, refluxing for dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate, and drying to obtain the final product of ginkgo leaf extract. The ginkgo leaf extract prepared by the invention has higher content of total flavonol glycoside, total terpene lactone and ginkgolide, can be used as an additive of health products, and can also be directly used as a medicament for treating hyperlipidemia and hypertension.

Description

Preparation method of ginkgo leaf extract
Technical Field
The invention belongs to the technical field of extraction of plant medicinal components, and particularly relates to a preparation method of a ginkgo leaf extract.
Background
Ginkgo leaf is a plant with high medicinal value, ginkgo tree is an ancient tree species, and is a magic medical tree, and ginkgo is one of the most flourishing plants when dwarf dinosaur masters the earth more than 2 hundred million for 5 thousand years. The folium Ginkgo contains flavonoids such as ginkgetin, isoginkgetin, demethylginkgetin, rutin, kaempferide-3-rhamnose glucoside, kaempferide, quercetin, isorhamnetin, etc.; further comprises bitter component ginkgolide A, B, C and bilobalide A; the acid component is toxic octaenoic acid, D-glycosyl acid, ginkgolic acid, bilobanone, octacosane, octacosanol, alpha-hexenal, beta-sitosterol, stigmasterol, vitamins, etc., and contains catechin and epicatechin.
The ginkgo leaf has the effects of astringing lung, relieving asthma, promoting blood circulation to remove blood stasis and relieving pain. Is used for treating cough and asthma due to lung deficiency; coronary heart disease, angina pectoris, hyperlipidemia, anticoagulation, and has certain probability of improving memory. The ginkgo leaf can promote blood circulation and prevent cardiovascular diseases, but other medicaments for treating cardiovascular diseases cannot be taken at the same time, and the ginkgo extract is concentrated granules, has strong free radical removal and antioxidation effects, and the yellow glycoside, amino acid and amino acid in the ginkgo leaf synthesize collagen components to beautify human bodies, inhibit melanin growth and maintain skin luster and elasticity.
The extraction of active ingredients from ginkgo has been reported more. Patent 201210353451.4 discloses a preparation method of ginkgo leaf extract. The method comprises the following steps: step 1) putting ginkgo leaves into an extraction tank, carrying out hot reflux extraction by using an ethanol aqueous solution, combining the extracting solutions, and concentrating to obtain a concentrate A with the density of 0.7-1.2 g/ml; step 2) dissolving the concentrate A with water, then putting the dissolved concentrate A into a DM130, AB-8, DM131 or D318 macroporous adsorption resin column for full adsorption, sequentially carrying out gradient elution by using ethanol water solution with volume fractions of 0%,10-30% and 40-80%, collecting 40-80% ethanol water eluent, and concentrating to obtain a concentrate B; dissolving the concentrate B with ethanol water solution, adding into D101, HPD100, LKY or DM2 macroporous adsorbent resin column, adsorbing thoroughly, eluting with 40-80% ethanol water solution, collecting eluate, concentrating under reduced pressure to density of 0.7-1.2g/ml to obtain concentrate C; and 3) adding ethanol water solution into the concentrate C for reflux dissolution, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate, and drying to obtain the finished ginkgo leaf extract product. The ginkgo active ingredient prepared by the method has low total flavonol glycoside content, total terpene lactone content and ginkgolide A content, ginkgolide B content and ginkgolide C content, and has no obvious medicinal effect.
Disclosure of Invention
The invention provides a preparation method of ginkgo leaf extract in order to overcome the defects in the prior art.
A preparation method of ginkgo leaf extract comprises the following steps:
(1) Sun drying folium Ginkgo and rhizoma Menispermi, grinding into powder, mixing 300-650 parts by weight of folium Ginkgo powder and 10-30 parts by weight of rhizoma Menispermi powder, adding 3-5 parts by weight of fermentation inoculant, adding 200-500 parts by weight of water, stirring, and fermenting at 25-30deg.C for 3-10 days to obtain fermented product;
(2) Putting the fermented product into an extraction tank, performing hot reflux extraction with ethanol water solution, mixing the extractive solutions, and concentrating to obtain concentrated solution with density of 0.8-1.2 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) by using water, then putting the dissolved concentrated solution into a macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent, 10-30 percent and 40-60 percent, collecting an ethanol water eluent with the volume fraction of 40-60 percent, and concentrating to obtain concentrated solution;
(4) Adding ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate, and drying to obtain the ginkgo leaf extract finished product.
The fermentation inoculant is a lactobacillus plantarum LP4 inoculant and an aspergillus niger T2 inoculant according to the mass ratio of 1:1, mixing mixed bacterial agents; the preservation number of the lactobacillus plantarum LP4 is CGMCC No.14533, and the aspergillus niger isThe preservation number of T2 is CGMCC No.2715; the content of viable bacteria in the fermentation inoculant is 1-9X10 8 cfu/g。
The volume fraction of the ethanol aqueous solution in the step (2) is 60-80%, and the dosage is 5-15 times of the mass of the fermented product.
The times of heat reflux extraction in the step (2) are 1-3 times, each time of extraction is 2-5 hours, and the extraction temperature is 60-80 ℃.
In the step (3), the concentrated solution is dissolved by adopting water with the volume of 3-8 times; concentrating 40-60% ethanol water eluent to 1/3-1/8 of original volume.
The macroporous adsorption resin column is D101, HPD100, LKY, 134 or DM2 macroporous adsorption resin column.
And (3) adding 10-30 times of ethanol water solution into the concentrated solution in the step (4) for reflux dissolution.
Concentrating the filtrate in the step (4) to 1/8-1/15 of the original volume.
The drying is spray drying, freeze drying or vacuum drying.
Compared with the prior art, the invention has the following beneficial effects: the ginkgo leaf extract prepared by the invention has higher content of total flavonol glycoside, total terpene lactone and ginkgolide, can be used as an additive of health products, and can also be directly used as a medicament for treating hyperlipidemia and hypertension.
Detailed Description
The following detailed description of specific embodiments of the invention is, but it should be understood that the invention is not limited to specific embodiments.
The lactobacillus plantarum LP4 adopted in the following examples has the preservation number of CGMCC No.14533 and the Aspergillus niger T2 has the preservation number of CGMCC No.2715, and are purchased from the China general microbiological culture Collection center.
Example 1
A preparation method of ginkgo leaf extract comprises the following steps:
(1) Sun drying folium Ginkgo and rhizoma Menispermi, grinding into powder, mixing 500 parts folium Ginkgo powder and 20 parts rhizoma Menispermi powder, and adding 2 partsAdding 300 parts of water into 2 parts of lactobacillus plantarum LP4 microbial inoculum and 2 parts of aspergillus niger T2 microbial inoculum, uniformly stirring, and fermenting for 6 days at 28 ℃ to obtain a fermented product; the content of viable bacteria in the lactobacillus plantarum LP4 microbial inoculum is 3X10 8 cfu/g; the content of viable bacteria in the Aspergillus niger T2 microbial inoculum is 3X10 8 cfu/g；
(2) Putting the fermented product into an extraction tank, adding an ethanol water solution with the volume fraction of 70% by weight for hot reflux extraction, wherein the times of hot reflux extraction are 2 times, each time of extraction is 3 hours, the extraction temperature is 70 ℃, and mixing the extracting solutions, and concentrating to obtain a concentrated solution with the density of 0.9 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) with 6 times of water, then putting the dissolved concentrated solution into a D101 macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent and 20 percent and 50 percent, collecting an ethanol water eluent with the volume fraction of 50 percent, and concentrating the ethanol water eluent to 1/5 of the original volume to obtain concentrated solution;
(4) Adding 20 times volume of ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate to 1/12 of the original volume, and spray drying to obtain the ginkgo leaf extract finished product.
The total flavonol glycoside content in the finished product is 34.9%, the total terpene lactone content is 18.2%, wherein the ginkgolide A content is 6.8%, the ginkgolide B content is 3.8%, and the ginkgolide C content is 6.2%.
Example 2
A preparation method of ginkgo leaf extract comprises the following steps:
(1) Sun-drying ginkgo leaf and asiatic moonseed rhizome, grinding into powder, uniformly mixing 300 parts of ginkgo leaf powder and 10 parts of asiatic moonseed rhizome powder according to parts by weight, adding 1.5 parts of lactobacillus plantarum LP4 microbial inoculum and 1.5 parts of aspergillus niger T2 microbial inoculum, then adding 200 parts of water, uniformly stirring, and fermenting for 3 days at 25 ℃ to obtain a fermented product; the content of viable bacteria in the lactobacillus plantarum LP4 microbial inoculum is 6X10 8 cfu/g; the content of viable bacteria in the Aspergillus niger T2 microbial inoculum is 6X10 8 cfu/g；
(2) Putting the fermented product into an extraction tank, adding 5 times of ethanol water solution with the volume fraction of 60% for hot reflux extraction, wherein the times of hot reflux extraction are 1 time, each time of extraction is 2 hours, the extraction temperature is 60 ℃, mixing the extracting solutions, and concentrating to obtain concentrated solution with the density of 0.8 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) with 3 times of water, then putting the dissolved concentrated solution into an HPD100 macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent and 10 percent and 40 percent, collecting an ethanol water eluent with the volume fraction of 40 percent, and concentrating the ethanol water eluent to 1/3 of the original volume to obtain concentrated solution;
(4) Adding 10 times volume of ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate to 1/8 of the original volume, and freeze-drying to obtain the ginkgo leaf extract finished product.
The total flavonol glycoside content in the finished product is 33.9%, the total terpene lactone content is 17.6%, wherein the ginkgolide A content is 6.4%, the ginkgolide B content is 3.7%, and the ginkgolide C content is 6.3%.
Example 3
A preparation method of ginkgo leaf extract comprises the following steps:
(1) Sun-drying ginkgo leaf and asiatic moonseed rhizome, grinding into powder, uniformly mixing 650 parts of ginkgo leaf powder and 30 parts of asiatic moonseed rhizome powder according to parts by weight, adding 2.5 parts of lactobacillus plantarum LP4 microbial inoculum and 2.5 parts of aspergillus niger T2 microbial inoculum, then adding 500 parts of water, uniformly stirring, and fermenting for 10 days at the temperature of 30 ℃ to obtain a fermented product; the content of viable bacteria in the lactobacillus plantarum LP4 microbial inoculum is 1X10 8 cfu/g; the content of viable bacteria in the Aspergillus niger T2 microbial inoculum is 1X10 8 cfu/g；
(2) Putting the fermented product into an extraction tank, adding an ethanol water solution with the volume fraction of 80% by weight of 15 times, performing hot reflux extraction for 3 times, extracting for 5 hours each time at the extraction temperature of 80 ℃, combining the extracting solutions, and concentrating to obtain a concentrated solution with the density of 1.2 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) with 8 times of water, then putting the dissolved solution into a LKY macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent and 30 percent and 60 percent, collecting an ethanol water eluent with the volume fraction of 60 percent, and concentrating the eluent to 1/8 of the original volume to obtain concentrated solution;
(4) Adding 30 times volume of ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate to 1/15 of the original volume, and vacuum drying to obtain the ginkgo leaf extract finished product.
The total flavonol glycoside content in the finished product is 36.5%, the total terpene lactone content is 17.8%, wherein the ginkgolide A content is 6.1%, the ginkgolide B content is 3.1%, and the ginkgolide C content is 6.1%.
Comparative example 1
A preparation method of ginkgo leaf extract comprises the following steps:
(1) Sun-drying folium ginkgo and rhizoma Menispermi, grinding into powder, mixing 500 parts of folium ginkgo powder and 20 parts of rhizoma Menispermi powder according to parts by weight, adding 4 parts of lactobacillus plantarum LP4 microbial inoculum, adding 300 parts of water, stirring uniformly, and fermenting at 28 ℃ for 6 days to obtain a fermented product; the content of viable bacteria in the lactobacillus plantarum LP4 microbial inoculum is 3X10 8 cfu/g；
(2) Putting the fermented product into an extraction tank, adding an ethanol water solution with the volume fraction of 70% by weight for hot reflux extraction, wherein the times of hot reflux extraction are 2 times, each time of extraction is 3 hours, the extraction temperature is 70 ℃, and mixing the extracting solutions, and concentrating to obtain a concentrated solution with the density of 0.9 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) with 6 times of water, then putting the dissolved concentrated solution into a D101 macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent and 20 percent and 50 percent, collecting an ethanol water eluent with the volume fraction of 50 percent, and concentrating the ethanol water eluent to 1/5 of the original volume to obtain concentrated solution;
(4) Adding 20 times volume of ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate to 1/12 of the original volume, and spray drying to obtain the ginkgo leaf extract finished product.
The total flavonol glycoside content in the finished product is 28.4%, the total terpene lactone content is 15.2%, wherein the ginkgolide A content is 5.1%, the ginkgolide B content is 3.1%, and the ginkgolide C content is 5.5%.
Comparative example 2
A preparation method of ginkgo leaf extract comprises the following steps:
(1) Sun-drying folium ginkgo and rhizoma Menispermi, grinding into powder, mixing 500 parts of folium ginkgo powder and 20 parts of rhizoma Menispermi powder according to parts by weight, adding 4 parts of Aspergillus niger T2 microbial inoculum, adding 300 parts of water, stirring uniformly, and fermenting at 28deg.C for 6 days to obtain fermented product; the content of viable bacteria in the Aspergillus niger T2 microbial inoculum is 3X10 8 cfu/g；
(2) Putting the fermented product into an extraction tank, adding an ethanol water solution with the volume fraction of 70% by weight for hot reflux extraction, wherein the times of hot reflux extraction are 2 times, each time of extraction is 3 hours, the extraction temperature is 70 ℃, and mixing the extracting solutions, and concentrating to obtain a concentrated solution with the density of 0.9 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) with 6 times of water, then putting the dissolved concentrated solution into a D101 macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent and 20 percent and 50 percent, collecting an ethanol water eluent with the volume fraction of 50 percent, and concentrating the ethanol water eluent to 1/5 of the original volume to obtain concentrated solution;
(4) Adding 20 times volume of ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate to 1/12 of the original volume, and spray drying to obtain the ginkgo leaf extract finished product.
The total flavonol glycoside content in the finished product is 23.6%, the total terpene lactone content is 11.2%, wherein the ginkgolide A content is 4.9%, the ginkgolide B content is 2.3%, and the ginkgolide C content is 4.8%.
Comparative example 3
A preparation method of ginkgo leaf extract comprises the following steps:
(1) Sun drying folium Ginkgo, grinding into powder, mixing 520 parts by weight of folium Ginkgo powder, adding 2 parts of Lactobacillus plantarum LP4 microbial inoculum and 2 parts of Aspergillus niger T2 microbial inoculum, adding 300 parts of water, stirring, and fermenting at 28deg.C for 6 days to obtain the final productTo the ferment; the content of viable bacteria in the lactobacillus plantarum LP4 microbial inoculum is 3X10 8 cfu/g; the content of viable bacteria in the Aspergillus niger T2 microbial inoculum is 3X10 8 cfu/g；
(2) Putting the fermented product into an extraction tank, adding an ethanol water solution with the volume fraction of 70% by weight for hot reflux extraction, wherein the times of hot reflux extraction are 2 times, each time of extraction is 3 hours, the extraction temperature is 70 ℃, and mixing the extracting solutions, and concentrating to obtain a concentrated solution with the density of 0.9 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) with 6 times of water, then putting the dissolved concentrated solution into a D101 macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent and 20 percent and 50 percent, collecting an ethanol water eluent with the volume fraction of 50 percent, and concentrating the ethanol water eluent to 1/5 of the original volume to obtain concentrated solution;
(4) Adding 20 times volume of ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate to 1/12 of the original volume, and spray drying to obtain the ginkgo leaf extract finished product.
Comparative example 4
A preparation method of rhizoma Menispermi extract comprises the following steps:
(1) Sun-drying rhizoma Menispermi, grinding into powder, taking 520 parts of rhizoma Menispermi powder according to parts by weight, adding 2 parts of lactobacillus plantarum LP4 microbial inoculum and 2 parts of Aspergillus niger T2 microbial inoculum, adding 300 parts of water, stirring uniformly, and fermenting at 28 ℃ for 6 days to obtain a fermented product; the content of viable bacteria in the lactobacillus plantarum LP4 microbial inoculum is 3X10 8 cfu/g; the content of viable bacteria in the Aspergillus niger T2 microbial inoculum is 3X10 8 cfu/g；
(2) Putting the fermented product into an extraction tank, adding an ethanol water solution with the volume fraction of 70% by weight for hot reflux extraction, wherein the times of hot reflux extraction are 2 times, each time of extraction is 3 hours, the extraction temperature is 70 ℃, and mixing the extracting solutions, and concentrating to obtain a concentrated solution with the density of 0.9 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) with 6 times of water, then putting the dissolved concentrated solution into a D101 macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent and 20 percent and 50 percent, collecting an ethanol water eluent with the volume fraction of 50 percent, and concentrating the ethanol water eluent to 1/5 of the original volume to obtain concentrated solution;
(4) Adding 20 times volume of ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate to 1/12 of the original volume, and spray drying to obtain rhizoma Menispermi extract product.
Experimental example:
the experimental animals were wintar rats, males, weighing about 200g, SPF grade, purchased from Chengdu experimental animals Inc.; the experimental animal feed adopts high-fat feed (73.8% basal feed, 15% lard, 10% yolk powder, 1% cholesterol and 0.2% sodium cholate) and rat growth maintenance material, which are purchased from the biological technology Co., ltd; yixintong, produced by Hainan Tianhuang pharmaceutical Co., ltd., lot number 1608211.
Modeling and grouping drug delivery: the rats are randomly grouped into groups of 10 animals, and each group is fed with 90mg/kg of the heart tonifying ketone group, and 50mg/kg of the ginkgo leaf extract prepared in the examples 1-3 and the comparative examples 1-4. Each of the other groups except the control group was given a high fat diet, and the control group was given a normal diet. The administration was performed 1 time per day by stomach irrigation, and 60 days after the administration, 10mL/kg purified water was administered by the control group and the model group. Weighing 1 time every 10 days, weighing 1 hour after administration on 60 days, taking the first 10 rats of each group, injecting 300mg/kg chloral hydrate into abdominal cavity for anesthesia, collecting blood from abdominal aorta, separating serum, taking heart, freezing with liquid nitrogen, and transferring to a refrigerator at-80 ℃ for standby.
And (3) index detection: measuring TC, TG, HDL-C, LDL-C content (mmol/L) in serum; SOD activity and MDA content (U/mgprot) in myocardial tissues are measured.
Statistical analysis using SPSS 24.0 software for metering data results(mean Standard deviation) means that data normalization test is performed by using a Kolmogorov-Smirnov test method, and for data conforming to normal distribution, the mean difference between two groups is compared by using a t test, so that the data is P<A difference of 0.05 is statistically significant.
The measurement results are shown in tables 1-2:
TABLE 1
Note that: * Represents p <0.05 compared to the control group; # represents p <0.05 compared to example 1.
TABLE 2
Note that: * Represents p <0.05 compared to the control group; # represents p <0.05 compared to example 1.
The above formulation ratios and the embodiments of the process are merely illustrative of the basic principles and characteristics of the present invention, which are not limited by the implementation of the above processes and ratios, and various modifications and changes can be made in the present invention without departing from the spirit and scope of the present invention. Such variations and modifications are therefore within the scope of the present invention.

Claims (9)

1. A preparation method of ginkgo leaf extract is characterized by comprising the following steps:
(1) Sun drying folium Ginkgo and rhizoma Menispermi, grinding into powder, mixing 300-650 parts by weight of folium Ginkgo powder and 10-30 parts by weight of rhizoma Menispermi powder, adding 3-5 parts by weight of fermentation inoculant, adding 200-500 parts by weight of water, stirring, and fermenting at 25-30deg.C for 3-10 days to obtain fermented product;
(2) Putting the fermented product into an extraction tank, performing hot reflux extraction with ethanol water solution, mixing the extractive solutions, and concentrating to obtain concentrated solution with density of 0.8-1.2 g/ml;
(3) Dissolving the concentrated solution prepared in the step (2) by using water, then putting the dissolved concentrated solution into a macroporous adsorption resin column for adsorption, sequentially carrying out gradient elution by using an ethanol water solution with the volume fraction of 0 percent, 10-30 percent and 40-60 percent, collecting an ethanol water eluent with the volume fraction of 40-60 percent, and concentrating to obtain concentrated solution;
(4) Adding ethanol water solution into the concentrated solution prepared in the step (3), refluxing and dissolving, filtering at normal temperature to remove insoluble impurities, concentrating the filtrate, and drying to obtain the ginkgo leaf extract finished product.
2. The preparation method of the ginkgo leaf extract according to claim 1, wherein the fermentation inoculant is lactobacillus plantarum LP4 inoculant and aspergillus niger T2 inoculant according to the mass ratio of 1:1, mixing mixed bacterial agents; the preservation number of the lactobacillus plantarum LP4 is CGMCC No.14533, and the preservation number of the aspergillus niger T2 is CGMCC No.2715; the content of viable bacteria in the fermentation inoculant is 1-9X10 8 cfu/g。
3. The method for preparing ginkgo leaf extract according to claim 1, wherein the volume fraction of the aqueous ethanol solution in the step (2) is 60-80% and the amount is 5-15 times the mass of the fermented product.
4. The method for preparing ginkgo leaf extract according to claim 1, wherein the number of times of thermal reflux extraction in the step (2) is 1-3, each time for 2-5 hours, and the extraction temperature is 60-80 ℃.
5. The method for preparing ginkgo leaf extract according to claim 1, wherein the concentrated solution in the step (3) is dissolved by 3-8 times of water; concentrating 40-60% ethanol water eluent to 1/3-1/8 of original volume.
6. The method for preparing ginkgo leaf extract according to claim 1, wherein the macroporous adsorbent resin column is D101, HPD100, LKY, 134 or DM2 macroporous adsorbent resin column.
7. The method for preparing ginkgo leaf extract according to claim 1, wherein 10-30 times of volume of ethanol aqueous solution is added to the concentrated solution in the step (4) for reflux dissolution.
8. The method for preparing ginkgo leaf extract according to claim 1, wherein the filtrate in the step (4) is concentrated to 1/8-1/15 of the original volume.
9. The method for preparing ginkgo leaf extract according to claim 1, wherein the drying is spray drying, freeze drying or vacuum drying.
CN202310619853.2A 2023-05-30 2023-05-30 Preparation method of ginkgo leaf extract Pending CN116637130A (en)

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