阅读说明：本技术 利用银杏外种皮制备高纯度银杏酚酸提取物的工艺方法 (Process for preparing high-purity ginkgolic acid extract by using ginkgo episperms ) 是由 杨宪勇 张爱丽 花丽 于 2021-08-15 设计创作，主要内容包括：本发明具体公开了一种利用银杏外种皮制备高纯度银杏酚酸提取物的工艺方法,该方法是将银杏外种皮经浸泡、磨成颗粒度在6.5～13μm的浆液；将浆液分别用乙醇和石油醚浸提,经回收溶剂后将浸提物合并,再将合并后的浸提物加入去离子水经充分搅拌加入聚酰胺进行吸附二次,再将吸附后的聚酰胺用乙醇洗脱三次,回收乙醇得稠膏状醇提物,将醇提物真空干燥、粉碎得浅棕黄色粉末,得到的最终产物银杏酚酸的含量＞90重量%。本发明高纯度的银杏酚酸提取物可作为药物制剂、化妆品制品、生物农药和生物杀虫剂制剂的用途。(The invention particularly discloses a process method for preparing a high-purity ginkgolic acid extract by utilizing ginkgo biloba sarcotesta, which comprises the steps of soaking the ginkgo biloba sarcotesta and grinding into slurry with the granularity of 6.5-13 mu m; respectively extracting the serous fluid with ethanol and petroleum ether, merging the extracts after recovering the solvent, adding deionized water into the merged extracts, fully stirring and adding polyamide for adsorption twice, eluting the adsorbed polyamide with ethanol for three times, recovering the ethanol to obtain thick paste-shaped ethanol extract, drying the ethanol extract in vacuum, and crushing to obtain light brown yellow powder, wherein the content of the ginkgolic acid of the final product is more than 90 percent by weight. The high-purity ginkgolic acid extract can be used as a pharmaceutical preparation, a cosmetic product, a biological pesticide and a biological insecticide preparation.)

1. The process method for preparing the high-purity ginkgolic acid extract by utilizing the ginkgo episperms is characterized by comprising the following steps of:

step one, selecting and cleaning ginkgo episperms and drying the ginkgo episperms according to the mass ratio of 1: 10, adding deionized water for soaking for 18-24 hours at the soaking water temperature of 40-43 ℃, and filtering out the soaking water after soaking is finished;

adding 1 time of deionized water into the soaked ginkgo biloba sarcotesta according to the weight of the dried ginkgo biloba sarcotesta, grinding the ginkgo biloba sarcotesta into coarse pulp by using a pulp grinder, and then finely grinding the coarse pulp by using a colloid mill to obtain fine grinding pulp, wherein the granularity of the fine grinding pulp is 6.5-13 mu m;

step three, extracting the fine grinding slurry obtained in the step two with ethanol with the content concentration of 98% V/V for one time, wherein the specific extraction conditions are that the using amount of the ethanol is 10 times of the weight of the fine grinding slurry, the extraction temperature is 42-45 ℃, the extraction time is 8-12 hours, filtering ethanol extract liquor, extracting filter residues with petroleum ether for one time, the specific extraction conditions are that the using amount of the petroleum ether is 10 times of the weight of the fine grinding slurry, the extraction temperature is 30-36 ℃, the extraction time is 18 hours, filtering the petroleum ether extract liquor, recovering the ethanol and the petroleum ether from the ethanol extract liquor and the petroleum ether extract liquor respectively, obtaining pasty ethanol extract and pasty petroleum ether extract respectively, combining the extracts, and obtaining mixed pasty extract;

step four, adding deionized water with the weight 30 times that of the mixed paste extract obtained in the step three into the mixed paste extract, fully stirring the mixture to be uniform, adding polyamide with the weight 5 times that of the mixed paste extract into the mixture under continuous stirring, stirring the mixture for 6 hours, standing and settling the mixture for 3 hours after the stirring is finished, and filtering the polyamide; adding polyamide 5 times the weight of the mixed paste extract into the filtrate, stirring for 6h, standing and settling for 3h after stirring is finished, and filtering to obtain polyamide; combining the polyamide obtained by two times of filtration in a filter, continuously adding deionized water from the filter until the filtered deionized water is basically colorless, and then controlling the water content of the polyamide;

step five, eluting the polyamide with water content controlled completely in the step four for three times by ethanol with the concentration of 98% V/V according to the weight of the polyamide of 10 times, 8 times and 5 times, wherein the eluting condition is that after the ethanol is added each time, the mixture is stirred for 3-5 hours at the temperature of 38-42 ℃ and the rotating speed of 30r/min, combining the eluates of the three times, and recovering the ethanol to obtain a thick paste-like ethanol extract;

putting the thick paste alcohol extract obtained in the step (five) into a vacuum drier, vacuum-drying at the temperature of not more than 50 ℃, and crushing to obtain light brown yellow ginkgolic acid extract powder; the obtained ginkgolic acid extract is detected to have the ginkgolic acid content of more than 90 percent by weight.

2. The use of the high purity ginkgolic acid extract of claim 1 as a pharmaceutical preparation for external use.

3. Use of the high purity ginkgolic acid extract of claim 1 as a cosmetic preparation.

4. The use of the high purity ginkgolic acid extract of claim 1 as a biopesticide or biopesticide formulation.

Technical Field

The invention belongs to the field of medicine and biological pesticide processing, relates to preparation of ginkgolic acid extracts contained in medicines and biological pesticides, and particularly relates to a process method for preparing a high-purity ginkgolic acid extract by utilizing ginkgo episperms.

Background

The ginkgolic acid is mainly from leaves, fruits, testa and rhizomes of ginkgo, wherein the content of the testa of ginkgo is the highest and accounts for about 3-5% of that of the testa of ginkgo. The ginkgolic acid has stronger killing or inhibiting effect on fungi and good killing effect on certain plant diseases and insect pests, so the ginkgolic acid is developed and prepared into a medicament in the field of medicine to be widely applied to the prevention and treatment of related diseases such as dermatology, gynecology and the like, has application in cosmetics, has better control effect on agricultural pathogenic bacteria and plant diseases and insect pests, and is prepared into biological pesticide for application; the application of ginkgolic acid in the field of agricultural diseases and insect pests is a technology well known by technicians in the field, and under the continuous efforts of researchers, the control effect of ginkgolic acid is continuously improved, and a favorable application effect is obtained. Although the ginkgolic acid can be chemically synthesized, the content of the active ingredients of the ginkgolic acid is stable, the ginkgolic acid has high production cost and high price, the production cost is increased, and even the price for producing the biopesticide is high, the burden of farmers is increased, so that the product cannot be popularized and applied. Meanwhile, China is a big country of the gingko planting industry and is a main production area of the world, Jiangsu, Twain and Shandong tancheng are main production areas of the domestic gingko industry, ginkgolic acid is widely existed in leaves, fruits, rhizomes, episperms and other parts of gingko, and the ginkgolic acid can be obtained from extracts of the parts of the gingko leaves, the fruits, the rhizomes, the episperms and the like or can be recovered from residual waste materials in the extraction and processing processes of the gingko leaves, the fruits, the rhizomes and the episperms. Particularly, in the harvesting season of the gingkoes, a large amount of ginkgo episperms are thrown away as waste materials every year, which also provides cheap resources for extracting the ginkgolic acid, thereby greatly reducing the production cost and being capable of becoming a continuous raw material source of the ginkgolic acid.

The method is characterized in that organic solutions such as ethanol, methanol and petroleum ether are generally used as solvents for extraction or refining to extract the ginkgolic acid in China, and processes such as ion exchange resin adsorption and elution are also used for extracting the ginkgolic acid, the extraction rate of the organic solutions is low, the impurity content is high, the content and purity of the ginkgolic acid are low, the content and purity of the ginkgolic acid prepared by an ion exchange method are relatively high, but the process is relatively complex, the production cost is high, a large amount of acid-base treatment is needed for resin regeneration, the operation is troublesome, and a large amount of acid liquor and alkali liquor can be used as waste liquor to flow out, so that the environmental pollution is caused. The search for a method which can prepare high-purity ginkgolic acid and has simple and feasible process is the direction of the current needs of intensive research and development.

Disclosure of Invention

The inventor continuously researches on the extraction of the ginkgolic acid from the ginkgo episperm serving as a raw material for many years to find an effective method which is simple and feasible in process, high in extraction rate and high in ginkgol content, so as to obtain better application of the ginkgolic acid in the preparation of external medicines and biological pesticides.

In order to achieve the purpose, the invention provides a process method for preparing a high-purity ginkgolic acid extract by utilizing ginkgo biloba sarcotesta. The aim of the invention is achieved by the following method:

the process method for preparing the high-purity ginkgolic acid extract by utilizing the ginkgo episperms is characterized by comprising the following steps of:

step one, selecting and cleaning ginkgo episperms and drying the ginkgo episperms according to the mass ratio of 1: 10, adding deionized water for soaking for 18-24 hours at the soaking water temperature of 40-43 ℃, and filtering out the soaking water after soaking is finished;

adding 1 time of deionized water into the soaked ginkgo biloba sarcotesta according to the weight of the dried ginkgo biloba sarcotesta, grinding the ginkgo biloba sarcotesta into coarse pulp by using a pulp grinder, and then finely grinding the coarse pulp by using a colloid mill to obtain fine grinding pulp, wherein the granularity of the fine grinding pulp is 6.5-13 mu m;

step three, extracting the fine grinding slurry obtained in the step two with ethanol with the content concentration of 98% V/V for one time, wherein the specific extraction conditions are that the using amount of the ethanol is 10 times of the weight of the fine grinding slurry, the extraction temperature is 42-45 ℃, the extraction time is 8-12 hours, filtering ethanol extract liquor, extracting filter residues with petroleum ether for one time, the specific extraction conditions are that the using amount of the petroleum ether is 10 times of the weight of the fine grinding slurry, the extraction temperature is 30-36 ℃, the extraction time is 18 hours, filtering the petroleum ether extract liquor, recovering the ethanol and the petroleum ether from the ethanol extract liquor and the petroleum ether extract liquor respectively, obtaining pasty ethanol extract and pasty petroleum ether extract respectively, combining the extracts, and obtaining mixed pasty extract;

step four, adding deionized water with the weight 30 times that of the mixed paste extract obtained in the step three into the mixed paste extract, fully stirring the mixture to be uniform, adding polyamide with the weight 5 times that of the mixed paste extract into the mixture under continuous stirring, stirring the mixture for 6 hours, standing and settling the mixture for 3 hours after the stirring is finished, and filtering the polyamide; adding polyamide 5 times the weight of the mixed paste extract into the filtrate, stirring for 6h, standing and settling for 3h after stirring is finished, and filtering to obtain polyamide; combining the polyamide obtained by two times of filtration in a filter, continuously adding deionized water from the filter until the filtered deionized water is basically colorless, and then controlling the water content of the polyamide;

step five, eluting the polyamide with water content controlled completely in the step four for three times by ethanol with the concentration of 98% V/V according to the weight of the polyamide of 10 times, 8 times and 5 times, wherein the eluting condition is that after the ethanol is added each time, the mixture is stirred for 3-5 hours at the temperature of 38-42 ℃ and the rotating speed of 30r/min, combining the eluates of the three times, and recovering the ethanol to obtain a thick paste-like ethanol extract;

putting the thick paste alcohol extract obtained in the step (five) into a vacuum drier, vacuum-drying at the temperature of not more than 50 ℃, and crushing to obtain light brown yellow ginkgolic acid extract powder; the obtained ginkgolic acid extract is detected to have the ginkgolic acid content of more than 90 percent by weight.

The invention has the beneficial effects that:

1. the extraction process for extracting the ginkgolic acid extract by utilizing the ginkgo episperm is simple, the solvent is recycled, the loss is low, the energy consumption is low, the process is simple and easy to implement, the extraction, elution and separation effects are good, the content of the ginkgolic acid in the extract is high, and the content of the ginkgolic acid in the ginkgolic acid extract obtained by the method is more than 90 wt%.

2. According to the invention, the ginkgolic acid can be maximally extracted by respectively leaching with ethanol and petroleum ether, and the solvent is completely recovered without environmental pollution; further utilizing the water-insoluble property of the ginkgolic acid, forming suspended turbid liquid in a large amount of deionized water under continuous stirring, utilizing the adsorption effect of polyamide on the ginkgolic acid, enabling the ginkgolic acid to be adsorbed by the polyamide in the stirring and time washing processes, then washing the adsorbed polyamide with deionized water to fully dissolve out water-soluble impurities, and then eluting the adsorbed polyamide with ethanol for three times, wherein the process is favorable for extracting the high-purity ginkgolic acid; the polyamide after elution is rinsed by water, then is soaked for more than 10 hours by ethanol, is filtered out, and can be repeatedly used or used for other purposes after being dried, thereby not causing waste and saving production cost.

3. The invention utilizes the characteristic that ginkgolic acid is dissolved in organic solutions such as alcohol, ether and the like and is not dissolved in water, firstly ethanol and petroleum ether are adopted for secondary extraction, then alcohol and ether extracts are diluted by a large amount of water, polyamide is adopted for adsorption, then water is repeatedly leached to further remove water-soluble impurities, and then the ginkgolic acid adsorbed by the polyamide is eluted by ethanol. The method solves the related problems of high extraction content of the ginkgolic acid, low energy consumption, simple and easy operation and no pollution.

4. The method for preparing the high-content ginkgolic acid extract also provides a more beneficial raw material guarantee for medical external pharmaceutical preparations, cosmetic products, biopesticide preparations and biopesticides, can ensure the good effects of low impurity content and high content of effective components of the preparations, and provides a practical, convenient and efficient method for efficiently utilizing the ginkgo episperms.

The ginkgolic acid extract prepared by the method can be used as a pharmaceutical external preparation and a cosmetic product; can also be used as biological pesticide and biological insecticide preparation.

The equipment adopted in the invention is conventional equipment.

In the invention, the content of the ginkgolic acid is detected by adopting a publicly known and common High Performance Liquid Chromatography (HPLC); professional qualified inspection institutions can also be entrusted to help with the detection.

Detailed Description

Example 1

Weighing 1kg of cleaned and dried ginkgo biloba testa, adding 10kg of deionized water, soaking for 24h at 41 ℃, and filtering out soaking water after soaking is finished;

step two, adding 1kg of deionized water into the ginkgo biloba sarcotesta soaked in the step one, grinding the ginkgo biloba sarcotesta into coarse pulp by using a pulp grinder, and then finely grinding the coarse pulp by using a colloid mill to obtain finely ground pulp, wherein the granularity of the finely ground pulp is 6.5-13 mu m;

step three, adding 20kg of ethanol with the content concentration of 98% V/V into the fine grinding slurry obtained in the step two, leaching for 12h at the temperature of 43 ℃, and filtering to obtain ethanol leaching liquor; adding 20kg of petroleum ether into filter residue, leaching for 18h at the temperature of 30 ℃, filtering to obtain petroleum ether extract, recovering ethanol and petroleum ether from the ethanol extract and the petroleum ether extract respectively to obtain pasty ethanol extract and pasty petroleum ether extract respectively, and combining the extracts to obtain 298g of mixed pasty extract;

adding 8.94kg of deionized water into the mixed paste extract obtained in the step (three), fully and uniformly stirring at the rotating speed of 30r/min, adding 1.49kg of polyamide under continuous stirring, continuously stirring for 6h, standing and settling for 3h after stirring is finished, filtering to obtain polyamide, adding 1.49kg of polyamide into the filtrate, stirring at the rotating speed of 30r/min for 6h, standing and settling for 3h after stirring is finished, filtering to obtain polyamide, combining the polyamide obtained by two times of filtering in a filter, continuously adding deionized water into the filter until the filtered deionized water is basically colorless, and then controlling the water content of the polyamide to be completely removed;

step five, adding 29.8kg of 98% V/V ethanol into the polyamide with water controlled completely in the step four, stirring and eluting for 5 hours at the temperature of 38 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, then adding 23.84kg of 98% V/V ethanol into the polyamide, stirring and eluting for 4 hours at the temperature of 40 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, adding 14.9kg of 98% V/V ethanol into the filtered polyamide, stirring and eluting for 3 hours at the temperature of 42 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, merging the three eluents, and recovering ethanol to obtain a thick paste ethanol extract;

and step six, putting the thick paste alcohol extract obtained in the step five into a vacuum drier, carrying out vacuum drying at the temperature of not more than 50 ℃, and crushing to obtain 49.6g of light brown yellow ginkgolic acid extract powder.

The content of ginkgolic acid in the extract powder was 92.74 wt% as determined by HPLC.

Example 2

Weighing 1kg of cleaned and dried ginkgo biloba testa, adding 10kg of deionized water, soaking for 20h at 42 ℃, and filtering out soaking water after soaking is finished;

step two, adding 1kg of deionized water into the ginkgo biloba sarcotesta soaked in the step one, grinding the ginkgo biloba sarcotesta into coarse pulp by using a pulp grinder, and then finely grinding the coarse pulp by using a colloid mill to obtain finely ground pulp, wherein the granularity of the finely ground pulp is 6.5-13 mu m;

step three, adding 20kg of ethanol with the content concentration of 98% V/V into the fine grinding slurry obtained in the step two, leaching for 10 hours at the temperature of 44 ℃, and filtering to obtain ethanol leaching liquor; adding 20kg of petroleum ether into filter residue, leaching for 18h at the temperature of 36 ℃, filtering to obtain petroleum ether leaching liquor, recovering ethanol and petroleum ether from the ethanol leaching liquor and the petroleum ether leaching liquor respectively to obtain pasty ethanol leaching matters and pasty petroleum ether leaching matters respectively, and combining the leaching matters to obtain 306g of mixed pasty leaching matters;

adding 9.18kg of deionized water into the mixed paste extract obtained in the step (three), fully and uniformly stirring at the rotating speed of 30r/min, adding 1.53kg of polyamide under continuous stirring, continuously stirring for 6h, standing and settling for 3h after stirring is finished, filtering to obtain polyamide, adding 1.53kg of polyamide into the filtrate, stirring at the rotating speed of 30r/min for 6h, standing and settling for 3h after stirring is finished, filtering to obtain polyamide, combining the polyamide obtained by two times of filtering in a filter, continuously adding deionized water into the filter until the filtered deionized water is basically colorless, and then controlling the water content of the polyamide to be completely removed;

step five, adding 30.6kg of 98% V/V ethanol into the polyamide with water controlled completely in the step four, stirring and eluting for 5 hours at the temperature of 39 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, then adding 24.48kg of 98% V/V ethanol into the polyamide, stirring and eluting for 4 hours at the temperature of 40 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, adding 15.3kg of 98% V/V ethanol into the filtered polyamide, stirring and eluting for 3 hours at the temperature of 41 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, merging the three eluents, and recovering ethanol to obtain a thick paste ethanol extract;

and step six, putting the thick paste alcohol extract obtained in the step five into a vacuum drier, carrying out vacuum drying at the temperature of not more than 50 ℃, and crushing to obtain 50.3g of light brown yellow ginkgolic acid extract powder.

The content of ginkgolic acid in the extract powder was 91.45 wt% by HPLC.

Example 3

Weighing 1kg of cleaned and dried ginkgo biloba testa, adding 10kg of deionized water, soaking for 18h at the temperature of 43 ℃, and filtering out soaking water after soaking is finished;

step two, adding 1kg of deionized water into the ginkgo biloba sarcotesta soaked in the step one, grinding the ginkgo biloba sarcotesta into coarse pulp by using a pulp grinder, and then finely grinding the coarse pulp by using a colloid mill to obtain finely ground pulp, wherein the granularity of the finely ground pulp is 6.5-13 mu m;

step three, adding 20kg of ethanol with the content concentration of 98% V/V into the fine grinding slurry obtained in the step two, leaching for 8 hours at the temperature of 45 ℃, and filtering to obtain ethanol leaching liquor; adding 20kg of petroleum ether into filter residue, leaching for 18h at the temperature of 33 ℃, filtering to obtain petroleum ether leaching liquor, recovering ethanol and petroleum ether from the ethanol leaching liquor and the petroleum ether leaching liquor respectively to obtain pasty ethanol leaching matters and pasty petroleum ether leaching matters respectively, and combining the leaching matters to obtain 310.3g of mixed pasty leaching matters;

adding 9.31kg of deionized water into the mixed paste extract obtained in the step (three), fully and uniformly stirring at the rotating speed of 30r/min, adding 1.55kg of polyamide under continuous stirring, continuously stirring for 6h, standing and settling for 3h after stirring is finished, filtering to obtain polyamide, adding 1.55kg of polyamide into the filtrate, stirring at the rotating speed of 30r/min for 6h, standing and settling for 3h after stirring is finished, filtering to obtain polyamide, combining the polyamide obtained by two times of filtering in a filter, continuously adding deionized water into the filter until the filtered deionized water is basically colorless, and then controlling the water content of the polyamide to be completely removed;

step five, adding 31kg of 98% V/V ethanol into the polyamide with water content controlled completely in the step four, stirring and eluting for 5 hours at the temperature of 39 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, then adding 24.8kg of 98% V/V ethanol into the polyamide, stirring and eluting for 4 hours at the temperature of 40 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, adding 15.5kg of 98% V/V ethanol into the filtered polyamide, stirring and eluting for 3 hours at the temperature of 42 ℃ and the rotation speed of 30r/min, filtering to obtain ethanol eluent, merging the three eluents, and recovering ethanol to obtain a thick paste ethanol extract;

and step (six), putting the thick paste alcohol extract obtained in the step (five) into a vacuum drier, carrying out vacuum drying at the temperature of not more than 50 ℃, and crushing to obtain 51g of light brown yellow ginkgolic acid extract powder.

The content of ginkgolic acid in the extract powder was 90.67 wt% by HPLC.