阅读说明：本技术 一种从银杏叶提取物生产废液中提取多种活性物质的方法 (Method for extracting multiple active substances from waste liquid of ginkgo leaf extract production ) 是由 何桥 曹春月 王黔阳 陈震 杨明武 陈伟 欧琴 于 2019-10-29 设计创作，主要内容包括：本发明公开了一种从银杏叶提取物生产废液中提取多种活性物质的方法,该方法包括如下步骤：(1)银杏叶提取物生产废液的醇沉；(2)醇沉物加工；(3)醇沉液加工；(4)莽草酸粗晶体再结晶。与现有技术相比较,本发明可有效地连续综合提取银杏叶中银杏叶提取物、银杏叶多糖、银杏叶蛋白及莽草酸4个产品,提高银杏叶药材的综合利用度；综合回收利用银杏叶多糖、银杏叶蛋白及莽草酸,从而降低生产过程中废水处理的成本；洗柱时采用阴离子及活性碳柱双柱串联,洗脱的同时进行脱色处理,减少结晶时二次脱色工序,提高生产效率。适用于银杏叶的综合加工利用。(The invention discloses a method for extracting various active substances from waste liquid in the production of ginkgo leaf extract, which comprises the following steps: (1) alcohol precipitation of waste liquid from ginkgo leaf extract production; (2) processing alcohol precipitation; (3) processing an alcohol precipitation solution; (4) recrystallizing shikimic acid crude crystal. Compared with the prior art, the invention can effectively and continuously extract 4 products of ginkgo leaf extract, ginkgo leaf polysaccharide, ginkgo leaf protein and shikimic acid in ginkgo leaves, and improve the comprehensive utilization degree of ginkgo leaf medicinal materials; comprehensively recycling the ginkgo biloba polysaccharides, the ginkgo biloba proteins and the shikimic acid, thereby reducing the cost of wastewater treatment in the production process; when the column is washed, double columns of anions and activated carbon columns are connected in series, and the decoloring treatment is carried out while the elution is carried out, so that the secondary decoloring procedure during the crystallization is reduced, and the production efficiency is improved. Is suitable for comprehensive processing and utilization of ginkgo leaves.)

1. A method for extracting a plurality of active substances from ginkgo leaf extract production waste liquid is characterized by comprising the following steps:

(1) alcohol precipitation of waste liquid from production of ginkgo leaf extract

Combining macroporous resin sample loading residual liquid and water washing liquid generated in the production process of the ginkgo biloba extract prepared according to Chinese pharmacopoeia, concentrating the mixture until the relative density of the material liquid is 1.1 to obtain a raw material liquid;

carrying out alcohol precipitation by adopting high-concentration ethanol to obtain a white precipitate and an alcohol precipitation solution for later use;

(2) processing alcohol precipitation

Dissolving the white precipitate in water, performing ultrafiltration on the obtained solution with the molecular weight of 1 ten thousand, further performing vacuum concentration on the concentrated solution to obtain about 150-200L, adding a Sevage reagent, mixing, oscillating, centrifuging to obtain a ginkgo leaf crude protein precipitate, and evaporating the centrifugate to dryness to obtain a ginkgo leaf crude polysaccharide;

(3) processing of alcohol precipitation liquid

Concentrating the alcohol precipitation solution until no alcohol smell exists, then carrying out column chromatography on the treated anion resin, slowly loading the extract above an adsorption column, adsorbing for a period of time, washing with distilled water to remove impurities, connecting an activated carbon column in series after removing the impurities, eluting the column in series with formic acid aqueous solution, collecting eluent, concentrating the eluent to a small volume, and placing the concentrated eluent in an environment at 0-25 ℃ for crystallization to obtain shikimic acid crude crystals;

(4) recrystallization of crude crystals of shikimic acid

Adding a proper amount of water into the shikimic acid crude crystal for dissolving, and then placing the solution in the temperature environment same as the crude crystal for secondary crystallization to obtain shikimic acid crystal with the purity of more than 98 percent.

2. The method according to claim 1, wherein the ethanol is present in an amount of 90 to 95% by weight in step (1).

3. The method according to claim 1, wherein in the step (2), the water is added in an amount such that the ratio of the feed to the liquid is 1: 10 to 1: 20; the vacuum degree of the vacuum concentration control is 0.06-0.09 MPa; the Sevage reagent is a mixture of chloroform and n-butanol, wherein the volume ratio of the chloroform to the n-butanol is 4: 1.

4. The method according to claim 1, wherein in the step (3), the sample loading speed is 1-2 times of column volume/h; the amount of the distilled water for washing and removing impurities is 2-4 times of the column volume; the mass fraction of the formic acid aqueous solution of the elution string column is 3% -8%, and the using amount of the formic acid aqueous solution is 3-5 times of the column volume.

5. The method according to claim 1, wherein in the step (4), the amount of the added water is 1/5-1/10% of the crude crystals; the same temperature as the crude crystal is 0 ℃ to 25 ℃.

Technical Field

The invention relates to multi-stage treatment of wastewater, in particular to a method for extracting ginkgo leaves and comprehensively utilizing waste liquid thereof to obtain various active substances.

Technical Field

As is well known, Ginkgo biloba is a leaf of Ginkgo biloba (Ginkgo biloba L.) belonging to the genus Ginkgo of the family Ginkgo. The ginkgo biloba extract has unique physiological and pharmacological effects and therapeutic values, and particularly, the ginkgolide is a PAF (platelet activating factor) receptor blocker with high specificity, so that the ginkgo biloba extract is well appreciated by people. The literature considers that the main pharmacological actions of the ginkgo biloba extract are as follows: has effects in treating central nervous system, resisting shock, and resisting allergy. The protective action for organ transplant rejection reaction, besides the above-mentioned action, has the inhibiting action for cell immunological reaction, eye pathological change and circulation change due to endotoxin, and also has the action for reproductive system. The ginkgo biloba extract is developed in China since the last 70 years, but the research on the comprehensive utilization of the ginkgo biloba extract waste liquid is not much.

At present, in the Chinese patent database, there are 2 patent application pieces relating to ginkgo leaf extraction wastewater: CN106495411A No. ICBB technology and device for treating wastewater in the extraction process of folium Ginkgo extract, and CN110002616A No. device and method for treating and recycling wastewater from folium Ginkgo extraction of ginkgetin, which are both treatment technologies for folium Ginkgo extraction wastewater. In addition, there are 3 patent applications relating to the preparation of shikimic acid from waste liquid of ginkgo leaf extraction: CN109593034A A method for preparing shikimic acid from waste liquid of ginkgo leaf extraction, CN108191639A A method for preparing shikimic acid extract from waste liquid of ginkgo leaf extract chromatography, and CN109369373A A method for preparing shikimic acid extract from waste liquid of ginkgo leaf extract chromatography are all technical schemes for preparing shikimic acid. At present, no patent application for comprehensive utilization of ginkgo leaf extraction waste liquid exists.

Disclosure of Invention

The invention aims to provide a method for extracting various active substances from ginkgo leaf extract production waste liquid, which not only changes the ginkgo leaf extract production waste liquid into valuable and reduces the pollution to the environment, but also can obtain ginkgo leaf polysaccharide, ginkgo leaf protein and shikimic acid products and realize comprehensive utilization.

The method for extracting various active substances from the production waste liquid of the ginkgo leaf extract provided by the inventor comprises the following steps:

(1) alcohol precipitation of waste liquid from production of ginkgo leaf extract

Macroporous resin sample loading residual liquid and water washing liquid generated in the production process of the ginkgo biloba extract prepared according to Chinese pharmacopoeia are concentrated to the relative density of the material liquid of about 1.1, thus obtaining the raw material liquid. Carrying out alcohol precipitation by adopting ethanol to obtain a white precipitate and an alcohol precipitation solution for later use;

(2) processing alcohol precipitation

Dissolving the white precipitate in water, ultrafiltering the obtained solution with molecular weight of 5000 or 1 ten thousand, vacuum concentrating the concentrated solution to 200L, adding Sevage reagent, mixing, oscillating, centrifuging to obtain folium Ginkgo crude protein precipitate, and evaporating the centrifugate to dryness to obtain folium Ginkgo crude polysaccharide;

(3) processing of alcohol precipitation liquid

Concentrating the alcohol precipitation solution until no alcohol smell exists, then carrying out column chromatography on the treated anion resin, slowly loading the extract above an adsorption column, adsorbing for a period of time, washing with distilled water to remove impurities, connecting an activated carbon column in series after removing the impurities, eluting the column in series with formic acid aqueous solution, collecting eluent, concentrating the eluent to a small volume, placing the concentrated eluent in an environment at 0-25 ℃ for crystallization, and obtaining shikimic acid crude crystals with the purity of more than 80%;

(4) recrystallization of crude crystals of shikimic acid

Adding methanol or water into the shikimic acid crude crystal for dissolving, and then placing the solution in the temperature environment the same as the temperature environment of the crude crystal for secondary crystallization to obtain the shikimic acid crystal with the purity of more than 98 percent.

In the step (1), the mass fraction of the ethanol is 90-95%.

In the step (2), the amount of the added water is controlled to enable the ratio of the materials to the liquid to be 1: 10-1: 20; the vacuum degree of the vacuum concentration control is 0.06-0.09 MPa; the Sevage reagent is a mixture of chloroform and n-butanol, wherein the volume ratio of the chloroform to the n-butanol is 4: 1.

In the step (3), the sample loading speed is 1-2 times of column volume/h; the amount of the distilled water for washing and removing impurities is 2-4 times of the column volume; the mass fraction of the formic acid aqueous solution of the elution string column is 3% -8%, and the using amount of the formic acid aqueous solution is 3-5 times of the column volume.

In the step (4), the amount of the added methanol or water is 1/5-1/10 of the amount of the shikimic acid crude crystals; the same temperature as the crude crystal is 0 ℃ to 25 ℃.

The inventor indicates that the ginkgo leaf crude protein precipitate and the ginkgo leaf crude polysaccharide prepared in the step (2) can be further processed and purified by adopting a conventional process according to the market demand to prepare a pure product.

The innovation points of the invention are as follows:

(1) the invention can effectively and continuously extract 4 products of ginkgo leaf extract, ginkgo leaf polysaccharide, ginkgo leaf protein and shikimic acid in ginkgo leaves, improves the comprehensive utilization rate of ginkgo leaf medicinal materials, lowers the comprehensive production cost of the products and effectively improves the market competitiveness of production plants.

(2) In the production process of the ginkgo leaf extract, ginkgo leaf polysaccharide, ginkgo leaf protein and shikimic acid are comprehensively recycled from waste wastewater, so that the wastewater treatment pressure in the production process is reduced.

(3) When the column is washed, double columns of anions and activated carbon columns are connected in series, so that the decolorization treatment is carried out while enrichment and elution are carried out, the secondary decolorization process during crystallization is reduced, and the production efficiency is improved.

The invention is suitable for comprehensive processing and utilization of ginkgo leaf extract produced by ginkgo leaves.

Drawings

FIG. 1 is a crystal pattern of shikimic acid,

FIG. 2 is a raw material liquid spectrum,

FIG. 3 is a map of shikimic acid in ginkgo leaves.

Detailed Description