阅读说明：本技术 一种银杏黄酮的纯化方法 (Purification method of ginkgo flavone ) 是由 王晓军 刘伶文 刘茜倩 胡世义 于 2019-09-23 设计创作，主要内容包括：本发明公开了一种银杏黄酮的纯化方法,具体按照以下步骤实施：步骤1、银杏黄酮粗提液的制备；步骤2、将银杏黄酮粗提液在乙醇-硫酸铵双水相法萃取并干燥处理得银杏黄酮粉末。本发明的有益效果是：采用乙醇-硫酸铵双水相体系萃取银杏黄酮,由于盐析作用,蛋白质等杂质溶解在盐相中,提高了有机相中的黄酮纯度；提取的有机溶剂选择乙醇,无毒无害无污染,并且成本较低,符合未来工业化朝着绿色可持续发展方向的要求；使用超声辅助提纯银杏黄酮粗提液,利用了超声波所产生的强烈机械振动及空化作用等效果,破碎细胞结构,便于溶剂进入细胞及黄酮的流出,从而达到缩短提取时间,提高提取效率的目的。(The invention discloses a method for purifying ginkgetin, which is implemented according to the following steps: step 1, preparing a crude extract of ginkgo flavone; and 2, extracting the crude extract of the ginkgetin in an ethanol-ammonium sulfate two-aqueous phase method, and drying to obtain ginkgetin powder. The invention has the beneficial effects that: the ethanol-ammonium sulfate aqueous two-phase system is adopted to extract the ginkgetin, and due to salting-out, impurities such as protein and the like are dissolved in a salt phase, so that the purity of the ginkgetin in an organic phase is improved; the extracted organic solvent is ethanol, so that the method is non-toxic, harmless and pollution-free, has low cost and meets the requirement of future industrialization on green sustainable development; the crude extract of ginkgo flavone is purified by the aid of ultrasound, the cell structure is broken by the aid of strong mechanical vibration and cavitation generated by the ultrasound, and a solvent can enter cells conveniently and the flavone flows out conveniently, so that the aims of shortening extraction time and improving extraction efficiency are fulfilled.)

1. The method for purifying ginkgetin is characterized by comprising the following steps:

step 1, preparing a crude extract of ginkgo flavone;

and 2, extracting the crude extract of the ginkgetin in an ethanol-ammonium sulfate two-aqueous phase method, and drying to obtain ginkgetin powder.

2. The method for purifying ginkgetin according to claim 1, wherein the step 1 is specifically performed according to the following steps:

step 1.1, Ginkgo biloba leaf treatment

Drying folium Ginkgo to constant weight, pulverizing, and sieving to obtain folium Ginkgo powder;

step 1.2, leaching with aqueous ethanol

Mixing folium Ginkgo powder with ethanol water solution to obtain mixture A, and leaching to obtain mixture B;

step 1.3, ultrasonic treatment

Carrying out ultrasonic treatment on the mixed solution B and then filtering to obtain a mixed solution C;

step 1.4, concentrating the mixed solution C to obtain a crude extract of ginkgo flavone.

3. The method for purifying ginkgetin according to claim 1, wherein the step 2 is specifically performed according to the following steps:

step 2.1, preparing ethanol-ammonium sulfate aqueous two-phase mixed liquor: mixing the ammonium sulfate aqueous solution and absolute ethyl alcohol according to a proportion to obtain an ethanol-ammonium sulfate double-aqueous phase mixed solution;

step 2.2 extraction: mixing the crude extract of ginkgetin with ethanol-ammonium sulfate aqueous two-phase mixed solution in proportion, and extracting to obtain ginkgetin extract;

step 2.3, drying treatment: evaporating and drying the ginkgo flavone extract to obtain ginkgo flavone powder.

4. The method for purifying ginkgetin according to claim 2, wherein in step 1.3, the ginkgo biloba leaf powder and the ethanol aqueous solution are mixed in a mass ratio of 1: 5-8 to obtain a mixed solution A, wherein the volume fraction of ethanol in the ethanol water solution is 70-90%, and the mixed solution A is leached for 15-20 min at 50-80 ℃ to obtain a mixed solution B.

5. The method for purifying ginkgetin according to claim 2, wherein in step 1.3, the mixed solution B is subjected to ultrasonic treatment at a power of 150-250 KW for 30-60 min.

6. The method for purifying the crude ginkgo flavone extracting solution according to claim 2, wherein the mixed solution C is treated in a rotary evaporator in the step 1.4, wherein the pressure of the rotary evaporator is 0.07-0.09 MPa, the rotating speed is 60-90 r/min, and the temperature is 55-70 ℃ to obtain the crude ginkgo flavone extracting solution.

7. The method for purifying ginkgetin according to claim 3, wherein the step 3.1 comprises the following steps:

the concentration is 1.8 to 2.4 mol.L^-1The ammonium sulfate aqueous solution and absolute ethyl alcohol are mixed according to the volume ratio of 0.4-1.3 to obtain the ethanol-ammonium sulfate double-aqueous phase mixed solution.

8. The method for purifying ginkgetin according to claim 3, wherein the step 3.2 comprises the following steps:

and adding the ginkgo biloba concentrated solution into the ethanol-ammonium sulfate double-aqueous phase mixed solution according to the volume ratio of 0.045-0.14, and extracting to obtain the ginkgo biloba extract.

9. The method for purifying ginkgetin according to claim 3, wherein the step 3.3 comprises the following steps:

and (3) treating the ginkgo flavone extract in a rotary evaporator, wherein the pressure of the rotary evaporator is 0.05-0.10 MPa, the rotating speed is 60-100 r/min, the temperature is 50-70 ℃, and drying the solid obtained by rotary evaporation to obtain ginkgo flavone powder.

Technical Field

The invention belongs to the technical field of plant functional component extraction, and particularly relates to a method for purifying ginkgetin.

Background

The ginkgetin is an important active ingredient in ginkgo leaf extract, and can increase cerebrovascular flow, improve cerebrovascular circulation, protect brain cells, dilate coronary artery, prevent angina pectoris and myocardial infarction, prevent thrombosis, and improve immunity. Is beneficial to patients with coronary heart disease, angina pectoris, cerebral arteriosclerosis, senile dementia and hypertension.

The ginkgo flavone has high medicinal value, and causes a plurality of scholars to carry out related research on the ginkgo flavone. At present, the extraction and purification methods of ginkgo flavone are organic solvent extraction, hot water extraction, supercritical fluid extraction, traditional two-aqueous phase extraction, resin purification and the like. Because the organic solvent extraction method has a plurality of steps, long period and high extraction rate of flavone, the residual organic solvent in the final product is more and more rejected by consumers; the hot water extraction method has the advantages that the extraction rate is low because the solubility of the flavonoid compounds in water is relatively low in ethanol, and the next separation and purification are difficult because the content of impurities such as water-soluble protein is high; the double water phase extraction method separates the sample to be separated by utilizing the solubility difference of each component in the sample to be separated in two phases so as to achieve the aim of quick separation and purification, and has the advantages of high separation efficiency, simple instrument and the like.

Disclosure of Invention

The invention aims to provide a purification method of ginkgo flavone, which solves the problem of low separation efficiency in the prior art.

The technical scheme adopted by the invention is that a method for purifying ginkgetin is implemented according to the following steps:

step 1, preparing a crude extract of ginkgo flavone;

and 2, extracting the crude extract of the ginkgetin by using an ethanol-ammonium sulfate two-aqueous phase method, and drying to obtain ginkgetin powder.

The invention is also characterized in that:

the step 1 is implemented according to the following steps:

step 1.1, Ginkgo biloba leaf treatment

Drying folium Ginkgo to constant weight, pulverizing, and sieving to obtain folium Ginkgo powder;

step 1.2, leaching with aqueous ethanol

Mixing folium Ginkgo powder with ethanol water solution to obtain mixture A, and leaching to obtain mixture B;

step 1.3, ultrasonic treatment

Carrying out ultrasonic treatment on the mixed solution B and then filtering to obtain a mixed solution C;

step 1.4, concentrating the mixed solution C to obtain the crude extract of ginkgo flavone.

The step 2 is implemented according to the following steps:

step 2.1, preparing ethanol-ammonium sulfate aqueous two-phase mixed liquor: mixing the ammonium sulfate aqueous solution and absolute ethyl alcohol according to a proportion to obtain an ethanol-ammonium sulfate double-aqueous phase mixed solution;

step 2.2 extraction: mixing the crude extract of ginkgetin with ethanol-ammonium sulfate aqueous two-phase mixed solution in proportion, and extracting to obtain ginkgetin extract;

step 2.3, drying treatment: evaporating and drying the ginkgo flavone extract to obtain ginkgo flavone powder.

Step 1.3, mixing the ginkgo leaf powder and an ethanol water solution according to a mass ratio of 1: 5-8 to obtain a mixed solution A, wherein the volume fraction of ethanol in the ethanol water solution is 70-90%, and the mixed solution A is leached for 15-20 min at 50-80 ℃ to obtain a mixed solution B.

And (1.3) carrying out ultrasonic treatment on the mixed solution B for 30-60min under the condition that the power is 150-250 KW.

And step 1.4, treating the mixed solution C in a rotary evaporator, wherein the pressure of the rotary evaporator is 0.07-0.09 MPa, the rotating speed is 60-90 r/min, and the temperature is 55-70 ℃, so as to obtain the crude ginkgo flavone extracting solution.

Step 2.1 the concrete steps are as follows:

the concentration is 1.8 to 2.4 mol.L^-1The ammonium sulfate aqueous solution and absolute ethyl alcohol are mixed according to the volume ratio of 0.4-1.3 to obtain the ethanol-ammonium sulfate double-aqueous phase mixed solution.

Step 2.2 the concrete steps are as follows:

and adding the ginkgo biloba concentrated solution into the ethanol-ammonium sulfate double-aqueous phase mixed solution according to the volume ratio of 0.045-0.14, and extracting to obtain the ginkgo biloba extract.

Step 2.3 the concrete steps are as follows:

and (3) treating the ginkgo flavone extract in a rotary evaporator, wherein the pressure of the rotary evaporator is 0.05-0.10 MPa, the rotating speed is 60-100 r/min, the temperature is 50-70 ℃, and drying the solid obtained by rotary evaporation to obtain ginkgo flavone powder.

The invention has the beneficial effects that: the ethanol-ammonium sulfate aqueous two-phase system is adopted to extract the ginkgetin, and due to salting-out, impurities such as protein and the like are dissolved in a salt phase, so that the purity of the ginkgetin in an organic phase is improved; the extracted organic solvent is ethanol, so that the method is non-toxic, harmless and pollution-free, has low cost and meets the requirement of future industrialization on green sustainable development; the crude extract of ginkgo flavone is purified by the aid of ultrasound, the cell structure is broken by the aid of strong mechanical vibration and cavitation generated by the ultrasound, and a solvent can enter cells conveniently and the flavone flows out conveniently, so that the aims of shortening extraction time and improving extraction efficiency are fulfilled.

Detailed Description

The present invention will be described in detail with reference to the following embodiments.

The invention relates to a method for purifying ginkgetin, which is implemented according to the following steps:

step 1, preparing the crude extract of ginkgo flavone, which is specifically implemented according to the following steps:

step 1.1, ginkgo leaf treatment, which is specifically implemented according to the following steps:

drying folium Ginkgo at 60-80 deg.C to constant weight, pulverizing, and sieving with 200 mesh sieve to obtain folium Ginkgo powder;

step 1.2, leaching with ethanol water, specifically comprising the following steps:

mixing ginkgo leaf powder and an ethanol water solution according to a mass ratio of 1: 5-8, mixing to obtain a mixed solution A, wherein the volume fraction of ethanol in the ethanol aqueous solution is 70-90%, and leaching the mixed solution A at 50-80 ℃ for 15-20 min to obtain a mixed solution B;

step 1.3, ultrasonic treatment, which is specifically implemented according to the following steps:

carrying out ultrasonic treatment on the mixed solution B for 30-60min under the condition that the power is 150-250 KW, and filtering to obtain mixed solution C;

step 1.4, evaporation treatment, which is specifically implemented according to the following steps:

and (3) treating the mixed solution C in a rotary evaporator, wherein the pressure of the rotary evaporator is 0.07-0.09 MPa, the rotating speed is 60-90 r/min, and the temperature is 55-70 ℃ to obtain the crude ginkgo flavone extracting solution.

Step 2, extracting the crude extract of ginkgo flavone in an ethanol-ammonium sulfate two aqueous phase method and drying to obtain ginkgo flavone powder, wherein the method is implemented according to the following steps:

step 2.1, preparing an ethanol-ammonium sulfate aqueous two-phase mixed solution, and specifically performing the following steps:

weighing 25-35g of ammonium sulfate, dissolving in 80-130 mL of water, and mixing with 100-180 mL of absolute ethanol to obtain an ethanol-ammonium sulfate double-water-phase mixed solution;

step 2.2, extraction is specifically carried out according to the following steps:

adding 15-25mL of the crude extract of ginkgo flavone into the ethanol-ammonium sulfate aqueous two-phase mixed solution, and extracting to obtain a ginkgo flavone extract;

step 2.3, drying treatment is specifically carried out according to the following steps:

and (3) treating the ginkgo flavone extract in a rotary evaporator, wherein the pressure of the rotary evaporator is 0.05-0.10 MPa, the rotating speed is 60-100 r/min, the temperature is 50-70 ℃, and drying the solid obtained by rotary evaporation to obtain ginkgo flavone powder.