阅读说明：本技术 一种低游离黄酮、低酸的银杏叶提取物的制备方法 (Preparation method of low-free-flavone and low-acid ginkgo leaf extract ) 是由 王允达 于 2018-06-25 设计创作，主要内容包括：本发明涉及一种低游离黄酮、低酸的银杏叶提取物的制备方法,包括以下步骤：粉碎、过筛、提取、浓缩、除杂、富集纯化、干燥、粉碎,采用的优化比例的大孔吸附树脂和无机吸附剂为层析柱填料,所得到的银杏叶提取物游离黄酮、银杏酸低,同时该制备方法现有设备即能满足要求,非常适合工业化生产,本发明设计合理,可大规模推广。(The invention relates to a preparation method of a ginkgo leaf extract with low free flavone and low acid, which comprises the following steps: the preparation method comprises the following steps of crushing, sieving, extracting, concentrating, impurity removing, enriching and purifying, drying and crushing, macroporous adsorption resin and inorganic adsorbent in optimized proportion are adopted as chromatographic column packing, the obtained ginkgo leaf extract has low free flavone and ginkgolic acid content, and meanwhile, the existing equipment of the preparation method can meet the requirements, so that the preparation method is very suitable for industrial production, and the preparation method is reasonable in design and can be popularized in a large scale.)

1. A method for preparing a low free flavone and low acid ginkgo leaf extract is characterized by comprising the following steps: pulverizing, sieving, extracting, concentrating, removing impurities, enriching, purifying, drying, and pulverizing.

2. The method for preparing the ginkgo biloba leaf extract with low free flavone and low acid content as claimed in claim 1, which comprises the following steps:

(1) pulverizing folium Ginkgo, and sieving with 20 ~ 30 mesh sieve;

(2) adding 55-75% ethanol 6-12 times the weight of folium Ginkgo into folium Ginkgo, extracting at 60-80 deg.C for 60-90 min each time, filtering, and mixing extractive solutions;

(3) concentrating the prepared extract until the density is 1.05-1.15, adding 1-2 times of purified water into the concentrated solution for dilution, stirring, standing overnight at room temperature, and collecting filtrate;

(4) and (2) taking the filtrate until a chromatographic column is saturated, wherein the chromatographic column is filled with macroporous adsorption resin and an inorganic adsorbent in a weight ratio of (1-5): 1;

(5) eluting with water, ethanol or mixture of ethanol and water as eluent at flow rate of 1-3BV/h, sequentially eluting with water, ethanol concentration of 15-25%, ethanol concentration of 60-80%, and eluent volume of 2-3BV, 3-5BV, and 2-4BV, collecting eluent with ethanol concentration of 60-80%, concentrating, drying, and pulverizing to obtain folium Ginkgo extract with low free flavone and low acid.

Technical Field

The invention relates to the field of ginkgo leaf extracts, in particular to a preparation method of a ginkgo leaf extract with low free flavone and low acid.

Background

The gingko is a gingko tree which is commonly known in China, is a common deciduous tree in China, is one of the most life-prolonging species known to people in the world at present, is located in Guizhou in China at present, and has the age of five to six thousand years. Gingko appears in 5 hundred million years ago in the stone charcoal period, and the movement of the fourth glacier causes the environment of the earth to become cold suddenly and is not suitable for the survival of most plants, most of gingko plants disappear in the period, and only in parts of China in the world survive due to the factors of the natural environment. Ginkgo biloba is the only species existing in the class of the gymnosperm class Ginkgoaceae and other species in the class have disappeared in the historical long rivers, and because of this, Ginkgo biloba is called "Actinolitum" in the plant kingdom.

Ginkgo biloba is one of the hot spots of research and development at home and abroad in recent years, and the Ginkgo Biloba Extract (GBE) is widely used for cardiovascular and cerebrovascular diseases, and a large number of researches show that the prevention and treatment effect of the ginkgo biloba extract on the cardiovascular and cerebrovascular diseases is closely related to the antioxidant activity of the ginkgo biloba extract. The main physiological active ingredients in GBE are flavonoids and ginkgo terpene lactones, wherein ginkgo flavonoids have multiple functions of scavenging free radicals, resisting oxidation, improving cardiovascular and cerebrovascular circulation, resisting fatigue, reducing blood fat, inhibiting bacteria and the like, the purity of the flavonoids in ginkgo leaves plays a role in the application of the flavonoids, namely, the ginkgo flavonoids can also be used as the ingredients of food additives to prepare health-care food for resisting fatigue and resisting aging, wherein the total flavonoid purity is required to be more than 24 percent when the ginkgo biloba extract is prepared into a medicament for preventing and treating cardiovascular and cerebrovascular diseases (generally, EGB761 produced by W.Svhwabe company in Germany is used as a standardized extract of ginkgo leaves, and the extract contains 24 percent of flavonoids). The extraction of flavonoids from ginkgo leaves has been patented in germany and france as early as 70 years, and from the common organic solvent extraction method of alcohol, ketone and the like to the supercritical fluid extraction method in recent years, the extraction method with high purity, high yield and environmental protection is always pursued.

The existing extraction method extracts the ginkgo leaf extract with high free flavone and high acid, which can not meet the requirements of high-end customers in European and American markets, and the extraction preparation method is improved.

Disclosure of Invention

In view of the current state of the prior art, the technical problem to be solved by the invention is to provide a preparation method of a low-free-flavone and low-acid ginkgo leaf extract which can meet the requirements of high-end customers in the markets of europe and america.

The technical scheme adopted by the invention for solving the technical problems is as follows: a method for preparing low free flavone and low acid folium Ginkgo extract comprises the following steps: pulverizing, sieving, extracting, concentrating, removing impurities, enriching, purifying, drying, and pulverizing.

Further, the method specifically comprises the following steps:

(1) pulverizing folium Ginkgo, and sieving with 20 ~ 30 mesh sieve;

(2) adding 55-75% ethanol 6-12 times the weight of folium Ginkgo into folium Ginkgo, extracting at 60-80 deg.C for 60-90 min each time, filtering, and mixing extractive solutions;

(3) concentrating the prepared extract until the density is 1.05-1.15, adding 1-2 times of purified water into the concentrated solution for dilution, stirring, standing overnight at room temperature, and collecting filtrate;

(4) and (2) taking the filtrate until a chromatographic column is saturated, wherein the chromatographic column is filled with macroporous adsorption resin and an inorganic adsorbent in a weight ratio of (1-5): 1;

(5) eluting with water, ethanol or mixture of ethanol and water as eluent at flow rate of 1-3BV/h, sequentially eluting with water, ethanol concentration of 15-25%, ethanol concentration of 60-80%, and eluent volume of 2-3BV, 3-5BV, and 2-4BV, collecting eluent with ethanol concentration of 60-80%, concentrating, drying, and pulverizing to obtain folium Ginkgo extract with low free flavone and low acid.

Compared with the prior art, the invention has the advantages that: the invention adopts the macroporous adsorption resin and the inorganic adsorbent with optimized proportion as the chromatographic column packing, the obtained ginkgo leaf extract has low free flavone and ginkgolic acid, and the existing equipment of the preparation method can meet the requirements at the same time, thereby being very suitable for industrial production.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail by the following specific embodiments.

EXAMPLE 1A method for preparing a Ginkgo biloba extract with low free flavone and low acid content

1. Pulverizing folium Ginkgo 150kg to 20 mesh. 900 liters of 55% ethanol heated to 65 ℃, starting an extraction tank for stirring, starting a steam valve, keeping the solvent at 65 ℃, extracting for 60 minutes, stopping stirring, filtering the extracting solution, transferring to a storage tank, adding 900 liters of 55% ethanol for secondary extraction, extracting for 30 minutes, filtering, transferring to the storage tank, and combining the extracting solutions; 2. concentrating the above extractive solution to obtain concentrated solution with density of 1.05, adding purified water 1 times the volume of the concentrated solution, stirring, standing at room temperature overnight, and filtering; 3. the method comprises the steps of adopting a chromatographic column with macroporous adsorption resin and inorganic adsorbent as fillers in a mass ratio of 1:1, loading filtrate into the column at a flow rate of 1.5BV/h, standing for 1 hour, then eluting, eluting with 2.0BV of water at a flow rate of 1.5BV/h, eluting with 3.5BV of 15% ethanol at a flow rate of 2.0BV/h, eluting with 2.5BV of 60% ethanol at a flow rate of 1.5BV/h, collecting 60% ethanol eluent, concentrating under reduced pressure, drying, and crushing to obtain 3.53Kg of ginkgo leaf extract with low free flavone and low acid.

EXAMPLE 2A method for preparing a Ginkgo biloba extract with low free flavone and low acid content

1. Pulverizing folium Ginkgo 150kg to 30 mesh. Heating 65% ethanol at 70 deg.C to 1200L, starting extraction tank, stirring, opening steam valve, maintaining solvent at 70 deg.C, extracting for 60 min, stopping stirring, filtering extractive solution, transferring to storage tank, extracting for 60 min with 55% ethanol 600L, filtering, transferring to storage tank, and mixing extractive solutions; 2. concentrating the above extractive solution to obtain concentrated solution with density of 1.10, adding 1.5 times of purified water, stirring, standing at room temperature overnight, and filtering; 3. the method comprises the steps of adopting a chromatographic column with macroporous adsorption resin and inorganic adsorbent as fillers in a mass ratio of 3:1, loading filtrate into the column at a flow rate of 2.5BV/h, standing for 1 hour, then eluting, eluting with 3.0BV of water at a flow rate of 2.5BV/h, eluting with 4.5BV of 15% ethanol at a flow rate of 3.0BV/h, eluting with 3.5BV of 60% ethanol at a flow rate of 3.0BV/h, collecting 60% ethanol eluent, concentrating under reduced pressure, drying, and crushing to obtain 3.78Kg of ginkgo leaf extract with low free flavone and low acid.

Comparative 1 preparation method of Ginkgo biloba leaf extract with low free flavone and low acid

The specific operation is the same as that of the embodiment 1, and the difference is that the chromatographic column packing in the step 3 is independently selected from macroporous adsorption resin.

Comparative 2 preparation method of Ginkgo biloba leaf extract with low free flavone and low acid

The specific operation is the same as that of the embodiment 2, and the difference is that the chromatographic column packing in the step 3 is independently selected from macroporous adsorption resin.

Data for examples 1, 2 and comparative examples 1, 2 are shown in table 1.

TABLE 1 data for examples 1, 2 and comparative examples 1, 2

Name (R)	Yield%	Flavonol glycosides%	Terpene lactones%	Rutin content	Free quercetin mg/g	Free kaempferol mg/g	Free isorhamnetin mg/g	Ginkgolic acid ppm
									EXAMPLE 1	2.35	25.67	7.01	1.53	0.54	0.23	0.14	0.9
EXAMPLE 2	2.52	26.12	7.21	1.47	0.63	0.32	0.18	0.6
									Comparative example 1	2.45	25.11	7.11	2.87	6.3	4.5	1.8	8.5
Comparative example 2	2.61	25.89	7.32	3.11	7.4	5.2	1.5	6.8

As can be seen from the results in Table 1, examples 1 and 2 showed comparable results to comparative examples 1 and 2, although the yields and contents of the resulting ginkgo biloba extracts were comparable; in the implementation 1 and 2 and the comparison 1 and 2, the macroporous resin and the inorganic adsorbent with optimized proportion are used as the chromatographic column packing, so that the enrichment and purification of the ginkgo leaf extract are greatly influenced, and the limit indexes of the implementation 1 and 2 are obviously lower than those of the comparison 1 and 2.

The calculation and measurement methods for each data in table 1 are as follows:

1. yield = (mass of ginkgo leaf extract/mass of ginkgo leaf) × 100%;

2. the measuring method of the contents of the flavonol glycosides and the terpene lactones is measured according to the inspection method of ginkgo biloba extract of the 2015 version of Chinese pharmacopoeia;

3. the content of free quercetin, free kaempferol and free isorhamnetin is determined by folium Ginkgo extract supplement test method;

4. the determination method of rutin content is determined according to the inspection method of folium Ginkgo extract in United states Pharmacopeia;

5. the determination method of ginkgolic acid content is determined according to the inspection method of ginkgo biloba extract in European pharmacopoeia.

Compared with the prior art, the invention has the advantages that: the invention adopts the macroporous adsorption resin and the inorganic adsorbent with optimized proportion as the chromatographic column packing, the yield and the content are not greatly different, but the difference of the yield and the content is obviously reduced for limited components, the obtained ginkgo leaf extract has low free flavone and ginkgolic acid, and meanwhile, the existing equipment of the preparation method can meet the requirements, thereby being very suitable for industrial production, having reasonable design and being capable of being popularized in a large scale.