阅读说明：本技术 一种复方银杏通脉有效部位、提取物及其制备方法和其应用 (Compound gingko effective part and extract for promoting blood circulation as well as preparation method and application thereof ) 是由 侯凤飞 戴鑫汶 邹奇 戴甲木 张豪 唐心文 邓声菊 于 2020-06-15 设计创作，主要内容包括：本发明涉及一种复方银杏通脉有效部位、复方银杏通脉提取物的醇提工艺及其制备条件,采用酸化乙醇溶液提取复方银杏通脉有效部位,并将其用于制备复方银杏通脉提取物或药物组合物,显著提高有效部位中总黄酮醇苷的含量和提取率,进而显著提高药品的质量和疗效,并显著提高资源利用率且显著降低成本。(The invention relates to an alcohol extraction process and preparation conditions of a compound gingko effective part for promoting blood circulation, a compound gingko effective part for promoting blood circulation extract, wherein the compound gingko effective part is extracted by adopting an acidified ethanol solution and is used for preparing the compound gingko effective part for promoting blood circulation or a pharmaceutical composition, so that the content and extraction rate of total flavonol glycosides in the effective part are obviously improved, the quality and curative effect of the medicine are further obviously improved, the resource utilization rate is obviously improved, and the cost is obviously reduced.)

1. The compound gingko effective part is characterized by being prepared from 180 parts of gingko leaves, 180 parts of prepared fleece-flower roots, 90 parts of glossy privet fruits, 90 parts of salvia miltiorrhiza and 60 parts of uncaria, and the preparation method of the effective part comprises the following steps: weighing required amount of folium Ginkgo, radix Polygoni Multiflori Preparata, fructus Ligustri Lucidi, Saviae Miltiorrhizae radix and ramulus Uncariae cum uncis, pulverizing into coarse powder, adding 30-95% acidified ethanol water solution 3-18 times (volume/weight ratio) of the medicinal materials, heating and reflux-extracting for 1-4 times (each time for 0.5-5 hr), filtering, collecting acidified ethanol extractive solution, concentrating, and drying the collected acidified ethanol extractive solution.

2. The active site of claim 1, wherein the acidified ethanol solution has a pH of 1 to 6, preferably the acidified ethanol aqueous solution has a pH of 2 to 5, more preferably the acidified ethanol aqueous solution has a pH of 3 to 4.

3. The active fraction according to any one of claims 1 to 2, wherein the content of total flavonol glycosides in the active fraction is not less than 0.2%, preferably not less than 0.25%, and the content of emodin in the active fraction is not less than 0.015%, preferably not less than 0.02%.

4. The preparation method of the effective part according to any one of claims 1 to 3, wherein the effective part is prepared from 180 parts by mass of ginkgo leaf, 180 parts by mass of prepared fleece-flower root, 90 parts by mass of glossy privet fruit, 90 parts by mass of red sage root and 60 parts by mass of uncaria, and the preparation method of the effective part comprises the following steps: weighing required amount of folium Ginkgo, radix Polygoni Multiflori Preparata, fructus Ligustri Lucidi, Saviae Miltiorrhizae radix and ramulus Uncariae cum uncis, pulverizing into coarse powder, adding 30-95% acidified ethanol water solution 3-18 times (volume/weight ratio) of the medicinal materials, heating and reflux-extracting for 1-4 times (each time for 0.5-5 hr), filtering, collecting acidified ethanol extractive solution, concentrating, and drying the collected acidified ethanol extractive solution.

5. The use of the compound gingko effective component of any one of claims 1 to 3 for preparing any one of a compound gingko vein relaxing extract and a compound gingko vein relaxing pharmaceutical composition.

6. The compound ginkgo biloba pulse-invigorating extract with high total flavonol glycoside content is prepared from 180 parts by weight of ginkgo biloba leaves, 180 parts by weight of prepared fleece-flower root, 90 parts by weight of glossy privet fruit, 90 parts by weight of salvia miltiorrhiza, 90 parts by weight of eucommia bark, 90 parts by weight of medicinal cyathula root and 60 parts by weight of uncaria, and the preparation method of the extract comprises the following steps:

1) weighing required amount of folium Ginkgo, radix Polygoni Multiflori Preparata, fructus Ligustri Lucidi, Saviae Miltiorrhizae radix and ramulus Uncariae cum uncis, pulverizing into coarse powder, adding 30-95% acidified ethanol water solution 3-18 times (volume/weight ratio) of the medicinal materials, heating and reflux-extracting for 1-4 times (0.5-5 hr per time), filtering, collecting acidified ethanol extractive solution, and concentrating the acidified ethanol extractive solution to obtain extract I;

2) weighing eucommia ulmoides and medicinal cyathula root according to the prescription amount, crushing the eucommia ulmoides and medicinal cyathula root into coarse powder, adding water which is 3-20 times (volume weight ratio) of the total amount of medicinal materials, decocting and extracting for 1-4 times, extracting for 0.5-4 hours each time, filtering, concentrating decoction, adding ethanol until the ethanol content reaches 30-90%, cooling the decoction to 4-35 ℃, standing for 6-56 hours, filtering, concentrating supernate and recovering ethanol to obtain an extract II;

(3) and mixing the prepared first extract and the second extract, and drying, or respectively drying the prepared first extract and the prepared second extract and mixing.

7. An extract as claimed in claim 6, wherein the total flavonol glycosides content of the extract is not less than 0.1%, preferably not less than 0.2%.

8. The preparation method of the compound ginkgo biloba extract containing the total flavonol glycoside effective part as claimed in any one of claims 6 to 7, wherein the extract is prepared from 180 parts by mass of ginkgo biloba leaves, 180 parts by mass of prepared fleece flower root, 90 parts by mass of glossy privet fruit, 90 parts by mass of red sage root, 90 parts by mass of eucommia bark, 90 parts by mass of medicinal cyathula root and 60 parts by mass of uncaria rhynchophylla, and the preparation method of the extract comprises the following steps:

1) weighing required amount of folium Ginkgo, radix Polygoni Multiflori Preparata, fructus Ligustri Lucidi, Saviae Miltiorrhizae radix and ramulus Uncariae cum uncis, pulverizing into coarse powder, adding 30-95% acidified ethanol water solution 3-18 times (volume/weight ratio) of the medicinal materials, heating and reflux-extracting for 1-4 times (0.5-5 hr per time), filtering, collecting acidified ethanol extractive solution, and concentrating the acidified ethanol extractive solution to obtain extract I;

2) weighing eucommia ulmoides and medicinal cyathula root according to the prescription amount, crushing the eucommia ulmoides and medicinal cyathula root into coarse powder, adding water which is 3-20 times (volume weight ratio) of the total amount of medicinal materials, decocting and extracting for 1-4 times, extracting for 0.5-4 hours each time, filtering, concentrating decoction, adding ethanol until the ethanol content reaches 30-90%, cooling the decoction to 4-35 ℃, standing for 6-56 hours, filtering, concentrating supernate and recovering ethanol to obtain an extract II;

(3) and mixing the prepared first extract and the second extract, and drying, or respectively drying the prepared first extract and the prepared second extract and mixing.

9. A compound ginkgo biloba pulse-invigorating pharmaceutical composition, which consists of the compound ginkgo biloba pulse-invigorating extract as claimed in any one of claims 6 to 7 and a pharmaceutically acceptable carrier.

10. The use of the compound ginkgo biloba tongmai extract as claimed in any one of claims 6 to 7 or the pharmaceutical composition as claimed in claim 9 for the preparation of a medicament for treating mild cerebral arteriosclerosis of the middle-aged and elderly people or any one of symptoms of dizziness, headache, tinnitus, deafness, blurred vision, hypomnesis, soreness and weakness of waist and knees and numbness of limbs caused by the mild cerebral arteriosclerosis or complications thereof.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a compound gingko effective part and extract for promoting blood circulation, and a preparation method and application thereof.

Background

The compound gingko oral liquid for promoting blood circulation consists of seven Chinese medicinal materials of ginkgo leaf, prepared fleece flower root, glossy privet fruit, red sage root, eucommia bark, medicinal cyathula root and uncaria stem with hooks. The medicine has effects of nourishing yin, invigorating kidney, dispersing stagnated liver qi, dredging collaterals, etc., and can be widely used for treating dizziness, headache, tinnitus, deafness, blurred vision, hypomnesis, soreness of waist and knees, and numbness of limbs caused by mild cerebral arteriosclerosis of middle aged and elderly people.

The research shows that the main active ingredients of the compound gingko vein-relaxing oral liquid comprise total flavonol glycosides, terpene lactones and the like. The total flavonol glycoside has the functions of promoting blood circulation to disperse blood clots, dredging meridian passage, relieving pain, astringing lung, relieving asthma, eliminating turbid pathogen, regulating fat, etc. and its effective components include quercetin, kaempferide, isorhamnetin, etc.

CN1596960A discloses the prescription composition of compound gingko vein-dredging oral liquid and the efficacy of the prescription components, including the efficacy of monarch drug ginkgo leaf, ministerial drug prepared fleece-flower root and other components and the interaction thereof, and verifies the clinical value of the compound gingko vein-dredging oral liquid in the aspects of reducing blood fat, softening cerebral vessels, increasing blood supply of brain tissues, improving the ischemia condition of the brain tissues, improving and relieving the clinical symptoms of patients with cerebral Arteriosclerosis (AS), improving the life quality and the like, and the compound gingko vein-dredging extract is prepared by the preparation steps of alcohol extraction, water decoction, alcohol precipitation and the like. However, the method has the defects of low extraction rate of total flavonol glycosides and low extraction rate of emodin, and the like.

CN104027428B discloses an improved preparation method of a compound gingko vein relaxing extract, which remarkably improves the treatment effect on various diseases caused by cerebral arteriosclerosis and has the prevention and treatment effect on senile dementia. However, the method has the defects of low extraction rate of total flavonol glycosides and low extraction rate of emodin, and the like.

Document 1 (zhang geng et al, high performance liquid chromatography for determining the content of total flavonol glycosides in compound ginkgo oral liquid for promoting blood circulation, 20 th of volume 27 of the Chinese pharmaceutical industry, 20 days of 10 months in 2018) establishes a High Performance Liquid Chromatography (HPLC) method for determining the content of total flavonol glycosides in compound ginkgo oral liquid for promoting blood circulation. The method has the advantages of simple operation, accurate result, good data repeatability and precision, strong specificity of the method, and the like, and is suitable for quality control of the compound gingko oral liquid for promoting blood circulation.

Disclosure of Invention

The invention aims to provide a compound gingko effective part for promoting blood circulation, which is prepared from 180 parts of gingko leaves, 180 parts of prepared fleece-flower roots, 90 parts of glossy privet fruits, 90 parts of salvia miltiorrhiza and 60 parts of uncaria, wherein the preparation method of the effective part comprises the following steps: weighing required amount of folium Ginkgo, radix Polygoni Multiflori Preparata, fructus Ligustri Lucidi, Saviae Miltiorrhizae radix and ramulus Uncariae cum uncis, pulverizing into coarse powder, adding 30-95% acidified ethanol water solution 3-18 times (volume/weight ratio) of the medicinal materials, heating and reflux-extracting for 1-4 times (each time for 0.5-5 hr), filtering, collecting acidified ethanol extractive solution, concentrating, and drying the collected acidified ethanol extractive solution.

In the preferred technical scheme of the invention, before heating reflux extraction, the medicinal materials are soaked after being added with acidified ethanol water solution, preferably for 1-12h, and more preferably for 2-10 h.

In the preferred technical scheme of the invention, the concentration of the acidified ethanol aqueous solution is 40-90%, and the concentration of the acidified ethanol aqueous solution is preferably 50-80%.

In a preferred technical scheme of the invention, the pH of the acidified ethanol solution is 1-6, preferably the pH of the acidified ethanol aqueous solution is 2-5, and more preferably the pH of the acidified ethanol aqueous solution is 3-4.

In a preferred technical scheme of the invention, the acid used for adjusting the pH of the acidified aqueous ethanol solution is any one or combination of hydrochloric acid, sulfuric acid, chloric acid, nitric acid, hydrobromic acid, hydrofluoric acid, phosphoric acid, sulfonic acid, malic acid, fumaric acid, citric acid, carboxylic acid, hydroxy acid, keto acid, acetic acid, oxalic acid, citric acid, succinic acid, formic acid, acetic acid, propionic acid, butyric acid, malonic acid, succinic acid, pyruvic acid, glutamic acid, tartaric acid, lactic acid, itaconic acid, ascorbic acid, fumaric acid, alpha-ketoglutaric acid and fruit acid.

In a preferred embodiment of the present invention, the acid concentration used to adjust the pH of the acidified aqueous ethanol solution is 0.5 to 8mol/L, preferably 0.8 to 6mol/L, more preferably 1 to 5 mol/L.

In the preferred technical scheme of the invention, the effective parts are extracted by adding acidified ethanol water solution into the medicinal materials and heating and refluxing for 2-3 times.

In the preferred technical scheme of the invention, the acidified ethanol water solution is added into the medicinal materials, and the heating reflux extraction time is 1.0-4 hours, preferably 1.0-3 hours.

In the preferred technical scheme of the invention, the adding amount of the acidified ethanol water solution in the medicinal materials is 4-15 times, preferably 5-12 times of the weight of the medicinal materials.

In the preferable technical scheme of the invention, the collected acidified ethanol extract or the combined collected acidified ethanol extract is cooled to 4-35 ℃, kept stand for 6-56h, filtered, supernatant is taken, concentrated and prepared into an extract with the relative density of 1.0-1.4 (measured at 60 ℃), preferably the relative density of the extract is 1.10-1.35 (measured at 60 ℃), and the prepared extract is preferably dried.

In a preferred embodiment of the present invention, the concentration is selected from any one of vacuum concentration, membrane concentration, and vacuum concentration, or a combination thereof.

In a preferred embodiment of the present invention, the drying is selected from any one or a combination of reduced pressure drying, vacuum drying, spray drying, atmospheric drying, and ebullient drying.

In the preferred technical scheme of the invention, the prepared dry extract is crushed, and the crushing method is preferably selected from any one or combination of a jet milling method, a fluid milling method, a colloid milling method, a jet milling method, a co-milling method and a ball milling method.

In the preferable technical scheme of the invention, the content of the total flavonol glycosides in the effective part is not less than 0.2%, preferably not less than 0.25%.

In the preferable technical scheme of the invention, the content of the emodin in the effective part is not less than 0.015 percent, and preferably not less than 0.02 percent.

The invention aims to provide a preparation method of an effective part with high total flavonol glycoside content, wherein the effective part is prepared from the following components in parts by mass, 180 parts of ginkgo leaf, 180 parts of prepared fleece-flower root, 90 parts of glossy privet fruit, 90 parts of red sage root and 60 parts of uncaria, and the preparation method of the effective part comprises the following steps: weighing required amount of folium Ginkgo, radix Polygoni Multiflori Preparata, fructus Ligustri Lucidi, Saviae Miltiorrhizae radix and ramulus Uncariae cum uncis, pulverizing into coarse powder, adding 30-95% acidified ethanol water solution 3-18 times (volume/weight ratio) of the medicinal materials, heating and reflux-extracting for 1-4 times (each time for 0.5-5 hr), filtering, collecting acidified ethanol extractive solution, concentrating, and drying the collected acidified ethanol extractive solution.

In the preferred technical scheme of the invention, before heating reflux extraction, the medicinal materials are soaked after being added with acidified ethanol water solution, preferably for 1-12h, and more preferably for 2-10 h.

In the preferred technical scheme of the invention, the concentration of the acidified ethanol aqueous solution is 40-90%, and the concentration of the acidified ethanol aqueous solution is preferably 50-80%.

In a preferred technical scheme of the invention, the pH of the acidified ethanol solution is 1-6, preferably the pH of the acidified ethanol aqueous solution is 2-5, and more preferably the pH of the acidified ethanol aqueous solution is 3-4.

In a preferred technical scheme of the invention, the acid used for adjusting the pH of the acidified aqueous ethanol solution is any one or combination of hydrochloric acid, sulfuric acid, chloric acid, nitric acid, hydrobromic acid, hydrofluoric acid, phosphoric acid, sulfonic acid, malic acid, fumaric acid, citric acid, carboxylic acid, hydroxy acid, keto acid, acetic acid, oxalic acid, citric acid, succinic acid, formic acid, acetic acid, propionic acid, butyric acid, malonic acid, succinic acid, pyruvic acid, glutamic acid, tartaric acid, lactic acid, itaconic acid, ascorbic acid, fumaric acid, alpha-ketoglutaric acid and fruit acid.

In a preferred embodiment of the invention, the acid concentration used to adjust the pH of the acidified aqueous ethanol solution is between 0.5 and 8mol/L, preferably between 0.8 and 6mol/L, more preferably between 1 and 5 mol/L.

In the preferred technical scheme of the invention, the effective parts are extracted by adding acidified ethanol water solution into the medicinal materials and heating and refluxing for 2-3 times.

In the preferred technical scheme of the invention, the acidified ethanol water solution is added into the medicinal materials, and the heating reflux extraction time is 1.0-4 hours, preferably 1.0-3 hours.

In the preferred technical scheme of the invention, the adding amount of the acidified ethanol water solution in the medicinal materials is 4-15 times, preferably 5-12 times of the weight of the medicinal materials.

In the preferable technical scheme of the invention, the collected acidified ethanol extract or the combined collected acidified ethanol extract is cooled to 4-35 ℃, kept stand for 6-56h, filtered, supernatant is taken, concentrated and prepared into an extract with the relative density of 1.0-1.4 (measured at 60 ℃), preferably the relative density of the extract is 1.10-1.35 (measured at 60 ℃), and the prepared extract is preferably dried.

In a preferred embodiment of the present invention, the concentration is selected from any one of vacuum concentration, membrane concentration, and vacuum concentration, or a combination thereof.

In a preferred embodiment of the present invention, the drying is selected from any one or a combination of reduced pressure drying, vacuum drying, spray drying, atmospheric drying, and ebullient drying.

In the preferred technical scheme of the invention, the prepared dry extract is crushed, and the crushing method is preferably selected from any one or combination of a jet milling method, a fluid milling method, a colloid milling method, a jet milling method, a co-milling method and a ball milling method.

In the preferable technical scheme of the invention, the content of the total flavonol glycosides in the effective part is not less than 0.2%, preferably not less than 0.25%.

In the preferable technical scheme of the invention, the content of the emodin in the effective part is not less than 0.015 percent, and preferably not less than 0.02 percent.

The invention also aims to apply the compound gingko effective part to prepare any one of compound gingko vein relaxing extract and compound gingko vein relaxing pharmaceutical composition.

In the preferred technical scheme of the invention, the pharmaceutical composition is a compound gingko vein-dredging oral preparation, and is preferably any one of compound gingko vein-dredging oral liquid, compound gingko vein-dredging granules, compound gingko vein-dredging capsules, compound gingko vein-dredging tablets, compound gingko vein-dredging powder and compound gingko vein-dredging soft capsules.

The invention aims to provide a compound ginkgo biloba extract with high content of total flavonol glycosides, which is prepared from 180 parts of ginkgo biloba leaves, 180 parts of prepared fleece-flower root, 90 parts of glossy privet fruit, 90 parts of salvia miltiorrhiza, 90 parts of eucommia bark, 90 parts of medicinal cyathula root and 60 parts of uncaria, wherein the preparation method of the extract comprises the following steps:

1) weighing required amount of folium Ginkgo, radix Polygoni Multiflori Preparata, fructus Ligustri Lucidi, Saviae Miltiorrhizae radix and ramulus Uncariae cum uncis, pulverizing into coarse powder, adding 30-95% acidified ethanol water solution 3-18 times (volume/weight ratio) of the medicinal materials, heating and reflux-extracting for 1-4 times (0.5-5 hr per time), filtering, collecting acidified ethanol extractive solution, and concentrating the acidified ethanol extractive solution to obtain extract I;

2) weighing eucommia ulmoides and medicinal cyathula root according to the prescription amount, crushing the eucommia ulmoides and medicinal cyathula root into coarse powder, adding water which is 3-20 times (volume weight ratio) of the total amount of medicinal materials, decocting and extracting for 1-4 times, extracting for 0.5-4 hours each time, filtering, concentrating decoction, adding ethanol until the ethanol content reaches 30-90%, cooling the decoction to 4-35 ℃, standing for 6-56 hours, filtering, concentrating supernate and recovering ethanol to obtain an extract II;

(3) and mixing the prepared first extract and the second extract, and drying, or respectively drying the prepared first extract and the prepared second extract and mixing.

In the preferable technical scheme of the invention, before the heating reflux extraction in the step 1), the medicinal materials are soaked after the acidified ethanol water solution is added, preferably soaked for 1-12h, and more preferably soaked for 2-10 h.

In the preferred technical scheme of the invention, the concentration of the acidified ethanol aqueous solution is 40-90%, and the concentration of the acidified ethanol aqueous solution is preferably 50-80%.

In a preferred technical scheme of the invention, the pH of the acidified ethanol solution is 1-6, preferably the pH of the acidified ethanol aqueous solution is 2-5, and more preferably the pH of the acidified ethanol aqueous solution is 3-4.

In a preferred technical scheme of the invention, the acid used for adjusting the pH of the acidified aqueous ethanol solution is any one or combination of hydrochloric acid, sulfuric acid, chloric acid, nitric acid, hydrobromic acid, hydrofluoric acid, phosphoric acid, sulfonic acid, malic acid, fumaric acid, citric acid, carboxylic acid, hydroxy acid, keto acid, acetic acid, oxalic acid, citric acid, succinic acid, formic acid, acetic acid, propionic acid, butyric acid, malonic acid, succinic acid, pyruvic acid, glutamic acid, tartaric acid, lactic acid, itaconic acid, ascorbic acid, fumaric acid, alpha-ketoglutaric acid and fruit acid.

In a preferred embodiment of the present invention, the acid concentration used to adjust the pH of the acidified aqueous ethanol solution is 0.5 to 8mol/L, preferably 0.8 to 6mol/L, more preferably 1 to 5 mol/L.

In the preferred technical scheme of the invention, the effective part is extracted by adding acidified ethanol water solution into the medicinal materials in the step 1) and heating and refluxing for 2-3 times.

In the preferable technical scheme of the invention, the acidified ethanol water solution is added into the medicinal materials in the step 1) for heating reflux extraction for 1.0-4 hours, and the preferable time is 1.0-3 hours.

In the preferred technical scheme of the invention, the adding amount of the acidified ethanol aqueous solution in the medicinal materials in the step 1) is 4-15 times, preferably 5-12 times of the weight of the medicinal materials.

In the preferable technical scheme of the invention, the acidified ethanol extract collected in the step 1) or the acidified ethanol extract combined and collected in the step is cooled to 4-35 ℃, kept stand for 4-56h, filtered, taken out of supernatant, concentrated and prepared into an extract with the relative density of 1.0-1.4 (measured at 60 ℃), and the relative density of the extract is preferably 1.10-1.35 (measured at 60 ℃).

In the preferable technical scheme of the invention, in the step 2), before heating, decocting and extracting, the medicinal materials are soaked in water for 1-12h, preferably for 2-10 h.

In the preferred technical scheme of the invention, the Chinese medicinal materials in the step 2) are added with water and decocted for 2-3 times.

In the preferable technical scheme of the invention, the time for decocting and extracting the medicinal materials in the step 2) is 1-3.5 hours, and preferably 1.5-3 hours.

In the preferred technical scheme of the invention, the collected water extract or the combined collected water extract is cooled to 4-35 ℃, kept stand for 4-56h, filtered, supernatant fluid is taken, concentrated and prepared into extract with the relative density of 1.0-1.4 (measured at 60 ℃), and the relative density of the extract is preferably 1.10-1.35 (measured at 60 ℃).

According to the preferable technical scheme of the invention, the relative density of the extract I and the extract II is 1.0-1.4 (measured at 60 ℃), and the preferred relative density is 1.1-1.35 (measured at 60 ℃).

In a preferred embodiment of the present invention, the concentration is selected from any one of vacuum concentration, membrane concentration, and vacuum concentration, or a combination thereof.

In a preferred embodiment of the present invention, the drying is selected from any one or a combination of reduced pressure drying, vacuum drying, spray drying, atmospheric drying, and ebullient drying.

In the preferred technical scheme of the invention, the prepared dry extract is crushed, and the crushing method is preferably selected from any one or combination of a jet milling method, a fluid milling method, a colloid milling method, a jet milling method, a co-milling method and a ball milling method.

In a preferred technical scheme of the invention, the content of the total flavonol glycosides in the extract is not less than 0.15%, preferably not less than 0.2%.

In a preferred technical scheme of the invention, the content of the emodin in the extract is not less than 0.01 percent, and preferably not less than 0.015 percent.

The invention also aims to provide a preparation method of the compound ginkgo biloba dredging extract containing the total flavonol glycoside effective part, the extract is prepared from 180 parts of ginkgo biloba leaves, 180 parts of prepared fleece-flower root, 90 parts of glossy privet fruit, 90 parts of salvia miltiorrhiza, 90 parts of eucommia bark, 90 parts of medicinal cyathula root and 60 parts of uncaria, and the preparation method of the extract comprises the following steps:

1) weighing required amount of folium Ginkgo, radix Polygoni Multiflori Preparata, fructus Ligustri Lucidi, Saviae Miltiorrhizae radix and ramulus Uncariae cum uncis, pulverizing into coarse powder, adding 30-95% acidified ethanol water solution 3-18 times (volume/weight ratio) of the medicinal materials, heating and reflux-extracting for 1-4 times (0.5-5 hr per time), filtering, collecting acidified ethanol extractive solution, and concentrating the acidified ethanol extractive solution to obtain extract I;

2) weighing eucommia ulmoides and medicinal cyathula root according to the prescription amount, crushing the eucommia ulmoides and medicinal cyathula root into coarse powder, adding water which is 3-20 times (volume weight ratio) of the total amount of medicinal materials, decocting and extracting for 1-4 times, extracting for 0.5-4 hours each time, filtering, concentrating decoction, adding ethanol until the ethanol content reaches 30-90%, cooling the decoction to 4-35 ℃, standing for 6-56 hours, filtering, concentrating supernate and recovering ethanol to obtain an extract II;

(3) and mixing the prepared first extract and the second extract, and drying, or respectively drying the prepared first extract and the prepared second extract and mixing.

In the preferable technical scheme of the invention, before the heating reflux extraction in the step 1), the medicinal materials are soaked after the acidified ethanol water solution is added, preferably soaked for 1-12h, and more preferably soaked for 2-10 h.

In the preferred technical scheme of the invention, the concentration of the acidified ethanol aqueous solution is 40-90%, and the concentration of the acidified ethanol aqueous solution is preferably 50-80%.

In a preferred technical scheme of the invention, the pH of the acidified ethanol solution is 1-6, preferably the pH of the acidified ethanol aqueous solution is 2-5, and more preferably the pH of the acidified ethanol aqueous solution is 3-4.

In a preferred technical scheme of the invention, the acid used for adjusting the pH of the acidified aqueous ethanol solution is any one or combination of hydrochloric acid, sulfuric acid, chloric acid, nitric acid, hydrobromic acid, hydrofluoric acid, phosphoric acid, sulfonic acid, malic acid, fumaric acid, citric acid, carboxylic acid, hydroxy acid, keto acid, acetic acid, oxalic acid, citric acid, succinic acid, formic acid, acetic acid, propionic acid, butyric acid, malonic acid, succinic acid, pyruvic acid, glutamic acid, tartaric acid, lactic acid, itaconic acid, ascorbic acid, fumaric acid, alpha-ketoglutaric acid and fruit acid.

In a preferred embodiment of the present invention, the acid concentration used to adjust the pH of the acidified aqueous ethanol solution is 0.5 to 8mol/L, preferably 0.8 to 6mol/L, more preferably 1 to 5 mol/L.

In the preferred technical scheme of the invention, the effective part is extracted by adding acidified ethanol water solution into the medicinal materials in the step 1) and heating and refluxing for 2-3 times.

In the preferable technical scheme of the invention, the acidified ethanol water solution is added into the medicinal materials in the step 1) for heating reflux extraction for 1.0-4 hours, and the preferable time is 1.0-3 hours.

In the preferred technical scheme of the invention, the adding amount of the acidified ethanol aqueous solution in the medicinal materials in the step 1) is 4-15 times, preferably 5-12 times of the weight of the medicinal materials.

In the preferable technical scheme of the invention, the acidified ethanol extract collected in the step 1) or the acidified ethanol extract combined and collected in the step is cooled to 4-35 ℃, kept stand for 4-56h, filtered, taken out of supernatant, concentrated and prepared into an extract with the relative density of 1.0-1.4 (measured at 60 ℃), and the relative density of the extract is preferably 1.10-1.35 (measured at 60 ℃).

In the preferable technical scheme of the invention, in the step 2), before heating, decocting and extracting, the medicinal materials are soaked in water for 1-12h, preferably for 2-10 h.

In the preferred technical scheme of the invention, the Chinese medicinal materials in the step 2) are added with water and decocted for 2-3 times.

In the preferable technical scheme of the invention, the time for decocting and extracting the medicinal materials in the step 2) is 1-3.5 hours, and preferably 1.5-3 hours.

In the preferred technical scheme of the invention, the collected water extract or the combined collected water extract is cooled to 4-35 ℃, kept stand for 4-56h, filtered, supernatant fluid is taken, concentrated and prepared into extract with the relative density of 1.0-1.4 (measured at 60 ℃), and the relative density of the extract is preferably 1.10-1.35 (measured at 60 ℃).

According to the preferable technical scheme of the invention, the relative density of the extract I and the extract II is 1.0-1.4 (measured at 60 ℃), and the preferred relative density is 1.1-1.35 (measured at 60 ℃).

In a preferred embodiment of the present invention, the concentration is selected from any one of vacuum concentration, membrane concentration, and vacuum concentration, or a combination thereof.

In a preferred embodiment of the present invention, the drying is selected from any one or a combination of reduced pressure drying, vacuum drying, spray drying, atmospheric drying, and ebullient drying.

In the preferred technical scheme of the invention, the prepared dry extract is crushed, and the crushing method is preferably selected from any one or combination of a jet milling method, a fluid milling method, a colloid milling method, a jet milling method, a co-milling method and a ball milling method.

In a preferred technical scheme of the invention, the content of the total flavonol glycosides in the extract is not less than 0.15%, preferably not less than 0.2%.

In a preferred technical scheme of the invention, the content of the emodin in the extract is not less than 0.01 percent, and preferably not less than 0.015 percent.

The invention also aims to provide a compound gingko vein relaxing pharmaceutical composition which consists of a compound gingko vein relaxing extract and a pharmaceutically acceptable carrier.

The pharmaceutical compositions of the present invention may be in various dosage forms well known in the art and may be prepared using formulation techniques conventional in the art. The preparation suitable for the invention is selected from oral preparation or external preparation, preferably oral preparation.

In a preferred technical scheme of the invention, the oral preparation is selected from any one of oral liquid preparation, tablets, capsules, granules, syrup, powder, distillate, effervescent agent, spray, suspension, pills, dripping pills, mixture, paste, emulsion and tea.

In a preferred technical scheme of the invention, the external preparation is any one selected from gels, ointments, emplastrums, creams, ointments, liniments, lotions, suppositories, smearing agents, jellies, ointments and atomizing agents.

In a preferred embodiment of the present invention, the pharmaceutically acceptable carrier is a conventional excipient or adjuvant known in the art for preparing a desired preparation. The common excipients or auxiliary materials of the oral preparation or the external preparation comprise but are not limited to a filler (also called a diluent), a lubricant (also called a glidant or an anti-sticking agent), a dispersing agent, a wetting agent, an adhesive, a regulator, a solubilizer, an antioxidant, a bacteriostatic agent, an emulsifier, a flavoring agent, an aromatizer and the like. Binders, such as syrup, acacia, gelatin, sorbitol, tragacanth, cellulose or derivatives thereof, gelatin slurry, starch slurry, polyvinylpyrrolidone, and the like, preferably the cellulose derivative is selected from microcrystalline cellulose, sodium carboxymethylcellulose, ethylcellulose, hydroxypropylmethylcellulose, and the like, preferably the starch derivative is selected from any one of sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch, or combinations thereof. Fillers, such as lactose, powdered sugar, dextrin, starch or derivatives thereof, cellulose or derivatives thereof, inorganic calcium salts, sorbitol, glycine and the like, preferably the inorganic calcium salts are selected from calcium chloride, calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate and the like, preferably the cellulose derivatives are selected from any one of microcrystalline cellulose, sodium carboxymethyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, preferably the starch derivatives are selected from any one of sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch or combinations thereof; lubricants such as aerosil, magnesium stearate, talc, colloidal silica, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol and the like; disintegrants, such as starch or its derivatives, polyvinylpyrrolidone, cross-linked polyvinylpyrrolidone, microcrystalline cellulose, etc., preferably starch derivatives selected from any one or combination of sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch; wetting agents such as sodium lauryl sulfate, water or alcohols, and the like. Antioxidants, such as sodium sulfite, sodium bisulfite, sodium metabisulfite, dibutylbenzoic acid, and the like. Bacteriostatic agents (bactericides), for example, 0.5% phenol, 0.3% cresol, 0.5% chlorobutanol, etc. Examples of the acid-base modifier (pH modifier) include hydrochloric acid, citric acid, sodium (potassium) hydroxide, sodium (potassium) citrate, sodium (potassium) malate, sodium (potassium) dihydrogen phosphate, and disodium (potassium) hydrogen phosphate. Emulsifiers such as polysorbate-80, sorbitan elatate, pluronic F-68, lecithin, soy lecithin, etc. Solubilizers, such as Tween-80, bile, glycerol, etc. Flavoring agents, such as honey, syrup, and the like.

In a preferred technical scheme of the invention, the compound ginkgo biloba extract is mixed with a pharmaceutically acceptable sustained release preparation carrier or controlled release preparation carrier according to the preparation requirements, and then the mixture is prepared into pellets, such as sustained release pellets or controlled release pellets, according to the preparation method of the sustained release preparation or controlled release preparation known in the art, such as adding a retardant coating or microencapsulating the compound ginkgo biloba extract. The slow release preparation carrier or the controlled release preparation carrier comprises but is not limited to an oil-fat doping agent, a hydrophilic colloid or a coating retarder and the like, wherein the oil-fat doping agent is selected from glyceryl monostearate, hydrogenated castor oil, mineral oil, polysiloxane or dimethyl siloxane and the like; the hydrophilic colloid is selected from any one or combination of sodium carboxymethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone (PVP), acacia gum, tragacanth gum or carbopol; the coating retarder is selected from Ethyl Cellulose (EC), hydroxypropyl methyl cellulose (HMPC), polyvinylpyrrolidone (PVP), Cellulose Acetate Phthalate (CAP), acrylic resin, etc.

In a preferred embodiment of the present invention, the amount or type of the pharmaceutically acceptable carrier in the pharmaceutical composition is determined by the physicochemical properties and content of the active ingredient in the pharmaceutical composition, the type of the preparation, the dissolution and bioavailability of the preparation, and other factors.

The invention also aims to provide application of the compound gingko vein relaxing extract or the pharmaceutical composition thereof in preparing a medicine for treating middle-aged and elderly people mild cerebral arteriosclerosis or any disease symptoms or complicating diseases thereof caused by the cerebral arteriosclerosis, such as dizziness, headache, tinnitus, deafness, blurred vision, hypomnesis, soreness and weakness of waist and knees and limb numbness.

The invention also aims to provide application of the compound gingko vein relaxing extract or the pharmaceutical composition thereof in preparing medicines for treating any one of symptoms caused by cerebral arteriosclerosis and cardiovascular and cerebrovascular diseases or complications thereof.

The invention also aims to provide application of the compound gingko vein relaxing extract or the pharmaceutical composition thereof in preparing medicines for preventing and treating senile dementia or complications thereof.

The invention detects the contents of total flavonol glycosides and emodin in the compound gingko effective part or the compound gingko vein relaxing extract or the compound gingko vein relaxing pharmaceutical composition by high performance liquid chromatography so as to control the quality and the curative effect.

Unless otherwise stated, the present invention detects the content of total flavonol glycosides in a sample (such as compound ginkgo effective part, compound ginkgo extract or compound ginkgo preparation for promoting blood circulation, etc.) according to a High Performance Liquid Chromatography (HPLC) method disclosed in document 1. The chromatographic column is Shimadzu C₁₈Column (250mm × 4.6.6 mm,5 μm) with mobile phase of methanol-0.4% phosphoric acid solutionThe flow rate of the solution (55:45) was 1.0mL/min, the column temperature was 25 ℃, the sample volume was 10. mu.l, the detection wavelength was 360nm, and the amount was determined by external standard method. The theoretical plate number is not lower than 2500 calculated according to the peak of quercetin, and the separation degree is more than 1.5 calculated according to the peak of kaempferide and isorhamnetin. The sample to be tested is heated and refluxed for 90min in water bath at 100 ℃ for acid hydrolysis.

1.1 instruments

LC-20AT high performance liquid chromatograph (including LC-20AT quaternary pump, SPD-M20A UV detector, SIL-20A autosampler, LCsolution chromatography workstation, Shimadzu corporation, Japan); HH-6 model digital display constant temperature water bath (temperature control range 0.0-100.0 deg.C, Changzhou Zhi Bo Rui apparatus manufacturing Co., Ltd.); one hundred thousand electronic analytical balances (mettler-toledo corporation) model XSE105 DU; Direct-Q3 model water purifier (Michibo, USA).

1.2 reagent

Quercetin reference substance (purity of 98.1%), kaempferol reference substance (purity of 93.2%), and isorhamnetin reference substance (purity of 100.0%), all provided by China food and drug testing research institute for content determination; compound gingko oral liquid (specification 1 ml/bottle) for promoting blood circulation; negative samples (1 ml/vial size); methanol (chromatographically pure, merckkkgaa); phosphoric acid (chromatografic, Tianjin Saifuri science and technology, Inc.); hydrochloric acid (super pure, chemical reagents of national drug group, ltd.); the water was ultrapure water prepared by a Millipore direct-Q3 type water purifier, and all other reagents were analytical grade.

1.3 sample content determination

Respectively calculating the contents of quercetin, kaempferide and isorhamnetin according to the peak areas calculated by an external standard method, and converting the contents into the contents of total flavonol glycosides according to the following formula: the total flavonol glycoside content (quercetin content + kaempferol content + isorhamnetin content) × 2.51.

Unless otherwise stated, the content of emodin in a sample (such as compound ginkgo blood circulation promoting effective parts, compound ginkgo blood circulation promoting extracts or compound ginkgo blood circulation promoting preparations) to be detected is detected according to an emodin detection method described in document CN 1596960A.

Compared with the prior art, the invention has the following beneficial effects:

1. the invention scientifically screens the effective parts of the compound gingko vein relaxing, the alcohol extraction process of the compound gingko vein relaxing extract and the preparation conditions thereof, adopts acidified ethanol solution to extract the effective parts of the compound gingko vein relaxing, and uses the effective parts of the compound gingko vein relaxing extract or the medicine composition to prepare the compound gingko vein relaxing extract or the medicine composition, thereby obviously improving the content and the extraction rate of total flavonol glycosides and emodin in the effective parts, further obviously improving the quality and the curative effect of the medicine, obviously improving the resource utilization rate and obviously reducing the cost.

2. The invention further optimizes the technological parameters and conditions in alcohol extraction, water extraction and alcohol precipitation, and obviously improves the content and extraction rate of total flavonol glycosides and emodin in the effective parts, thereby obviously improving the quality and curative effect of the medicine, obviously improving the resource utilization rate and obviously reducing the cost.

3. The preparation method has the advantages of simple and convenient operation, contribution to comprehensive utilization of resources, contribution to industrial production and the like.

Detailed Description

The technical solution of the present invention is further illustrated by the following examples.

Comparative example 1Preparation of compound gingko effective part for promoting blood circulation

Comparative example 1 the preparation of the compound ginkgo effective part for promoting blood circulation comprises the following steps:

1) weighing 18g of ginkgo leaf, 18g of prepared fleece-flower root, 9g of glossy privet fruit, 9g of salvia miltiorrhiza and 6g of uncaria, crushing the ginkgo leaf, the prepared fleece-flower root, the glossy privet fruit, the salvia miltiorrhiza and the uncaria into coarse powder, adding 70% ethanol water solution (pH is 6.8), heating, carrying out reflux extraction for 1 hour, filtering, and collecting an extracting solution, wherein the adding amount of the ethanol water solution is 10 times (volume to weight ratio) of the total amount of the medicinal materials;

2) cooling the extracting solution collected in the step 1) to 25 ℃, standing for 12 hours, filtering, taking supernatant, concentrating the supernatant under reduced pressure, recovering ethanol until no alcohol smell exists, and performing spray drying to obtain the extract.

Comparative example 2Preparation of compound gingko effective part for promoting blood circulation

Comparative example 2 the preparation of the compound ginkgo effective part for promoting blood circulation comprises the following steps:

1) weighing 18g of ginkgo leaves, 18g of prepared fleece-flower roots, 9g of glossy privet fruits, 9g of salvia miltiorrhiza and 6g of uncaria rhynchophylla, crushing the ginkgo leaves, the processed ginkgo leaves, the prepared fleece-flower roots, the glossy privet fruits and the uncaria rhynchophylla into coarse powder, adding an alkalized ethanol aqueous solution (pH 8) with the concentration of 70%, heating, carrying out reflux extraction for 1 hour, filtering, and collecting an extracting solution, wherein the adding amount of the alkalized ethanol aqueous solution (pH 8) is 10 times (volume to weight ratio) of the total amount of medicinal materials, and the alkalized ethanol aqueous solution (pH 8) is adjusted by sodium hydroxide with;

2) cooling the extracting solution collected in the step 1) to 25 ℃, standing for 12 hours, filtering, taking supernatant, concentrating the supernatant under reduced pressure, recovering ethanol until no alcohol smell exists, and performing spray drying to obtain the extract.