阅读说明：本技术 治疗冠心病的药物组合物及其制备方法 (Medicinal composition for treating coronary heart disease and preparation method thereof ) 是由 雷晓琳 张春礼 王海珍 于 2021-09-27 设计创作，主要内容包括：本申请涉及中药的技术领域,具体公开了一种治疗冠心病的药物组合物及其制备方法。药物组合物的制备方法包括以下步骤：S1、将鹿衔草以60～70Vol.%的乙醇溶液提取,经处理得到鹿衔草清膏和鹿衔草药渣；将淫羊藿以65～75Vol.%的乙醇溶液提取,经处理得到淫羊藿清膏和淫羊藿药渣；S2、将鹿衔草药渣、淫羊藿药渣和其他除绞股蓝总苷外的四味药混合后水提,最终得到混合清膏；S3、将混合清膏、鹿衔草清膏以及淫羊藿清膏混合后经醇沉、浓缩,得稠膏；S4、将绞股蓝总苷加至稠膏后加水稀释、静置、过滤并将滤液灌装,即得。本申请的药物组合物具有滋阴补肾效果以及优异的对心肌损伤后的保护效果。(The application relates to the technical field of traditional Chinese medicines, and particularly discloses a pharmaceutical composition for treating coronary heart disease and a preparation method thereof. The preparation method of the pharmaceutical composition comprises the following steps: s1, extracting the pyrola with 60-70 Vol.% ethanol solution, and processing to obtain pyrola clear paste and pyrola herb residues; extracting epimedium with 65-75 Vol.% of ethanol solution, and processing to obtain epimedium clear paste and epimedium herb residues; s2, mixing herba Pyrolae dregs, herba Epimedii dregs and other four medicines except herba Gynostemmatis total glycosides, and extracting with water to obtain mixed fluid extract; s3, mixing the mixed clear paste, the Chinese pyrola herb clear paste and the epimedium clear paste, precipitating with ethanol, and concentrating to obtain thick paste; s4, adding gypenosides into the soft extract, diluting with water, standing, filtering, and packaging the filtrate. The pharmaceutical composition has the effects of nourishing yin and tonifying kidney and excellent effect of protecting the myocardium after injury.)

1. The preparation method of the pharmaceutical composition for treating coronary heart disease is characterized in that the raw materials for preparing the pharmaceutical composition for treating coronary heart disease comprise ginkgo leaf, salvia miltiorrhiza, gypenoside, corydalis tuber, hawthorn, pyrola and epimedium herb;

the preparation method comprises the following steps:

s1, soaking the pyrola in 60-70 Vol.% ethanol solution, performing reflux extraction, taking pyrola extracting solution, removing an extracting agent to obtain pyrola clear paste, and taking pyrola herb residue for later use;

soaking the epimedium in 65-75 Vol.% ethanol solution, performing reflux extraction, taking an epimedium extracting solution, removing an extracting agent to obtain an epimedium clear paste, and taking epimedium herb residues for later use;

s2, crushing folium Ginkgo, Saviae Miltiorrhizae radix, rhizoma corydalis and fructus crataegi respectively to obtain crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi;

decocting the herba Pyrolae residue, herba Epimedii residue, crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi in water to obtain mixed extractive solution; removing water from the mixed extractive solution to obtain mixed fluid extract;

s3, mixing the mixed clear paste, the pyrola clear paste and the epimedium clear paste, precipitating with ethanol, filtering, and concentrating the solid substances to obtain thick paste;

s4, adding the gypenosides into the thick paste, adding water for dilution, stirring uniformly, standing, filtering, and filling the filtrate to obtain the gynostemma pentaphylla extract.

2. The preparation method according to claim 1, wherein the process of obtaining the mixed fluid extract in step S2 comprises the following steps:

s21, crushing folium Ginkgo, Saviae Miltiorrhizae radix, rhizoma corydalis and fructus crataegi respectively to obtain crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi;

s22, decocting the crushed hawthorn with water to obtain a primary hawthorn extracting solution and a primary hawthorn extracting filter residue;

adding the primary extract of hawthorn into the crushed ginkgo leaf, decocting and extracting, adding water, decocting and extracting, and removing water from the extract to obtain ginkgo leaf clear paste;

mixing the primary extraction residue of fructus crataegi with the pulverized Saviae Miltiorrhizae radix and rhizoma corydalis, decocting with water, and removing water to obtain initial mixed fluid extract;

s23, mixing the initial mixed fluid extract and the ginkgo leaf fluid extract to obtain mixed fluid extract.

3. The preparation method according to claim 1, wherein the process of obtaining the mixed fluid extract in step S2 comprises the following steps:

s21, crushing folium Ginkgo, Saviae Miltiorrhizae radix, rhizoma corydalis and fructus crataegi respectively to obtain crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi;

s22, decocting the crushed hawthorn with water to obtain a primary hawthorn extracting solution and a primary hawthorn extracting filter residue;

decocting the crushed red sage root in water to obtain red sage root primary extract and red sage root primary extraction filter residue;

adding fructus crataegi primary extract and Saviae Miltiorrhizae radix primary extract into folium Ginkgo crushed material, decocting, adding water, decocting, and removing water to obtain folium Ginkgo fluid extract;

mixing the primary extraction residue of fructus crataegi, the primary extraction residue of Saviae Miltiorrhizae radix, and the crushed material of rhizoma corydalis, decocting in water, and removing water to obtain initial mixed fluid extract;

s23, mixing the initial mixed fluid extract and the ginkgo leaf fluid extract to obtain mixed fluid extract.

4. The preparation method according to claim 3, wherein the time for decocting the crushed Danshen root is 2.5 to 4.5 hours.

5. The method according to claim 1, wherein in step S1, the wintergreen is soaked in 60-70 Vol.% ethanol solution for 25-32 h, and the reflux extraction time is 1-2.5 h; the time for soaking herba epimedii in 65-75 Vol.% ethanol solution is 25-32 hours, and the time for reflux extraction is 1-2.5 hours.

6. The preparation method according to claim 1, wherein in step S1, when the pyrola is soaked in 60-70 Vol.% ethanol solution, the weight ratio of the feed liquid is 1: 9-12; when the epimedium is soaked in 65-75 Vol.% ethanol solution, the weight ratio of the feed liquid is 1: 9-12.

7. The preparation method of claim 1, wherein the pharmaceutical composition for treating coronary heart disease comprises the following raw materials in parts by weight:

18-22 parts of ginkgo leaves, 8-12 parts of salvia miltiorrhiza, 0.01-0.02 part of gynostemma pentaphylla total glycosides, 1.4-1.9 parts of corydalis tuber, 13-17 parts of hawthorn, 13-17 parts of pyrola and 13-17 parts of epimedium herb.

8. A pharmaceutical composition for treating coronary heart disease, characterized by being prepared by the preparation method of any one of claims 1 to 7.

9. The pharmaceutical composition for treating coronary heart disease according to claim 8, wherein the content of 2' -0-galloylhyperin in the pharmaceutical composition for treating coronary heart disease is 3.34-4.25 mg/mL, the content of monotropein is 2.74-3.44 mg/mL, the content of icariin is 2.23-2.94 mg/mL, the content of total flavonol is 2.38-4.58 mg/mL, and the content of terpene lactone is 3.30-6.35 mg/mL.

Technical Field

The application relates to the technical field of traditional Chinese medicines, in particular to a pharmaceutical composition for treating coronary heart disease and a preparation method thereof.

Background

In the preparation of the pharmaceutical composition for treating coronary heart disease, the preparation raw materials comprise gypenoside, ginkgo leaves, salvia miltiorrhiza, corydalis tuber, hawthorn, pyrola and epimedium, and the preparation method comprises the following steps: crushing folium Ginkgo, Saviae Miltiorrhizae radix, rhizoma corydalis, fructus crataegi, herba Pyrolae and herba Epimedii, extracting with water, concentrating the extractive solution to obtain soft extract, precipitating with ethanol, mixing with herba Gynostemmatis total glycosides, adding sugar, diluting, and packaging to obtain the pharmaceutical composition for treating coronary heart disease.

Wherein the herba Pyrolae extract contains effective medicinal components including 2' -0-galloyl hyperoside and crystal orchid glycoside, and the herba Epimedii effective medicinal components including icariin; 2 '-0-galloylhyperin, monothioloside and icariin have the effect of nourishing qi and blood, and related researches show that 2' -0-galloylhyperin also has an obvious protective effect on myocardial ischemia reperfusion injury of rats.

However, the medicinal composition for treating coronary heart disease prepared by the method has insufficient content of effective components in pyrola and epimedium herb, so that the medicinal composition has certain effects of treating kidney qi deficiency, heart vessel stasis type coronary heart disease and angina pectoris, but has poor effects of nourishing qi and blood and protecting myocardial damage.

Disclosure of Invention

In order to further improve the content of 2' -0-galloyl hyperin, monothioloside and icariin in the prepared pharmaceutical composition for treating coronary heart disease, thereby improving the qi and blood nourishing effect and the protection effect after myocardial damage, the application provides the pharmaceutical composition for treating coronary heart disease and the preparation method thereof.

In a first aspect, the application provides a preparation method of a pharmaceutical composition for treating coronary heart disease, which adopts the following technical scheme:

the preparation method of the pharmaceutical composition for treating coronary heart disease comprises the following raw materials of ginkgo leaf, salvia miltiorrhiza, gynostemma pentaphylla total glycoside, corydalis tuber, hawthorn, pyrola and epimedium;

the preparation method comprises the following steps:

s1, soaking the pyrola in 60-70 Vol.% ethanol solution, performing reflux extraction, taking pyrola extracting solution, removing an extracting agent to obtain pyrola clear paste, and taking pyrola herb residue for later use;

soaking the epimedium in 65-75 Vol.% ethanol solution, performing reflux extraction, taking an epimedium extracting solution, removing an extracting agent to obtain an epimedium clear paste, and taking epimedium herb residues for later use;

s2, crushing folium Ginkgo, Saviae Miltiorrhizae radix, rhizoma corydalis and fructus crataegi respectively to obtain crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi;

decocting the herba Pyrolae residue, herba Epimedii residue, crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi in water to obtain mixed extractive solution; removing water from the mixed extractive solution to obtain mixed fluid extract;

s3, mixing the mixed clear paste, the pyrola clear paste and the epimedium clear paste, precipitating with ethanol, filtering, and concentrating the solid substances to obtain thick paste;

s4, adding the gypenosides into the thick paste, adding water for dilution, stirring uniformly, standing, filtering, and filling the filtrate to obtain the gynostemma pentaphylla extract.

By adopting the technical scheme, when the ethanol solutions with different concentrations are independently used as the extracting agents to extract the effective components in the pyrola, the extracting agents are 45-55 Vol.% of ethanol solutions, so that the extraction rate of the 2' -0-galloyl hyperin and the crystal orchid glycoside is excellent; when the effective components in the epimedium are extracted by refluxing by taking ethanol solutions with different concentrations as extracting agents, the extraction rate of the icariin is the highest when the extracting agent is 45-55 Vol. However, when the pyrola extract obtained by soaking and extracting in 45-55 Vol.% ethanol solution and the epimedium extract obtained by soaking and extracting in 45-55 Vol.% ethanol solution are used for preparing the pharmaceutical composition for treating coronary heart disease, the components in the pyrola extract and the epimedium extract can affect each other: the existing stability of the 2 '-0-galloylhyperin, the monotropein and the icariin in the system is poor, so that the 2' -0-galloylhyperin, the monotropein and the icariin in the finished product of the pharmaceutical composition for treating coronary heart disease are obviously lost, and finally the pharmaceutical effect of the finished product of the pharmaceutical composition for treating coronary heart disease is reduced.

However, when the pyrola extract obtained by soaking and extracting pyrola with 60-70 Vol.% ethanol solution and the epimedium extract obtained by soaking and extracting epimedium with 65-75 Vol.% ethanol solution are used for preparing the pharmaceutical composition for treating coronary heart disease, the following results are found: the content loss of the 2' -0-galloyl hyperin, the monotropein and the icariin in the finished medicinal composition for treating the coronary heart disease is obviously reduced, and the content of the active ingredients in the finished medicinal composition for treating the coronary heart disease is obviously improved, so that the effect of nourishing yin and tonifying kidney of the medicinal composition for treating the coronary heart disease is obviously improved, and the protective effect of myocardial damage is further improved, thereby achieving the aim of assisting in improving the treatment effect of the coronary heart disease.

Preferably, the process for obtaining the mixed clear paste in the step S2 comprises the following steps:

s21, crushing folium Ginkgo, Saviae Miltiorrhizae radix, rhizoma corydalis and fructus crataegi respectively to obtain crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi;

s22, decocting the crushed hawthorn with water to obtain a primary hawthorn extracting solution and a primary hawthorn extracting filter residue;

adding the primary extract of hawthorn into the crushed ginkgo leaf, decocting and extracting, adding water, decocting and extracting, and removing water from the extract to obtain ginkgo leaf clear paste;

mixing the primary extraction residue of fructus crataegi with the pulverized Saviae Miltiorrhizae radix and rhizoma corydalis, decocting with water, and removing water to obtain initial mixed fluid extract;

s23, mixing the initial mixed fluid extract and the ginkgo leaf fluid extract to obtain mixed fluid extract.

By adopting the technical scheme, if the ginkgo leaf, the salvia miltiorrhiza, the hawthorn and the corydalis tuber are extracted by water together, the ginkgo leaf extract is easily influenced by the hawthorn extract, so that part of substances in the ginkgo leaf extract are damaged, and the drug effect of the ginkgo leaf extract is reduced. The scheme of the application is as follows: the ginkgo leaf extract is obtained by adopting a mode of combining water extraction and primary extraction of hawthorn extract. Wherein, the primary hawthorn extracting solution obtained by extracting hawthorn for one time contains a certain amount of hawthorn extract; when the primary hawthorn extracting solution is used for extracting the ginkgo leaf extract, the primary hawthorn extracting solution can obviously improve the change of cell permeability of ginkgo leaf cells on one hand, so that the extraction effect of the ginkgo leaf extract is improved; meanwhile, because the content of the hawthorn extract in the primary hawthorn extracting solution is not high, the effective medicinal components in the ginkgo leaf extract are difficult to destroy to a large extent; in the pharmaceutical composition for treating coronary heart disease, the content of total flavonol and terpene lactone in the ginkgo leaf extract is obviously improved, which is beneficial to improving the treatment effect on coronary heart disease.

Preferably, the process for obtaining the mixed clear paste in the step S2 comprises the following steps:

s21, crushing folium Ginkgo, Saviae Miltiorrhizae radix, rhizoma corydalis and fructus crataegi respectively to obtain crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi;

s22, decocting the crushed hawthorn with water to obtain a primary hawthorn extracting solution and a primary hawthorn extracting filter residue;

decocting the crushed red sage root in water to obtain red sage root primary extract and red sage root primary extraction filter residue;

adding fructus crataegi primary extract and Saviae Miltiorrhizae radix primary extract into folium Ginkgo crushed material, decocting, adding water, decocting, and removing water to obtain folium Ginkgo fluid extract;

mixing the primary extraction residue of fructus crataegi, the primary extraction residue of Saviae Miltiorrhizae radix, and the crushed material of rhizoma corydalis, decocting in water, and removing water to obtain initial mixed fluid extract;

s23, mixing the initial mixed fluid extract and the ginkgo leaf fluid extract to obtain mixed fluid extract.

By adopting the technical scheme, a water extraction mode is further adopted, and the primary extracting solution of the hawthorn and the primary extracting solution of the salvia miltiorrhiza are combined to extract the ginkgo leaf extract, so that the extraction effect of effective medicinal substances of the ginkgo leaves is further improved, and the treatment effect of the pharmaceutical composition on the coronary heart disease can be further improved.

Preferably, when the crushed salvia miltiorrhiza bunge is decocted and extracted by water, the decoction time is 2.5-4.5 h.

By adopting the technical scheme, the control of the extraction degree of the salvia miltiorrhiza is realized by controlling the decoction time of the salvia miltiorrhiza, the content of the salvia miltiorrhiza extract in the primary extract of the salvia miltiorrhiza is ensured to be in a proper range, so that a more excellent ginkgo leaf extraction effect is finally obtained, and the drug effect of the pharmaceutical composition is finally and effectively improved.

Preferably, in step S1, the gerba pyrolae is soaked in 60-70 Vol.% ethanol solution for 25-32 hours, and the reflux extraction time is 1-2.5 hours; the time for soaking herba epimedii in 65-75 Vol.% ethanol solution is 25-32 hours, and the time for reflux extraction is 1-2.5 hours.

Preferably, in step S1, when the pyrola is soaked in 60-70 Vol.% ethanol solution, the weight ratio of the feed liquid is 1: 9-12; when the epimedium is soaked in 65-75 Vol.% ethanol solution, the weight ratio of the feed liquid is 1: 9-12.

Preferably, the pharmaceutical composition for treating coronary heart disease is prepared from the following raw materials in parts by weight:

18-22 parts of ginkgo leaves, 8-12 parts of salvia miltiorrhiza, 0.01-0.02 part of gynostemma pentaphylla total glycosides, 1.4-1.9 parts of corydalis tuber, 13-17 parts of hawthorn, 13-17 parts of pyrola and 13-17 parts of epimedium herb.

In a second aspect, the present application provides a pharmaceutical composition for treating coronary heart disease, which adopts the following technical scheme:

the pharmaceutical composition for treating coronary heart disease is prepared by the preparation method.

By adopting the technical scheme, when the pharmaceutical composition for treating coronary heart disease is prepared, the related external extracting agents are only ethanol and water, and other extracting agents are not introduced too much, so that the pharmaceutical composition for treating coronary heart disease is more green and healthy; meanwhile, the cost of the extractant is lower, so that the preparation cost of the product is low. In addition, the pharmaceutical composition for treating coronary heart disease prepared by the method has better curative effect on coronary heart disease, and effects of nourishing qi and blood and protecting cardiac muscle.

Preferably, the content of 2' -0-galloyl hyperin in the pharmaceutical composition for treating coronary heart disease is 3.34-4.25 mg/mL, the content of monotropein is 2.74-3.44 mg/mL, the content of icariin is 2.23-2.94 mg/mL, the content of total flavonol is 2.38-4.58 mg/mL, and the content of terpene lactone is 3.30-6.35 mg/mL.

By adopting the technical scheme, the content of each effective substance in the finished medicinal composition for treating the coronary heart disease is effectively improved, so that the content of the components with the functions of activating blood circulation, nourishing and protecting cardiac muscle of the finally prepared medicinal composition for treating the coronary heart disease is obviously improved, and the more obvious medicinal effect is obtained.

In summary, the present application has the following beneficial effects:

1. according to the application, the pyrola is soaked and refluxed with 60-70 Vol.% of ethanol solution and the epimedium is soaked and refluxed with 65-75 Vol.% of ethanol solution to extract the epimedium, so that the pyrola extract and the epimedium extract with proper components and contents are obtained, the loss of 2 '-0-galloyl hyperoside and monotropein in the active medicinal ingredients in the pyrola extract is effectively reduced, the loss of the icariin serving as the active medicinal ingredients in the epimedium is also effectively reduced, and finally, the contents of the 2' -0-galloyl hyperoside, the monotropein and the icariin with the nourishing effect in the finished product of the pharmaceutical composition for treating the coronary heart disease are obviously improved, so that the pharmaceutical composition for treating the coronary heart disease has excellent nourishing and myocardial protecting effects.

2. In the application, the folium ginkgo is preferably extracted by adopting a primary hawthorn extracting solution, so that the content of effective medicinal components in the folium ginkgo extract is obviously improved; the content of the total flavonol and the terpene lactone in the finished product of the pharmaceutical composition for treating the coronary heart disease is obviously improved, and the content is respectively as follows: 4.05-4.35 mg/mL, 5.05-5.91 mg/mL. Finally, a better coronary heart disease treatment effect is obtained.

3. According to the method, the ginkgo leaf is preferably extracted by taking the mixed solution of the primary hawthorn extract and the primary salvia miltiorrhiza extract as an extracting agent, so that the content of effective medicinal components in the ginkgo leaf extract is further improved; the content of the total flavonol and the terpene lactone in the finished product of the pharmaceutical composition for treating coronary heart disease is further improved, and the content is respectively as follows: 4.45-4.65 mg/mL, 6.05-6.65 mg/mL.

Detailed Description

The present application will be described in further detail with reference to examples.

The Pyrolae is purchased from Pyrolae in Taibai county of Shaanxi, contains 28 +/-0.5 mg/g of 2' -0-galloyl hyperin, is higher than the internal control requirement by 3.0mg/g, and can be used; the content of the monotropein is 24 + -0.5 mg/g, which is higher than 1.5mg/g of the internal control requirement, and can be used.

The content of icariin in herba Epimedii is 20 + -0.5 mg/g, which is higher than 6.0mg/g required for internal control.

Preparation example of herba Pyrolae clear paste

Preparation example 1 of herba Pyrolae clear paste

The preparation method of the herba pyrolae clear paste comprises the following steps:

pulverizing herba Pyrolae raw materials, and completely using in the preparation of herba Pyrolae fluid extract. Adding water 10 times the weight of herba Pyrolae into pulverized herba Pyrolae, soaking for 30h, heating and reflux-extracting for 1 time for 1.5h, filtering the extractive solution, retaining herba Pyrolae residue, concentrating the filtrate under reduced pressure to obtain herba Pyrolae fluid extract with relative density of 1.15 (50 deg.C), and keeping.

Preparation example 2 of herba Pyrolae fluid extract

Different from the preparation example 1 of the pyrola herb clear paste, the extractant is selected to be 30Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the pyrola herb clear paste.

Preparation example 3 of herba Pyrolae fluid extract

Different from the preparation example 1 of the pyrola herb clear paste, the extractant is selected to be 50Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the pyrola herb clear paste.

Preparation example 4 of herba Pyrolae fluid extract

Different from the preparation example 1 of the pyrola herb clear paste, the extractant is selected to be 60Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the pyrola herb clear paste.

Preparation example 5 of herba Pyrolae fluid extract

Different from the preparation example 1 of the pyrola herb clear paste, the extractant is selected to be 70Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the pyrola herb clear paste.

Preparation example 6 of herba Pyrolae fluid extract

Different from the preparation example 1 of the pyrola herb clear paste, the extractant is selected to be 90Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the pyrola herb clear paste.

Preparation example of Epimedium herb clear paste

Preparation example 1 of Epimedium herb extract

The preparation method of the epimedium herb clear paste comprises the following steps:

pulverizing herba Epimedii raw material, and completely using in preparation of herba Epimedii fluid extract. Adding water 10 times the weight of herba Epimedii into pulverized herba Epimedii, soaking for 30h, heating and reflux extracting for 1 time for 1.5h, filtering the extractive solution, retaining herba Epimedii residue, and concentrating the filtrate under reduced pressure to obtain herba Epimedii fluid extract with relative density of 1.15-1.20 (50 deg.C) for use.

Preparation example 2 of Epimedium herb extract

Different from the preparation example 1 of the epimedium clear paste, the extractant is selected to be 25Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the epimedium clear paste.

Preparation example 3 of Epimedium herb extract

Different from the preparation example 1 of the epimedium clear paste, the extractant is selected to be 50Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the epimedium clear paste.

Preparation example 4 of Epimedium herb extract

Different from the preparation example 1 of the epimedium clear paste, the extractant is selected to be 65Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the epimedium clear paste.

Preparation example 5 of Epimedium herb extract

Different from the preparation example 1 of the epimedium clear paste, the extractant is selected to be 75Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the epimedium clear paste.

Preparation example 6 of Epimedium herb extract

Different from the preparation example 1 of the epimedium clear paste, the extractant is selected to be 95Vol.% ethanol solution, and the other steps are the same as the preparation example 1 of the epimedium clear paste.

Examples

Example 1

The preparation raw materials of the pharmaceutical composition for treating coronary heart disease are as follows: 180g of ginkgo leaf, 130g of pyrola, 130g of epimedium, 80g of salvia miltiorrhiza, 0.1g of gypenoside, 14g of corydalis tuber and 130g of hawthorn.

The preparation method of the pharmaceutical composition for treating coronary heart disease comprises the following steps:

s1, taking all the pyrola, preparing the pyrola clear paste and the pyrola dregs by the method of the pyrola clear paste preparation example 4 for standby;

and taking all the epimedium herbs, and preparing the epimedium herb clear paste and the epimedium herb dregs by the method of the epimedium herb clear paste preparation example 4.

S2, preparing the mixed clear paste

S21, crushing the ginkgo biloba, the salvia miltiorrhiza, the corydalis tuber and the hawthorn respectively to obtain crushed ginkgo biloba, crushed salvia miltiorrhiza, crushed corydalis tuber and crushed hawthorn;

s22, mixing herba Pyrolae residue, herba Epimedii residue, crushed folium Ginkgo, crushed Saviae Miltiorrhizae radix, crushed rhizoma corydalis and crushed fructus crataegi, adding 3000mL of water, and soaking for 30 min. Then decocting with water for three times: adding water for the first time, wherein the water is 8 times of the total weight of the soaked medicinal materials (the total weight of herba Pyrolae crushed material, herba Epimedii crushed material, folium Ginkgo crushed material, Saviae Miltiorrhizae radix crushed material, rhizoma corydalis crushed material and fructus crataegi crushed material), and decocting for 1.5 hr; adding water for the second time, wherein the weight of the water is 5 times of the total weight of the soaked medicinal materials, and decocting for 1 h; adding water 4 times the total weight of the soaked materials for the third time, decocting for 45min, mixing the decoctions, filtering, concentrating the filtrate under reduced pressure to obtain mixed fluid extract with relative density of 1.25 (60 deg.C), and cooling.

S3, mixing the mixed clear paste prepared in the step S2, the Chinese pyrola herb clear paste prepared in the step S1 and the epimedium clear paste, adding ethanol to ensure that the ethanol content reaches 65wt%, then standing, filtering, recovering the ethanol, and concentrating to form thick paste with the relative density of 1.30 (60 ℃).

S4, adding 10mL of water into the gypenosides to dissolve the gypenosides, adding the dissolved gypenosides into the thick paste obtained in the step S3, then adding 167g of 50wt% simple syrup, adding water to 1000mL, uniformly stirring, and standing for 10 days at 1-5 ℃; removing the filter residue after the filtrate is removed, and filling the filtrate to obtain the final product.

Example 2

The preparation raw materials of the pharmaceutical composition for treating coronary heart disease are as follows: 200g of ginkgo leaf, 150g of pyrola, 150g of epimedium, 100g of salvia miltiorrhiza, 0.167g of gypenosides, 16.7g of corydalis tuber and 150g of hawthorn.

The preparation method of the pharmaceutical composition for treating coronary heart disease comprises the following steps:

s1, taking all the pyrola, preparing the pyrola clear paste and the pyrola dregs by the method of the pyrola clear paste preparation example 4 for standby;

and taking all the epimedium herbs, and preparing the epimedium herb clear paste and the epimedium herb dregs by the method of the epimedium herb clear paste preparation example 5.

S2, same as example 1.

S3, mixing the mixed clear paste prepared in the step S2, the Chinese pyrola herb clear paste prepared in the step S1 and the epimedium clear paste, adding ethanol to ensure that the ethanol content reaches 65wt%, then standing, filtering, recovering the ethanol, and concentrating to form thick paste with the relative density of 1.30 (60 ℃).

S4, adding 10mL of water into the gypenosides to dissolve the gypenosides, adding the dissolved gypenosides into the thick paste obtained in the step S3, then adding 167g of 50wt% simple syrup, adding water to 1000mL, uniformly stirring, and standing for 10 days at 1-5 ℃; removing the filter residue after the filtrate is removed, and filling the filtrate to obtain the final product.

Example 3

The preparation raw materials of the pharmaceutical composition for treating coronary heart disease are as follows: 220g of ginkgo leaves, 170g of pyrola, 170g of epimedium, 120g of salvia miltiorrhiza, 0.2g of gypenoside, 19g of corydalis tuber and 170g of hawthorn.

The preparation method of the pharmaceutical composition for treating coronary heart disease comprises the following steps:

s1, taking all the pyrola, preparing the pyrola clear paste and the pyrola dregs by the method of the pyrola clear paste preparation example 5 for standby;

and taking all the epimedium herbs, and preparing the epimedium herb clear paste and the epimedium herb dregs by the method of the epimedium herb clear paste preparation example 5.

S2, same as example 1.

S3, mixing the mixed clear paste prepared in the step S2, the Chinese pyrola herb clear paste prepared in the step S1 and the epimedium clear paste, adding ethanol to ensure that the ethanol content reaches 65wt%, then standing, filtering, recovering the ethanol, and concentrating to form thick paste with the relative density of 1.30 (60 ℃).

S4, adding 10mL of water into the gypenosides to dissolve the gypenosides, adding the dissolved gypenosides into the thick paste obtained in the step S3, then adding 167g of 50wt% simple syrup, adding water to 1000mL, uniformly stirring, and standing for 10 days at 1-5 ℃; removing the filter residue after the filtrate is removed, and filling the filtrate to obtain the final product.

Example 4

The difference between this embodiment and embodiment 2 is that, when preparing the pharmaceutical composition for treating coronary heart disease, step S2 is different, specifically:

s21, crushing the ginkgo leaves, the salvia miltiorrhiza, the corydalis tuber and the hawthorns with the same weight as that in the embodiment 2 respectively to obtain crushed ginkgo leaves, crushed salvia miltiorrhiza, crushed corydalis tuber and crushed hawthorn;

s22, adding 10 times of water into the crushed hawthorn, soaking for 30min, decocting and extracting for 1.5h, and then filtering to obtain a primary hawthorn extracting solution and a primary hawthorn extracting filter residue.

Adding 800mL of water into the crushed ginkgo leaves, and then soaking for 30 min. Adding the fructus crataegi primary extractive solution into crushed folium Ginkgo, and decocting for 1.5 hr. After filtration, the filter residue is extracted with water twice: adding water 5 times the weight of crushed folium Ginkgo for the first time, decocting for 1 hr, and filtering; adding water 4 times the weight of crushed folium Ginkgo for the second time, decocting for 45min, and filtering; mixing the three decoctions, filtering, concentrating the filtrate under reduced pressure to obtain folium Ginkgo fluid extract with relative density of 1.20 (60 deg.C), and cooling.

Mixing the residue after the primary extraction of hawthorn, the crushed red sage root and the crushed corydalis tuber, adding 1600mL of water, and soaking for 30 min. Then decocting with water for three times: adding water in an amount of 8 times of the total weight of the soaked materials (the total weight of fructus crataegi crushed material, Saviae Miltiorrhizae radix crushed material and rhizoma corydalis crushed material) for the first time, and decocting for 1.5 hr; adding water for the second time, wherein the weight of the water is 5 times of the total weight of the soaked medicinal materials, and decocting for 1 h; adding water 4 times the total weight of the soaked materials for the third time, decocting for 45min, mixing the decoctions, filtering, concentrating the filtrate under reduced pressure to obtain initial mixed fluid extract with relative density of 1.20 (60 deg.C), and cooling.

S23, mixing the initial mixed fluid extract and the ginkgo leaf fluid extract, and stirring uniformly to obtain mixed fluid extract.

The rest is the same as example 2.

Example 5

The difference between this embodiment and embodiment 2 is that, when preparing the pharmaceutical composition for treating coronary heart disease, step S2 is different, specifically:

s21, crushing the ginkgo leaves, the salvia miltiorrhiza, the corydalis tuber and the hawthorns with the same weight as that in the embodiment 2 respectively to obtain crushed ginkgo leaves, crushed salvia miltiorrhiza, crushed corydalis tuber and crushed hawthorn;

s22, adding 10 times of water into the crushed hawthorn, soaking for 30min, decocting and extracting for 1.5h, and then filtering to obtain a primary hawthorn extracting solution and a primary hawthorn extracting filter residue.

Adding 10 times of water into the crushed red sage root for soaking for 30min, decocting and extracting for 1.5h, and then filtering to obtain a red sage root primary extract and a red sage root primary extraction filter residue.

Adding 800mL of water into the crushed ginkgo leaves, and then soaking for 30 min. Adding the crushed folium Ginkgo extract into the primary extractive solution of fructus crataegi and Saviae Miltiorrhizae radix, and decocting for 1.5 hr. After filtration, the filter residue is extracted with water twice: adding water 5 times the weight of crushed folium Ginkgo for the first time, decocting for 1 hr, and filtering; adding water 4 times the weight of crushed folium Ginkgo for the second time, decocting for 45min, and filtering; mixing the three decoctions, filtering, concentrating the filtrate under reduced pressure to obtain folium Ginkgo fluid extract with relative density of 1.15 (60 deg.C), and cooling.

Mixing the primary extraction filter residue of hawthorn, the primary extraction filter residue of salvia miltiorrhiza and the crushed material of corydalis tuber, adding 800mL of water, and soaking for 30 min. Then decocting twice with water: adding water 5 times the total weight of the soaked materials (the total weight of fructus crataegi crushed material, Saviae Miltiorrhizae radix crushed material and rhizoma corydalis crushed material) for the first time, and decocting for 1 hr; adding water 4 times the total weight of the soaked materials for the second time, decocting for 45min, mixing the decoctions, filtering, concentrating the filtrate under reduced pressure to obtain initial mixed fluid extract with relative density of 1.20 (60 deg.C), and cooling.

S23, mixing the initial mixed fluid extract and the ginkgo leaf fluid extract, and stirring uniformly to obtain mixed fluid extract.

The rest is the same as example 2.

Comparative example 1

The difference between the comparative example and the example 2 is that in the step S1, the method of the preparation example 1 of the epimedium clear paste is selected for preparation of the epimedium clear paste.

The rest is the same as example 2.

Comparative example 2

The difference between the comparative example and the comparative example 1 is that in the step S1, the method of the preparation example 2 of the epimedium clear paste is selected for preparation.

The rest is the same as in comparative example 1.

Comparative example 3

The difference between the comparative example and the comparative example 1 is that in the step S1, the method of the preparation example 3 of the epimedium clear paste is selected for preparation.

The rest is the same as in comparative example 1.

Comparative example 4

The difference between the comparative example and the comparative example 1 is that in the step S1, the method of the preparation example 6 of the epimedium clear paste is selected for preparation.

The rest is the same as in comparative example 1.

Comparative example 5

The difference between the comparative example and example 2 is that in step S1, when preparing the herba pyrolae fluid extract, the method of preparation example 1 of herba pyrolae fluid extract is selected.

The rest is the same as example 2.

Comparative example 6

The difference between the comparative example and the example 2 is that in the step S1, when the pyrola fluid extract is prepared, the method of the pyrola fluid extract preparation example 2 is selected for preparation.

The rest is the same as example 2.

Comparative example 7

The difference between the comparative example and example 2 is that in step S1, when preparing the herba pyrolae fluid extract, the method of preparation example 3 of the herba pyrolae fluid extract is selected.

The rest is the same as example 2.

Comparative example 8

The difference between the comparative example and example 2 is that in step S1, when preparing the herba pyrolae fluid extract, the method of preparation example 6 of the herba pyrolae fluid extract is selected.

The rest is the same as example 2.

Comparative example 9

The preparation raw materials of the pharmaceutical composition for treating coronary heart disease are as follows: 200g of ginkgo leaf, 150g of pyrola, 150g of epimedium, 100g of salvia miltiorrhiza, 0.167g of gypenosides, 16.7g of corydalis tuber and 150g of hawthorn.

The preparation method of the pharmaceutical composition for treating coronary heart disease comprises the following steps:

s1, crushing the pyrola, the epimedium herb, the ginkgo leaf, the salvia miltiorrhiza, the corydalis tuber and the hawthorn, adding 3000mL of water, and soaking for 30 min. Then decocting with water for three times: adding water for the first time, wherein the water is 8 times of the total weight of the soaked medicinal materials (the total weight of herba Pyrolae crushed material, herba Epimedii crushed material, folium Ginkgo crushed material, Saviae Miltiorrhizae radix crushed material, rhizoma corydalis crushed material and fructus crataegi crushed material), and decocting for 1.5 hr; adding water for the second time, wherein the weight of the water is 5 times of the total weight of the soaked medicinal materials, and decocting for 1 h; adding water 4 times the total weight of the soaked materials for the third time, decocting for 45min, mixing the decoctions, filtering, concentrating the filtrate under reduced pressure to obtain mixed fluid extract with relative density of 1.25 (60 deg.C), and cooling.

S2, adding ethanol into the mixed clear paste prepared in the step S1 to ensure that the ethanol content reaches 65wt%, then standing, filtering, recovering ethanol, and concentrating to thick paste with the relative density of 1.30 (60 ℃).

S3, adding 10mL of water into the gypenosides to dissolve the gypenosides, adding the dissolved gypenosides into the thick paste obtained in the step S3, then adding 167g of 50wt% simple syrup, adding water to 1000mL, uniformly stirring, and standing for 10 days at 1-5 ℃; removing the filter residue after the filtrate is removed, and filling the filtrate to obtain the final product.

Performance detection

Content detection of active ingredients of finished pharmaceutical composition for treating coronary heart disease

1.1, 2' -0-galloylhyperin content detection

Determining 2' -0-galloyl hyperin (C) by high performance liquid chromatography₅₂H₂₄O₂₅）。

Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile-0.05 wt% of monopotassium phosphate-0.05 wt% of phosphoric acid solution (wherein the volume ratio of the acetonitrile, the 0.05wt% of monopotassium phosphate and the 0.05wt% of phosphoric acid solution is 20:30: 85) is used as a mobile phase; the detection wavelength is as follows: 263 nm. Column temperature: 25 ℃; flow rate: 1 mL/min; the number of theoretical plates is 2' -0-galloylhyperin (C)₅₂H₂₄O₂₅) The calculation should be no less than 4000.

Preparation of control solutions: precisely weighing 2' -0-galloylhyperin (C)₅₂H₂₄O₂₅) Adding appropriate amount of reference substance into 25mL volumetric flask, dissolving with methanol to obtain a solution containing 2' -0-galloylhyperin (C) per 1mL methanol₅₂H₂₄O₂₅) 0.25mg of the solution.

Preparation of a test solution: precisely measuring 2mL of the product, placing into a 10mL measuring flask, adding methanol to dilute to scale, shaking, and filtering with microporous membrane (0.45 um).

The determination method comprises the following steps: precisely sucking 10 μ L of each of the reference solution and the sample solution, injecting into a liquid chromatograph, measuring, and calculating.

And detecting the content of the crystal orchid glycoside

The content of the monotropein is determined by high performance liquid chromatography.

Chromatographic conditions and system applicability test: the chromatographic column is selected from Agilent Zorbax SB-Aq chromatographic column (250 mm × 4.6 mm, 5 μm); taking methanol-0.1 wt% phosphoric acid solution (wherein, the volume ratio of the methanol to the 0.1% phosphoric acid solution is 5: 95) as a mobile phase; the detection wavelength is as follows: 235 nm. Column temperature: 25 ℃; flow rate: 1 mL/min.

Preparation of control solutions: precisely weighing a proper amount of the monoterpin, placing the monoterpin in a volumetric flask with 25mL, adding methanol for dissolving, and preparing a solution containing 0.25mg of monoterpin per 1mL of methanol.

Preparation of a test solution: precisely measuring 2mL of the product, placing into a 10mL measuring flask, adding methanol to dilute to scale, shaking, and filtering with microporous membrane (0.45 um).

The determination method comprises the following steps: precisely sucking 10 μ L of each of the reference solution and the sample solution, injecting into a liquid chromatograph, measuring, and calculating.

And detecting the content of icariin

Determining icariin content by high performance liquid chromatography.

Chromatographic conditions and system applicability test: the chromatographic column was selected as Agilent Zorbax SB-C18 column (150 mm. times.4.6 mm, 5 μm); acetonitrile-water (wherein the volume ratio of the acetonitrile to the water is 3: 7) is used as a mobile phase; the detection wavelength is as follows: 270 nm. Column temperature: 25 ℃; flow rate: 1 mL/min.

Preparation of control solutions: precisely weighing appropriate amount of icariin reference substance, placing in 25mL volumetric flask, adding methanol to dissolve, and making into solution containing icariin 0.25mg per 1mL methanol.

Preparation of a test solution: precisely measuring 2mL of the product, placing into a 10mL measuring flask, adding methanol to dilute to scale, shaking, and filtering with microporous membrane (0.45 um).

The determination method comprises the following steps: precisely sucking 10 μ L of each of the reference solution and the sample solution, injecting into a liquid chromatograph, measuring, and calculating.

Content detection of total ginkgolic acid

And (3) determining the content of total negative acid by adopting a high performance liquid chromatography.

Chromatographic conditions and system applicability test: the column was chosen as Asahi ultimate LP-C18 column (150 mm. times.4.6 mm, 5 μm) packed with octadecylsilane chemically bonded silica. Using 0.1wt% trifluoroacetic acid acetonitrile solution-0.1 wt% trifluoroacetic acid water solution as mobile phase; wherein, 0.1wt% trifluoroacetic acid acetonitrile solution is phase A, and 0.1wt% trifluoroacetic acid water solution is phase B. Gradient elution procedure at 0min, 75% phase A and 25% phase B; when the time is 0-30 min, the phase A is changed from 75% to 90%; maintaining the phase A at 90% when the time is 30-35 min; when 35-36 min is reached, the phase A is changed from 90% to 75%; maintaining the phase A at 75% when the time is 36-45 min; both phases a and B above are expressed as volume percent in the mobile phase. The detection wavelength is as follows: 310 nm. The number of theoretical plates is not less than 4000 according to the peak of neoacidity of ginkgo.

Preparation of control solutions: precisely weighing appropriate amount of neoacid ginkgolide reference, placing in 25mL volumetric flask, adding methanol to dissolve, and making into solution containing neoacid ginkgolide 5 μ g per 1mL methanol.

Preparation of a test solution: precisely measuring 2mL of the product, placing into a 10mL measuring flask, adding methanol to dilute to scale, shaking, and filtering with microporous membrane (0.45 um).

The determination method comprises the following steps: precisely sucking 10 μ L of each of the reference solution and the sample solution, injecting into a liquid chromatograph, measuring, and calculating.

Content detection of total flavonol glycoside

The content of total flavonol glycosides is determined by high performance liquid chromatography.

Chromatographic conditions and system applicability test: selecting a chromatographic column as an ODS C18 column, and taking octadecylsilane chemically bonded silica as a filler; taking methanol-0.4 wt% phosphoric acid solution (wherein, the volume ratio of the methanol to the 0.4% phosphoric acid solution is 1: 1) as a mobile phase; the detection wavelength is as follows: 360 nm. The number of theoretical plates should not be lower than 2500 calculated according to the peak of quercetin.

Preparation of control solutions: accurately weighing appropriate amount of quercetin reference substance, kaempferide reference substance and isorhamnetin reference substance, placing in a 25mL volumetric flask, adding methanol for dissolving, and making into mixed solution containing quercetin, kaempferide and isorhamnetin 30 μ g, 30 μ g and 20 μ g respectively per 1mL methanol.

Preparation of a test solution: precisely measuring 2mL of the product, placing into a 10mL measuring flask, adding methanol to dilute to scale, shaking, and filtering with microporous membrane (0.45 um).

The determination method comprises the following steps: precisely sucking 10 μ L of each of the reference solution and the sample solution, injecting into a liquid chromatograph, measuring, and calculating.

Total flavonol glycoside content = (quercetin content + kaempferide content + isorhamnetin content) × 2.51.

Content detection of terpene lactones

Determining terpene lactone content by high performance liquid chromatography.

Chromatographic conditions and system applicability test: selecting a chromatographic column as an ODS C18 column, and taking octadecylsilane chemically bonded silica as a filler; taking n-propanol-tetrahydrofuran-water (wherein the volume ratio of n-propanol to tetrahydrofuran to water is 1:15: 84) as a mobile phase; detected with an evaporative light scattering detector. The number of theoretical plates is not less than 2500 calculated according to the peak of bilobalide.

Preparation of control solutions: precisely weighing appropriate amount of bilobalide reference substance, bilobalide A reference substance, bilobalide B reference substance and bilobalide C reference substance, placing in a 25mL volumetric flask, adding methanol for dissolving, and making into mixed solution containing 2mg, 1mg and 1mg of bilobalide, bilobalide A, bilobalide B and bilobalide C in each 1mL methanol.

Preparation of a test solution: precisely measuring 2mL of the product, placing into a 10mL measuring flask, adding methanol to dilute to scale, shaking, and filtering with microporous membrane (0.45 um).

The determination method comprises the following steps: precisely sucking 10 μ L of reference solution and sample solution respectively, injecting into liquid chromatograph, measuring, and calculating bilobalide, bilobalide A, bilobalide B and bilobalide C content respectively by using external standard two-point logarithmic equation.

Terpene lactone content = bilobalide content + bilobalide a content + bilobalide B content + bilobalide C content.

The contents of the active ingredients 2' -0-galloylhyperin, monothioloside and icariin were measured for the pharmaceutical compositions for treating coronary heart disease prepared in examples 1 to 3 and comparative examples 1 to 8, and the results are shown in Table 1.

TABLE 1

As shown in the data results in Table 1, the pharmaceutical composition for treating coronary heart disease (example 1-5) prepared in the present application has a 2' -0-galloylhyperin content of 3.34-4.25 mg/mL, a monotropein content of 2.74-3.44 mg/mL, and an icariin content of 2.23-2.94 mg/mL. The contents of the three effective medicinal substances are far higher than those of the medicinal compositions for treating coronary heart disease in the comparative examples 1-9.

Analyzing from the data of comparative examples 1-4 and examples 1-2, it was found that when 65-75% ethanol solution was used as the extractant for extracting icariin, and when the extractants for extracting 2 '-0-galloylhyperin and monotropein were 60% ethanol solution, the conversion rates of icariin, 2' -0-galloylhyperin and monotropein were high: respectively accounting for 82.3-85.6%, 91.8-93.2% and 87.9-90.1%, and the content of the three active ingredients in the prepared pharmaceutical composition for treating coronary heart disease is high. In comparative example 3, the extractant for extracting icariin was a 50% ethanol solution; although the 50% ethanol solution has the best effect on extracting icariin, the components in the pyrola extractive bring adverse effects on the stability of the icariin, and finally the content of the icariin in the pharmaceutical composition for treating coronary heart disease is reduced, and the extraction rate is not high.

Analyzing the data of comparative examples 5-8 and examples 2-3 to obtain that 60-70% ethanol solution is selected as an extracting agent when 2' -0-galloyl hyperin and monotropein are extracted; meanwhile, when the extracting agent for extracting the icariin is a 75% ethanol solution, the conversion rates of the icariin, the 2' -0-galloylhyperin and the monotropein are higher: respectively accounting for 82.3-86.4%, 89.3-93.2% and 84.4-90.1%, and the content of the three active ingredients in the prepared pharmaceutical composition for treating coronary heart disease is high. In comparative example 7, the extractant for extracting 2' -0-galloylhyperin and monotropein was a 50% ethanol solution; although 50% ethanol solution has the best effect on the extraction of 2 ' -0-galloylhyperin and monotropein, the components in the epimedium extract have an adverse effect on the stable presence of 2 ' -0-galloylhyperin and monotropein, which finally results in a reduced content of 2 ' -0-galloylhyperin and monotropein in the pharmaceutical composition for treating coronary heart disease and a low extraction rate.

Therefore, when the method is selected to prepare the pharmaceutical composition for treating coronary heart disease, the content of epimedium, 2' -0-galloyl hyperin and crystal orchid glycoside in the pharmaceutical composition for treating coronary heart disease is higher.

The pharmaceutical compositions for treating coronary heart disease prepared in examples 1 to 5 and comparative example 9 were tested for the contents of the effective components total flavonol glycosides and terpene lactones, and the content of total ginkgolic acid was determined, the results are shown in table 2.

TABLE 2

As seen from the data results in Table 2, in the pharmaceutical composition for treating coronary heart disease, the content of total flavonol is 2.38-4.58 mg/mL, and the content of terpene lactone is 3.30-6.35 mg/mL.

By comparing the example 4 with the examples 1-3, when the ginkgo biloba extract is extracted in a way of combining the primary extraction solution extraction and the water extraction of the hawthorn, the content of the total flavonol in the pharmaceutical composition for treating coronary heart disease is increased from 2.38-2.81 mg/mL to 4.15mg/mL, which is increased by 47.69-74.37%; the content of terpene lactones in the pharmaceutical composition for treating coronary heart disease is increased from 3.30-4.00 mg/mL to 5.45mg/mL, and is increased by 36.25-65.15%. Therefore, when the ginkgo leaf extract is extracted in a mode of combining primary extract extraction and water extraction of the hawthorn, the content of the total flavonol and the terpene lactone in the pharmaceutical composition for treating the coronary heart disease can be obviously improved.

By comparing the example 5 with the examples 1-3, when the ginkgo leaf extract is extracted by the mixed solution of the primary hawthorn extract and the primary salvia miltiorrhiza extract and combining the water extraction mode, the content of the total flavonol in the pharmaceutical composition for treating coronary heart disease is increased from 2.38-2.81 mg/mL to 4.58mg/mL, and is increased from 62.99-92.44%; the content of terpene lactones in the pharmaceutical composition for treating coronary heart disease is increased from 3.30-4.00 mg/mL to 6.35mg/mL, and is increased by 58.75-92.42%. Therefore, when the ginkgo leaf extract is extracted by the mixed solution of the primary hawthorn extract and the primary salvia miltiorrhiza extract and combining the water extraction mode, the content of the total flavonol and the terpene lactone in the pharmaceutical composition for treating the coronary heart disease can be obviously improved.

The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.