CN114224922A - Preparation method and application of anti-depression ginkgo leaf extract - Google Patents
Preparation method and application of anti-depression ginkgo leaf extract Download PDFInfo
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- CN114224922A CN114224922A CN202111361003.4A CN202111361003A CN114224922A CN 114224922 A CN114224922 A CN 114224922A CN 202111361003 A CN202111361003 A CN 202111361003A CN 114224922 A CN114224922 A CN 114224922A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/16—Ginkgophyta, e.g. Ginkgoaceae (Ginkgo family)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/24—Antidepressants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention provides a preparation method and application of an anti-depression ginkgo leaf extract, belonging to the technical field of medicines and comprising the following steps: (1) drying and pulverizing folium Ginkgo, ultrasonic extracting in alkaline water, filtering, and concentrating to obtain folium Ginkgo alkaline water solution; (2) mixing alkaline water solution of folium Ginkgo with ethanol solution, adjusting pH of the solution to neutral, extracting, filtering, extracting the filtrate with petroleum ether, concentrating under reduced pressure, and drying to obtain folium Ginkgo extract. The preparation method can extract the effective components of ginkgo flavone, bilobalide and ginkgo polysaccharide in the ginkgo leaves to the maximum extent. The ginkgo leaf extract has a remarkable treatment effect on depression, has small side effect on a human body, is high in safety, and has a good prospect in developing and preparing anti-depression drugs.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a preparation method and application of an anti-depression ginkgo leaf extract.
Background
Depression is an affective disorder that severely affects physical and mental health, with patients presenting with persistent mood depression and cognitive impairment. With the acceleration of modern life rhythm and the increasingly fierce social competition, depression becomes a common disease and a high disease incidence in the modern society, the incidence rate rises rapidly, according to statistics, 25-30% of severe patients may die from suicide finally, and the depression seriously harms the physical and mental health, families and society of people, so that people pay high attention to the depression.
In recent years, the dominance of drug therapy has been established with the progress of research on the pathogenesis of depression. Clinically, the commonly used anti-depression drugs mainly comprise monoamine reuptake inhibition dosage forms, monoamine oxidase inhibitors and botanical drugs. Modern pathology studies have shown that: the pathogenesis of depression is very complex, the induction causes are more, and most of the synthesized antidepressants have the defects of narrow antidepressant spectrum, large side effect, high price, easy relapse and the like. Therefore, the research and development of the antidepressant at home and abroad increasingly pay attention to the traditional plant medicine.
The ginkgo leaf is dry leaf of ginkgo of Ginkgoaceae, the main chemical components of the ginkgo leaf are polysaccharide, flavonoid and terpene lactone compounds, the content of toxic components is low, meanwhile, the ginkgo leaf is rich in nutrient components, high in protein content, and rich in vitamin C, vitamin E, carotene, calcium, phosphorus, boron, selenium and the like. Researches show that the ginkgo biloba extract has pharmacological effects of expanding blood vessels, regulating blood fat, resisting oxidation, resisting aging, improving memory, resisting tumors and the like, and has already been clinically used for treating angina pectoris, coronary heart disease, myocardial ischemia, hypertension, hyperlipidemia, diabetes, Alzheimer disease, atherosclerosis, gastric cancer and other diseases, so the ginkgo biloba extract has good development prospect when being used for developing medicaments.
Disclosure of Invention
The invention aims to provide a preparation method and application of an anti-depression ginkgo leaf extract, and the ginkgo leaf extract can effectively treat depression and has high safety.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of an anti-depression ginkgo leaf extract, which comprises the following steps:
(1) drying and pulverizing folium Ginkgo, ultrasonic extracting in alkaline water, filtering, and concentrating to obtain folium Ginkgo alkaline water solution;
(2) mixing alkaline water solution of folium Ginkgo with ethanol solution, adjusting pH of the solution to neutral, extracting, filtering, extracting the filtrate with petroleum ether, concentrating under reduced pressure, and drying to obtain folium Ginkgo extract.
Preferably, the crushed particle size is 40-60 meshes.
Preferably, the base water is sodium bicarbonate solution.
Preferably, the mass volume ratio of the ginkgo leaves to the alkaline water is 1: 10-15.
Preferably, the ultrasonic power is 200-300W, the ultrasonic temperature is 40-50 ℃, the ultrasonic time is 15-25 min, and the ultrasonic extraction frequency is 2-3.
Preferably, the volume percentage of the ethanol solution is 60-80%, and the mass volume ratio of the ginkgo leaf alkaline water solution to the ethanol solution is 1: 6-8.
Preferably, in the step (2), the extraction temperature is 60-75 ℃, the extraction time is 1.5-3 h, and the extraction times are 1-2.
Preferably, in the step (2), the volume ratio of the petroleum ether to the filtrate is 1: 20-30.
Preferably, the petroleum ether extraction times are 1-2.
The invention also provides application of the ginkgo leaf extract prepared by any one of the preparation methods in preparing an anti-depression medicament.
Compared with the prior art, the invention has the following beneficial effects:
the preparation method of the ginkgo leaf extract is simple and convenient, and can extract the effective components of ginkgo flavone, ginkgolide and ginkgo polysaccharide in the ginkgo leaves to the maximum extent by using alkaline water, ultrasonic treatment, ethanol extraction and petroleum ether extraction. The ginkgo leaf extract has a remarkable treatment effect on depression, has small side effect on a human body, is high in safety, and has a good prospect in developing and preparing anti-depression drugs.
Detailed Description
The invention provides a preparation method of an anti-depression ginkgo leaf extract, which comprises the following steps:
(1) drying and pulverizing folium Ginkgo, ultrasonic extracting in alkaline water, and filtering to obtain folium Ginkgo alkaline water solution;
(2) mixing alkaline water solution of folium Ginkgo with ethanol solution, adjusting pH of the solution to neutral, extracting, filtering, extracting the filtrate with petroleum ether, concentrating under reduced pressure, and drying to obtain folium Ginkgo extract.
In the invention, after the ginkgo leaves are dried and crushed, the ginkgo leaves are dried to constant weight in a constant-temperature drying oven at 50-60 ℃, and the crushed particle size is preferably 40-60 meshes, and more preferably 50 meshes. The invention increases the contact area of the ginkgo leaf and the alkaline water by crushing, and can shorten the time for extracting the effective components.
In the invention, the ginkgo leaf alkaline water solution is obtained by ultrasonic extraction in alkaline water, filtration and concentration. The alkaline water is preferably sodium bicarbonate solution; the mass-volume ratio of the ginkgo leaves to the alkaline water is preferably 1: 10-15 (g/mL); the ultrasonic power is preferably 200-300W, more preferably 250W, the ultrasonic temperature is preferably 40-50 ℃, more preferably 45 ℃, the ultrasonic time is preferably 15-25 min, more preferably 20min, the ultrasonic extraction frequency is preferably 1-2, and the concentration can be reduced pressure concentration. The invention can extract the active substances of ginkgetin, bilobalide and polysaccharide in the ginkgo leaves to the maximum extent and effectively by carrying out ultrasonic extraction on the ginkgo leaves in alkaline water.
In the invention, the ginkgo leaf alkaline water solution is mixed with the ethanol solution, and the solution is adjusted to be neutral, extracted and filtered. The volume percentage of the ethanol solution is preferably 60-80%, and more preferably 70%; the mass volume ratio of the ginkgo leaf alkaline water solution to the ethanol solution is preferably 1: 6-8, more preferably 1: 7; the mode of adjusting to be neutral can be selected from adjusting by hydrochloric acid; the extraction temperature is 60-75 ℃, the extraction time is 1.5-3 h, and the extraction times are 1-2. The invention can further improve the content of effective components by ethanol extraction.
In the invention, the ginkgo leaf extract is obtained by extracting the filtrate with petroleum ether, concentrating under reduced pressure and drying. The volume ratio of the petroleum ether to the filtrate is preferably 1: 20-30, more preferably 1:25, and the number of times of petroleum ether extraction is preferably 1-2; the reduced pressure concentration can be carried out at a temperature of 50-55 ℃, and the drying can be carried out in a constant-temperature drying oven at a temperature of 60-70 ℃ to constant weight. According to the invention, impurities such as chlorophyll in the extract can be effectively removed after petroleum ether extraction, and the effect of the finally obtained ginkgo leaf extract is ensured to be exerted.
The invention also provides an application of the ginkgo leaf extract obtained by any one of the preparation methods in preparing an anti-depression medicament.
In the invention, the medicine takes the ginkgo biloba extract as an active ingredient of the medicine, and the medicine can be prepared into tablets, capsules, injections, sustained release agents, granules, pills, solutions and syrups. The medicine of the invention can be applied by oral administration, abdominal cavity, intragastric administration, intravenous administration and intramuscular administration.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A method for preparing an anti-depression ginkgo leaf extract comprises the following steps:
(1) drying 100g folium Ginkgo in a constant temperature drying oven at 55 deg.C to constant weight, pulverizing to 50 mesh, extracting with 1400mL sodium bicarbonate solution at 45 deg.C under 250W ultrasonic for 20min, filtering to obtain filtrate and residue, adding 1100mL sodium bicarbonate solution into the residue, extracting at 45 deg.C under 250W ultrasonic for 15min, mixing filtrates, and concentrating under reduced pressure to 1/3 to obtain folium Ginkgo alkaline water solution;
the preparation method of the sodium bicarbonate solution comprises the steps of adding 250g of sodium bicarbonate into 10kg of sodium bicarbonate, and stirring and mixing uniformly to obtain the sodium bicarbonate solution.
(2) Mixing the above folium Ginkgo alkaline water solution with 7 times of 70% ethanol solution, adjusting pH to neutral with hydrochloric acid, extracting at 70 deg.C for 2.5h, filtering to obtain filtrate and residue, extracting the residue repeatedly for 1 time according to the above operation, extracting the filtrate with 1/25 volume of petroleum ether for 2 times, concentrating under reduced pressure at 50 deg.C, and drying in 65 deg.C constant temperature drying oven to constant weight to obtain folium Ginkgo extract.
Example 2
A method for preparing an anti-depression ginkgo leaf extract comprises the following steps:
(1) drying 100g folium Ginkgo in a constant temperature drying oven at 50 deg.C to constant weight, pulverizing to 40 mesh, extracting with 1500mL sodium bicarbonate solution at 50 deg.C under 200W ultrasound for 15min, filtering to obtain filtrate and residue, adding 1000mL sodium bicarbonate solution into the residue, extracting at 50 deg.C under 200W ultrasound for 15min, mixing filtrates, and concentrating under reduced pressure to 1/3 to obtain folium Ginkgo alkaline water solution;
the preparation method of the sodium bicarbonate solution comprises the steps of adding 250g of sodium bicarbonate into 10kg of sodium bicarbonate, and stirring and mixing uniformly to obtain the sodium bicarbonate solution.
Mixing the above folium Ginkgo alkaline water solution with 8 times of 60% ethanol solution, adjusting pH to neutral with hydrochloric acid, extracting at 75 deg.C for 1.5h, filtering to obtain filtrate and residue, extracting the filtrate with 1/30 volume of petroleum ether, concentrating under reduced pressure at 55 deg.C, and drying in 70 deg.C constant temperature drying oven to constant weight to obtain folium Ginkgo extract.
Example 3
A method for preparing an anti-depression ginkgo leaf extract comprises the following steps:
(1) drying 100g folium Ginkgo in a constant temperature drying oven at 60 deg.C to constant weight, pulverizing to 60 mesh, performing ultrasonic extraction at 40 deg.C and 300W for 25min in 1300mL sodium bicarbonate solution, filtering to obtain filtrate and residue, adding 1000mL sodium bicarbonate solution into the residue, performing ultrasonic extraction at 45 deg.C and 300W for 15min, mixing filtrates, and concentrating under reduced pressure to 1/3 to obtain folium Ginkgo alkaline water solution;
the preparation method of the sodium bicarbonate solution comprises the steps of adding 250g of sodium bicarbonate into 10kg of sodium bicarbonate, and stirring and mixing uniformly to obtain the sodium bicarbonate solution.
Mixing the above folium Ginkgo alkaline water solution with 6 times of 80% ethanol solution, adjusting pH to neutral with hydrochloric acid, extracting at 60 deg.C for 3 hr, filtering to obtain filtrate and residue, extracting the residue according to the above operation for 1 time, extracting the filtrate with 1/30 volume of petroleum ether for 2 times, concentrating under reduced pressure at 55 deg.C, and drying in 60 deg.C constant temperature drying oven to constant weight to obtain folium Ginkgo extract.
Test examples
Test animals: 150 healthy ICR mice are male, have the weight of 18-22 g, are purchased from the experimental animal center of Fujian Chinese medicinal university, and are adaptive to the environment. In the whole test process, the mice are fed with water freely, the room temperature is 20-25 ℃, the relative humidity is 50-60%, and the mice are raised in 12h of illumination and 12h of darkness.
Test drugs:
positive control group: dissolving fluoxetine in normal saline containing 0.1% Tween-80 to obtain 2mg/mL liquid medicine;
adding normal saline into the ginkgo biloba extract prepared in the example 1 to prepare 100mg/mL, 200mg/mL and 400mg/mL of low-dose ginkgo biloba extract, medium-dose ginkgo biloba extract and high-dose ginkgo biloba extract;
negative control group: 0.1% Tween-80 in physiological saline was used as a negative control.
The model making of the depression mice: suspending the mouse tail tip portion 1/3 in a tail suspension box by using non-stick glue, enabling the head of the mouse to be opposite to a lens and be 10cm away from the bottom of the box, suspending for 6min, and recording the incubation period of the animal in an immobile state within 6min and the duration of the immobile state within 4min later by using a camera system, wherein the immobile state means that the animal gives up actively struggling, and the body is in a suspended and untwisted state.
1. Tail suspension test for mice
Screening 50 mice with immobility time of 100-160 s from the model building mice, randomly dividing the mice into 5 groups, 10 mice in each group, respectively comprising a negative control group, a positive control group, a low-dose ginkgo leaf extract group, a medium-dose ginkgo leaf extract group and a high-dose ginkgo leaf extract group, performing gastric lavage administration according to the weight of 10mL/kg of the mice, administering once per day for 7 days continuously, performing tail suspension experiment 1h after administration on the 7 th day, suspending the mouse tail tip part 1/3 in a tail suspension box, keeping the head part opposite to a lens 10cm away from the bottom of the box, suspending for 6min, recording the whole process by using a camera system, wherein an actor is a double-blind experimenter who is trained and has no knowledge on animal grouping and administration conditions, and keeping the mice in an immobility state within 4min after observation and recording, and the immobility time refers to the time when the mice stop struggling in water or are suspended statically, Or only a small amplitude motion such as a slight flexing of the forefoot. The results of the immobility time of the mouse tail suspension experiment from different treatment groups are shown in table 1.
Table 1 results of different treatment groups on immobility time of tail suspension experiment of mice
| Group of | Immobility time/s within 4min |
| Negative control group | 128±15 |
| Positive control group | 100±20 |
| Low dose ginkgo biloba extract | 85±16 |
| Ginkgo biloba leaf extract of medium dosage | 78±19 |
| High dose ginkgo biloba extract | 65±18 |
The results in table 1 show that the low, medium and high dose of ginkgo biloba leaves extract of the present invention showed significantly shorter immobility time and dose-dependency in the tail suspension test compared to the positive control group.
2. Forced swimming test for mice
50 mice with immobility time of 100-160 s are screened out from the model-making mice, the model-making mice are randomly divided into 5 groups, 10 mice in each group are respectively a negative control group, a positive control group, a low-dose ginkgo leaf extract group, a medium-dose ginkgo leaf extract group and a high-dose ginkgo leaf extract group, the model-making mice are administrated by gastric irrigation according to 10mL/kg of the weight of the mice, the administration is carried out once a day for 7 days continuously, forced mouse swimming is carried out 1h after the administration on the 7 th day, the mice immediately start observation after swimming for 2min, the observation time lasts for 4min, the struggling or floating state of the mice in water within 4min or the duration time of only micro limb movement to keep the head floating on the water surface is recorded, and the results of the immobility time of the forced swimming test of the mice in different treatment groups are shown in a table 2.
Table 2 results of immobility time of forced swimming test of mice by different treatment groups
The results in Table 2 show that the low, medium and high doses of the ginkgo biloba extract according to the present invention showed significantly shorter immobility time and dose-dependency in the forced swimming test than the positive control group.
In conclusion, the ginkgo leaf extract has obvious antidepressant effect, so that the ginkgo leaf extract can be used for preparing antidepressant medicines.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A preparation method of an anti-depression ginkgo leaf extract is characterized by comprising the following steps:
(1) drying and pulverizing folium Ginkgo, ultrasonic extracting in alkaline water, filtering, and concentrating to obtain folium Ginkgo alkaline water solution;
(2) mixing alkaline water solution of folium Ginkgo with ethanol solution, adjusting pH of the solution to neutral, extracting, filtering, extracting the filtrate with petroleum ether, concentrating under reduced pressure, and drying to obtain folium Ginkgo extract.
2. The method according to claim 1, wherein the pulverized particles have a particle size of 40 to 60 mesh.
3. The method according to claim 1, wherein the alkali water is a sodium bicarbonate solution.
4. The preparation method of claim 1, wherein the mass volume ratio of the ginkgo leaves to the alkaline water is 1: 10-15.
5. The preparation method according to claim 1, wherein the ultrasonic power is 200-300W, the ultrasonic temperature is 40-50 ℃, the ultrasonic time is 15-25 min, and the ultrasonic extraction frequency is 2-3.
6. The method according to claim 1, wherein the ethanol solution is 60 to 80 vol%, and the ratio of the ginkgo biloba leaf alkaline aqueous solution to the ethanol solution is 1:6 to 8 by mass/volume.
7. The preparation method according to claim 1, wherein in the step (2), the extraction temperature is 60-75 ℃, the extraction time is 1.5-3 h, and the extraction times are 1-2.
8. The preparation method according to claim 1, wherein in the step (2), the volume ratio of the petroleum ether to the filtrate is 1: 20-30.
9. The method according to claim 1, wherein the number of times of the petroleum ether extraction is 1 to 2.
10. Use of the ginkgo biloba extract prepared by the method of any one of claims 1 to 9 in the preparation of an anti-depression drug.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116712468A (en) * | 2023-05-31 | 2023-09-08 | 神威药业集团有限公司 | Extraction method of ginkgo leaf extract and preparation method of Shuxuening injection |
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| CN101904877A (en) * | 2010-08-06 | 2010-12-08 | 合肥工业大学 | New preparation method of low-phenolic acid gingko flavone extractive |
| CN105412167A (en) * | 2015-11-27 | 2016-03-23 | 陕西嘉禾生物科技股份有限公司 | Method for extracting and separating ginkgo flavone and ginkgolide from ginkgo leaves |
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2021
- 2021-11-17 CN CN202111361003.4A patent/CN114224922A/en active Pending
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| CN116712468A (en) * | 2023-05-31 | 2023-09-08 | 神威药业集团有限公司 | Extraction method of ginkgo leaf extract and preparation method of Shuxuening injection |
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