CN111035670A - Phyllanthus emblica extract and preparation method and application thereof - Google Patents
Phyllanthus emblica extract and preparation method and application thereof Download PDFInfo
- Publication number
- CN111035670A CN111035670A CN201911366808.0A CN201911366808A CN111035670A CN 111035670 A CN111035670 A CN 111035670A CN 201911366808 A CN201911366808 A CN 201911366808A CN 111035670 A CN111035670 A CN 111035670A
- Authority
- CN
- China
- Prior art keywords
- emblic
- extract
- ethanol
- feed liquid
- reduced pressure
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/47—Euphorbiaceae (Spurge family), e.g. Ricinus (castorbean)
-
- 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
-
- 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
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- 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
-
- 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
- A61K2236/55—Liquid-liquid separation; Phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Neurosurgery (AREA)
- Pain & Pain Management (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Neurology (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Psychiatry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an emblic extract and a preparation method and application thereof, wherein the preparation method comprises the following steps: (1) and (3) extraction and concentration: crushing raw material emblic leafflower fruit medicinal material, adding 70-95% ethanol, extracting at 50-80 ℃, filtering, concentrating under reduced pressure, centrifuging, taking precipitate, adding ethanol for dissolving to obtain feed liquid a; (2) kaolin adsorption: adding kaolin into the feed liquid a, carrying out water bath, stirring, centrifuging, taking supernate, concentrating under reduced pressure, and supplementing water until no alcohol smell exists to obtain feed liquid b; (3) column chromatography: and (3) adjusting the pH value of the feed liquid b to 8.5-10, then purifying by strong base anion resin, washing with water to remove impurities, resolving by NaCl with the concentration of 5-10%, concentrating under reduced pressure, centrifuging, taking precipitate, and freeze-drying to obtain the phyllanthus emblica extract. The polyphenol content of the emblic leafflower fruit extract is more than 98 percent. The emblic extract prepared by the invention has antidepressant activity. The method has simple process, easy operation and easy industrial production.
Description
Technical Field
The invention relates to the technical field of plant extraction, and particularly relates to an emblic leafflower fruit extract and a preparation method and application thereof.
Background
Phyllanthus emblica L is Phyllanthus genus of Euphorbiaceae (Euphorbiaceae), originated from Burma and India, mainly distributed in India, China, Burma and Malaysia, etc., with the most yield in China, and wild Phyllanthus in Yunnan, Guangxi and Hainan, etc. Emblic leafflower fruit is a common Tibetan medicine, and is loaded in Chinese pharmacopoeia in 1990, and mainly used for treating blood heat and blood stasis, dyspepsia, akabane disease, leukemia and hypertension. The results of recent researches show that the emblic leafflower fruit has the effects of resisting inflammation, resisting mutation, resisting hypertension, reducing blood pressure, reducing blood fat, protecting liver and the like, the main components of the emblic leafflower fruit are polyphenol and polysaccharide substances, and the antidepressant activity of the polyphenol substances is not reported.
Depression is also called depressive disorder, and is a mental disorder disease which seriously troubles the mental activities of human beings. The main clinical features are marked and persistent mood depression, and the method has the defects of lack of interest in things, suicide tendency and the like. The accelerated pace of social life and the excessive mental stress are considered as social factors inducing depression. In some cases, the anxiety and the motility are obvious, and in severe cases, the psychopathic symptoms such as hallucinations and delusions can appear. Each episode lasts at least two weeks and up to several years. In most cases, there is a tendency for recurrent episodes, with most of each episode remitting and some remaining symptoms or becoming chronic. According to statistics, the lifetime prevalence rate of depression reaches 5.2% -16.2% in the global population background, wherein the female morbidity is particularly prominent. The prevalence of depression in the mainland areas of China is presumed to be 7-8%, and the trend of the depression is obvious. According to the statistics and predictions of the world health organization, depression is predicted to become the second largest disease burden following coronary heart disease by 2030. Currently, drug therapy is the main treatment for depression attacks of more than moderate degree. The traditional tricyclic and tetracyclic antidepressants and monoamine oxidase inhibitors have larger adverse reactions and obviously reduced application. The current first-line clinical antidepressants mainly comprise selective 5-hydroxytryptamine reuptake inhibitors, 5-hydroxytryptamine and norepinephrine reuptake inhibitors and the like, but the drugs have slow effect, narrow action spectrum and easy relapse after drug withdrawal. At present, researchers are researching and developing more effective therapeutic drugs with small side effects.
In the prior patent CN101032322A, the emblic extract and the preparation method and application thereof are that crude extract obtained by extracting emblic by microwave-assisted solvent is treated by macroporous resin, and then extracted by petroleum ether, ether and ethyl acetate to obtain the emblic extract, wherein the polyphenol content of the emblic extract reaches about 94 percent. The patent is complicated in operation, and the biological activity of the patent is not researched.
In the prior patent CN103768130A, an extraction method of phyllanthus emblica polyphenol is provided, in which fresh phyllanthus emblica fruits are added with a pre-cooled acetone solution for extraction, the extract is filtered by a hollow fiber membrane, and then purified by a polyamide chromatographic column, so as to obtain the phyllanthus emblica polyphenol finished product with a content of 99%. The hollow fiber membrane used in this patent is expensive, resulting in high cost, and the number of times of repeated use of the polyamide column is small.
In the prior patent CN109876031A, an enrichment method of phyllanthus emblica tannin fraction is to filter the phyllanthus emblica tannin extract centrifugally to obtain supernatant, to be absorbed by macroporous absorption resin, and to be eluted with ethanol water in a gradient way to obtain the phyllanthus emblica tannin fraction with the tannin content of about 56%. The emblic leafflower fruit compound prepared by the patent contains a mixture of tannin and polyphenol, and has low purity.
Disclosure of Invention
The invention aims to overcome the technical defects of the background technology and provides an emblic leafflower fruit extract and a preparation method and application thereof. The invention takes Phyllanthus emblica (Phyllanthus emblica) as a raw material, and the refined Phyllanthus emblica extract is obtained after extraction and purification, and the polyphenol content of the Phyllanthus emblica extract is more than 98 percent. The emblic extract prepared by the invention has antidepressant activity. The invention has the advantages of simple process, easy operation, new application of the emblic leafflower fruit extract activity and the like, and the process of the invention is easy for industrial production.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a preparation method of an emblic extract comprises the following steps:
(1) and (3) extraction and concentration: crushing raw material emblic leafflower fruit medicinal material, adding 70-95% ethanol, extracting at 50-80 ℃, filtering, concentrating under reduced pressure, centrifuging, taking precipitate, adding ethanol for dissolving to obtain feed liquid a;
(2) kaolin adsorption: adding kaolin into the feed liquid a, carrying out water bath, stirring, centrifuging, taking supernate, concentrating under reduced pressure, and supplementing water until no alcohol smell exists to obtain feed liquid b;
(3) column chromatography: and (3) adjusting the pH value of the feed liquid b to 8.5-10, then purifying by strong base anion resin, washing with water to remove impurities, resolving by NaCl with the concentration of 5-10%, concentrating under reduced pressure, centrifuging, taking precipitate, and freeze-drying to obtain the phyllanthus emblica extract.
Preferably, in the step (1), the adding amount of the 70-95% ethanol is 6-12 times of the weight of the raw material emblic leafflower fruit medicinal material according to the mass-volume ratio; the mass volume ratio of 1 means that 1kg of the raw material emblic leafflower fruit medicinal material and 1L of 70-95% ethanol are added.
Preferably, in the step (1), the number of times of extraction is 2-3, and the time for each extraction is 1-2 hours.
Preferably, in the step (1), the filtration is performed by using a filter cloth.
More preferably, in the step (1), the mesh number of the filter cloth is 200 meshes.
Preferably, in the step (1), the filtrates are combined after the filtration.
Preferably, in the step (1), the concentration under reduced pressure is carried out at 58-60 ℃ and-0.09 MPa.
Preferably, in the step (1), the volume of the material liquid is reduced and concentrated to be 1 time of the mass of the raw material emblic leafflower fruit medicinal material according to the mass-to-volume ratio; the mass volume ratio of 1 means that the raw material emblic leafflower fruit medicinal material is 1kg, and the volume of the feed liquid is 1L.
Preferably, in step (1), the centrifugation is performed while hot.
Preferably, in the step (1), the rotation speed during centrifugation is 6000 r/min.
Preferably, in the step (1), the concentration of the ethanol when the ethanol is added for dissolution is 60%.
Preferably, in the step (1), the dissolving is performed by using ultrasound.
Preferably, in the step (2), the adding amount of the kaolin is 1 to 5 percent of the amount of the raw material emblic leafflower fruit medicinal material according to the mass ratio.
Preferably, in the step (2), the temperature of the water bath is 40-55 ℃.
Preferably, in the step (2), the stirring time is 0.5-1 hour.
Preferably, in step (2), the centrifugation is performed while hot.
Preferably, in the step (2), the rotation speed during centrifugation is 6000 r/min.
Preferably, in the step (2), the concentration under reduced pressure is carried out at 58-60 ℃ and-0.09 MPa.
Preferably, in the step (3), NaOH with a concentration of 5% is added to the feed liquid b to adjust the pH.
Preferably, in the step (3), the strong base anion resin is any one of D201, 7170, D204, HZ202, ZGA307, 201x 7.
More preferably, in the step (3), the strong base anion resin is any one of D201, 7170, and D204.
Preferably, in the step (3), the water is purified water.
Preferably, in the step (3), the addition amount of the water is 1-3 BV.
Preferably, in the step (3), the concentration under reduced pressure is carried out at 58-60 ℃ and-0.09 MPa.
Preferably, in the step (3), the volume of the liquid is reduced to 1/10 times of the mass of the raw material emblic leafflower fruit medicinal material according to the mass-to-volume ratio; the mass volume ratio of 1/10 means that the raw material emblic leafflower fruit medicinal material is 1kg, and the volume of the material liquid is 100 ml.
Preferably, in the step (3), the rotation speed during centrifugation is 6000 r/min.
An extract of Phyllanthus emblica is prepared by the above preparation method.
Preferably, the phyllanthus emblica extract has a polyphenol content of greater than 98%.
More preferably, the phyllanthus emblica extract has a polyphenol content of more than 99%.
The application of the emblic leafflower fruit extract in the aspect of depression resistance.
In the technical scheme, the percentage is mass percentage.
The basic principle of the invention is as follows:
through a large number of experiments, the invention finds that the specific technical effect of the invention can be realized only by adopting the specific technical scheme of the invention, and other technical schemes can not realize the specific technical effect of the invention.
Compared with the prior art, the invention has the beneficial effects that:
the invention takes Phyllanthus emblica (Phyllanthus emblica) as a raw material, and the refined Phyllanthus emblica extract is obtained after extraction and purification, and the polyphenol content of the Phyllanthus emblica extract is more than 98 percent. The emblic extract prepared by the invention has antidepressant activity. The invention has the advantages of simple process, easy operation, new application of the emblic leafflower fruit extract activity and the like, and the process of the invention is easy for industrial production.
Detailed Description
For a better understanding of the present invention, reference is made to the following examples. It is to be understood that these examples are for further illustration of the invention and are not intended to limit the scope of the invention. In addition, it should be understood that the invention is not limited to the above-described embodiments, but is capable of various modifications and changes within the scope of the invention.
Example 1
Pulverizing 1kg raw material fructus Phyllanthi, adding 95% ethanol, reflux extracting for 3 times, wherein the amount of 95% ethanol added each time is 6 times of the weight of fructus Phyllanthi, heating and refluxing at 70 deg.C for 1 hr, filtering with 200 mesh filter cloth, mixing filtrates, controlling temperature at 60 deg.C, concentrating under reduced pressure of 0.09Mpa to 1L, centrifuging at 6000r/min, dissolving precipitate with 500ml 60% ethanol under ultrasound, adding 25g Kaolin, stirring in water bath at 50 deg.C for 30 min, centrifuging to remove Kaolin, controlling temperature at 59 deg.C, concentrating supernatant under reduced pressure of 0.09Mpa, adding water to ethanol-free solution, adding 5% sodium hydroxide into 400ml feed liquid to adjust pH to 9.1, performing column chromatography with strong base anion D201 (Techno resin Co., Techno technology corporation) with column volume of 0.5L, purifying water washing with impurity at flow rate of 3 s/drop, and resolving sodium chloride solution of 1L 5%, collecting eluate, concentrating under-0.09 Mpa to 100ml at 59 deg.C, centrifuging by the same method, collecting precipitate, and freeze drying to obtain 13.6g fructus Phyllanthi extract with yield of 1.36% and polyphenol content of 99.02%.
Example 2
Pulverizing 2kg of fructus Phyllanthi, extracting with 90% ethanol under reflux for 3 times, wherein the amount of 90% ethanol added is 8 times of the weight of fructus Phyllanthi, heating and reflux extracting at 65 deg.C for 3 times of 1.5, 1 and 1 hr, filtering with 200 mesh filter cloth, mixing filtrates, controlling temperature at 58 deg.C, -0.09Mpa, concentrating to 2L, centrifuging at 6000r/min, dissolving precipitate with 1L 60% ethanol under ultrasonic, adding 60g Kaolin, stirring in water bath at 45 deg.C for 40 min, centrifuging to remove Kaolin, controlling temperature at 59 deg.C, concentrating supernatant at 0.09Mpa, adding water to no alcohol, adding 5% sodium hydroxide into 800ml of feed liquid, adjusting pH to 8.9, separating with column chromatography with 1L of strong alkali anion 7170 (Anhui Samsung resin Co., Ltd.), eluting with 1L water at 3 s per second, eluting with 1L of purified impurities, and resolving 1.5L of 7% sodium chloride solution, collecting eluate, concentrating under-0.09 Mpa to 200ml at 60 deg.C, centrifuging, collecting precipitate, and freeze drying to obtain 27.2g fructus Phyllanthi extract with yield of 1.36% and polyphenol content of 99.08%.
Example 3
Pulverizing 2kg of fructus Phyllanthi, extracting with 85% ethanol under reflux for 3 times, wherein the amount of 85% ethanol added is 10 times of the weight of fructus Phyllanthi, extracting under reflux at 70 deg.C for 1.5 hr, filtering with 200 mesh filter cloth, mixing filtrates, controlling the temperature at 59 deg.C, concentrating under 0.09Mpa to 2L, centrifuging at 6000r/min, dissolving precipitate with 800ml of 60% ethanol under ultrasonic, adding 55g of Kaolin, stirring at 40 deg.C in water bath, centrifuging to remove Kaolin, controlling the temperature at 58 deg.C, concentrating the supernatant under 0.09Mpa, adding water to no alcohol, adding 5% sodium hydroxide into 900ml to adjust pH to 9.0, purifying by column chromatography with 1L strong base anion D204 column chromatography (Anhui Samsung resin Co., Ltd.), eluting with 1L of purified water at flow rate of 3 s and 1.5L of 6% sodium chloride solution, resolving, collecting eluate, concentrating under-0.09 Mpa to 200ml at 58 deg.C, centrifuging, collecting precipitate, and freeze drying to obtain 27.0g fructus Phyllanthi extract with yield of 1.35% and polyphenol content of 99.16%.
Comparative example 1
Pulverizing 2kg of fructus Phyllanthi, extracting with 85% ethanol under reflux for 3 times, wherein the amount of 85% ethanol added is 10 times of the weight of fructus Phyllanthi, extracting under reflux at 70 deg.C for 1.5 hr, filtering with 200 mesh filter cloth, mixing filtrates, controlling the temperature at 59 deg.C, concentrating under reduced pressure of 0.09Mpa to 2L, centrifuging at 6000r/min, dissolving precipitate with 800ml of 60% ethanol under ultrasonic, adding 55g of active carbon, stirring at 40 deg.C in water bath, centrifuging to remove active carbon, controlling the temperature at 58 deg.C, concentrating the supernatant under reduced pressure of 0.09Mpa, adding water to no alcohol, adding 5% sodium hydroxide into 900ml of the solution, adjusting pH to 9.0, performing column chromatography with column volume of 1L of strong base anion D204 (Asahi Samson resin Co., Ltd.), eluting with 1L of purified water at flow rate of 3 s per drop, eluting with 1.5L of 6% sodium chloride solution, collecting eluate, concentrating under-0.09 Mpa to 200ml at 58 deg.C, centrifuging, collecting precipitate, and freeze drying to obtain 39.0g fructus Phyllanthi extract with yield of 1.95% and polyphenol content of 69.11%.
Comparative example 2
Pulverizing 2kg of fructus Phyllanthi, extracting with 85% ethanol under reflux for 3 times, wherein the amount of 85% ethanol added is 10 times of the weight of fructus Phyllanthi, extracting under reflux at 70 deg.C for 1.5 hr, filtering with 200 mesh filter cloth, mixing filtrates, controlling temperature at 59 deg.C, concentrating under reduced pressure of 0.09Mpa to 2L, centrifuging at 6000r/min, dissolving precipitate with 800ml of 60% ethanol under ultrasonic, adding 55g of Kaolin, stirring at 40 deg.C in water bath, centrifuging to remove Kaolin, controlling temperature at 58 deg.C, concentrating the supernatant under reduced pressure of 0.09Mpa, adding water to no alcohol, adding 5% sodium hydroxide into 900ml to adjust pH to 9.0, separating with column volume of 1L of macroporous resin AB-8 (Xian blue, science and technology, Ltd.), purifying with water at flow rate of 3 s per drop and 1L, resolving with 2L 60% ethanol solution, collecting resolved solution, concentrating under reduced pressure of-0.09 Mpa at 58 deg.C to 200ml, centrifuging by the same method, collecting precipitate, and freeze drying to obtain 50.14g fructus Phyllanthi extract with yield of 2.57% and polyphenol content of 52.34%.
Effects of the embodiment
Firstly, the yield of the emblic leafflower fruit extract is equal to the mass of a sample/the mass of medicinal materials multiplied by 100 percent in examples 1 to 3 and comparative examples 1 to 2.
Secondly, the polyphenol content of the emblic leafflower fruit extract is measured by referring to a Folin-Ciocalteu method according to the detection method for the polyphenol content of the emblic leafflower fruit extract disclosed in the embodiments 1 to 3 and the comparative examples 1 to 2.
Thirdly, the emblic leafflower fruit extract in the embodiment 1 to 3 and the comparative example 1 to 2 is subjected to animal experiments, and the method comprises the following steps:
the method adopts forced swimming of mice in a pharmacological experimental methodology (the pharmacological experimental methodology, people's health press, 2005: 807-808) and antidepressant bioactivity research of intragastric administration.
SPF male SD rats weighing 180-200 g are purchased from Schlekschada laboratory animals Co., Ltd, Hunan, and the license numbers are as follows: SCXK (Xiang) 2019-.
Establishment of chronic unpredictable stress depression rat model: 100 rats were randomly divided into 10 groups of 10 rats each. The rat model of chronic unpredictable stress depression is established by single-cage breeding in the following stimulation modes: swimming in cold water at 4 ℃ for 5 minutes, keeping water for 24 hours, fasting for 24 hours, clamping tail for 1 minute, shaking the squirrel cage for 5 minutes 1 time per second, wetting padding for 12 hours, reversing day and night for 12 hours, and binding for 5 minutes; each rat used 1 stimulation regimen daily, and the same stimulation regimen was not applied to the same rat consecutively, making the stimulation unpredictable. The molding time was three weeks.
The administration scheme is as follows: each group of rats was gavaged from day 8 after the start of stress. The 10 groups of rats were: (1) a model control group; (2) imipramine (Sigma) positive control group; (3) example 1 emblic leafflower fruit polyphenol low dose group; (4) example 1 emblic leafflower fruit polyphenol high dose group; (5) example 2 low dose group of emblic leafflower fruit polyphenols; (6) example 2 emblic leafflower fruit polyphenol high dose group; (7) example 3 emblic polyphenol low dose group; (8) example 3 emblic leafflower fruit polyphenol high dose group; (9) comparative example 1 emblica officinalis extract high dose group; (10) comparative example 2 emblica officinalis extract high dose group.
The model control group was perfused with normal saline 1 time a day for 14 days.
The imipramine positive control group is perfused with normal saline of imipramine 1 time a day for 14 days continuously, and each rat is perfused with 15mg/kg daily.
Example 1 Low dose group of Phyllanthus emblica polyphenols A physiological saline mixture of Phyllanthus emblica polyphenols was gavaged 1 time a day for 14 consecutive days with 5mg/kg per rat per day.
Example 1 high dose group of emblic leafflower fruit polyphenols Normal saline mixture of emblic leafflower fruit polyphenols was gavaged 1 time a day for 14 consecutive days, each rat was gavaged 15mg/kg a day.
Example 2 Low-dose administration group of Phyllanthus emblica polyphenols Normal saline mixture of Phyllanthus emblica polyphenols 1 time a day, continuous administration for 14 days, and administration of 5mg/kg per rat a day.
Example 2 high dose group of emblic leafflower fruit polyphenols Normal saline mixture of emblic leafflower fruit polyphenols was gavaged 1 time a day for 14 consecutive days, each rat was gavaged 15mg/kg a day.
Example 3 Low dose group of Phyllanthus emblica polyphenols A physiological saline mixture of Phyllanthus emblica polyphenols was gavaged 1 time a day for 14 consecutive days with 5mg/kg per rat per day.
Example 3 high dose group of emblic leafflower fruit polyphenols Normal saline mixture of emblic leafflower fruit polyphenols was intragastrically administered 1 time a day for 14 consecutive days, each rat was intragastrically administered 15mg/kg a day.
Comparative example 1 emblic polyphenol high dose group was intragastrically administered 1 time a day with a physiological saline mixture of emblic polyphenol for 14 consecutive days, 15mg/kg per rat per day.
Comparative example 2 emblic polyphenol high dose group was intragastrically infused with a physiological saline mixture of emblic polyphenol 1 time a day for 14 consecutive days, 15mg/kg per rat per day.
On day 22, i.e. on day 3 after the completion of the stimulation, i.e. on day 3 after the completion of the gavage, the immobility time of each group of rats in the swimming pool was observed by forced swimming test, and the immobility time was determined: the animal stops struggling in water or is in a floating state, only the nostril is exposed to keep breathing, and only the small limb moves to keep the head floating on the water. The self-made glass tank is used as a swimming pool, the depth is 60 cm, the length is 2 m, the water adding depth is 30 cm, and the water temperature is controlled to be 23-26 ℃. Rats were placed in water and recorded for 5 minutes from the time the floating body gave up swimming, and the immobility time of the forced swimming experiment was observed. The immobility time of each group of rats is expressed as mean and standard deviation. The comparison between groups was performed by t-test. The results of the experiments are shown in the following table.
TABLE 1 antidepressant test data for various groups of rats
Note: compared with the model control group: p < 0.05; denotes p < 0.01.
The forced swimming immobility time is a typical index for inspecting depression rats. The experimental results in table 1 show that the experiments in the high-dose groups in examples 1, 2 and 3 can significantly reduce the immobility time of rats, have very significant antidepressant effect, and the effect is better than that of the traditional antidepressant drug imipramine, and the mice in the high-dose groups in examples 1, 2 and 3 have good states, smooth weight growth and no obvious toxic or side effect. Examples 1, 2 and 3 the low dose group rats had a significant antidepressant effect at immobility time, and the effect was concentration dependent; both groups of comparative examples 1 and 2 had no significant antidepressant effect.
The above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Those skilled in the art should also realize that changes, modifications, additions and substitutions can be made without departing from the true spirit and scope of the invention.
Claims (9)
1. A preparation method of an emblic extract is characterized by comprising the following steps:
(1) and (3) extraction and concentration: crushing raw material emblic leafflower fruit medicinal material, adding 70-95% ethanol, extracting at 50-80 ℃, filtering, concentrating under reduced pressure, centrifuging, taking precipitate, adding ethanol for dissolving to obtain feed liquid a;
(2) kaolin adsorption: adding kaolin into the feed liquid a, carrying out water bath, stirring, centrifuging, taking supernate, concentrating under reduced pressure, and supplementing water until no alcohol smell exists to obtain feed liquid b;
(3) column chromatography: and (3) adjusting the pH value of the feed liquid b to 8.5-10, then purifying by strong base anion resin, washing with water to remove impurities, resolving by NaCl with the concentration of 5-10%, concentrating under reduced pressure, centrifuging, taking precipitate, and freeze-drying to obtain the phyllanthus emblica extract.
2. The preparation method of the emblic extract according to claim 1, wherein in the step (1), the addition amount of the 70-95% ethanol is 6-12 times of the weight of the raw material emblic.
3. The method for preparing the emblic extract according to claim 1, wherein in the step (1), the concentration under reduced pressure is carried out until the volume of the material liquid is 1 time of the mass of the raw material emblic.
4. The method for preparing the emblic extract according to claim 1, wherein the concentration of the ethanol in the step (1) is 60% when the ethanol is added for dissolution.
5. The method according to claim 1, wherein in the step (2), the amount of the kaolin added is 1 to 5% of the amount of the raw material emblic leafflower fruit.
6. The method for preparing the emblic leafflower fruit extract according to claim 1, wherein the temperature of the water bath in the step (2) is 40 to 55 ℃.
7. The method for preparing the emblic extract according to claim 1, wherein in the step (3), the strong base anion resin is any one of D201, 7170, D204, HZ202, ZGA307, 201x 7.
8. An extract of emblic leafflower fruit, characterized by being prepared by the process for preparing an extract of emblic leafflower fruit according to any one of claims 1 to 7.
9. Use of an extract of Phyllanthus emblica as claimed in claim 8 for anti-depression.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911366808.0A CN111035670A (en) | 2019-12-26 | 2019-12-26 | Phyllanthus emblica extract and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911366808.0A CN111035670A (en) | 2019-12-26 | 2019-12-26 | Phyllanthus emblica extract and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111035670A true CN111035670A (en) | 2020-04-21 |
Family
ID=70240181
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911366808.0A Pending CN111035670A (en) | 2019-12-26 | 2019-12-26 | Phyllanthus emblica extract and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111035670A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115606734A (en) * | 2022-09-15 | 2023-01-17 | 滇西科技师范学院 | Elaeagnus conferta roxb antioxidant extract and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007014334A2 (en) * | 2005-07-27 | 2007-02-01 | Natreon, Inc. | Method of treatment or management of stress |
| CN106617060A (en) * | 2016-12-27 | 2017-05-10 | 云南中烟工业有限责任公司 | Fructus phyllanthi polyphenols enriching and purifying method |
| CN109662967A (en) * | 2019-02-19 | 2019-04-23 | 孙益虎 | A kind of antidepressant and application thereof |
-
2019
- 2019-12-26 CN CN201911366808.0A patent/CN111035670A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007014334A2 (en) * | 2005-07-27 | 2007-02-01 | Natreon, Inc. | Method of treatment or management of stress |
| CN106617060A (en) * | 2016-12-27 | 2017-05-10 | 云南中烟工业有限责任公司 | Fructus phyllanthi polyphenols enriching and purifying method |
| CN109662967A (en) * | 2019-02-19 | 2019-04-23 | 孙益虎 | A kind of antidepressant and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| 尤文挺: "多酚类化合物抗抑郁及抑郁相关学习记忆障碍研究进展", 《国际药学研究杂志》 * |
| 曹广尚: "阴离子交换树脂分离纯化绵马贯众总多酚的工艺研究", 《中草药》 * |
| 郝利平: "《食品添加剂》", 31 July 2016, 中国农业大学出版社 * |
| 郭炳春等: "余甘子总多酚提取工艺优化研究", 《热带作物学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115606734A (en) * | 2022-09-15 | 2023-01-17 | 滇西科技师范学院 | Elaeagnus conferta roxb antioxidant extract and preparation method and application thereof |
| CN115606734B (en) * | 2022-09-15 | 2024-04-30 | 滇西科技师范学院 | Antioxidant extract of elaeagnus conferta roxb as well as preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1754541A (en) | Steroid saponin pharmaceutical composition and its preparation method and uses | |
| CN101463061A (en) | Ginseng saponin Rg1 and Rb1 in pseudo-ginseng and preparation of total saponin thereof | |
| CN112870236B (en) | Flavone effective part of abelmoschus manihot and preparation method and application thereof | |
| WO2014131213A1 (en) | Process for preparing epigallocatechin gallate and application thereof | |
| CN110283220B (en) | Separation and purification method of crocin-1 and application thereof | |
| CN107412430A (en) | A kind of radix scrophulariae water extract and its application | |
| CN111035670A (en) | Phyllanthus emblica extract and preparation method and application thereof | |
| CN101214250B (en) | Use of hederagenin and its derivatives in preparing antidepressant product | |
| JP2020534357A (en) | Flavonoid extract of Fusilier gynura, its preparation method and its use for treating alcoholic fatty liver | |
| CN109280090B (en) | Extraction method and application of coridius stinkbug family insect polysaccharide with kidney warming and yang tonifying activities | |
| CN1947736A (en) | Prepn. method and application of injection contg. Erigeron breviscapus | |
| CN1199650C (en) | Medicine composition of red peony total glucoside for curing cardio-cerebrovascular diseases and its preparation method | |
| CN1857622A (en) | Medicine composition and preparation containing effective components of gastrodia tuber and Chuanxiong rhizome | |
| CN112107639B (en) | Application of fructus viticis extract in preparation of antidepressant drugs | |
| TWI438001B (en) | Plant extract for treating diabetes and process for making same | |
| CN111358826B (en) | Total flavone extract with antidepressant effect, and preparation method and application thereof | |
| CN1698717A (en) | Chinese medicinal compound fat emulsion injection and its preparation method | |
| CN109810153B (en) | Preparation method and analgesic application of aromatic substituted glucose compound and pharmaceutical composition thereof | |
| CN1193766C (en) | Gardenia total glycoside composite for curing hepatitis and its preparation method | |
| CN106265956B (en) | Traditional Chinese medicine extract and composition for resisting cerebral ischemia, and preparation and application thereof | |
| CN107412393B (en) | Preparation method and application of clausena lansium leaf polyphenol extract | |
| CN116509921B (en) | Application of rice flower sesterterpene extract K01 in preparation of medicines for treating sepsis | |
| JP4995727B2 (en) | Use of winter savory (SATUREJAMMONTANA) or its extract for the manufacture of a drug for the treatment of premature ejaculation | |
| CN114588149B (en) | Application of petasites tricholobus total lactone and petasites lactone compounds in preparation of drugs for improving exercise capacity | |
| CN105712982A (en) | Dihydro-beta-agarofuran sesquiterpenoids, and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200421 |