CN104666474B - A kind of Wild jujube seeds extract and the brain tonic sleeping oral liquid containing Wild jujube seeds extract - Google Patents
A kind of Wild jujube seeds extract and the brain tonic sleeping oral liquid containing Wild jujube seeds extract Download PDFInfo
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Abstract
The invention belongs to biomedicine field, and in particular to a kind of Wild jujube seeds extract and the brain tonic sleeping oral liquid containing Wild jujube seeds extract.The preparation method of described Wild jujube seeds extract comprises the following steps：Jujube kernel crushing, active material water extraction, semi-bionic extraction method, concentration, alcohol deposition method decontamination, obtain Wild jujube seeds extract.The brain tonic sleeping oral liquid containing Wild jujube seeds extract is prepared with other components hook tunes such as honey further using Wild jujube seeds extract as raw material by the present invention.The oral liquid contains spina date seed sleep aiding and nerve soothing active ingredient, including spina date seed flavonoid substance, spina date seed alkaloid, jujuboside D and E, have calm the nerves, brain tonic, hypnosis, effect of benefiting intelligence.
Description
Technical field
The invention belongs to biomedicine field, and in particular to a kind of Wild jujube seeds extract and being good for containing Wild jujube seeds extract
Brain sleeping oral liquid.
Background technology
Insomnia is a kind of most common sleep-disorder, has significant damage to function of human body, intelligence and immunity.It is long-term to lose
Sleep, not only damage the thinking activities of people, and the immune system of people can be influenceed, eventually shorten the lifetime.Clinically commonly use
Chemical synthetic drug treatment insomnia with sedative-hypnotic effect.Questionnaire result shows that about 44% patient takes benzene phenodiazine for a long time
Tall and erect class (BDZ) and barbiturate hypnosis, but these medicines because with side effect and it is additive make its application be restricted.
Spina date seed is rhamnaceae plant wild jujube Ziziphus jujuba Mill var.spinosa Huex.H.F.Chou
Drying and ripening kernel,《Chinese Pharmacopoeia》Nineteen ninety-five version is recorded, and is had tonifying liver, calming heart, arrest sweating, the function of promoting the production of body fluid, is used for
The diseases such as restlessness of asrhenia type and insomnia, horrified to much dream, injury thirst, it is a kind of conventional Chinese medicine.Its main composition of calming the nerves is probably acid therein
Jujube kernel saponin, wild jujube flavone c-glycoside and swertisin etc..Traditional Chinese medicine has good sedation of calming the nerves, and adverse reaction is few,
With not habituation, the remarkable advantages such as dependence, tolerance will not be also produced, there is broad prospect of application.
The content of the invention
For overcome the deficiencies in the prior art and shortcoming, primary and foremost purpose of the invention is to provide a kind of Wild jujube seeds extract
Preparation method.
The Wild jujube seeds extract being prepared another object of the present invention is to provide above-mentioned preparation method.
It is still another object of the present invention to provide a kind of brain tonic sleeping oral liquid containing above-mentioned Wild jujube seeds extract.
The purpose of the present invention is achieved through the following technical solutions：
A kind of preparation method of Wild jujube seeds extract, is comprised the following steps：
(1) spina date seed crushes, and obtains spiny jujuba seed powder；Spiny jujuba seed powder hot water extraction's active material, filtering, obtains filtrate 1
With filter residue 1；
(2) filter residue 1 that step (1) is prepared is soaked with pH 1~5 acid solution, filtering, obtains filtrate 2 and filter
Slag 2；Filter residue 2 is soaked with pH 8~10 alkaline solution again, filtering, obtains filtrate 3 and filter residue 3；
(3) filtrate 1,2 and 3 that combining step (1) and step (2) are prepared, concentration, obtains concentrate；
(4) concentrate being prepared in step (3) is subjected to alcohol precipitation with ethanol, except desizing, mucilaginous substance, natural gum, egg
The impurity such as white matter, grease, fat-soluble pigment, polysaccharide, inorganic salts, tannin；Centrifugation, filtrate is collected, obtains Wild jujube seeds extract；
The temperature of hot water extraction described in step (1) is preferably 75~80 DEG C, and time of hot water extraction is preferably 2~
4h；
The time of acid solution immersion described in step (2) is preferably 2~4h；
The pH value of acid solution described in step (2) is preferably 5；
Acid solution described in step (2) is preferably hydrochloric acid solution；
The time of alkaline solution immersion described in step (2) is preferably 2~4h；
The pH value of alkaline solution described in step (2) is preferably 8.2；
Alkaline solution described in step (2) is preferably sodium hydroxide solution；
Concentration described in step (3) is preferably concentrated under reduced pressure or freeze concentration；
The volume fraction of ethanol described in step (4) is preferably 75%~80%；
The time of alcohol precipitation described in step (4) is preferably 3~4h；
A kind of Wild jujube seeds extract, it is prepared by above-mentioned preparation method；
Described Wild jujube seeds extract includes 8 kinds of Flavonoid substances, spina date seed alkaloid, jujuboside class material；(figure
1~4)
8 kinds of described Flavonoid substances are respectively swertisin (Fig. 1 Chinese styles I), apigenin -6-C- β-D- glucopyras
Glucoside (Fig. 1 Chinese styles II), Puerarin (Fig. 1 Chinese styles III), 6 " '-forulic acid wild jujube flavone c-glycoside (Fig. 1 Chinese styles IV), wild jujube flavones
Carbon glucoside glycosides (Fig. 1 Chinese styles V), isovitexin (VI formula in Fig. 1), different wild jujube flavone c-glycoside (Fig. 1 Chinese styles VII) and 6 " '-forulic acid
Different wild jujube flavone c-glycoside (Fig. 1 Chinese styles VIII)；
The structural formula of described spina date seed alkaloid is as shown in Figure 2；
Described jujuboside class material is jujuboside D and jujuboside E；(Fig. 3 and Fig. 4)
In described Wild jujube seeds extract, the contents of Flavonoid substances is 0.576~0.594mg/g, jujuboside class
The content of material is 0.365~0.382mg/g；
A kind of brain tonic sleeping oral liquid containing Wild jujube seeds extract, includes component as follows by mass percentage：
Above-mentioned Wild jujube seeds extract 20%~22%；
Honey 8%~10%；
Water supplies 100%；
The preparation method of the described brain tonic sleeping oral liquid containing Wild jujube seeds extract, is comprised the following steps：
By each component hook tune of the above-mentioned brain tonic sleeping oral liquid containing Wild jujube seeds extract, sterilizing, obtain containing wild jujube
The brain tonic sleeping oral liquid of benevolence extract；
The condition of described sterilizing is preferably the 25min that sterilized at 78~80 DEG C；
There are the mainly two class materials, i.e. Flavonoid substances and jujuboside class thing of syngignoscism of calming the nerves in spina date seed
Matter, concrete structure are shown in Fig. 1, Fig. 2, Fig. 3 and Fig. 4.The present invention is to having the active principle for syngignoscism of calming the nerves to carry out in spina date seed
Separation and purifying, obtain Wild jujube seeds extract, and further science is configured to brain tonic sleeping oral liquid, and by measuring perfusion mouse
The mode Test extraction thing of righting reflex loss and recovery time activity and toxicological effect, it was demonstrated that the nontoxic secondary work of Wild jujube seeds extract
With curative for effect.The brain tonic sleeping oral liquid can be used for caused by overstrain one's nerves, nervous, operating pressure are big insomnia,
The symptoms such as anxiety, have calm the nerves, brain tonic, hypnosis, intelligence development and other effects.
The present invention is had the following advantages relative to prior art and effect：
(1) preparation method of Wild jujube seeds extract provided by the invention is simple to operation：Crushed by jujube kernel, active material
Water extraction, semi-bionic extraction method, concentration, alcohol deposition method decontamination, obtain Wild jujube seeds extract；Active ingredient retains more；Whole extraction
The organic solvent that process is related to only has ethanol, safely controllable, has the advantages of harmless to the human body.
(2) yield of product is significantly improved made from the preparation method of Wild jujube seeds extract provided by the invention.
(3) it is effective to contain spina date seed sleep aiding and nerve soothing for the brain tonic sleeping oral liquid provided by the invention containing Wild jujube seeds extract
Composition, including spina date seed flavonoid substance, spina date seed alkaloid, jujuboside D and E, have no toxic side effect, tranquilizing and allaying excitement curative effect is true
Cut, can be used for the symptom such as insomnia, anxiety caused by overstrain one's nerves, nervous, operating pressure are big.
Brief description of the drawings
Fig. 1 is the Flavonoid substances structural formula schematic diagram that the Wild jujube seeds extract that the present invention is prepared includes.
Fig. 2 is the spina date seed alkaloid monomer structural formula schematic diagram that the Wild jujube seeds extract that the present invention is prepared includes.
Fig. 3 is the jujuboside D structural formula schematic diagrames that the Wild jujube seeds extract that the present invention is prepared includes.
Fig. 4 is the jujuboside E structural formula schematic diagrames that the Wild jujube seeds extract that the present invention is prepared includes.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.
Embodiment 1
(1) spina date seed after air-dried or airing is crushed with the pulverizer by sterilization, obtains spiny jujuba seed powder；Wild jujube
Benevolence powder filtering, obtains filtrate 1 and filter residue 1 with after 80 DEG C of hot-water soak 3 hours, wherein, the Cord blood of filtrate 1 is standby；
(2) semi-bionic extraction method：The filter residue 1 that step (1) is prepared is soaked with pH 5 hydrochloric acid solution, is filtered after 3 hours,
Filtrate 2 and filter residue 2 are obtained, wherein, the Cord blood of filtrate 2 is standby；After filter residue 2 is soaked 3 hours with pH8.2 sodium hydroxide solution
Filtering, obtains filtrate 3 and filter residue 3, wherein, the Cord blood of filtrate 3 is standby；
(3) concentrate：The filtrate 1,2 and 3 that combining step (1) and step (2) are prepared, using being evaporated under reduced pressure, progress is dense
Contracting, obtains concentrate；
(4) alcohol precipitation：The concentrate that step (3) is prepared is subjected to alcohol precipitation with the ethanol that volume fraction is 80%, removed
The impurity such as starch, mucilaginous substance, natural gum, protein, grease, fat-soluble pigment, polysaccharide, inorganic salts, tannin.Alcohol precipitation is carried out 3 hours
Afterwards, filtrate is collected by centrifugation, obtains wild jujube extract, recovery rate 4%.
Embodiment 2
(1) spina date seed after air-dried or airing is crushed with the pulverizer by sterilization, obtains spiny jujuba seed powder；Wild jujube
Benevolence powder filtering, obtains filtrate 1 and filter residue 1 with after 75 DEG C of hot-water soak 4 hours, wherein, the Cord blood of filtrate 1 is standby；
(2) semi-bionic extraction method：The filter residue 1 that step (1) is prepared is soaked with pH 1 hydrochloric acid solution, is filtered after 2 hours,
Filtrate 2 and filter residue 2 are obtained, wherein, the Cord blood of filtrate 2 is standby；After filter residue 2 is soaked 2 hours with pH10 sodium hydroxide solution
Filtering, obtains filtrate 3 and filter residue 3, wherein, the Cord blood of filtrate 3 is standby；
(3) concentrate：The filtrate 1,2 and 3 that combining step (1) and step (2) are prepared, using being evaporated under reduced pressure, progress is dense
Contracting, obtains concentrate；
(4) alcohol precipitation：The concentrate that step (3) is prepared is subjected to alcohol precipitation with the ethanol that volume fraction is 75%, removed
The impurity such as starch, mucilaginous substance, natural gum, protein, grease, fat-soluble pigment, polysaccharide, inorganic salts, tannin.It is small that alcohol precipitation carries out 3.5
Shi Hou, filtrate is collected by centrifugation, obtains wild jujube extract, recovery rate 4%.
Embodiment 3
(1) spina date seed after air-dried or airing is crushed with the pulverizer by sterilization, obtains spiny jujuba seed powder；Wild jujube
Benevolence powder filtering, obtains filtrate 1 and filter residue 1 with after 78 DEG C of hot-water soak 2 hours, wherein, the Cord blood of filtrate 1 is standby；
(2) semi-bionic extraction method：The filter residue 1 that step (1) is prepared is soaked with pH 4 hydrochloric acid solution, is filtered after 4 hours,
Filtrate 2 and filter residue 2 are obtained, wherein, the Cord blood of filtrate 2 is standby；Mistake after filter residue 2 is soaked 4 hours with pH9 sodium hydroxide solution
Filter, obtains filtrate 3 and filter residue 3, wherein, the Cord blood of filtrate 3 is standby；
(3) concentrate：The filtrate 1,2 and 3 that combining step (1) and step (2) are prepared, using being evaporated under reduced pressure, progress is dense
Contracting, obtains concentrate；
(4) alcohol precipitation：The concentrate that step (3) is prepared is subjected to alcohol precipitation with the ethanol that volume fraction is 78%, removed
The impurity such as starch, mucilaginous substance, natural gum, protein, grease, fat-soluble pigment, polysaccharide, inorganic salts, tannin.Alcohol precipitation is carried out 4 hours
Afterwards, filtrate is collected by centrifugation, obtains wild jujube extract, recovery rate 4%.
Embodiment 4
(1) jujuboside D in Wild jujube seeds extract, E it is qualitative
Using methanol as solvent, concentration 1mg/mL jujuboside D and jujuboside E (in being purchased from are configured to respectively
State's medicine biological products assay institute) solution, as reference substance solution 1 and reference substance solution 2.
According to thin-layered chromatography (《Chinese Pharmacopoeia》2005 editions one) experiment, drawing need testing solution, (embodiment 1,2 and 3 is made
Standby obtained Wild jujube seeds extract) and each 10 μ L of blank control liquid (methanol), reference substance solution 1,2 each 5 μ L, put respectively in same
On silica gel g thin-layer plate, using water saturated butanol solution as solvent, deploy, take out, dry, spray 1% (mass percent)
Vanillin-sulfuric acid solution, inspect immediately.In test sample chromatogram, on position corresponding with reference substance chromatogram, identical face is shown
The spot of color, it is jujuboside so as to demonstrate the active ingredient in the Wild jujube seeds extract that embodiment 1,2 and 3 is prepared
D、E。
(2) in Wild jujube seeds extract saponins material quantitative experiment：
The preparation of standard curve：Precision weighs ginsenoside Rb1's reference substance of dry constant weight (purchased from Chinese medicine biology system
Institute is determined in product examine) 5.0mg, 10mL volumetric flasks are moved to, with absolute ethyl alcohol constant volume, are configured to 0.5mg/mL reference substance solution.It is accurate
The general Rb1 reference substance solutions (C=0.5mg/mL) 0 of ginseng soap, 0.1,0.2,0.3,0.4,0.5,0.6ml are drawn in 10mL tool plug mills
In oral examination pipe, reagent is volatilized in an oven, adds 5% (mass percent) vanillic aldehyde-glacial acetic acid solution 0.2mL and perchloric acid
0.8mL, shake up, 60 DEG C of heating water bath 20min, 3min is cooled down in ice-water bath, add glacial acetic acid 5mL, shake up, retinue reagent is made empty
In vain, absorbance is determined at 545nm with spectrophotometer, using ginsenoside Rb1's concentration as abscissa, it is vertical to absorb angle value
Coordinate, standard curve is drawn, as a result find that ginsenoside Rb1 is in good linear relation in the range of 0.01-0.05mg/mL, obtain
Regression equation y=17.925x-0.0498, R2=0.9988.
Each 0.5mL of Wild jujube seeds extract that accurate extraction embodiment 1,2 and 3 is prepared, it is placed in 10mL tool plug ground examinations
Guan Zhong, operated according to " preparation of standard curve ", by above-mentioned standard curve, try to achieve the content of saponins material：Embodiment 1,2
It is respectively 0.382mg/g, 0.365mg/g, 0.370mg/g with 3 contents that saponins material in Wild jujube seeds extract is prepared.
(3) in Wild jujube seeds extract Flavonoid substances quantitative experiment：
The preparation of standard curve：Precision draws rutin standard liquid (being purchased from Nat'l Pharmaceutical ＆ Biological Products Control Institute) (C=
0.2mg/mL) 0,2,4,6,8,10mL adds distilled water diluting then to add 5% (quality hundred to 10mL in 50mL volumetric flask
Divide ratio) NaNO2Solution 2mL, shakes up, and stands 6min；Again plus 10% (mass percent) Al (NO3)3Solution 2mL, shakes up, and stands
6min；4% (mass percent) NaOH solution 20mL is eventually adding, scale is settled to distilled water, shakes up, stands 15min, is used
Spectrophotometer determines absorbance at 510nm.Using the concentration of control substance of Rutin as abscissa, absorbance is ordinate,
Standard curve is drawn, obtains regression equation y=13.113x-0.0177, R2=0.9994.
Precision draws each 2mL of Wild jujube seeds extract that above-described embodiment 1,2 and 3 is prepared, and is placed in 50mL volumetric flasks,
Operated according to " preparation of standard curve ", by above-mentioned standard curve, try to achieve the content of Flavonoid substances：Embodiment 1,2 and 3 is made
The standby content for obtaining Flavonoid substances in Wild jujube seeds extract is respectively 0.594mg/g, 0.576mg/g, 0.580mg/g.
The preparation of brain tonic sleeping oral liquid of the embodiment 5 containing Wild jujube seeds extract
(1) hook tune：Wild jujube seeds extract, honey and the mixing of water hook tune that embodiment 1 is prepared；Wherein, spina date seed
The mass percent of extract, honey and water is respectively 20%, 8%, 72%；
(2) dispense and sterilize：Mixed liquor after step (1) hook tune is subjected to pipe dress, it is every small bottled 10 milliliters.Use Bath
Moral sterilizer is carried out disinfection, and 25min is sterilized at 80 DEG C, then is carried out bacterial number sanitary index and examined that (reference standard is food
Product sanitary quality standard), qualified labeling.Every 10 fill a box, every 50 box-packed case.
Storage of finished products is at low temperature, ventilation, drying, or is stored in 4 DEG C of refrigerators, and long shelf-life was up to more than 6 months.
The preparation of brain tonic sleeping oral liquid of the embodiment 6 containing Wild jujube seeds extract
(1) hook tune：Wild jujube seeds extract, honey and the mixing of water hook tune that embodiment 1 is prepared；Wherein, spina date seed
The mass percent of extract, honey and water is respectively 21%, 9%, 70%；
(2) dispense and sterilize：Mixed liquor after step (1) hook tune is subjected to pipe dress, it is every small bottled 10 milliliters.Use Bath
Moral sterilizer is carried out disinfection, and 24min is sterilized at 78 DEG C, then is carried out bacterial number sanitary index and examined that (reference standard is food
Product sanitary quality standard), qualified labeling.Every 10 fill a box, every 50 box-packed case.
Storage of finished products is at low temperature, ventilation, drying, or is stored in 4 DEG C of refrigerators, and long shelf-life was up to more than 6 months.
The preparation of brain tonic sleeping oral liquid of the embodiment 7 containing Wild jujube seeds extract
(1) hook tune：Wild jujube seeds extract, honey and the mixing of water hook tune that embodiment 1 is prepared；Wherein, spina date seed
The mass percent of extract, honey and water is respectively 22%, 10%, 68%；
(2) dispense and sterilize：Mixed liquor after step (1) hook tune is subjected to pipe dress, it is every small bottled 10 milliliters.Use Bath
Moral sterilizer is carried out disinfection, and 20min is sterilized at 79 DEG C, then is carried out bacterial number sanitary index and examined that (reference standard is food
Product sanitary quality standard), qualified labeling.Every 10 fill a box, every 50 box-packed case.
Storage of finished products is at low temperature, ventilation, drying, or is stored in 4 DEG C of refrigerators, and long shelf-life was up to more than 6 months.
The direct sleep effect of the Wild jujube seeds extract of embodiment 8
Mouse (being purchased from Guangzhou Experimental Animal Center) 40, male and female half and half, is randomly divided into 4 groups.Wherein 3 groups of test group,
I.e. brain tonic sleeping oral liquid high (30mL/kg), in (20mL/kg), low (10mL/kg) dosage group.Remaining one group is normal control
Group, brain tonic sleeping oral liquid are prepared by embodiment 5.It was up to more than 40 seconds with mouse righting reflex loss after gavage oral liquid
Fall asleep, revival is reverted to righting reflex, record and calculate the number of elements of every group of mice sleep, sleep rate, time for falling asleep and sleep
Duration.As a result it is as shown in table 1.
As a result show, Normal group mouse is not slept, and activity is normal.Each dosage brain tonic sleeping oral liquid can make
There is sleep (table 1) in mouse.
The direct sleep laboratory of brain tonic sleeping oral liquid of table 1
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (7)
1. a kind of preparation method of Wild jujube seeds extract, it is characterised in that comprise the following steps：
(1) spina date seed crushes, and obtains spiny jujuba seed powder；Spiny jujuba seed powder hot water extraction's active material, filtering, obtain filtrate 1 and filter
Slag 1；
(2) filter residue 1 that step (1) is prepared is soaked with pH 5 acid solution, filtering, obtains filtrate 2 and filter residue 2；Filter residue 2
Soaked again with pH 8~10 alkaline solution, filtering, obtain filtrate 3 and filter residue 3；
(3) filtrate 1,2 and 3 that combining step (1) and step (2) are prepared, concentration, obtains concentrate；
(4) concentrate being prepared in step (3) is subjected to alcohol precipitation with ethanol, centrifuged, collect filtrate, obtain spina date seed extraction
Thing；
The temperature of hot water extraction described in step (1) is 75~80 DEG C, and the time of hot water extraction is 2~4h；
The time of acid solution or alkaline solution immersion described in step (2) is 2~4h.
2. the preparation method of Wild jujube seeds extract according to claim 1, it is characterised in that：
The pH value of alkaline solution described in step (2) is 8.2.
3. the preparation method of Wild jujube seeds extract according to claim 1, it is characterised in that：
Concentration described in step (3) is is concentrated under reduced pressure or freeze concentration.
4. the preparation method of Wild jujube seeds extract according to claim 1, it is characterised in that：
The volume fraction of ethanol described in step (4) is 75%~80%；
The time of alcohol precipitation described in step (4) is 3~4h.
5. a kind of Wild jujube seeds extract, it is characterised in that be prepared into by the preparation method described in any one of Claims 1 to 4
Arrive.
6. a kind of brain tonic sleeping oral liquid containing Wild jujube seeds extract, it is characterised in that comprising as follows by mass percentage
Component：
Wild jujube seeds extract 20%~22% described in claim 5；
Honey 8%~10%；
Water supplies 100%.
7. the preparation method of the brain tonic sleeping oral liquid containing Wild jujube seeds extract described in claim 6, it is characterised in that bag
Containing following steps：
The each component of the brain tonic sleeping oral liquid containing Wild jujube seeds extract described in claim 6 is subjected to hook tune, sterilizing, obtained
To the brain tonic sleeping oral liquid containing Wild jujube seeds extract.
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| CN106620314A (en) * | 2016-11-11 | 2017-05-10 | 李成义 | Oral sorrow-forgetting and sleep-promoting solution for improving sub-health state |
| CN106858596A (en) * | 2017-02-10 | 2017-06-20 | 天津金匮堂生物科技有限公司 | A kind of probiotic acid jujube oral liquid and preparation method thereof |
| CN114304603A (en) * | 2020-09-29 | 2022-04-12 | 拜尔普兰（厦门）生物药业有限公司 | Sublingual instant granules of spina date seeds and preparation method thereof |
| CN113214349B (en) * | 2021-05-14 | 2022-07-26 | 华南理工大学 | Spina date seed saponin, preparation thereof and application thereof in preparing hypoglycemic drugs |
| CN113142344A (en) * | 2021-05-18 | 2021-07-23 | 北京晚安科技有限责任公司 | Sleep-aiding solid beverage and preparation method thereof |
| CN114504105A (en) * | 2022-02-15 | 2022-05-17 | 江中食疗科技有限公司 | Preparation method and application of wild jujube kernel oil extract |
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| CN103272018A (en) * | 2013-05-10 | 2013-09-04 | 华南理工大学 | Sedative and hypnotic active saponin components of spina date seed, as well as separation and purification method and application of sedative and hypnotic active saponin components |
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| CN103272018A (en) * | 2013-05-10 | 2013-09-04 | 华南理工大学 | Sedative and hypnotic active saponin components of spina date seed, as well as separation and purification method and application of sedative and hypnotic active saponin components |
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