CN103272018A - 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 - Google Patents
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 Download PDFInfo
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Abstract
The invention discloses sedative and hypnotic active saponin components of a spina date seed, as well as separation and purification method and application of the sedative and hypnotic active saponin components, and belongs to the field of separation of natural products. The invention designs a new and relatively safe extracting route without introducing obviously toxic solvents and agents. The separation and purification method is characterized by comprising the following steps of: extracting a spina date seed sedative and hypnotic active saponin sample from a spina date seed sample by the alcohol extraction method and n-butanol extraction method; and then separating spina date seed sedative and hypnotic active saponin components I and II from the saponin sample by the column chromatography and thin-layer chromatography. The two spina date seed saponin components separated from the spina date seed show obvious hypnotic activity; and the natural sedative and hypnotic active components extracted and separated from traditional Chinese medicine can be used for solving the problems of multiple side effects and addiction of chemical synthetic drugs and is beneficial to human health and contributed to all the society.
Description
Technical field
The invention belongs to the Separation of Natural Products field, particularly calm the nerves hypnotic activity saponin component and isolation and purification method and application of Semen Ziziphi Spinosae.
Background technology
Along with the development of society, the quickening of rhythm of life, more fear and anxieties to failure lie dormant in people's the life.Unexpectedly strive and piece together richly, brought huge pressure for people's spirit, insomnia, anxiety neurosis sickness rate sharply rise.Insomnia is modal a kind of sleep disorder, and function of human body, intelligence and immunity are had significant damage.Long-term insomnia not only damages people's thinking activities, and can influence people's immune system, finally can shorten life.Use the chemical synthetic drug Cure for insomnia with sedative-hypnotic effect clinically always.Investigation finds that about 44% patient takes Benzodiazepines (BDZ) and barbiturates hypnotic drug for a long time, but these medicines are restricted its application because having side effect and addiction.Barbital sodium class medicine increases gradually with dosage and produces calmness, hypnosis, drowsiness, convulsion and anesthetic action, and toxic dose can cause respiratory paralysis and death.Chinese medicine has the sedation of well calming the nerves, and untoward reaction is few, has not addiction, also can not produce remarkable advantages such as dependency, toleration, has broad application prospects.
Existing forefathers carry out the research of hypnotic activity respectively to the flavone in the Semen Ziziphi Spinosae, saponin, oils and fats and alkaloid etc.But have some differences between each result of study, for example the Guo Sheng people wait (Chinese crude drug .1998, (21) 11:578～579; Northwest medical journal .1996, (8) 11:166～168) total flavones, total saponins all have hypnotic activity in the report Semen Ziziphi Spinosae, (Phytochemistry.1978 such as Woo.Siekwoo, 18:353～355) think that also jujuboside A is one of effective ingredient, but (CHINA JOURNAL OF CHINESE MATERIA MEDICA .1991 such as Wu Shuxun, 16 (7): 435～437) discover that jujuboside A does not have the central nerve inhibition effect, the opposite excitation that but can strengthen amfetamine thinks that jujuboside A may not be the effective ingredient of sedative action; (bulletin of Chinese materia medica .1987 such as Yuan Changlu, 12 (3): 34～35) point out that by experiment flavones ingredient also is one of effective ingredient of tranquilizing soporific, and the fatty oil of Petroleum ether extraction does not have this effect, but (the journal .1995 of Xian Medical Univ such as Zhao Qiuxian, (16) 4:421～423) studied the influence of Semen Ziziphi Spinosae fatty oil to the central nervous system, thought that Semen Ziziphi Spinosae oil also may be the effective ingredient of tranquilizing soporific.But evaluation and the separation method of the hypnotic activity composition of calming the nerves in the Semen Ziziphi Spinosae have great significance to human beings'health, still have a lot of needs to study.
Summary of the invention
For overcoming the shortcoming and defect of prior art, primary and foremost purpose of the present invention is to provide a kind of Semen Ziziphi Spinosae isolation and purification method of hypnotic activity saponin component of calming the nerves.This method be one new, do not introduce and have natural product extraction separation method overt toxicity solvent and reagent, comparatively safe.
Another object of the present invention is to provide the Semen Ziziphi Spinosae of the above-mentioned isolation and purification method preparation hypnotic activity saponin component of calming the nerves.
A further object of the present invention is to provide the application of hypnotic activity saponin component of calming the nerves of described Semen Ziziphi Spinosae.
Purpose of the present invention is achieved through the following technical solutions: the calm the nerves isolation and purification method of hypnotic activity saponin component of a kind of Semen Ziziphi Spinosae comprises the steps:
(1) the Semen Ziziphi Spinosae preparation of hypnotic activity saponin sample of calming the nerves:
Semen Ziziphi Spinosae is crossed 40 mesh sieves after parch, pulverizing, 60 ℃ of dry 24h prepare spiny jujuba seed powder; With spiny jujuba seed powder by cable-styled extraction method with the petroleum ether of 10 times of quality at 60 ℃～65 ℃ defat 4h, acquisition residue and Semen Ziziphi Spinosae oil; Take out residue, dry, obtain the defat spiny jujuba seed powder after grinding, cross 40 mesh sieves; Be ethanol reflux, extract, 3h under 85 ℃～90 ℃ conditions of 70% with 25 times of volume fractions with the defat spiny jujuba seed powder, sucking filtration obtains residue I and pure liquid; With pure liquid 60 ℃ of following evaporated under reduced pressure to paste, be dissolved in water, and obtain aqueous solution behind the filtering and impurity removing; The water-saturated n-butanol that adds 1 times of volume to aqueous solution extracts, and abandons water layer, obtains n-butanol layer I; With the KOH solution basification of n-butanol layer I with the 0.1M of 1 times of volume, obtain aqueous alkali layer and n-butanol layer II; With n-butanol layer II evaporated under reduced pressure, namely obtain thick saponin; Thick saponin through three dissolving-filtration-drying at room temperature, is namely obtained the Semen Ziziphi Spinosae hypnotic activity saponin sample of calming the nerves.
(2) the Semen Ziziphi Spinosae isolation and purification method of hypnotic activity saponin component of calming the nerves:
1) column chromatographic isolation and purification
The saponin samples with water of step (1) preparation is mixed with the last sample saponin sample liquid that concentration is 25mg/ml, in the chromatographic column that adsorbent is housed, go up sample 10ml, volume parts is that 60% alcoholic solution is made eluent 40ml, it is 0.5ml/min that adjusting peristaltic pump rotating speed makes flow velocity, and every 10ml one pipe is collected eluent; Every pipe is got 1ml from eluent, and adding the 1ml mass fraction is 2% vanillin solution, the 2ml concentrated sulphuric acid, and through ultraviolet light spectrophotometer scanning discovery the eluent of maximum absorption band being arranged at the 507nm place is the main peak eluent, collects the main peak eluent standby.
2) thin layer chromatography separation and purification
The main peak eluant component that step 1) is obtained is concentrated to 12.5mg/ml, developing agent is n-butyl alcohol-glacial acetic acid-water (4:5:1), mass fraction is that 2% vanillin alcoholic solution is made developer, at lamellae lower end point sample 30 μ l, sampling point is vertically launched onboard.Sample liquid is after lamellae launches 20cm, with two some colour developings on the lamellae, the silica gel of exhibition apart from nearer navy blue colour developing point and exhibition apart from aeruginous colour developing point far away collects respectively, after being 70% alcohol solution dipping 24h with volume fraction respectively, stir 30min, centrifugal 5 minutes of reuse 5000r/min obtains colourless transparent solution; 60 ℃ are evaporated to 1/5 of original volume, obtain light yellow transparent concentrated solution, are labeled as Semen Ziziphi Spinosae calm the nerves hypnotic activity saponin component I and component I I respectively.
Dissolving-filtration described in the step (1)-drying at room temperature concrete operations are the n-butyl alcohol dissolving that thick saponin samples with water is saturated, filter with filter paper, obtain solution, at room temperature n-butyl alcohol are slowly volatilized;
Adsorbent described in the step 1) is a kind of in SA aluminium oxide or the silica gel; Be preferably silica gel; Because saponin polarity is big, generally adopts SA aluminium oxide or silica gel to make adsorbent, use the solvent of opposed polarity to carry out eluting according to different saponin again.
The dress column method of the chromatographic column that adsorbent is housed described in the step 1) is: the adsorbent that 100～200 orders is used for column chromatography is dried 1h under 110 ℃ of conditions, impurity is removed in the abundant drip washing of reuse distilled water; The reuse volume fraction is 95% soak with ethanol 24h; After the vacuum outgas, the wet method chromatographic column (36mm * 80mm) that packs into;
Pack into the method for chromatographic column of described wet method is: add a certain amount of water earlier in post, to be with water absorbent to pour in the post then, excessive water will be emitted at the bottom of by post, and keep the water surface to be higher than more than the silica gel aspect 3cm, up to layer of silica gel identity distance capital 7cm～8cm place, standby;
Step 2) it is 4:5:1 that the developing agent described in is preferably n-butyl alcohol-glacial acetic acid-water volume ratio;
The preparation method of the lamellae step 2) is: 100～200 purpose thin layer silica gel G75g, adding 200ml mass fraction is 0.5% CMC-Na solution, grind to form pasty state, the shop is applied on 20cm * 10cm thin layer of glass plate, make the thick plate of 1mm for every, behind the natural drying, in 105 ℃ of activation 60min, it is standby to make lamellae.
Step 2) exhibition described in is opened up apart between 3.5cm～4cm apart from nearer navy blue colour developing point;
Step 2) exhibition described in is opened up apart between 4.5cm～5cm apart from aeruginous colour developing point far away.
The described Semen Ziziphi Spinosae hypnotic activity saponin component of calming the nerves comprises Semen Ziziphi Spinosae calm the nerves hypnotic activity saponin component I and component I I, separates acquisition by above-mentioned preparation method from Semen Ziziphi Spinosae.
The described Semen Ziziphi Spinosae hypnotic activity saponin component of calming the nerves is used in the hypnotic activity medicine is calmed the nerves in preparation.
Principle of the present invention is:
Be defat reagent with the petroleum ether, ethanol is for extracting reagent, n-butyl alcohol is extraction agent, and Semen Ziziphi Spinosae is carried out defat, extracts, extraction, the impurity that makes itself and other be insoluble to organic solvent separates, and then handles with alkali liquor, and organic layer solution is obtained thick saponin after solvent is sloughed in distilling under reduced pressure, again thick saponin is obtained the main peak eluent by column chromatographic isolation and purification, obtain Semen Ziziphi Spinosae calm the nerves hypnotic activity saponin component I and component I I by the further separation and purification of thin layer chromatography again.
The present invention has following advantage and effect with respect to prior art:
1, the present invention's extraction separation from the conventional Chinese medicine Semen Ziziphi Spinosae that is used for the treatment of the disease of sleeping hypnotic activity saponin of calming the nerves has overcome the side effect that the Cure for insomnia chemical synthetic drug exists and has reached problems such as addiction more.
2, the present invention combines the former study method, designed one new, do not introduce and have extraction route overt toxicity solvent and reagent, comparatively safe, adopt alcohol extracting method, n-butanol extraction from having extracted the saponin sample a collection of Semen Ziziphi Spinosae sample.
3, the organic solvent extractionprocess of the present invention's employing is dissolved among the organic solvent saponin component, makes it separate with other components that are insoluble to organic solvent.Outside desolventizing, the organic solvent extraction process do not add other chemical agents, the saponin purity height that obtains, and content of ashes is low, does not contain flavone and polysaccharose substance.
4, the saponins component sample content of ashes that obtains of organic solvent extractionprocess of the present invention is low, the purity height, the hypnotic activity height of calming the nerves, the active group that has kept the natural soap glycoside preferably, its hypnosis performance of calming the nerves is not subjected to the influence of preparation method, can be used for the saponins material that the extraction separation Semen Ziziphi Spinosae has the hypnotic activity of calming the nerves.
5, the present invention has carried out further separation and purification with thick saponin through column chromatography and thin layer chromatography, has obtained the hypnotic activity component.
6, the present invention is by the spontaneous activity (walk about time and two upper limb lift number of times) to mice, the coordination exercise test, barbital sodium test (change of dosage barbital sodium induced mice length of one's sleep, sub-threshold dose barbital sodium induced mice sleep number of elements above threshold) is to the component sample of gained carried out the calming the nerves mensuration of hypnotic activity.
Description of drawings
Fig. 1 is the calm the nerves flow chart of separation method of hypnotic activity saponin sample of Semen Ziziphi Spinosae.
Fig. 2 is the thin-layer chromatogram of sample liquid after lamellae launches 20cm.
The specific embodiment
The present invention is described in further detail below in conjunction with embodiment and accompanying drawing, but embodiments of the present invention are not limited thereto.
The calm the nerves preparation of hypnotic activity saponin sample of embodiment 1 Semen Ziziphi Spinosae
Spiny jujuba seed powder sample (Semen Ziziphi Spinosae is crossed 40 mesh sieves, 60 ℃ of dry 24h after parch, pulverizing) is put apparatus,Soxhlet's with petroleum ether defat 4h under 60 ℃ of conditions of 10 times of volumes, obtains residue and Semen Ziziphi Spinosae oil; Residue takes out, and is dry, obtain the defat spiny jujuba seed powder after grinding, cross 40 mesh sieves, is 90 ℃ of water-bath reflux, extract, of ethanol 3h of 70% with the defat spiny jujuba seed powder with the volume fraction of 25 times of volumes, sucking filtration acquisition residue I and pure liquid; With pure liquid 60 ℃ of following evaporated under reduced pressure to paste, be dissolved in water, and obtain aqueous solution behind the filtering and impurity removing, add the water-saturated n-butanol extraction of 1 times of volume in the aqueous solution, abandon water layer, obtain n-butanol layer I; With the KOH solution basification of n-butanol layer I with the 0.1M of 1 times of volume, obtain aqueous alkali layer and n-butanol layer II; With n-butanol layer II evaporated under reduced pressure, namely obtain thick saponin; The n-butyl alcohol dissolving that thick saponin samples with water is saturated, filter with filter paper, obtain solution, at room temperature n-butyl alcohol is slowly volatilized the saponin sample after obtaining being further purified, triplicate namely obtains the Semen Ziziphi Spinosae hypnotic activity saponin sample (flow chart of preparation such as Fig. 1) of calming the nerves.
The calm the nerves separation and purification of hypnotic activity saponin component of embodiment 2 Semen Ziziphi Spinosaes
Because saponin polarity is big, generally adopts SA aluminium oxide or silica gel to make adsorbent, use the solvent of opposed polarity to carry out eluting according to different saponin again.The silica gel that 100～200 orders is used for column chromatography is dried 1h under 110 ℃ of conditions, impurity is removed in the abundant drip washing of reuse distilled water; Reuse 95% soak with ethanol 24h; After the vacuum outgas, the wet method chromatographic column (36mm * 80mm) that packs into.The method of dress post: in post, add a certain amount of water earlier, will be with water silica gel to pour in the post then, excessive water is emitted by post is low, keep the water surface to be higher than more than the silica gel aspect 3cm, up to layer of silica gel identity distance capital 7cm～8cm place, standby.Last sample saponin sample liquid concentration 25mg/ml, applied sample amount 10ml, 60% alcoholic solution is made eluent, and it is 0.5ml/min that adjusting peristaltic pump rotating speed makes flow velocity, and every 10ml collects a pipe.After collecting about 40 pipes, every pipe is got 1ml, and adding the 1ml mass fraction is 2% vanillin solution, and the 2ml concentrated sulphuric acid all has absorption maximum through the ultraviolet spectral photometer scanning discovery about 507nm.Eluent sulfuric acid layer under uviol lamp of collecting main peak is green, illustrates and contains saponin constituent.
Above-mentioned main peak eluant component is concentrated, with saponin sample concentration simmer down to 12.5mg/ml, be developing agent with n-butyl alcohol-glacial acetic acid-water (4:5:1), mass fraction is that 2% vanillin alcoholic solution is made developer, applied sample amount is 30 μ l, place the chromatography cylinder that fills developing solvent to launch to take out behind 8cm～9cm and dry, the spray developer, the hair dryer hot blast blows to colour developing.All present two points on the lamellae, first some exhibition is navy blue apart from 3.5cm～4cm; Second some exhibition is aeruginous (thin-layer chromatogram such as Fig. 2) apart from 4.5cm～5cm.Adopt the sample liquid of 1/2 concentration to do thin layer chromatography and not only can separate each component, and the area of its colour developing point is bigger.So in the later experiment sample liquid being diluted to 1/2 concentration separates.
With 20cm * 10cm thin layer of glass plate lower end point sample, in large beaker, vertically place, sampling point is vertically launched onboard.Sample liquid still has only two some colour developings after lamellae launches 20cm, illustrate and have only two components in this saponin sample.The silica gel of the each point position on the lamellae is collected respectively, behind 70% alcohol solution dipping 24h, stir 30min, centrifugal 5 minutes of reuse 5000r/min obtains colourless transparent solution; 60 ℃ of concentrating under reduced pressure obtain light yellow transparent concentrated solution, obtain Semen Ziziphi Spinosae calm the nerves hypnotic activity component I and component I I respectively.
Embodiment 3 hypnotic activity measurings
(1) to the influence of mice autonomic activities and coordination exercise
(1) autonomic movement
Calm the nerves hypnotic activity saponin components I, II of Semen Ziziphi Spinosae is made into the medicinal liquid of 10mg/ml respectively.Get 30 of mices, male and female half and half are divided into three groups at random, 10 every group.Each group is irritated the medicinal liquid (be equivalent to crude drug 17g/kgd, be about 50 times of clinical adult's dosage) that stomach gives 0.15ml/10gd respectively, and blank group gives with the volume normal saline, 3d continuously once a day, and before the last perfusion fasting 24h.Behind the last administration 45min, mice is placed carton, make it adapt to 5min after, movable number of times in the record 5min (measure that mice walk about time and two forelimbs upwards praise number of times).The components I group has utmost point significant difference with respect to the blank group in the time of walking about, and shortens to 21.47 seconds by 68.56 seconds of blank group, and lifts no significant difference on the number of times at two upper limb, is 16 times; The composition group is lifted at the time of walking about and two upper limb with respect to the blank group all notable difference on the number of times, shorten to 39.26 seconds by 68.56 seconds of blank group, and two upper limb lift number of times and are reduced to 6 times by 16 times of blank group.
(2) coordination exercise
Mice is the same.Irritate stomach 6d.Measure mice coordination exercise situation behind last administration 45min, method is placed on the glass plate upper end for tilting 42.5 ° with smooth sliding glass plate with mice, records it and slides the time.There were significant differences in coordination exercise with respect to the blank group for the components I group, shortened to 1.16 seconds of sample I group by 2.41 seconds of blank group from the glass plate time of sliding; The composition group does not have significant difference with respect to the blank group in coordination exercise.
(2) barbital sodium is caused the influence of mouse sleep time
(1) barbital sodium above threshold reaches determining of the dosage of sleeping under the threshold
Other gets 40 of mices, and male and female half and half are divided into 4 groups at random, 10 every group.Each administration group is lumbar injection barbital sodium 35,40,45,50mg/kg respectively; Blank group gives the normal saline with volume.The number of mice of falling asleep in the record administration 60min (righting reflex loss surpasses 1min as sleeping index).The threshold value that can determine barbital sodium thus is 45mg/kg.So in the zoopery afterwards, sleep dosage adopts 40mg/kg under the barbital sodium threshold, the dosage of above threshold sleeping adopts 50mg/kg.
(2) to the influence of dosage pentobarbital sodium induced mice time for falling asleep above threshold
Mice is the same.Irritate stomach 9d.Behind last administration 30min, all by the dosage lumbar injection barbital sodium of 50mg/kg, mice sleep incubation period and prolonged sleep time (righting reflex loss is to the time of righting reflex recovery) respectively organized in record to each group.Above threshold the length of one's sleep, the influence experiment showed that the components I group has the utmost point significant difference length of one's sleep with respect to the blank group above threshold to sample to the barbital sodium of mice, was extended for 115.8 seconds of sample I group by 91.9 seconds of blank group; Also there were significant differences on the length of one's sleep above threshold with respect to the blank group for the composition group, is extended for 121.3 seconds of component I I group by 91.9 seconds of blank group.
(3) to the influence of the sleeping number of elements of sub-threshold dose pentobarbital sodium induced mice
Mice is the same.Irritate stomach 12d.Behind last administration 30min, the mice number of elements that sleep occurs is observed and recorded to each group all by the dosage lumbar injection barbital sodium of 40mg/kg.Sample shows the influence experiment of the sleeping number of elements of sub-threshold dose pentobarbital sodium induced mice, and the components I group has increased by 30% with respect to the blank group number of elements of sleeping under threshold; The composition group has increased by 20% with respect to the blank group number of elements of sleeping under threshold.
By mice autonomic activities experiment, coordination exercise experiment and barbital sodium experiment, can find out that calm the nerves hypnotic activity components I, II of Semen Ziziphi Spinosae all has remarkable hypnotic activity, and components I is better than composition.
Above-described embodiment is preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. the Semen Ziziphi Spinosae isolation and purification method of hypnotic activity saponin component of calming the nerves is characterized in that comprising the steps:
(1) the Semen Ziziphi Spinosae preparation of hypnotic activity saponin sample of calming the nerves:
Semen Ziziphi Spinosae is crossed 40 mesh sieves after parch, pulverizing, 60 ℃ of dry 24h prepare spiny jujuba seed powder; With spiny jujuba seed powder by cable-styled extraction method with the petroleum ether of 10 times of quality at 60 ℃～65 ℃ defat 4h, acquisition residue and Semen Ziziphi Spinosae oil; Take out residue, dry, obtain the defat spiny jujuba seed powder after grinding, cross 40 mesh sieves; Be ethanol reflux, extract, 3h under 85 ℃～90 ℃ conditions of 70% with the defat spiny jujuba seed powder with the volume fraction of 25 times of quality, sucking filtration obtains residue I and pure liquid; With pure liquid 60 ℃ of following evaporated under reduced pressure to paste, be dissolved in water, and obtain aqueous solution behind the filtering and impurity removing; The water-saturated n-butanol that adds 1 times of volume to aqueous solution extracts, and abandons water layer, obtains n-butanol layer I; With the KOH solution basification of n-butanol layer I with the 0.1M of 1 times of volume, obtain aqueous alkali layer and n-butanol layer II; With n-butanol layer II evaporated under reduced pressure, namely obtain thick saponin; Thick saponin through three dissolving-filtration-drying at room temperature, is namely obtained the Semen Ziziphi Spinosae hypnotic activity saponin sample of calming the nerves;
(2) the Semen Ziziphi Spinosae isolation and purification method of hypnotic activity saponin component of calming the nerves:
1) column chromatographic isolation and purification
The saponin samples with water of step (1) preparation is mixed with the last sample saponin sample liquid that concentration is 25mg/ml, in the chromatographic column that adsorbent is housed, go up sample 10ml, volume parts is that 60% alcoholic solution is made eluent 40ml, it is 0.5ml/min that adjusting peristaltic pump rotating speed makes flow velocity, and every 10ml one pipe is collected eluent; Every pipe is got 1ml from eluent, and adding the 1ml mass fraction is 2% vanillin solution, the 2ml concentrated sulphuric acid, and through ultraviolet light spectrophotometer scanning discovery the eluent of maximum absorption band being arranged at the 507nm place is the main peak eluent, collects the main peak eluent standby;
2) thin layer chromatography separation and purification
The main peak eluant component that step 1) is obtained is concentrated to 12.5mg/ml, developing agent is the mixed solution of n-butyl alcohol-glacial acetic acid-water, mass fraction is that 2% vanillin alcoholic solution is made developer, at lamellae lower end point sample 30 μ l, sampling point is vertically launched onboard; Sample liquid is after lamellae launches 20cm, with two some colour developings on the lamellae, the silica gel of exhibition apart from nearer navy blue colour developing point and exhibition apart from aeruginous colour developing point far away collects respectively, after being 70% alcohol solution dipping 24h with volume fraction respectively, stir 30min, centrifugal 5 minutes of reuse 5000r/min obtains colourless transparent solution; 60 ℃ are evaporated to 1/5 of original volume, obtain light yellow transparent concentrated solution, are labeled as Semen Ziziphi Spinosae calm the nerves hypnotic activity saponin component I and component I I respectively.
2. the Semen Ziziphi Spinosae according to claim 1 isolation and purification method of hypnotic activity saponin component of calming the nerves, it is characterized in that: the dissolving-filtration described in the step (1)-drying at room temperature concrete operations are the n-butyl alcohol dissolving that thick saponin samples with water is saturated, filter with filter paper, obtain solution, at room temperature n-butyl alcohol is slowly volatilized.
3. the Semen Ziziphi Spinosae according to claim 1 isolation and purification method of hypnotic activity saponin component of calming the nerves is characterized in that: the adsorbent described in the step 1) is a kind of in SA aluminium oxide or the silica gel.
4. the Semen Ziziphi Spinosae according to claim 1 isolation and purification method of hypnotic activity saponin component of calming the nerves, it is characterized in that: the dress column method of the chromatographic column that adsorbent is housed described in the step 1) is: the adsorbent that 100～200 orders is used for column chromatography is dried 1h under 110 ℃ of conditions, impurity is removed in the abundant drip washing of reuse distilled water; The reuse volume fraction is 95% soak with ethanol 24h; After the vacuum outgas, the wet method chromatographic column of packing into.
5. the Semen Ziziphi Spinosae according to claim 4 isolation and purification method of hypnotic activity saponin component of calming the nerves, it is characterized in that: pack into the method for chromatographic column of described wet method is: add a certain amount of water earlier in post, to be with water absorbent to pour in the post then, excessive water is emitted at the bottom of by post, keep the water surface to be higher than silica gel aspect 3cm, up to layer of silica gel identity distance capital 7cm～8cm place, standby.
6. the Semen Ziziphi Spinosae according to claim 1 isolation and purification method of hypnotic activity saponin component of calming the nerves is characterized in that: step 2) described in developing agent be that the volume ratio of n-butyl alcohol-glacial acetic acid-water is 4:5:1.
7. the Semen Ziziphi Spinosae according to claim 1 isolation and purification method of hypnotic activity saponin component of calming the nerves, it is characterized in that: step 2) described in the preparation method of lamellae be: 100～200 purpose thin layer silica gel G75g, adding 200ml mass fraction is 0.5% CMC-Na solution, grind to form pasty state, the shop is applied on 20cm * 10cm thin layer of glass plate, makes the thick plate of 1mm for every, behind the natural drying, in 105 ℃ of activation 60min, it is standby to make lamellae.
8. the Semen Ziziphi Spinosae according to claim 1 isolation and purification method of hypnotic activity saponin component of calming the nerves is characterized in that: step 2) described in exhibition apart from nearer navy blue colour developing point exhibition apart between 3.5cm～4cm;
Step 2) exhibition described in is opened up apart between 4.5cm～5cm apart from aeruginous colour developing point far away.
9. the Semen Ziziphi Spinosae hypnotic activity saponin component I of calming the nerves is separated acquisition with component I I by each described isolation and purification method of claim 1～8 from Semen Ziziphi Spinosae.
10. calm the nerves hypnotic activity saponin component I and component I I of the described Semen Ziziphi Spinosae of claim 9 uses in the hypnotic activity medicine is calmed the nerves in preparation.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104435295A (en) * | 2014-11-06 | 2015-03-25 | 辽宁中医药大学 | Medicine composition containing active ingredients of jujube kernel tranquilizing pellets |
| CN104666474A (en) * | 2015-01-27 | 2015-06-03 | 华南理工大学 | Spina date seed extract and brain-strengthening and sleeping-promoting oral liquid containing spina date seed extract |
| CN107412371A (en) * | 2017-09-01 | 2017-12-01 | 安徽省芬格欣生物药业有限公司 | It is a kind of that there is mental-tranquilization, the jujuboside extracting method of tranquilizing soporific function |
| CN108465102A (en) * | 2018-04-28 | 2018-08-31 | 山东星之诚生物科技有限公司 | A kind of tranquilizing the mind patch and preparation method thereof and application method |
| CN109521138A (en) * | 2018-12-07 | 2019-03-26 | 浙江工业大学 | A kind of method for quick identification of semen ziziphi spinosae and Semen Ziziphi Spinosae (parched) |
| CN113109495A (en) * | 2021-05-24 | 2021-07-13 | 北京大学 | Quality detection method of jujube kernel nerve-soothing capsules based on thin-layer chromatography |
| CN115326965A (en) * | 2022-08-16 | 2022-11-11 | 周至年青保制药有限公司 | Quality control method of nerve-soothing Shenmu capsule for improving deep sleep |
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| CN104435295A (en) * | 2014-11-06 | 2015-03-25 | 辽宁中医药大学 | Medicine composition containing active ingredients of jujube kernel tranquilizing pellets |
| CN104666474A (en) * | 2015-01-27 | 2015-06-03 | 华南理工大学 | Spina date seed extract and brain-strengthening and sleeping-promoting oral liquid containing spina date seed extract |
| CN104666474B (en) * | 2015-01-27 | 2017-12-01 | 华南理工大学 | A kind of Wild jujube seeds extract and the brain tonic sleeping oral liquid containing Wild jujube seeds extract |
| CN107412371A (en) * | 2017-09-01 | 2017-12-01 | 安徽省芬格欣生物药业有限公司 | It is a kind of that there is mental-tranquilization, the jujuboside extracting method of tranquilizing soporific function |
| CN108465102A (en) * | 2018-04-28 | 2018-08-31 | 山东星之诚生物科技有限公司 | A kind of tranquilizing the mind patch and preparation method thereof and application method |
| CN109521138A (en) * | 2018-12-07 | 2019-03-26 | 浙江工业大学 | A kind of method for quick identification of semen ziziphi spinosae and Semen Ziziphi Spinosae (parched) |
| CN109521138B (en) * | 2018-12-07 | 2020-11-13 | 浙江工业大学 | Rapid identification method for spina date seeds and fried spina date seeds |
| CN113109495A (en) * | 2021-05-24 | 2021-07-13 | 北京大学 | Quality detection method of jujube kernel nerve-soothing capsules based on thin-layer chromatography |
| CN115326965A (en) * | 2022-08-16 | 2022-11-11 | 周至年青保制药有限公司 | Quality control method of nerve-soothing Shenmu capsule for improving deep sleep |
| CN115326965B (en) * | 2022-08-16 | 2023-09-15 | 周至年青保制药有限公司 | Quality control method of Shenmu tranquilization capsule for improving deep sleep |
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