CN108498526A - A kind of pharmaceutical composition and the preparation method and application thereof of prevention blahs aypnia disease - Google Patents
A kind of pharmaceutical composition and the preparation method and application thereof of prevention blahs aypnia disease Download PDFInfo
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- CN108498526A CN108498526A CN201810468386.7A CN201810468386A CN108498526A CN 108498526 A CN108498526 A CN 108498526A CN 201810468386 A CN201810468386 A CN 201810468386A CN 108498526 A CN108498526 A CN 108498526A
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Abstract
The invention discloses a kind of pharmaceutical composition and the preparation method and application thereof of prevention blahs aypnia disease, which includes that active constituent magnoflorine is formed with spinosin proportioning.The present invention also provides a kind of method that separation prepares magnoflorine and spinosin from spina date seed using high-speed countercurrent chromatography, the separation condition is：Solvent system is n-butanol ethyl acetate water=1~3:1~5:2~7, upper phase is stationary phase, and lower phase is mobile phase.The preparation method of the present invention has the characteristics that good separating effect, preparation amount are big, lose less, is easy to operate, economic and environment-friendly.Show that the composition is significantly improved and therapeutic effect depression, insomnia disease tool through animal pharmacodynamics experimental study.
Description
Technical field
The invention belongs to medicine, field of health care products, it is related to a kind of pharmaceutical composition and preparation method thereof and presses down in treatment
Application in terms of strongly fragrant disease, insomnia disease.
Background technology
With the quickening pace of modern life, the incidence of insomnia and depression gradually rises.Patients with depression 90% is with mistake
The somatizations such as dormancy disease, sleep disturbance, and it is to cause depression onset, the key factor of recurrence to have a sleepless night.But insomnia at present
Disease and depression there is no specific diagnosis, therapy, and disease time is long, and easily repeatedly, patient needs long-time medication.Perhaps
Although more antidepression Western medicine drug effects are definite, target spot is single, cannot improve depression and occur together insomnia, and its side effect
Certain somatizations can also be aggravated.Chinese medicine can multi-level, multiple target effect, it is excellent in terms for the treatment of depression and the insomnia to occur together
Gesture protrudes, and bad hair answers incidence low, and therapeutic effect is obvious, the compound medicine especially developed with native compound ingredient
Increasingly paid attention to by market.
Spina date seed cures mainly diseases such as " restlessness of asrhenia type and insomnia ", has both antidepression and tranquilizing soporific as traditional resolving stagnation for tranquilization Chinese medicine
Effect is one of food and medicament dual-purpose Chinese medicine as defined in the Ministry of Public Health of China, has no toxic side effect, wherein containing saponin(e, flavones, alkaloid etc.
Multiple types ingredient.Due to spina date seed complicated component, play antidepression have a sleepless night effect active ingredient and mechanism of action not yet
It illustrates completely clear.Internal and external test is the results show that spina date seed alkaloid cooperates with antidepression insomnia effect with flavones part combination
Significantly, effect is significantly stronger than other part combinations, and magnoflorine (magnoflorine) and spinosin (spinosin) divide
Not Wei alkaloid and flavones position principle active component, both there is antidepression, sedative-hypnotic effect, and show significantly
Synergistic function.Separate active ingredients are extracted from spina date seed at present mostly uses traditional side such as column chromatography, recrystallization
Method, not only time-consuming and laborious, fixed relative sample have irreversibility suction-operated, low yield.
Invention content
The comparative compositions that an object of the present invention is to provide two monomers in a kind of spina date seed occur together in prevention depression
Application in insomnia, the monomer component are respectively magnoflorine and spinosin.
The method for preparing magnoflorine and spinosin the second object of the present invention is to provide high speed adverse current chromatogram, this method
Pure magnoflorine and spinosin can be extracted from spina date seed medicinal material, to overcome extraction separation spina date seed work in the prior art
Property ingredient is time-consuming and laborious, fixed relative sample has the problem of irreversibility suction-operated, low yield.
The invention is realized by the following technical scheme：
A kind of pharmaceutical composition of prevention blahs aypnia disease, using magnoflorine and spinosin as main active,
The wherein described magnoflorine is alkaloids, molecular formula C20H24NO4, molecular structural formula is as follows：
Spinosin is flavonoid glycoside, molecular formula C28H32O15, molecular structural formula is as follows：
The magnoflorine and spinosin press the gauge of substance, and the weight ratio of magnoflorine and spinosin is 1:1~
26。
The active constituent is that extraction separation is similar either by chemical synthesis process or selected from having from spina date seed
The natural drug of pharmacological action.
A kind of drug of prevention blahs aypnia disease, using magnoflorine and spinosin as main active, addition medicine
Learn acceptable auxiliary material be made injection, tablet, capsule, granule, oral solution, powder, dripping pill, sublingual formulation, pellet or
Spray.
A kind of preparation method of the pharmaceutical composition of prevention blahs aypnia disease, the described method comprises the following steps：
(1) preparation of crude extract：
Spina date seed crushes, and is sieved by 40 mesh, and after petroleum ether degreasing, the concentration expressed in percentage by volume that 6~12 mass times are added is
60%~80% ethanol water, heating and refluxing extraction 2~3 times, 1~3 hour every time, filtration merged extracting solution, filtrate
It concentrates and measures water dissolutions with 2~6 times after ethyl alcohol to no alcohol taste, with extracting n-butyl alcohol 4-6 times, extracting solution reduced pressure, recycling design
After obtain n-butanol crude extract, i.e., sample to be separated；
(2) high-speed countercurrent chromatography separation magnoflorine and spinosin are used：
It is 1~3 to take the volume ratio of ethyl acetate, n-butanol and water:1~5:2~7, it is placed in separatory funnel and prepares two-phase
Solvent system, it is stationary phase to take phase, and lower phase is mobile phase；Stationary phase is full of to the chromatographic column of adverse current chromatogram, at 25~35 DEG C
Under, host rotates forward, and rotating speed is 300~900r/min, then enters mobile phase with the flow pump of 1.5~10ml/min, until two mix
Agent reaches equilibrium state in column；The mixing crude extract for taking step (1) to prepare, with phase, lower phase volume ratio 1:1 mixed solvent
Sample introduction after dissolving collects the efflux of magnoflorine and spinosin, concentration, drying respectively under UV detector, you can point
Magnoflorine and spinosin are not obtained.
In step (2), the volume ratio of n-butanol-ethyl acetate-water is：2:3:5.
In step (2), at 25 DEG C, host rotates forward, then rotating speed 500r/min is become a mandarin with the flow pump of 7ml/min
Dynamic phase.
In the step (2), the wavelength of the UV detector is 254~325nm.
It is a kind of prevention blahs aypnia disease pharmaceutical composition production prevention and treatment depression, insomnia disease drug in
Using.
In the step (2), the appearance time of the magnoflorine and spinosin be followed successively by 72~84min, 86~
100min and 176~202.5min.
The method provided by the invention for isolating and purifying this skin Northey of alkaloid monomer magnoflorine and chromocor compound, relatively
In the prior art, the present invention can disposably isolate and purify a kind of alkaloid monomer and flavonoid glycoside compound list from spina date seed
Body, isolate and purify monomer is efficient, preparation amount is big, loss less, product purity it is high, easy to operate, economic and environment-friendly.The present invention is primary
The alkaloid and flavonoid glycoside monomer that property isolates and purifies reach 90% or more through high performance liquid chromatography (HPLC) method purity.
The present invention detaches new skill according to the property of magnoflorine in spina date seed and spinosin using new medicine extraction
Art-high-speed countercurrent chromatography, detaches it, easy to operate, the rate of recovery is high, and fractional dose is big and separative efficiency is high, can reach
The purpose of quick separating spina date seed ingredient magnoflorine and spinosin simultaneously.
Description of the drawings
Fig. 1 is magnoflorine and Si Pi Northeys, 6 made from embodiment 1, and " '-asafoetide acyl spinosin mixes crude extract
HPLC collection of illustrative plates；
Fig. 2 is high speed adverse current chromatogram (HSCCC) figure of embodiment 1；
Fig. 3 is the HPLC collection of illustrative plates of 1 magnoflorine of compound made from embodiment 1；
Fig. 4 is the HPLC collection of illustrative plates of this skin Northey of compound 2 made from embodiment 1；
Fig. 5 is " the HPLC collection of illustrative plates of '-asafoetide acyl spinosin of compound 36 made from embodiment 1.
Specific implementation mode
It is further illustrated the present invention below by way of specific embodiment, not limitation of the invention, according to known in this field
The prior art, embodiments of the present invention are not limited to specific embodiment.In the premise without prejudice to spirit of that invention and principle
Under, it is fallen in the scope of the present invention to inventing any modifications or changes that individual technical steps carry out.
Embodiment 1
1. the preparation of crude extract：
Spina date seed 250g is crushed, and is sieved by 40 mesh, after petroleum ether degreasing, adds 1200ml, 800ml, 600ml percentage respectively
A concentration of 70% ethanol water, heating and refluxing extraction 3 times, 1 hour every time, filtration merged extracting solution, and filtrate concentrates ethyl alcohol
With 2 times of amount water dissolutions after to no alcohol taste, with extracting n-butyl alcohol 5 times, extracting solution is concentrated under reduced pressure, and it is thick to obtain n-butanol after recycling design
Extract, i.e., sample to be separated, HPLC collection of illustrative plates are as shown in Figure 1.
2. using high-speed countercurrent chromatography separation magnoflorine and the " '-asafoetide acyl spinosin of spinosin, 6：
It is 2 to take the volume ratio of n-butanol, ethyl acetate and water:3:5, it is placed in preparation two-phase solvent system in separatory funnel,
It is stationary phase to take phase, and lower phase is mobile phase；Stationary phase is full of to the chromatographic column of adverse current chromatogram, at 25 DEG C, host rotates forward, and turns
Speed is 500r/min, then enters mobile phase with the flow pump of 7ml/min, until two-phase solvent reaches equilibrium state in column；Take step
Suddenly the mixing crude extract that prepared by (1), with phase, lower phase volume ratio 1:Sample introduction after 1 mixed solvent dissolving, with wavelength 280nm's
UV detector detection efflux is shown in Fig. 2 according to high speed adverse current chromatogram figure, to 72min to 84min, 86min to 100min and
The corresponding effluxes of 176min to 202.5min merge collection, concentration, drying to constant weight, you can respectively obtain lily magnolia respectively
Flower alkali (flow point I) and " '-asafoetide acyl spinosin (flow point III) of spinosin (flow point II), 6.
3. monomer purity measures and structure determination
Gained monomeric compound is detected using high performance liquid chromatograph to what is obtained respectively.Chromatographic column：Agilent
TC-C18Column (4.6mm × 250mm, 5 μm)；- 0.1% aqueous acetic acid of acetonitrile, gradient elution；Run time 40min；0min
10% acetonitrile, 20% acetonitriles of 5min, 5~40min, 20%~30% acetonitriles；Flow velocity：1mL/min；Column temperature：30℃；Sample size：
20μL；Detection wavelength 280nm.Measuring compound magnoflorine, spinosin and 6, " purity of '-asafoetide acyl spinosin is distinguished
Reach 95.0%, 98.2% and 97.3% (Fig. 3, Fig. 4 and Fig. 5).Each monomer respectively through mass spectrum (MS),1H-NMR and13C-NMR
Analysis, compares, gained monomeric compound is respectively magnoflorine 250mg and spinosin through structure elucidation and with data in literature
" '-asafoetide acyl spinosin the 15mg of 150mg, 6.
Test example 1
The experimental study that PC12 cellular damages influence is caused on glutamic acid
1. test material
1.1 drugs and reagent
Drug：Magnoflorine, spinosin, magnoflorine+spinosin
Cell line：PC12 cell lines (source：Ratt μ s norvegic μ s Adrenal Pheochromocytomas, Nanjing Keygen Biotech
Development in science and technology Co., Ltd provides).
Reagent：(it is limited to match silent your biochemistry product Beijing of winged generation by Hyclone for H-DMEM cell culture mediums, fetal calf serum
Company)；Pancreatin cell dissociation buffer (the green skies), penicillin streptomycin mixed liquor is dual anti-(BI companies of Israel), L-sodium
(Klonetech, Japan).Tetramethyl azo azoles salt (MTT), dimethyl sulfoxide (DMSO) (DMSO) (pokeweed mitogen,
Sigma companies).
1.2 test apparatus
Steam pressure sterilizer (YXDG02, Xinhua Medical Apparatus and instruments Factory, Shandong)；(SW-CJ-2G, Suzhou are net for superclean bench
Change)；Carbon dioxide incubator (HF160W, Heal Force companies)；Desk centrifuge (B600, Bai Yang centrifuge factory)；Enzyme mark
Instrument (Promega, GloMax-Multi)；Inverted microscope (CKX41, Japanese Olympus companies)；Pipettor (Gilson,
Eppendorf)；Culture bottle (Nest)；96 orifice plates (Costar, USA)
2. test method
PC12 cytochemistry damage models are prepared with sodium glutamate, absorption is measured at 490nm using tetrazolium bromide (MTT) method
Value, repair of the observation drug candidate to PC12 cellular damages.
Modeling：25mmol/L sodium glutamates (to be loaded final concentration, being dissolved in incomplete culture medium) act on PC12 cells
24h。
Grouping：Blank group, chemical damage model group, chemical damage+medicine group
3. content of the test
3.1 cell culture
From the PC12 cell recoveries frozen are taken in ultra low temperature freezer in culture bottle, with the H-DMEM containing 10% fetal calf serum
Medium culture.When PC12 cell growths to 80% fusion, with 0.25% pancreatin cell dissociation buffer (containing 0.02%EDTA)
It is digested, with 1 × 105A cell per well is inoculated in 96 orifice plates, and zeroing hole is not added with cell.
3.2 cellular damage
After the cell in 96 orifice plates covers with single layer, culture medium is discarded, final concentration of 25mmol/L is added per hole for model group
200 μ L (being dissolved in incomplete culture medium) of glutamic acid in being cultivated in incubator for 24 hours, that is, cause PC12 cellular damage models.
3.3 dosing reparations
Culture medium is discarded, administration group is separately added into the drug and damage liquid of the final concentration of 20mmol/L of drug containing screening per hole
200 μ L of mixed liquor, 5 multiple holes of each concentration.Using the DMEM culture mediums of not drug containing as blank control group.Then cell is set
Enter 37 DEG C, 5%CO2Continue culture in incubator for 24 hours.
3.4 mtt assay measure cell inhibitory rate
100 μ L plasma-free DMEM mediums are added per hole after discarding culture medium, 5mg/mL MTT solution is added then at every hole
20 μ L are in being protected from light 4h in incubator.It discards culture medium and 100 μ L DMSO microplate reader is added in 490nm measurement OD values, meter
Calculate cell survival rate.
4. test result
The influence of the PC12 cell survival rates of 4.1 pairs of damages
1 each administration group of table to the results of PC12 cell survival rates (N=5)
Note：Compared with model group, * * P<0.01；Compared with blank group,##P<0.01
PC12 cells after giving 20 μM of different pharmaceuticals, calculate each group after the processing for 24 hours of 25mM L-sodiums
The survival rate of cell can reflect protective effect of the drug to PC12 damaging cells indirectly.Table 1 is the results show that model group cell
After the processing for 24 hours of 25mM L-sodiums, compared with blank group, cell survival rate is only 45.06%, cell damage significantly (P
<0.01) after, giving drug therapy, magnoflorine group, 20 μM of groups of magnoflorine+spinosin cell survival rate rise respectively
To 49.33%, 66.93% (P<0.01).It prompts magnoflorine+spinosin that there is collaboration to play nerve under 20 μM of concentration to protect
Shield acts on.
Test example 2
Pharmacodynamic experiment of the pharmaceutical composition to CUMS blahs aypnia mouse models
1 experiment material
1.1 experimental animal
Healthy male ICR mouse 100 (20 ± 2g of weight) is purchased from Institute of Radiation Medicine, Chinese Academy of Medical Sciences's experiment
Animal center；Raising adapts to environment in 3 days before experiment.22 ± 2 DEG C of room temperature, relative humidity 65%~70%.Day illumination 12h, freely
Diet takes the photograph water.
1.2 reagents and drug：
Magnoflorine, spinosin (self-control of this laboratory)；Venlafaxine hydrochloride slow-release capsule (Chengdu Kang Hong medicine companies group
Limited liability company)
2 experimental methods
2.1 animal packets and administration
Mouse is randomly divided into 8 groups, every group 10, respectively blank group, model group, Western medicine positive group (Venlafaxine), in
Patent medicine positive group (resolving stagnation for tranquilization granule), decocting group, magnoflorine group, spinosin, magnoflorine+spinosin group.It removes
Outside blank group, remaining each group mouse presses " 2.2 " method modeling；From daystart every morning 8 of modeling:00-9:00 fills
Stomach administration continues 21 days, and dosage regimen is shown in Table 2.
2 animal packet of table and dosage regimen
The method for building up of 2.2 CUMS blahs aypnia mouse models
10 kinds of stress factors of random arrangement are pressed in 21d, arbitrary two kinds of stimulations are given once daily, to avoid mouse from generating adaptation
Mild-natured each stimulation is given 4 times.
(1) moist raising：Suitable quantity of water is added in mouse bedding and padding makes its humidity, and survive 12h in a humid environment.
(2) fasting：12h is jejunitas.
(3) prohibit water：12h cuts off the water supply.
(4) raising is tilted：Mouse cage is tilted into 45 ° of raising mouse 12h.
(5) tail is pressed from both sides：Mouse is put into fixed cage, is clamped away from lasting 1min at root of the tail 1cm with haemostatic clamp.
(6) it is raised without bedding and padding：Mouse, each 12h are raised in the mouse cage of no bedding and padding.
(7) sleep deprivation：Sleep deprivation is for 24 hours.Using chain-wales water environment method, in container bottom setting diameter 3cm, height
The platform of 2cm, periphery fill water (depth of water 0.5cm), can fall into the water if mice sleep.
(8) cold-wate swimming：Animal is placed on to (depth of water 5cm) 5min in the container for filling 10 DEG C of water.
(9) hot water is swum：Animal is placed on to (depth of water 5cm) 5min in the container for filling 45 DEG C of water.
(10) it overturns round the clock：In early 7:The cage of mouse is placed on dark space when 00 not turn on light, until evening 19:It will when 00
Mouse is placed in the room turned on light until next day early 7:When 00.
2.3 observation indexs and method
(1) weight
The changes of weight of measurement mouse during the experiment, i.e., the 1st day, the 3rd day, the 10th day, the 17th day, the 21st day body
Weight, and calculate each group mouse weight increment.
(2) tail-suspention test (TST)
Mouse tail end is attached on a horizontal waddy, waddy is liftoff to make animal in vacantly projecting shape, hangs both sides partition board
Block mouse sight.Animal is to overcome abnormal position and struggle activity, but activity is after a certain period of time, discontinuity " motionless " occurs
" disappointment " state of display.It tests and carries out 6min, the dead time after record in 4min.
(3) forced swim test (FST)
Mouse is placed individually into the transparent glass cylinder for filling deep 10cm or more, temperature 45 C water, experiment carries out 6min.It is small
Mouse is to struggle to escape to after feeling hopeless to stop struggling, only by head above water four limbs in motionless state, after record in 4min
The dead time of mouse.
(4) yellow Jackets synergistic effect experiment
Sub-threshold dose yellow Jackets, dosage 28mg/kg are injected in the 8th day gavage 0.5h pneumoretroperitoneum of zoopery.
There is the number of elements slept in record each group mouse.Gavage 0.5h pneumoretroperitoneums inject above threshold dose of sodium pentobarbitone, dosage 38 within 9th day
mg/kg.Start timing after injection, records Sleep latency and sleeping time.Using when righting reflex loss as sleep onset time,
Righting reflex reverts to the sleep end time.To sleep onset time it is Sleep latency after intraperitoneal injection, incubation period is more than
15min is recorded by 15min.
3 results
3.1 each administration groups test CUMS mouse TST the influence of dead time
Model group obviously increases compared with the naive mice TST dead times.The TST of magnoflorine group, spinosin group
Dead time is below model group, and difference has statistical significance (P ＜ 0.05)；Magnoflorine+spinosin group, Venlafaxine
Group, resolving stagnation for tranquilization granule group the TST dead times be substantially less than model group, there is extremely significant difference (P ＜ 0.01),
It the results are shown in Table 4.
4 each administration group of table to the CUMS blahs aypnia mouse tail suspension dead times influence (N=10)
* the P compared with model group<0.05** P compared with model group<0.01
3.2 each administration groups test CUMS mouse FST the influence of dead time
The FST dead times of magnoflorine group, spinosin group, magnoflorine+spinosin group are compared with model group
It is substantially reduced, there is notable significant difference (P ＜ 0.01), positive drug group, decocting the group difference compared with model group to have statistics
Learn meaning (P ＜ 0.05).It the results are shown in Table 5.
5 each administration group of table to the mouse forced swimming test dead time influence (N=10)
* the P compared with model group<0.05** P compared with model group<0.01
The influence that 3.3 each administration groups test mouse sub-threshold dose yellow Jackets
Indicate that the sleep number of elements percentage of each administration group is significantly raised compared with model group by 6 experimental result of table, wherein
Magnoflorine+spinosin group sleep number of elements high percentage is identical as blank group in magnoflorine, spinosin group.
The influence that 6 each administration group of table tests mouse sub-threshold dose yellow Jackets
The influence that 3.4 each administration groups act synergistically to above threshold dose of sodium pentobarbitone
It is shown by 7 data of table, each administration group Sleep latency no difference of science of statistics.Each administration group sleeping time is longer than mould
Type group, wherein magnoflorine+spinosin group, Venlafaxine group sleeping time are considerably longer than model group and magnoflorine, this skin
Promise element group.
7 each administration group of table to mouse, above threshold test by dose of sodium pentobarbitone
* the P compared with model<0.05** P compared with model group<0.01.
Claims (9)
1. a kind of pharmaceutical composition of prevention blahs aypnia disease, which is characterized in that with magnoflorine and spinosin be main
Active constituent, wherein the magnoflorine is alkaloids, molecular formula C20H24NO4 +, molecular structural formula is as follows：
Spinosin is flavonoid glycoside, molecular formula C28H32O15, molecular structural formula is as follows：
2. according to claim 1 prevent blahs aypnia disease pharmaceutical composition, which is characterized in that the magnoflorine and
Spinosin presses the gauge of substance, and the weight ratio of magnoflorine and spinosin is 1:1~26.
3. preventing the pharmaceutical composition of blahs aypnia disease according to claim 1, which is characterized in that the active constituent is
Extraction separation is either by chemical synthesis process or selected from the natural drug with similar pharmacological action from spina date seed.
4. a kind of drug of prevention blahs aypnia disease, which is characterized in that using magnoflorine and spinosin as chief active at
Point, pharmaceutically acceptable auxiliary material is added, injection, tablet, capsule, granule, oral solution, powder, dripping pill, sublingual system is made
Agent, pellet or spray.
5. a kind of preparation method of the pharmaceutical composition of prevention blahs aypnia disease, which is characterized in that the method includes following
Step：
(1) preparation of crude extract：
Spina date seed crushes, and is sieved by 40 mesh, and after petroleum ether degreasing, the concentration expressed in percentage by volume that 6~12 mass times are added is 60%
~80% ethanol water, heating and refluxing extraction 2~3 times, 1~3 hour every time, filtration merged extracting solution, and filtrate concentrates second
With 2~6 times of amount water dissolutions after alcohol to no alcohol taste, with extracting n-butyl alcohol 4-6 time, extracting solution reduced pressure obtains after recycling design
N-butanol crude extract, i.e., sample to be separated；
(2) high-speed countercurrent chromatography separation magnoflorine and spinosin are used：
It is 1~3 to take the volume ratio of ethyl acetate, n-butanol and water:1~5:2~7, it is placed in separatory funnel and prepares two-phase solvent
System, it is stationary phase to take phase, and lower phase is mobile phase；Stationary phase is full of to the chromatographic column of adverse current chromatogram, it is main at 25~35 DEG C
Machine rotates forward, and rotating speed is 300~900r/min, then enters mobile phase with the flow pump of 1.5~10ml/min, until two-phase solvent is in column
In reach equilibrium state；The mixing crude extract for taking step (1) to prepare, with phase, lower phase volume ratio 1:After 1 mixed solvent dissolving
Sample introduction collects the efflux of magnoflorine and spinosin, concentration, drying, you can respectively obtain respectively under UV detector
Magnoflorine and spinosin.
6. the preparation method of the pharmaceutical composition of prevention blahs aypnia disease according to claim 5, it is characterised in that：Step
(2) in, the volume ratio of n-butanol-ethyl acetate-water is：2:3:5.
7. the preparation method of the pharmaceutical composition of prevention blahs aypnia disease according to claim 5, it is characterised in that：Step
(2) in, at 25 DEG C, host rotates forward, then rotating speed 500r/min enters mobile phase with the flow pump of 7ml/min.
8. the preparation method of the pharmaceutical composition of prevention blahs aypnia disease according to claim 5, which is characterized in that described
In step (2), the wavelength of the UV detector is 254~325nm.
9. a kind of pharmaceutical composition of prevention blahs aypnia disease answering in production prevention and treatment depression, insomnia disease drug
With.
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| CN114129580A (en) * | 2021-12-24 | 2022-03-04 | 西安荷默奇医药科技有限公司 | Traditional Chinese medicine-based component composition and application thereof in treating depression |
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| CN109632995A (en) * | 2018-12-21 | 2019-04-16 | 广东方制药有限公司 | A kind of method for building up of semen ziziphi spinosae flavones ingredient UPLC finger-print and application |
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| CN114129580A (en) * | 2021-12-24 | 2022-03-04 | 西安荷默奇医药科技有限公司 | Traditional Chinese medicine-based component composition and application thereof in treating depression |
| CN114129580B (en) * | 2021-12-24 | 2024-05-10 | 海南云泽医药科技有限公司 | Traditional Chinese medicine-based component composition and application thereof in treating depression |
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