CN107629108A - Saikosaponin a, c, d extracting method in a kind of Radix Bupleuri - Google Patents
Saikosaponin a, c, d extracting method in a kind of Radix Bupleuri Download PDFInfo
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Abstract
The present invention relates to Radix Bupleuri technical field, more specifically, is related to the extracting method of saikosaponin a, c, d in a kind of Radix Bupleuri；Comprise the following steps：Using single factor test and orthogonal test, it is determined that extracting saikosaponin a, c, d method simultaneously；Chinese Thorowax Root is placed in conical flask with cover, 10% ammoniacal liquor methanol solution is added, ultrasonic extraction 40min, repeats extraction twice, obtain comprising saikosaponin a, c, d extract solution；And saikosaponin a in extract solution, c, d content are determined with high performance liquid chromatography；Method that is provided by the invention while extracting saikosaponin a, c, d, using radix bupleuri as research object, the relatively all higher extracting method of saikosaponin a, c, d content is obtained using single factor test and orthogonal test, theoretical reference is provided for industrial abstract saikosaponin a, c, d.
Description
Technical field
The present invention relates to Radix Bupleuri technical field, more specifically, is related to saikosaponin a, c, d in a kind of Radix Bupleuri
Extracting method.
Background technology
Radix bupleuri is the conventional traditional Chinese medicine in China, cold nature, acrid flavour, hardship.Return liver, courage, lung channel.It can be dredged with dispelling wind and heat from the body
Irritability solution pent-up, yang invigorating.For treating fevers and chills alternate, cold headache heating, shaoyang disease, chest and abdomen swelling and pain, metroptosis, take off
The illnesss such as anus.Prove that there are radix bupleuri many kinds to act on through pharmaceutical research, such as radix bupleuri can solve body heat, calmness, analgesia, town
Cough, Anti-bacterium, antiviral, strengthen immunity and the pharmacological action such as antitumor.2015 editions《Chinese Pharmacopoeia》Regulation radix bupleuri is umbrella shape
The dry root of section's plant radix bupleuri or radix bupleuri scorzoneraefolii.It is different by character, practise claim " Bupleurum Chinese " and " RADIX BUPLEURI SCORZONERAEFOLII " respectively.
Now with much on the extraction conditions of saikoside in radix bupleuri and the report of assay method, they employ difference
Extracting method (ultrasound, backflow) and different Extraction solvent (70% methanol, 5% ammoniacal liquor methanol etc.), but these extracting methods
It is different, so that assay result has larger difference.
The content of the invention
In the presence of overcoming the shortcomings of prior art, the present invention provides saikosaponin a, c, d in a kind of Radix Bupleuri
Extracting method, this method preferably gone out by single factor test and orthogonal test while extracts saikosaponin a in Radix Bupleuri, c, d
The best approach.
In order to solve the above-mentioned technical problem, the technical solution adopted in the present invention is：
Saikosaponin a, c, d extracting method, comprise the following steps in a kind of Radix Bupleuri：
（1）Single factor experiment is carried out, draws influence of each factor to saikosaponin a, c, d extraction effect in same Radix Bupleuri；
（2）Carry out orthogonal test, according to extracted simultaneously in same Radix Bupleuri saikosaponin a, c, d these three saponin contents it is big
It is small, determine extraction conditions；
（3）According to the extraction conditions of above-mentioned determination, Radix Bupleuri is ground into powder, crosses No. four sieves, precision weighs 0.5g；
（4）Powder after weighing is placed in conical flask with cover, adds ammoniacal liquor methanol solution；
（5）Conical flask with cover is put into supersonic extractors and carries out ultrasonic extraction, the solution after ultrasonic extraction is filtered, obtained
To filtrate；
（6）Repeat step（5）Extraction；
（7）Gained filtrate is merged, obtained comprising saikosaponin a, c, d extract solution.
The step（1）In, single factor experiment includes extracting method, solvent load, extraction time, extraction time and solvent
Species.
In the single factor experiment, extracting method selects ultrasonic extraction, extraction time 20min-40min, extraction time 1-3
Secondary, solvent species selects 5%-15% ammoniacal liquor methanol solutions, when solvent load is 10ml-30ml, each factor extraction effect influence compared with
Greatly.
The step（2）In, orthogonal test specifically refers to, according to single factor experiment result, 3 respectively be selected under each factor
Individual level, factor level table is formulated, using orthogonal arrage L9(34) tested, according to the extreme difference value of each factor, determine each factor point
The other extraction effect on saikosaponin a, c, d influences, and obtains optimum extraction condition.
Extraction conditions are the ammoniacal liquor methanol solution using 30ml 10%, and the ultrasonic extraction time is 40min, are extracted 2 times.
Step（1）In single factor experiment and step（2）In orthogonal test, using high performance liquid chromatography to extraction
Mixed solution comprising saikosaponin a, c, d is measured, and calculates the concrete content of three kinds of saponin(es, so as to more each extraction conditions
Quality.
Performance liquid chromatographic column in high performance liquid chromatography is Waters XTerra®MSC18Chromatographic column, a length of 150mm are interior
Footpath is 4.6mm, and packing material size is 5 μm；Mobile phase A is acetonitrile, and Mobile phase B is ultra-pure water, flow velocity 1ml/min；Chromatographic column post
Temperature is 30 DEG C；Detector is UVD, Detection wavelength 210nm.
High performance liquid chromatography measure saponin(e a, c, d content comprise the following steps：
（1）Prepare reference substance solution
Precision weighs radix bupleuri reference substance saikosaponin a 2.6mg, saikoside c 1.77mg, saikoside d 2.64mg, is placed in
In 5ml measuring bottles, add methanol ultrasonic dissolution and constant volume, obtain saikoside reference substance mixed solution, concentration be followed successively by 0.520mg/ml,
0.354mg/ml, 0.528mg/ml, it is standby；
（2）Prepare need testing solution
Extract solution comprising saikosaponin a, c, d is volatilized filtrate with thermostat water bath, with appropriate methanol ultrasound after volatilizing
Dissolved residue, in the measuring bottle of last constant volume to 5ml, through filtering with microporous membrane, need testing solution can be obtained；
（3）It is accurate respectively to draw in the μ l of the reference substance solution 10 and μ l of need testing solution 10 injection high performance liquid chromatographs, determine peak
Area, saikosaponin a, c, d content are calculated using one point external standard method.
Step（2）The temperature of middle thermostat water bath is 60 DEG C, and methanol is chromatographically pure, and the aperture of miillpore filter is 0.45 μm.
Compared with prior art, the advantageous effect of present invention is that：
Saikosaponin a, c, d extracting method in a kind of Radix Bupleuri provided by the invention, using radix bupleuri as research object, using list
Factor and orthogonal test preferably go out while extract saikosaponin a, c, d best approach：Ultrasonic extracting method is good, and method letter
It is single, it is more easy to operate；Extraction solvent amount is in 30ml, saikosaponin a, c, d content highest；During extraction time 40min, radix bupleuri soap
The basic dissolution of glycosides；Extraction time is 2 times, saikosaponin a, c, d content highest；Good, the ammoniacal liquor first with ammoniacal liquor methanol extraction comparison
Alcohol is at 10%, saikosaponin a, c, d content highest.Saponin(e a, c, d content are specifically have detected with high performance liquid chromatography, intuitively
Provide some theoretical reference foundations for industrial abstract saikoside.
Brief description of the drawings
Fig. 1 is influence of the quantity of solvent to saikoside recovery rate；
Fig. 2 is influence of the extraction time to saikoside recovery rate；
Fig. 3 is influence of the ultrasonic number to saikoside recovery rate；
Fig. 4 is influence of the solvent species to saikoside recovery rate.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation describes, it is clear that described embodiment is only part of the embodiment of the present invention, rather than whole embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art obtained on the premise of creative work is not made it is all its
His embodiment, belong to the scope that the present invention is protected.
1st, instrument and reagent
1.1 reagent
Reference substance：Saikosaponin a reference substance（Chengdu it is general have to Bioisystech Co., Ltd, lot number 16122101）, saikoside c
Reference substance（Chengdu it is general have to Bioisystech Co., Ltd, lot number 16061305）, saikoside d reference substance（Chengdu is general to give birth to
Thing Technology Co., Ltd., lot number 160513）, radix bupleuri（Hebei Quan Tai pharmaceutcal corporation, Ltds, lot number 1612203）, acetonitrile（Tianjin
Ke Miou chemical reagent Co., Ltd, chromatographically pure, lot number 20151214）, methanol（The limited public affairs of Tianjin Ke Miou chemical reagent
Department, chromatographically pure, lot number 20090210）, water be ultra-pure water；Ammoniacal liquor（Tianjin recovery development in science and technology Co., Ltd, lot number
20141216）, methanol（Tianjin Kermel Chemical Reagent Co., Ltd., analyze pure, lot number 20151019）.
1.2 instrument
KQ5200V type ultrasonic cleaners（Kunshan Ultrasonic Instruments Co., Ltd.）；Waters2695 high performance liquid chromatographs（It is beautiful
State）；HH-2 digital display thermostat water baths（Jie Ruier Electrical Appliances Co., Ltd of Jintan City）；FA224 electronic balances（Shanghai Shun Yuhengping sections
Skill Instrument Ltd.）；The a ten thousandth electronic balances of AB135-S ten.
1.3 chromatographic condition
Chromatographic column：Waters XTerra®MSC18Chromatographic column（4.6mm × 150mm, 5 μm）；Mobile phase A：Acetonitrile, Mobile phase B：
Ultra-pure water, gradient elution is carried out by the regulation in table 1；Flow velocity：1ml/min；Chromatographic column column temperature：30℃；Detector：UVD, detection
Wavelength is 210nm.
The condition of gradient elution of table 1
2nd, solution preparation and assay method
2.1 reference substance solution
Precision weighs radix bupleuri reference substance saikosaponin a 2.6mg, saikoside c 1.77mg, saikoside d 2.64mg, is placed in
In 5mL measuring bottles, add methanol ultrasonic dissolution and constant volume, obtain saikoside reference substance mixed solution, concentration be followed successively by 0.520mg/mL,
0.354mg/mL, 0.528mg/mL, it is standby.
2.2 need testing solution
Precision weighs Radix Bupleuri powder (cross No. four sieve) about 0.5 g, is placed in conical flask with cover, according to single factor test and orthogonal
Various extracting conditions in experiment, solvent needed for precision addition, are extracted, and are filtered, and putting 60 DEG C with thermostat water bath temperature regulating will
Filtrate volatilizes, with appropriate methanol after volatilizing（Chromatographically pure）Ultrasonic dissolution residue, in the volumetric flask of last constant volume to 5mL, mistake
0.45 μm of miillpore filter, test liquid can be obtained.
2.3 determination method
Accurate draw in the μ l and μ l of the need testing solution 10 injection high performance liquid chromatographs of reference substance mixed solution 10 determines respectively, adopts
Calculated with one point external standard method.
3rd, extraction saikosaponin a, c, d best practice are determined
3.1 single factor experiment
（1）The influence of ultrasound and backflow to saikoside recovery rate
Precision weighs Radix Bupleuri powder 0.5g（Cross No. four sieves）, 2 parts, place in conical flask with cover, add 10% ammoniacal liquor first respectively
Alcoholic solution 25ml, a copy of it ultrasonic extraction 40min, 2 times, another refluxing extraction 40min, 2 times, merging filtrate, puts 60 DEG C
Volatilized in water bath with thermostatic control, the appropriate methanol ultrasonic dissolution of residue, be placed in 5ml volumetric flask and be settled to scale, finally use
0.45 μm of filtering with microporous membrane.Using 2.3 determination method measure saikosaponin a, c, d peak area, radix bupleuri soap is calculated
Glycosides a, c, d content（%）, saikoside c, a, d content are respectively 0.125%, 0.389%, 0.987% in circumfluence method, ultrasonic method
Middle saikoside c, a, d content are respectively 0.141%, 0.405%, 1.014%, and by comparing, ultrasound is good compared with refluxing extraction effect
Some, and method is simple, it is more easy to operate, as shown in table 2.
The ultrasound of table 2 and influence of the backflow to saikoside recovery rate
（2）Influence of the quantity of solvent to saikoside recovery rate
Precision weighs Chinese Thorowax Root 0.5g, 5 parts, puts in conical flask with cover, respectively plus 10% ammoniacal liquor methanol solution 10ml, 20ml,
30ml, 40ml, 50ml, ultrasonic extraction 40min, 2 times, merging filtrate, put in 60 DEG C of waters bath with thermostatic control and volatilize, the appropriate first of residue
Alcohol ultrasonic dissolution, it is placed in 5ml volumetric flask and is settled to scale, before use with 0.45 μm of filtering with microporous membrane.Using
2.3 determination method determines saikosaponin a, c, d peak area, calculates saikosaponin a, c, d content（%）.Saikoside c's contains
Amount is respectively 0.132%, 0.138%, 0.145%, 0.143%, 0.138%, the content of saikosaponin a is respectively 0.312%,
0.398%th, 0.431%, 0.418%, 0.417%, the content of saikoside d is respectively 0.802%, 1.018%, 1.078%, 1.060%,
1.040%.For Extraction solvent amount in 10ml~30ml, saikosaponin a, c, d changes of contents are consistent, are all used with solvent
The increase of amount and increase, then decline slightly during more than 30ml, as shown in Figure 1.
（3）Influence of the extraction time to saikoside recovery rate
Precision weighs Chinese Thorowax Root 0.5g, 5 parts, puts in conical flask with cover, adds 10% ammoniacal liquor methanol solution 25ml, respectively ultrasound
10min, 20min, 30min, 40min, 50min are extracted, 2 times, merging filtrate, puts in 60 DEG C of waters bath with thermostatic control and volatilizes, residue is used suitable
The methanol ultrasonic dissolution of amount, is placed in 5ml volumetric flask and is settled to scale, is with 0.45um filtering with microporous membrane before use
Can.Using 2.3 determination method measure saikosaponin a, c, d peak area, saikosaponin a, c, d content are calculated（%）, radix bupleuri soap
Glycosides c content is respectively 0.116%, 0.140%, 0.141%, 0.145%, 0.143%, and the content of saikosaponin a is respectively
0.368%th, 0.380%, 0.387%, 0.405%, 0.404%, the content of saikoside d is respectively 0.956%, 0.991%, 1.043%,
1.051%、1.03%.The content of saikoside is with the increase of extraction time in Radix Bupleuri, and increases, bavin when having arrived 40min
The basic dissolution of Hu saponin(e.Continue to extend the time, saikoside may degrade, as shown in Figure 2.
（4）Influence of the ultrasonic number to saikoside recovery rate
Precision weighs Chinese Thorowax Root 0.5g, 3 parts, puts in conical flask with cover, adds 10% ammoniacal liquor methanol solution 25ml, ultrasonic extraction
40 min, extracting 1 time respectively, 2 times, 3 merging filtrates, put in 60 DEG C of waters bath with thermostatic control and volatilize, residue is dissolved with appropriate methanol,
It is placed in 5ml volumetric flask and is settled to scale, before use with 0.45 μm of filtering with microporous membrane.Using 2.3 determination method
Saikosaponin a, c, d peak area are determined, calculates saikosaponin a, c, d content（%）, saikoside c content is respectively
0.166%th, 0.176%, 0.178%, the content of saikosaponin a is respectively 0.480%, 0.495%, 0.494%, and saikoside d contains
Amount is respectively 1.247%, 1.310%, 1.311%.Saikosaponin a, c, d content are with the increase of extraction time, content increase,
As shown in Figure 3.
（5）Influence of the solvent species to saikoside recovery rate
Precision weighs the g of Chinese Thorowax Root 0.5,5 parts, puts in conical flask with cover, adds 5% ammoniacal liquor methanol solution, 10% ammoniacal liquor respectively
Methanol solution, 15% ammoniacal liquor methanol solution, 90% methanol solution, 70% methanol solution 25ml, ultrasonic extraction 40 min, 2
It is secondary, merging filtrate, put in 60 DEG C of waters bath with thermostatic control and volatilize, the appropriate methanol ultrasonic dissolution of residue, be placed in 5 ml volumetric flask
Scale is settled to, before use with 0.45 μm of filtering with microporous membrane.Using 2.3 determination method measure saikosaponin a, c, d
Peak area, calculate saikosaponin a, c, d content（%）, saikoside c content is respectively 0.135%, 0.145%,
0.139%th, 0.145%, 0.121%, the content of saikosaponin a is respectively 0.391%, 0.432%, 0.405%, 0.220%, 0.182%,
The content of saikoside d is respectively 0.993%, 1.110%, 1.051%, 0.314%, 0.259%.Through analysis, alone methanol extraction
The content of radix bupleuri is smaller, good with ammoniacal liquor methanol extraction comparison, ammoniacal liquor methanol at 5%~10%, saikosaponin a, c, d content with
The amount of ammoniacal liquor increases and increased, and between 10%~15%, content is as the amount of ammoniacal liquor increases and reduces, as shown in Figure 4.
3.2 orthogonal test
It can be seen that each factor has different degrees of shadow to the extraction effect of bupleurum total saponin from above-mentioned single factor experiment
Ring.According to single factor experiment result, 3 levels are respectively selected under superincumbent wherein 4 factors, factor level table is formulated, is shown in Table
3。
The saikoside extraction process factor level table of table 3
Using orthogonal arrage L9(34) tested, extracted according to the condition of orthogonal arrage, extract solution volatilizes, and is then surpassed with methanol
Sound is settled to 5ml volumetric flask, crosses 0.45 μm of miillpore filter, determines peak area using 2.3 determination method, calculates saikoside
A, c, d mass fraction.Test arrangement and 4 are the results are shown in Table, table 5.
The saikoside extraction process L of table 49（34）Orthogonal experiments
The analysis of variance table of table 5
3.3 interpretation of result
By the extreme difference value of listed each factor in comparison sheet 3, as can be seen from Table 4, each of saikoside c extraction effects is influenceed
Factor order is A ＞ B ＞ C ＞ D, influences saikosaponin a, each factor order of d extraction effects is A ＞ D ＞ B ＞ C.From the side of table 5
Poor analysis shows, factor A have significant difference, and there was no significant difference by factor B, C, D.With reference to single factor exploration result, solvent
Dosage and the species of solvent influence slightly larger on extraction effect.10% ammoniacal liquor methanol solution, the ml of quantity of solvent 30,40 min of ultrasound,
Extraction 2 times, the saikosaponin a extracted, c, d composition are all relatively more, so optimal extracting factor is A3B2C3D2。
4th, checking test
As shown in table 6,3 batches of samples are taken, are extracted according to above-mentioned optimum extraction process, extract obtained middle saikoside c's
Mass fraction is respectively 0.152%, 0.146%, 0.149%, RSD 2.01%.The mass fraction of saikosaponin a is respectively
0.432%, 0.445%, 0.450%, RSD 2.1%.The mass fraction of saikoside d is respectively 1.105%, 1.098%, 1.056%
, RSD 2.4%.As a result show that the process conditions are simple, reliable, reproducible.
The checking test of table 6
The present invention investigates influence of each factor to bupleurum total saponin extraction effect by single factor experiment, from extraction saikoside
3 levels are selected in 4 influence factors, then optimize saikosaponin a, c, d optimal extract process using orthogonal design.
As a result 10% ammoniacal liquor methanol solution is shown, 30 ml, 40 min of ultrasound, 2 times extraction effect is best.With the increase of extraction time
With increasing for extraction time, the content of saikoside declines on the contrary, and this is relevant with the stability of saikoside.Saikoside indissoluble
Ethanol is soluble in water, in acid condition, it may occur that secondary biochemistry, cause its activity to reduce, compare under weak basic condition
It is stable.Therefore, using the methanol containing ammoniacal liquor as Extraction solvent.Saikosaponin a, d have stronger pharmacological activity, its content
Height determines the power of drug effect.According to the literature, saikosaponin a, d can become with the extension of heat time.For maximum limit
Degree ground plays the drug effect of radix bupleuri, improves saikosaponin a, d content in bupleurum extract, suitable temperature should be selected in extraction process
Degree, extraction time are unsuitable long.
Only presently preferred embodiments of the present invention is explained in detail above, but the present invention is not limited to above-described embodiment,
In those of ordinary skill in the art's possessed knowledge, it can also be made on the premise of present inventive concept is not departed from each
Kind change, various change should be included in the scope of the protection.
Claims (9)
1. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri, it is characterised in that comprise the following steps：
Single factor experiment is carried out, draws influence of each factor to saikosaponin a, c, d extraction effect in same Radix Bupleuri；
Orthogonal test is carried out, according to the size for extracting saikosaponin a, c, d these three saponin contents in same Radix Bupleuri simultaneously,
Determine extraction conditions；
According to the extraction conditions of above-mentioned determination, Radix Bupleuri is ground into powder, crosses No. four sieves, precision weighs 0.5g；
Powder after weighing is placed in conical flask with cover, adds ammoniacal liquor methanol solution；
Conical flask with cover is put into supersonic extractors and carries out ultrasonic extraction, the solution after ultrasonic extraction is filtered, obtained
Filtrate；
Repeat step（5）Extraction；
Gained filtrate is merged, obtained comprising saikosaponin a, c, d extract solution.
2. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri as claimed in claim 1, it is characterised in that：It is described
Step（1）In, single factor experiment includes extracting method, solvent load, extraction time, extraction time and solvent species.
3. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri as claimed in claim 2, it is characterised in that：It is described
In single factor experiment, extracting method selects ultrasonic extraction, extraction time 20min-40min, extraction time 1-3 times, solvent species
From 5%-15% ammoniacal liquor methanol solutions, when solvent load is 10ml-30ml, each factor extraction effect has a great influence.
4. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri as claimed in claim 1, it is characterised in that：It is described
Step（2）In, orthogonal test is specifically referred to according to single factor experiment result, respectively selects 3 levels under each factor, formulate because
Plain water-glass, using orthogonal arrage L9(34) tested, according to the extreme difference value of each factor, determine each factor respectively to saikoside
A, c, d extraction effect influence, and obtain optimum extraction condition.
5. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri as claimed in claim 4, it is characterised in that：Extraction
Condition is the ammoniacal liquor methanol solution using 30ml 10%, and the ultrasonic extraction time is 40min, is extracted 2 times.
6. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri as claimed in claim 1, it is characterised in that：Step
（1）In single factor experiment and step（2）In orthogonal test, saikoside is included to extraction using high performance liquid chromatography
A, c, d mixed solution are measured, and calculate the concrete content of three kinds of saponin(es, so as to the quality of more each extraction conditions.
7. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri as claimed in claim 6, it is characterised in that：Efficiently
Performance liquid chromatographic column in liquid chromatogram is Waters XTerra®MSC18Chromatographic column, a length of 150mm, internal diameter 4.6mm, fill out
It is 5 μm to expect particle diameter；Mobile phase A is acetonitrile, and Mobile phase B is ultra-pure water, flow velocity 1ml/min；Chromatographic column column temperature is 30 DEG C；Detection
Device is UVD, Detection wavelength 210nm.
8. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri as claimed in claim 6, it is characterised in that：Efficiently
Liquid chromatogram measuring saponin(e a, c, d content comprise the following steps：
（1）Prepare reference substance solution
Precision weighs radix bupleuri reference substance saikosaponin a 2.6mg, saikoside c 1.77mg, saikoside d 2.64mg, is placed in
In 5ml measuring bottles, add methanol ultrasonic dissolution and constant volume, obtain saikoside reference substance mixed solution, concentration be followed successively by 0.520mg/ml,
0.354mg/ml, 0.528mg/ml, it is standby；
（2）Prepare need testing solution
Extract solution comprising saikosaponin a, c, d is volatilized filtrate with thermostat water bath, with appropriate methanol ultrasound after volatilizing
Dissolved residue, in the measuring bottle of last constant volume to 5ml, through filtering with microporous membrane, need testing solution can be obtained；
（3）It is accurate respectively to draw in the μ l of the reference substance solution 10 and μ l of need testing solution 10 injection high performance liquid chromatographs, determine peak
Area, saikosaponin a, c, d content are calculated using one point external standard method.
9. saikosaponin a, c, d extracting method in a kind of Radix Bupleuri as claimed in claim 8, it is characterised in that：Step
（2）The temperature of middle thermostat water bath is 60 DEG C, and methanol is chromatographically pure, and the aperture of miillpore filter is 0.45 μm.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114225001A (en) * | 2021-12-31 | 2022-03-25 | 北京橘井健康科技集团有限公司 | Preparation method of improved bupleuri decoction |
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2017
- 2017-09-22 CN CN201710867932.XA patent/CN107629108A/en active Pending
Non-Patent Citations (1)
| Title |
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| 刘伟等: "正交试验法优选柴胡中柴胡早苷提取工艺", 《中国医院药学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114225001A (en) * | 2021-12-31 | 2022-03-25 | 北京橘井健康科技集团有限公司 | Preparation method of improved bupleuri decoction |
| CN114225001B (en) * | 2021-12-31 | 2024-04-19 | 北京橘井健康科技集团有限公司 | Preparation method of improved major bupleurum decoction |
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Application publication date: 20180126 |