CN102579529A - Half-bionic combined ultrasonic preparation method of standardized extract of Bupleurum chinense DC. - Google Patents
Half-bionic combined ultrasonic preparation method of standardized extract of Bupleurum chinense DC. Download PDFInfo
- Publication number
- CN102579529A CN102579529A CN2012100522888A CN201210052288A CN102579529A CN 102579529 A CN102579529 A CN 102579529A CN 2012100522888 A CN2012100522888 A CN 2012100522888A CN 201210052288 A CN201210052288 A CN 201210052288A CN 102579529 A CN102579529 A CN 102579529A
- Authority
- CN
- China
- Prior art keywords
- extract
- saikoside
- radix bupleuri
- ethanol
- extraction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a preparation method of a Chinese medicine extract, particularly a half-bionic combined ultrasonic preparation method of standardized extract of Bupleurum chinense DC. The method comprises the following steps: crushing Bupleurum chinense DC. into coarse powder, adding a proper amount of 60 % ethanol, adjusting the pH value to 8, infiltrating for 8h, carrying out ultrasonic extraction for three times at 40 DEG C, wherein 40 minutes are taken at a time, 8 times amount of ethanol is added at a time; adjusting the pH value to 8, filtering, merging the filtrates, recovering ethanol until no alcohol taste, condensing at 60 DEG C until the relative density is 1.10, and then carrying out spray drying or microwave vacuum drying. According to the invention, the extraction is carried out in a half-bionic environment, the total content of main effective components (Saikosaponin a, b, c, and d) is no less than 1.874 %, the average flour yield is 15.35 %, the average yield is 0.288 %, and the extraction rate is 84.706 %; and the qualitative and quantitative assessment of the quality of the extract of Bupleurum chinense DC. can be realized, the unification and controllability of quality of the extract of Bupleurum chinense DC., and the invention is beneficial for raising the medicine quality and regulating the production and market operation.
Description
Technical field
The present invention relates to a kind of method for preparing of Chinese crude drug standardized extract, i.e. Radix Bupleuri standardized extract half bionical combined ultrasonic method for preparing.
Background technology
In the prior art, the dry root of umbelliferae bupleurum Bupleurum chinense DC. through being processed into extract, is the yellowish-brown powder, and feeble QI is fragrant, mildly bitter flavor.The function of Radix Bupleuri extract with cure mainly for: evacuate and to bring down a fever dispersing the stagnated live-QI to relieve the stagnation of QI, elevate a turnable ladder yang-energy.Be used for cold, fever, alternate attack of chill and fever, sternal rib pain, menoxenia, menoxenia uterine prolapse, proctoptosis.Sell the Radix Bupleuri extract market mostly is water extract, but its effective ingredient does not have the qualitative, quantitative standard, only provides specification: 5: 1,10: 1,20: 1 (getting the powder rate), can not guarantee every batch of product quality, as crude drug, influence the medicine product quality.Radix Bupleuri mainly contains
SaikosideSaponin such as (a, b, c, d),
Sterol,
Volatile oil(bupleurumol,
EugenolDeng),
Fatty acid(oleic acid, linoleic acid, Palmic acid, stearic acid etc.) and polysaccharide etc., what wherein mainly play therapeutical effect is
Saikoside(a, b, c, d) has to evacuate and brings down a fever, dispersing the stagnated live-QI to relieve the stagnation of QI, and the effect of elevate a turnable ladder yang-energy can be treated cold, fever, alternate attack of chill and fever, sternal rib pain, menoxenia, the menoxenia uterine prolapse, diseases such as proctoptosis, medical value is very high.Water extraction particularly when routine is extracted; Can make the fracture of primary glycosides epoxy ehter bond; The extraction ratio of main pharmacodynamics composition saikoside (a, b, c, d) is very low, and thin layer is inspected and HPLC detects, and does not almost have display dot and the absworption peak of saikoside (a, b, c, d); The main pharmacodynamics composition is lost, and does not have obvious therapeutic effect.
Summary of the invention
The objective of the invention is provides the higher Radix Bupleuri standardized extract of a kind of saikoside (a, b, c, d) extraction ratio half bionical combined ultrasonic method for preparing to above-mentioned deficiency.
Technical solution of the present invention is: Radix Bupleuri standardized extract half bionical combined ultrasonic method for preparing: get Radix Bupleuri and be ground into powder (coarse powder), it is an amount of to add 60% ethanol, transfers pH=8 with stabilizing agent (1.5%KOH); Soak into 8h, under 40 ℃ of conditions of temperature, ultrasonic (power 250W, 50kHz) extracts 3 times; Each 40 minutes, add 8 times of amount ethanol at every turn, transfer pH=8 with stabilizing agent (1.5%KOH); Filter, merging filtrate reclaims ethanol to there not being the alcohol flavor; Under 60 ℃, be concentrated into relative density 1.10 spray dryinges or microwave vacuum drying, promptly get.The used Radix Bupleuri of this process is Radix Bupleuri.FGC-TQ/4/1.6/3.6 still pot type ultrasonic extraction, vacuum concentration complete set of equipments are selected in batch process for use.
Through detecting; Radix Bupleuri extract contains four saikoside monomers: saikoside a, saikoside b, saikoside c, saikoside d sum >=1.874%; Demonstrate fully the strict quality standard requirement of Chinese medicine standardized extract; Except that having former plant same function function, also can be used for doing the crude drug of preparation.
Semi-biomimetic method prepares Radix Bupleuri standardized extract principle, mainly is under the condition of constant body temperature of bionical object and organism large intestine ecological environment pH value, in conjunction with other process conditions factor, makes the orthogonal experiment environment set under half bionical environment, extract screening.On the optimum process condition basis of screening, formulate method for preparing.
Leaching process under this half bionical environmental condition; Can avoid the fracture of primary glycosides epoxy ehter bond; Primary glycosides epoxy ehter bond is constantly split; Can make a large amount of acetyl group saikoside (2-0-acetyl group saikoside a, 2-0-acetyl group saikoside b2,3-0-acetyl group saikoside b2 etc.) deacetylate again, make it become corresponding saikoside, can increase substantially the yield of main component like this.Can make the extraction rate reached to 84.706% of main pharmacodynamics composition saikoside a bcd.And the acid impurities in the viscous polysaccharide and alkali generate deposition and accelerate filtyration velocity, reduce impurity, thereby reach the purpose of improving the quality of products with the yield of product.
Advantage of the present invention is: 1, this project is formed with the principal element of semi-biomimetic method principle design orthogonal test, and combined ultrasonic extracts the method that adopts the quadrature screening experiment to combine with single factor screening test; The screening optimum process condition is formulated " Radix Bupleuri GMP production technology standard. ", makes main effective ingredient (saikoside a, b, c, d) yield height in the Radix Bupleuri; Extraction ratio is good, and energy-conserving and environment-protective are with low cost; Be convenient to suitability for industrialized production, quality standard is higher than common extract standard.2, saikoside a, b, c, d (its minimum content >=1.3%) have been guaranteed to contain in the Radix Bupleuri extract of the present invention; Can carry out the qualitative, quantitative evaluation to the Radix Bupleuri extract quality; Guaranteed that the Radix Bupleuri extract quality is unified, controlled; Help improving drug quality, the standard production and the market operation improve international competitiveness.
To combine embodiment that embodiment of the present invention is described in further detail below.
Description of drawings
Fig. 1 is that test sample is measured collection of illustrative plates in the assay,
Fig. 2 is " a contrast characteristic spectrum ",
Fig. 3 is " characteristic peak coincide collection of illustrative plates ",
Fig. 4 is a TLC thin layer chromatic graph.
The specific embodiment
1 three batches of stable technology amplification tests of embodiment (every crowd of 250g)
Radix Bupleuri standardized extract half bionical combined ultrasonic method for preparing: get Radix Bupleuri 250g and be ground into coarse powder, it is an amount of to add 60% ethanol, transfers pH=8 with stabilizing agent (1.5%koH); Soak into 8h, under 40 ℃ of conditions of temperature, ultrasonic (power 250W, 50kHz) extracts 3 times; Each 40 minutes, add 8 times of amount ethanol at every turn, transfer pH=8 with stabilizing agent (1.5%koH); Filter, merging filtrate reclaims ethanol to there not being the alcohol flavor; Be concentrated into relative density 1.10 (60 ℃) spray drying or microwave vacuum drying, promptly get (concrete technological parameter sees three batches of stable amplification test process parameter tables 2 for details).FGC-TQ/4/1.6/3.6 still pot type ultrasonic extraction, vacuum concentration complete set of equipments are selected in batch process for use.
The average flour extraction of the three batches of amplification tests 15.35%, with saikoside a bcd total content calculate that its extract average content is 1.874%, average yield is 0.288%, extraction ratio is 84.706%.See table 2
Three batches of amplification test main technologic parameters of table 2:
Table 1 orthogonal test main technologic parameters:
2, method design principle and target,
According to orthogonal test and three batches of preferred process conditions of stable amplification test, formulate Radix Bupleuri standardized extract method for preparing.
Semi-biomimetic method prepares Radix Bupleuri standardized extract principle, mainly is under the condition of constant body temperature of bionical object and organism large intestine ecological environment pH value, in conjunction with other process conditions, makes the orthogonal experiment environment, is set under the half bionical environment and extracts screening.
The leaching process of purpose of design under this environmental condition can be avoided the fracture of primary glycosides epoxy ehter bond; Primary glycosides epoxy ehter bond is constantly split and a large amount of acetyl group saikoside (2-0-acetyl group saikoside a, 2-0-acetyl group saikoside b2,3-0-acetyl group saikoside b2 etc.) deacetylate can be made; Make it become corresponding saikoside; Can increase substantially the yield of main component like this; And routine is extracted particularly water extraction, inspects and the HPLC detection at thin layer, does not almost have display dot and the absworption peak of saikoside a bcd.The main pharmacodynamics composition is lost.Under half bionical environment, extract; Can make the extraction rate reached to 84.706% of main pharmacodynamics composition saikoside a bcd; And acid impurities in the viscous polysaccharide and alkali generate deposition quickening filtyration velocity; Reduce the water-solubility impurity in the filtrating, thereby reach the purpose of improving the quality of products with the yield of product.
The content's index control of Radix Bupleuri standardized extract: with saikoside a bcd sum >=1.3% is the content controlling index.
Embodiment 2
The Radix Bupleuri extract quality standard comprises the projects such as the identical collection of illustrative plates mensuration of discriminating, saikoside assay, characteristic spectrum and characteristic peak of Radix Bupleuri extract.
Radix Bupleuri extract differentiates and content assaying method that its step is following:
(1) Radix Bupleuri extract preparation: get Radix Bupleuri 250g and be ground into coarse powder, it is an amount of to add 60% ethanol, transfers pH=8 with stabilizing agent (1.5%KOH), soaks into 8h; Under 40 ℃ of conditions of temperature, ultrasonic (power 250W, 50kHz) extracts 3 times, each 40 minutes, adds 8 times of amount ethanol at every turn; Transfer pH=8 with stabilizing agent (1.5%KOH), filter merging filtrate; Reclaim ethanol to there not being the alcohol flavor, be concentrated into relative density 1.10 (60 ℃) spray drying or microwave vacuum drying, promptly get (seeing table 2).
(2) differentiate:
The preparation of need testing solution: get Radix Bupleuri extract 0.6g, add methanol 10ml, supersound process 2min filters, as need testing solution;
The preparation of saikoside reference substance solution: get saikoside a reference substance, saikoside d reference substance, saikoside b reference substance, saikoside c reference substance, add methanol and process the mixed solution that every 1ml contains 0.4mg, as reference substance solution;
" each 10 μ l of above-mentioned solution are drawn in an appendix VI of Chinese pharmacopoeia version in 2010 B test, put respectively on same silica gel g thin-layer plate according to thin layer chromatography; With ethyl acetate, ethanol, water=8: 2: 1 was developing solvent, launched, and took out; Dry; It is clear that spray is heated to the speckle colour developing with 40% sulfuric acid solution of 2% pair of dimethylbenzaldehyde at 60 ℃, under daylight and ultra-violet lamp 365n m, inspects respectively, in the test sample chromatograph; Be on the reference substance chromatograph relevant position, show the speckle or the fluorescence speckle of same color;
(3) saikoside assay: " appendix VID of Chinese pharmacopoeia version in 2010 measures according to HPLC; Chromatographic condition and system suitability test are carried out according to assay under the Radix Bupleuri item;
The preparation of reference substance solution: get the saikoside a reference substance, saikoside d reference substance, saikoside c reference substance, saikoside b reference substance are an amount of, accurately claim surely, add methanol and process the solution that every 1ml contains each 0.4mg of saikoside, shake up, and promptly get;
The preparation of need testing solution: get the about 0.6g of these article powder, the accurate title, decide, and puts in the 20ml measuring bottle, adds methanol to scale, and ultrasonic 10 minutes, filter, get subsequent filtrate, promptly get;
Algoscopy: accurate respectively reference substance solution 10ul and the need testing solution 10ul of drawing, inject chromatograph of liquid, measure, promptly get, see Fig. 1;
These article are pressed dry product and are calculated, and contain saikoside a (C
42H
68O
13), saikoside d (C
42H
68O
13) total amount of saikoside c, saikoside b must not be less than 1.3%.
(4) " characteristic spectrum " " characteristic peak coincide collection of illustrative plates " measured: " appendix VID of Chinese pharmacopoeia version in 2010 measures according to HPLC;
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With the acetonitrile is mobile phase A, is Mobile phase B with water, and the regulation in the according to the form below is carried out gradient elution; The detection wavelength is 210nm, and number of theoretical plate calculates by the saikoside a peak should be not less than 10000;
The preparation of object of reference solution: it is an amount of to get saikoside a reference substance, saikoside b reference substance, saikoside c reference substance, saikoside d reference substance; The accurate title, decide; Put in the 10ml measuring bottle; Add dissolve with methanol and be diluted to and contain the mixed solution of getting saikoside a 0.4mg, saikoside b 0.5mg, saikoside c 3mg, saikoside d 0.5mg among every 1ml, shake up, promptly get;
The preparation of need testing solution: get the about 0.6g of these article, the accurate title, decide, and puts in the 20ml measuring bottle, adds methanol to scale, and ultrasonic 10 minutes, filter, get subsequent filtrate, promptly get;
Accurate respectively object of reference solution and each 20ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, measures, and writes down 55 minutes chromatogram, promptly gets characteristic spectrum, sees Fig. 2,3.
Should present 4 characteristic peaks in the test sample characteristic spectrum; With the corresponding peak of object of reference be the S peak; Calculate each characteristic peak and the corresponding peak-to-peak RRT of contrast characteristic spectrum; 3 peaks should be identical with corresponding object of reference peak retention time respectively therein, its RRT should setting ± 5% within; Setting is: 17.133 (peaks 1), 20.950 (peaks 2), 21.167 (peaks 3), 26.467 (peaks 4).
The Radix Bupleuri standardized extract differentiates, adopts " characteristic peak coincide collection of illustrative plates ", " characteristic peak contrast collection of illustrative plates ", " TLC chromatogram " to carry out quality control.All accomplish good reproducibility, specificity is strong, has accomplished 4 peaks of saikoside, fits like a glove, and retention time is identical.
Detection method is stable, accurately, can repeat.Test detects the method adopted and is " the official method that Chinese pharmacopoeia version in 2010 is recorded.
(5) foundation of TLC thin layer chromatic graph
According to " appendix a VI of Chinese pharmacopoeia version in 2010 B test.
It is that reference substance and extract contrast that this thin layer is differentiated with saikoside a bcd, adopts the contrast of saikoside a bcd mixed solution and extract simultaneously, takes the dual contrast of thin layer like this, favorable reproducibility, and specificity is strong.The selection of thin layer condition is according to " appendix a VI of Chinese pharmacopoeia version in 2010 B test.
Claims (3)
1. Radix Bupleuri standardized extract half a bionical combined ultrasonic method for preparing is characterized in that step is following: get Radix Bupleuri and be ground into powder, add 60% ethanol, transfer pH=8 with stabilizing agent; Soak into 8h, under 40 ℃ of conditions of temperature, supersound extraction 3 times, each 40 minutes; Add 8 times of amount ethanol at every turn, transfer pH=8, filter merging filtrate with stabilizing agent; Reclaim ethanol to there not being the alcohol flavor, under 60 ℃, be concentrated into relative density 1.10 spray dryinges or microwave vacuum drying, promptly get.
2. according to the described Radix Bupleuri standardized extract of claim 1 half bionical combined ultrasonic method for preparing, it is characterized in that: with saikoside a, b, c, d total content calculate that its extract average content is 1.874%, average flour extraction 15.35%, average yield be 0.288% or extraction ratio be 84.706%.
3. according to claim 1 or 2 described Radix Bupleuri standardized extract, half bionical combined ultrasonic method for preparing, it is characterized in that: supersound extraction power 250W, frequency 50kHz.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100522888A CN102579529A (en) | 2012-03-01 | 2012-03-01 | Half-bionic combined ultrasonic preparation method of standardized extract of Bupleurum chinense DC. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100522888A CN102579529A (en) | 2012-03-01 | 2012-03-01 | Half-bionic combined ultrasonic preparation method of standardized extract of Bupleurum chinense DC. |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102579529A true CN102579529A (en) | 2012-07-18 |
Family
ID=46469132
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012100522888A Pending CN102579529A (en) | 2012-03-01 | 2012-03-01 | Half-bionic combined ultrasonic preparation method of standardized extract of Bupleurum chinense DC. |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102579529A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104530180A (en) * | 2015-01-21 | 2015-04-22 | 黑龙江中医药大学 | Method for extracting saikosaponin D from radix bupleuri |
| CN108445139A (en) * | 2018-03-26 | 2018-08-24 | 河南省洛正药业有限责任公司 | A kind of discrimination method of Chinese medicine preparation that treating synovitis |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101084948A (en) * | 2006-06-08 | 2007-12-12 | 天津天士力之骄药业有限公司 | Method for preparing saikosaponin |
-
2012
- 2012-03-01 CN CN2012100522888A patent/CN102579529A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101084948A (en) * | 2006-06-08 | 2007-12-12 | 天津天士力之骄药业有限公司 | Method for preparing saikosaponin |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104530180A (en) * | 2015-01-21 | 2015-04-22 | 黑龙江中医药大学 | Method for extracting saikosaponin D from radix bupleuri |
| CN104530180B (en) * | 2015-01-21 | 2016-06-08 | 黑龙江中医药大学 | From radix bupleuri, extract the method for saikosaponin D |
| CN108445139A (en) * | 2018-03-26 | 2018-08-24 | 河南省洛正药业有限责任公司 | A kind of discrimination method of Chinese medicine preparation that treating synovitis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113791163B (en) | Method for detecting quality of bunge cherry seed standard decoction | |
| CN105467059A (en) | Quality detecting method for traditional Chinese medicine composition for treating hematuresis | |
| CN102353732B (en) | Quality detection method of Zhenlong brain-refreshment preparation | |
| CN104306745A (en) | Quality control method for rhizoma gastrodiae capsule | |
| CN102565272A (en) | Quality standard of standardized bupleurum extract | |
| CN104914199A (en) | Determining method for contents of twelve components in traditional Chinese medicine composition preparation | |
| CN109142588B (en) | HPLC (high performance liquid chromatography) characteristic spectrum of Lianzhi anti-inflammation capsules as well as construction method and application thereof | |
| CN115144507B (en) | Method for simultaneously measuring active ingredients in Sang Ren lung-heat clearing oral liquid | |
| CN109668970B (en) | Ultra-high performance liquid chromatography detection method for traditional Chinese medicine composition | |
| CN110907550B (en) | Method for simultaneously detecting contents of six components in infant lung-ventilating and cough-relieving granules | |
| CN100437112C (en) | Method for inspecting Chinese medicinal preparation quality in treatment of old man eyes dieases | |
| CN102218122A (en) | Quality control and detection method for sea dragon and gecko oral liquid | |
| CN100541195C (en) | A kind of detection method of Chinese medicine preparation | |
| CN101543545B (en) | Traditional Chinese medicine preparation for curing rhinitis and quality control method thereof | |
| CN104383370A (en) | Jianqu and its preparation method and quality detection method | |
| CN108037200B (en) | Quality detection method of kidney nourishing and tranquilizing pills | |
| CN102579529A (en) | Half-bionic combined ultrasonic preparation method of standardized extract of Bupleurum chinense DC. | |
| CN102707006B (en) | Quality detection method of cudrania tricuspidata formula granules | |
| CN104922587A (en) | Preparation method of swelling-removing and pain-relieving paste and quality-detecting method of swelling-removing and pain-relieving paste | |
| CN102389445A (en) | Angelica dahurica root dispensing granule and quality control thereof | |
| Wang et al. | Development and validation of a high-performance thin-layer chromatographic fingerprint method for the evaluation of QiYi capsules with the reference of myotonin | |
| CN102335333A (en) | Method for detecting traditional Chinese medicine preparation of compound nutritions paste | |
| CN115266975A (en) | Method for measuring content of genistin in endothelium corneum gigeriae galli and processed decoction pieces, standard decoction and formula granules thereof | |
| CN104111295A (en) | Method for controlling quality of Chinese herbal preparation | |
| CN103983735A (en) | Detection method for preparing Gongyanping (brand) capsules |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120718 |