CN103115975A - Method for detecting quality of salvianolic acid B in yellow spot removing capsules - Google Patents
Method for detecting quality of salvianolic acid B in yellow spot removing capsules Download PDFInfo
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- CN103115975A CN103115975A CN201310029346XA CN201310029346A CN103115975A CN 103115975 A CN103115975 A CN 103115975A CN 201310029346X A CN201310029346X A CN 201310029346XA CN 201310029346 A CN201310029346 A CN 201310029346A CN 103115975 A CN103115975 A CN 103115975A
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- tanshin polyphenolic
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Abstract
The invention discloses a method for detecting the quality of salvianolic acid B in yellow spot removing capsules, which belongs to the technical field of medicine quality control. The method carries out identification and content detection through high efficiency liquid chromatography by taking the salvianolic acid B as a comparison product and the contents of the yellow spot removing capsules as a tested product. The high efficiency liquid chromatography is implemented under the conditions that octadecyl silane bonded silica gel is taken as a filler, methanol-acetonitrile-formic acid-water (in a ratio of 23:10:1:66) are taken as mobile phases, a detection wavelength is 286 nanometers, and the number of theoretical plates is not less than 2000 according to the peak of the salvianolic acid B. The method disclosed by the invention can be used for carrying out the identification and the content detection on the salvianolic acid B by the high efficiency liquid chromatography, is more scientific, more advanced in detection means and more accurate in qualitative and quantitative detection, and can not only better and more comprehensively reflect the effective components of the yellow spot removing capsules, but also prevent the condition that other medicinal materials substitute for radix salviae miltiorrhizae for production, thereby ensuring the quality, the safety and the effectiveness of the medicine.
Description
Technical field
The present invention relates to the quality determining method of active component in a kind of pharmaceutical preparation, specifically, what relate to is the quality determining method of tanshin polyphenolic acid B in a kind of Chinese medicinal capsule preparation for the treatment of chloasma, belongs to drug quality control technology field.
Background technology
The Chinese medicinal capsule preparation of the treatment chloasma that the present invention is specifically related to is namely caught up with yellow freckle-diminishing capsule, after catching up with yellow careless 835g and red sage root 300g to make thick paste, through adding suitable auxiliary material to prepare, its concrete Preparation method and use has been recorded in the patent of invention that publication number is CN101700276, said preparation has the effect of removing jaundice, stagnation resolvation, nti-freckle, intends being used for the treatment of clinically the treatment of the women's chloasma that causes due to stagnation of liver qi.
Catch up with to contain in yellow freckle-diminishing capsule and catch up with yellow grass and the red sage root, proof, catch up with the main effective constituent-Quercetin of yellow grass to have nti-freckle, anti-inflammatory, antiallergy after deliberation, strengthens capillary of skin permeability function; Principal ingredient in the red sage root has Tanshinone II
AAnd tanshin polyphenolic acid B, wherein: Tanshinone II
AFunction with anti-inflammatory, tanshin polyphenolic acid B have powerful oxidation resistant effect.Main effective constituent also find this product in to the analysis of catching up with yellow freckle-diminishing capsule in is Quercetin, Tanshinone II
AAnd tanshin polyphenolic acid B.
Find only have independent mensuration to catch up with the Quercetin in yellow careless preparation or measure separately Tanshinone II in red sage formulation by inquiring about present pertinent literature both domestic and external
AWith the report of tanshin polyphenolic acid B, also not about to by the Quercetin in the compound preparation of catching up with yellow grass, the red sage root two flavor medicines to form, Tanshinone II
AThe bibliographical information that detects with tanshin polyphenolic acid B.In order better to control the quality of catching up with yellow freckle-diminishing capsule, guarantee the security of clinical application, be necessary to set up the complete quality control standard of a cover with the quality of better control this product.
Catch up with the thick paste preparation method who relates generally in the preparation technology of yellow freckle-diminishing capsule to be: red sage root 300g, add 90% alcohol reflux 1.5 hours, filter, filtrate recycling ethanol is to thick paste; The red sage root dregs of a decoction with catch up with yellow careless 835g, boiling 2 times, each 2 hours, collecting decoction filters, and it is 1.15～1.18(60～65 ℃ that filtrate is condensed into relative density) clear cream, cooling, add ethanol and make and contain alcohol amount and reach 60%, stir, standing, filter, precipitation merges washing lotion and filtrate with 60% ethanol washing three times, and recovery ethanol also is condensed into the thick paste of relative density 1.30～1.32; Merge above-mentioned two kinds of thick pastes, standby.
Get thick paste, add appropriate amount of starch, microcrystalline cellulose, talcum powder, dolomol etc., mixing breaks into the powder direct packaging after granulate dry rear packing or medicinal extract convection drying, and get final product.
What obtain here is the content of capsule in fact, can learn by analysis, in whole production technology quality control is fairly simple and coarse, may be because of production batch different, perhaps just at last same batch also can produce the product that content is different, streaking is even.
If complete or in the content preparation process, just content carried out suitable discriminating and content detection in the content preparation, this stablizes to steady quality and the curative effect of product the vital effect that plays undoubtedly.
Summary of the invention
The objective of the invention is does not also have the pharmaceutical preparation of listing to catch up with yellow freckle-diminishing capsule at present domestic, and a kind of quality determining method that can differentiate comprehensively and detect one of said preparation effective constituent tanshin polyphenolic acid B is provided, and has guaranteed the safe and effective of this pharmaceutical preparation.
For achieving the above object, the concrete technical scheme of the present invention's employing is as follows:
Catch up with the quality determining method of tanshin polyphenolic acid B in yellow freckle-diminishing capsule, it is characterized in that: take tanshin polyphenolic acid B as reference substance, take the potpourri of methyl alcohol, acetonitrile, formic acid and water as mobile phase, methyl alcohol: acetonitrile: formic acid: water v/v/v/v=23:10:1:66, differentiate and content detection with high performance liquid chromatography.
The condition of described high performance liquid chromatography is: be filling agent with octadecylsilane chemically bonded silica, take methyl alcohol-acetonitrile-formic acid-water (23:10:1:66) as mobile phase, the detection wavelength is 286nm; Number of theoretical plate should be not less than 2000 by tanshin polyphenolic acid B peak calculating.
The concrete grammar that described discriminating detects is as follows:
The preparation of A reference substance solution: it is appropriate that precision takes the tanshin polyphenolic acid B reference substance, makes the reference substance solution that contains 0.1mg in every 1ml with 75% methanol solution;
The preparation of B need testing solution: accurately weighedly catch up with yellow freckle-diminishing capsule content 0.5g, put in conical flask, add methyl alcohol: the solution 50ml of water v/v=75:25, accurately weighed after, ultrasonic extraction 30 minutes, take out, let cool, more accurately weighed and employing methyl alcohol: the solution of water v/v=75:25 is supplied the weight that loses, and shakes up, filter, filtrate is as need testing solution;
C liquid chromatogram measuring chromatographic peak: precision measures reference substance solution and each 20 μ l of need testing solution respectively, in the injection liquid chromatography, detects chromatographic peak consistent with reference substance chromatographic peak retention time in the test sample chromatogram.
The concrete steps of described content detection are as follows:
The preparation of A reference substance solution: it is appropriate that precision takes the tanshin polyphenolic acid B reference substance, makes the reference substance solution that contains 0.1mg in every 1ml with 75% methanol solution;
The preparation of B need testing solution: accurately weighedly catch up with yellow freckle-diminishing capsule content 0.5g, put in conical flask, add methyl alcohol: the solution 50ml of water v/v=75:25, accurately weighed after, ultrasonic extraction 30 minutes, take out, let cool, more accurately weighed and employing methyl alcohol: the solution of water v/v=75:25 is supplied the weight that loses, and shakes up, filter, filtrate is as need testing solution;
C liquid chromatogram measuring content: precision measures reference substance solution and each 20 μ l of need testing solution respectively, in the injection liquid chromatography, measures, and get final product.
The inventor has carried out checking research to the feasibility of discrimination method and content assaying method; Concrete experimental data is as follows:
1, the high efficiency liquid phase of tanshin polyphenolic acid B is differentiated research
Method: high performance liquid chromatography
Sample and reference substance disposal route: it is appropriate that precision takes the tanshin polyphenolic acid B reference substance, adds 75% methanol solution and make the reference substance solution liquid that contains 0.1mg in every 1ml.Need testing solution preparation: get test sample 0.5g, accurately weighed, put in conical flask, add 50ml methyl alcohol: the solution of water v/v=75:25, accurately weighed after, ultrasonic extraction 30 minutes, take out, let cool, more accurately weighed and employing methyl alcohol: the solution of water v/v=75:25 is supplied the weight that loses, shake up, filter.As need testing solution.Accurate reference substance solution and each 20ul of need testing solution of drawing, in the injection liquid chromatography, measure, and get final product respectively.
Chromatographic column: month rising sun XB-C
18(250mm * 4.6mm, 5 μ m)
Mobile phase: methyl alcohol-acetonitrile-formic acid-water (23:10:1:66)
Detect wavelength: 286nm
Flow velocity: 1.0ml/min
Result: drafting under condition, the chromatographic peak consistent with the reference substance retention time arranged in sample.
2, the high efficiency liquid phase assay of tanshin polyphenolic acid B research
Method: high performance liquid chromatography
1. instrument and experimental drug
Alltech-426 type high efficiency liquid phase, the CQX25-06 ultrasonic cleaner.Methyl alcohol, acetonitrile are chromatographic grade, and formic acid is pure for analyzing.The tanshin polyphenolic acid B reference substance provides for Nat'l Pharmaceutical ﹠ Biological Products Control Institute, catches up with yellow freckle-diminishing capsule to be provided by Chengdu Li Site Remedy Research Limited.
2. method and result
2 .1 chromatographic conditions:
Chromatographic column: month rising sun XB-C
18(250mm * 4.6mm, 5 μ m)
Mobile phase: methyl alcohol-acetonitrile-formic acid-water (23:10:1:66)
Detect wavelength: 286nm
Flow velocity: 1.0ml/min
2.2 solution preparation
2.2.1 it is appropriate that precision takes the tanshin polyphenolic acid B reference substance, adds 75% methanol solution and make the reference substance solution liquid that contains 0.1mg in every 1ml.
2.2.2 need testing solution preparation: get approximately 0.5g of test sample, accurately weighed, put in conical flask, add 50ml methyl alcohol: the solution of water v/v=75:25, after accurately weighed, ultrasonic extraction 30 minutes is taken out, and lets cool, accurately weighed and adopt methyl alcohol again: the solution of water v/v=75:25 is supplied the weight that loses, shake up, filter, filtrate is as need testing solution.
2.3 system suitability experiment: precision is drawn reference substance solution and each 20 μ l of need testing solution respectively, inject high performance liquid chromatograph, record chromatogram, the retention time of tanshin polyphenolic acid B is 17.214min as a result, the degree of separation of tanshin polyphenolic acid B peak and front and back chromatographic peak is all greater than 1.5, and theoretical tray is calculated as 5474. with the tanshin polyphenolic acid B peak
2.4 linear relationship: precision is drawn reference substance solution 2.5 under the 2.2.1 item respectively, 5,7.5,10,15,20 μ l, in the injection liquid chromatography, measure peak area according to above-mentioned chromatographic condition, A returns sample size C with the peak area integrated value, the typical curve equation: A=1014741C-10244 r=0.9991, result shows that tanshin polyphenolic acid B sample size peak area and sample size in the 2.5-20ng scope have good linear relationship.
2.5 precision research, reference substance solution 20 μ l under accurate absorption 2.2.1 item in the injection liquid chromatography, measure peak area according to above-mentioned chromatographic condition, repeat sample introduction 6 times, and recording tanshin polyphenolic acid B peak area RSD is 1.32%, shows that instrument precision is good.
2.6 stability study, reference substance solution 20 μ l under accurate absorption 2.2.2 item are respectively 0,2, in 4,6,8 hours injection liquid chromatographies, measure peak area according to above-mentioned chromatographic condition, recording tanshin polyphenolic acid B peak area RSD is 0.99%, shows that need testing solution is stable in 8 hours.
2.7 sample solution 20 μ l under the accurate absorption of repeated experiment 2.2.2 item in the injection liquid chromatography, measure peak area according to above-mentioned chromatographic condition, repeat sample introduction 6 times, recording tanshin polyphenolic acid B peak area RSD is 1.09%, shows that instrument precision is good.
2.8 recovery experiment: precision takes 9 parts of this product of known content, every part of 0.5g, be divided into 3 groups, every group 3 parts, closely add respectively the tanshin polyphenolic acid B contrast solution appropriate, according to legal system available test sample solution below the 2.2.2 item, measure peak area according to above-mentioned chromatographic condition, calculate recovery rate, the recovery are 99.05%, RSD=0.69%.Experimental result shows that this product has the good recovery.
2.9 sample size is measured: precision is drawn reference substance solution and each 20 μ l of need testing solution respectively, in the injection liquid chromatography, measures, and get final product.The every 1g of this product contains tanshin polyphenolic acid B (C
36H
30O
16) must not be less than 8.6mg.
Advantage of the present invention is to adopt high performance liquid chromatography that discriminating and the content of tanshin polyphenolic acid B are detected, method is more scientific, detection means is more advanced, qualitative, quantitative is more accurate, not only can better more fully reflect the effective constituent of catching up with yellow freckle-diminishing capsule, but also can stop to adopt other medicinal materials to replace the situation of catching up with yellow grass to feed intake and produce, thus guaranteeing the quality of this medicine, the civilian medicine of underwriter is safe and effective.
Another advantage of the present invention is, by discriminating and the content detection to tanshin polyphenolic acid B, can just carry out quality control before capsule-filling, in order to produce up-to-standard product, simultaneously, also can provide for commercially available circulation product a kind of scientific and reasonable detection method, so that from producing and field of circulation control product quality.
Embodiment
Below in conjunction with embodiment, foregoing of the present invention is described in further detail again, but this should be interpreted as technical solution of the present invention only limits to following examples.
The discriminating of embodiment 1 tanshin polyphenolic acid B
Be filling agent with octadecylsilane chemically bonded silica; Take methyl alcohol-acetonitrile-formic acid-water (23:10:1:66) as mobile phase; The detection wavelength is 286nm; Number of theoretical plate should be not less than 2000 by tanshin polyphenolic acid B peak calculating.It is appropriate that precision takes the tanshin polyphenolic acid B reference substance, makes the solution that contains 0.1mg in every 1ml with 75% methanol solution, in contrast product solution.Get approximately 0.5g of this product content powder, accurately weighed, be placed in conical flask, add 50ml methyl alcohol: the solution of water v/v=75:25, after precise weighing, ultrasonic extraction 30 minutes is taken out, and lets cool, accurately weighed and adopt methyl alcohol again: the solution of water v/v=75:25 is supplied the weight that loses, shake up, filter, filtrate is as need testing solution.Precision measures reference substance solution and each 20 μ l of need testing solution respectively, in the injection liquid chromatography, detects, and should have the chromatographic peak consistent with reference substance chromatographic peak retention time in the test sample chromatogram.
The assay of embodiment 2 tanshin polyphenolic acid Bs
Be filling agent with octadecylsilane chemically bonded silica; Take methyl alcohol-acetonitrile-formic acid-water (23:10:1:66) as mobile phase; The detection wavelength is 286nm; Number of theoretical plate should be not less than 2000 by tanshin polyphenolic acid B peak calculating.It is appropriate that precision takes the tanshin polyphenolic acid B reference substance, adds 75% methanol solution and make the solution that contains 0.1mg in every 1ml, in contrast product solution liquid.Get this product content powder 0.5g, accurately weighed, be placed in conical flask, add 50ml methyl alcohol: the solution of water v/v=75:25, after accurately weighed, ultrasonic extraction 30 minutes is taken out, and lets cool, accurately weighed and adopt methyl alcohol again: the solution of water v/v=75:25 is supplied the weight that loses, shake up, filter, filtrate is as need testing solution.Precision measures reference substance solution and each 20 μ l of need testing solution respectively, in the injection liquid chromatography, measures and get final product.
Claims (4)
1. catch up with the quality determining method of tanshin polyphenolic acid B in yellow freckle-diminishing capsule, it is characterized in that: take tanshin polyphenolic acid B as reference substance, take the potpourri of methyl alcohol, acetonitrile, formic acid and water as mobile phase, methyl alcohol: acetonitrile: formic acid: water v/v/v/v=23:10:1:66, differentiate and content detection with high performance liquid chromatography.
2. the quality determining method of catching up with tanshin polyphenolic acid B in yellow freckle-diminishing capsule according to claim 1, it is characterized in that: the condition of described high performance liquid chromatography is: be filling agent with octadecylsilane chemically bonded silica, take methyl alcohol-acetonitrile of volume ratio 23:10:1:66-formic acid-water as mobile phase, the detection wavelength is 286nm, and number of theoretical plate is not less than 2000 by tanshin polyphenolic acid B peak calculating.
3. the quality determining method of catching up with tanshin polyphenolic acid B in yellow freckle-diminishing capsule according to claim 1 is characterized in that: the concrete grammar that described discriminating detects is as follows:
The preparation of A reference substance solution: it is appropriate that precision takes the tanshin polyphenolic acid B reference substance, makes the reference substance solution that contains 0.1mg in every 1ml with 75% methanol solution;
The preparation of B need testing solution: accurately weighedly catch up with yellow freckle-diminishing capsule content 0.5g, be placed in conical flask, add methyl alcohol: the solution 50ml of water v/v=75:25, accurately weighed after, ultrasonic extraction 30 minutes, take out, let cool, more accurately weighed and employing methyl alcohol: the solution of water v/v=75:25 is supplied the weight that loses, and shakes up, filter, filtrate is as need testing solution;
C liquid chromatogram measuring chromatographic peak: precision measures reference substance solution and each 20 μ l of need testing solution respectively, in the injection liquid chromatography, detects chromatographic peak consistent with reference substance chromatographic peak retention time in the test sample chromatogram.
4. the quality determining method of catching up with tanshin polyphenolic acid B in yellow freckle-diminishing capsule according to claim 1, it is characterized in that: the concrete steps of described content detection are as follows:
The preparation of A reference substance solution: it is appropriate that precision takes the tanshin polyphenolic acid B reference substance, makes the reference substance solution that contains 0.1mg in every 1ml with 75% methanol solution;
The preparation of B need testing solution: accurately weighedly catch up with yellow freckle-diminishing capsule content 0.5g, be placed in conical flask, add methyl alcohol: the solution 50ml of water v/v=75:25, accurately weighed after, ultrasonic extraction 30 minutes, take out, let cool, more accurately weighed and employing methyl alcohol: the solution of water v/v=75:25 is supplied the weight that loses, and shakes up, filter, filtrate is as need testing solution;
C liquid chromatogram measuring content: precision measures reference substance solution and each 20 μ l of need testing solution respectively, in the injection liquid chromatography, measures, and get final product.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101703583A (en) * | 2009-11-30 | 2010-05-12 | 贵阳新天药业股份有限公司 | Method for detecting quality of Xinning capsule |
| CN101780170A (en) * | 2010-03-18 | 2010-07-21 | 辽宁正鑫药物研究有限公司 | Compound capsule containing root of red-rooted salvia and preparation method and quality determination method thereof |
-
2013
- 2013-01-25 CN CN201310029346XA patent/CN103115975A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101703583A (en) * | 2009-11-30 | 2010-05-12 | 贵阳新天药业股份有限公司 | Method for detecting quality of Xinning capsule |
| CN101780170A (en) * | 2010-03-18 | 2010-07-21 | 辽宁正鑫药物研究有限公司 | Compound capsule containing root of red-rooted salvia and preparation method and quality determination method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 李晓红 等: "高效液相色谱法测定丹芪养血颗粒中丹酚酸B的含量", 《中南药学》 * |
| 李红磊: "香丹注射液中丹酚酸B的HPLC法测定", 《分析测试学报》 * |
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Application publication date: 20130522 |