CN104678036A - Method for detecting quality of woman dysmenorrheal granules - Google Patents
Method for detecting quality of woman dysmenorrheal granules Download PDFInfo
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Abstract
The invention discloses a method for detecting the quality of woman dysmenorrheal granules, and belongs to the technical field of traditional Chinese medicine quality detection. The method comprises the following steps: (1) determining the content of tetrahydropalmatine; (2) determining the content of tanshinone IIA; and (3) determining the content of protocatechuic acid. According to the method, the original quality standard is improved, and the contents of the tetrahydropalmatine, the tanshinone IIA and the protocatechuic acid in the woman dysmenorrheal granules are determined. The original quality standard is improved, so that the product quality detection and stability in the industrial production are ensured.
Description
Technical field
The invention belongs to traditional Chinese medicine quality detection technique field, be specifically related to a kind of quality determining method of dysmenorrhes particle.
Technical background
Dysmenorrhes particle is Chinese herbal granules, and by traditional Chinese medicine corydalis tuber, the red sage root, excrementum pteropi, cattail pollen four taste Chinese medicine forms, and have and invigorate blood circulation, menstruation regulating, the effect of pain relieving, is used for the treatment of qi and blood stagnation, little abdominal distension pain, bellyache in menstrual period.
In dysmenorrhes particle Chinese medicine excrementum pteropi promoting blood circulation and removing blood stasis have improve the effect such as Hemorheology, anti-freezing platelet aggregation-against, protocatechuic acid is one of important activity element monomers of traditional Chinese medicine excrementum pteropi.In dysmenorrhes particle, salviamiltiorrhizabung has stasis-dispelling and pain-killing, activating blood to promote menstruation, the effect of the relieving restlessness that clears away heart-fire.For irregular menstruation, through closing dysmenorrhoea, a lump in the abdomen causing distension and pain, chest ventral spine pain, hot numbness pain, and sore swells and ache, dysphoria and insomnia; Hepatosplenomegaly, angina pectoris, wherein principle active component tanshinone IIA suppresses ADP induced platelet aggregation, expansion animal arteriole, makes the open increasing number of capillary network, accelerates velocity of blood flow, flow, thus improve microcirculation disorder, wait effect.Corydalis tuber in prescription, the red sage root, the consumption of excrementum pteropi is 1:1:1, all has therapeutic action to main disease.And only assay has been carried out to tetrahydropalmatine in proper mass standard, carried out TLC distinguish to the red sage root, not to the red sage root, in excrementum pteropi, index composition carries out assay, is unfavorable for the detection ensureing medicine inherent quality in process of production.
Summary of the invention
The object of the invention is for described defect above, there is provided a kind of quality determining method of dysmenorrhes particle, the method establishes main ingredient corydalis tuber in compound preparation, the red sage root, the content assaying method of index components in excrementum pteropi, ensures that compound preparation has higher quality level.
The object of the invention is to be achieved by the following technical programs.
A quality determining method for dysmenorrhes particle, is characterized in that, the method is made up of following steps:
(1) Content determination of dl-tetrahydropalmatine measures:
A, chromatographic condition and system suitability test: be filling agent with octadecylsilane chemically bonded silica, with 0.1% phosphoric acid solution (diethylamine adjust pH to 6.0)-methyl alcohol for mobile phase, the volume ratio of phosphoric acid solution and methyl alcohol is 35 ~ 45: 65 ~ 55; Prioritizing selection: with 0.1% phosphoric acid solution (diethylamine adjust pH to 6.0)-methyl alcohol for mobile phase, the volume ratio of phosphoric acid solution and methyl alcohol is 40 ~ 45: 60 ~ 55; Determined wavelength is 280nm; Number of theoretical plate calculates should be not less than 3000 by tetrahydropalmatine;
The preparation of b, reference substance solution: it is appropriate that precision takes tetrahydropalmatine reference substance, adds methyl alcohol and makes the solution of every 1ml containing 50ug, to obtain final product;
The preparation of c, need testing solution: get this product and be about 1g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, enriching ammonia 2ml soaks, and places 15 minutes, precision adds solvent (ether or chloroform) 50ml, weighed weight, extract (ultrasonic, backflow or apparatus,Soxhlet's extract 60 minutes, prioritizing selection is ultrasonic), let cool, weighed weight again, supplies the weight of less loss, shakes up, filters with solvent (ether or chloroform); Precision measures subsequent filtrate 25ml, and low temperature volatilizes solvent, and residue adds methyl alcohol and dissolves, and is transferred in 10ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product;
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product;
The every g sample of this product in tetrahydropalmatine (C21H25NO4), must not be less than 0.2mg containing corydalis tuber;
(2) determation of tanshinone ⅡA:
A, chromatographic condition and system suitability test: take octadecylsilane chemically bonded silica as filling agent; Take methanol-water as mobile phase, the volume ratio of methyl alcohol and water is 65 ~ 85: 35 ~ 15; Prioritizing selection take methanol-water as mobile phase, and the volume ratio of methyl alcohol and water is 75 ~ 85: 25 ~ 15; Determined wavelength is 270nm; Number of theoretical plate calculates should be not less than 2000 by tanshinone IIA peak;
The preparation of b, reference substance solution: get tanshinone IIA reference substance appropriate, accurately weighed, put in brown measuring bottle, add methyl alcohol and make the solution of every 1ml containing tanshinone IIA 16 μ g, to obtain final product;
The preparation of c, need testing solution: get this product and be about 1g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, precision adds solvent (one in methyl alcohol, ethanol or normal butyl alcohol, prioritizing selection methyl alcohol) 50ml, weighed weight, ultrasonic process 30 minutes, let cool, more weighed weight, the weight of less loss is supplied with solvent (one in methyl alcohol, ethanol or normal butyl alcohol), shake up, filter; Precision measures subsequent filtrate 25ml, evaporate to dryness, and residue adds methyl alcohol and dissolves, and is transferred in 10ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product;
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product;
The every g sample of this product in tanshinone IIA, must not be less than 0.3mg containing the red sage root;
(3) protocatechuic acid assay:
A, chromatographic condition and system suitability test: be filling agent with octadecylsilane chemically bonded silica, with 0.1% phosphoric acid solution-methyl alcohol for mobile phase, the volume ratio of phosphoric acid solution and methyl alcohol is 95 ~ 85: 5 ~ 15; Prioritizing selection is with 0.1% phosphorus acetum-methyl alcohol for mobile phase, and the volume ratio of phosphoric acid solution and methyl alcohol is 92 ~ 87: 8 ~ 13; Determined wavelength is 260nm, and number of theoretical plate calculates should be not less than 3000 by protocatechuic acid;
The preparation of b, reference substance solution: it is appropriate that precision takes protocatechuic acid reference substance, adds methyl alcohol and makes the solution of every 1ml containing 50ug, to obtain final product;
The preparation of c, need testing solution: get this product and be about 3g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, precision adds solvent (60% ~ 90% methyl alcohol or 60% ~ 90% ethanol) 100ml, weighed weight, ultrasonic process 30 minutes (power 250W, frequency 50KHz), let cool, more weighed weight, the weight of less loss is supplied with solvent (60% ~ 90% methyl alcohol or 60% ~ 90% ethanol), shake up, filter; Precision measures subsequent filtrate 50ml, evaporate to dryness, and residue adds methyl alcohol and dissolves, and is transferred in 5ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product;
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product;
The every g sample of this product in protocatechuic acid (C7H6O4), must not be less than 0.01mg containing excrementum pteropi.
The invention has the beneficial effects as follows: present invention improves over initial quality standard, add the assay of tetrahydropalmatine, tanshinone IIA, protocatechuic acid in dysmenorrhes particle.Invention increases initial quality standard, the quality testing and the constant product quality that are conducive to product in commercial production ensure.
Accompanying drawing explanation
Accompanying drawing 1: embodiment tetrahydropalmatine reference substance collection of illustrative plates.
Accompanying drawing 2: embodiment Content determination of dl-tetrahydropalmatine working sample collection of illustrative plates.
Accompanying drawing 3: embodiment tanshinone IIA contrast collection of illustrative plates.
Accompanying drawing 4: embodiment determation of tanshinone ⅡA sample collection of illustrative plates.
Accompanying drawing 5: embodiment protocatechuic acid reference substance collection of illustrative plates.
Accompanying drawing 6: embodiment protocatechuic acid assay sample collection of illustrative plates.
Embodiment
In order to better explain enforcement of the present invention, provide the embodiment of following assay method, the present invention is described further.
Embodiment.
(1) Content determination of dl-tetrahydropalmatine measures.
A, chromatographic condition and system suitability test: be filling agent with octadecylsilane chemically bonded silica, with 0.1% phosphoric acid solution (diethylamine adjust pH to 6.0)-methyl alcohol (45: 55) for mobile phase; Determined wavelength is 280nm.Number of theoretical plate calculates should be not less than 3000 by tetrahydropalmatine.
The preparation of b, reference substance solution: it is appropriate that precision takes tetrahydropalmatine reference substance, adds methyl alcohol and makes the solution of every 1ml containing 50ug, to obtain final product.
The preparation of c, need testing solution: get this product and be about 1g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, enriching ammonia 2ml soaks, place 15 minutes, precision adds ether 50ml, weighed weight, ultrasonic extraction 60min, let cool, more weighed weight, ether supplies the weight of less loss, shakes up.Filter.Precision measures subsequent filtrate 25ml, and low temperature volatilizes solvent, and residue adds methyl alcohol, ethanol or n-Butanol soluble, is transferred in 10ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product.
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product.
The every g sample of this product in tetrahydropalmatine (C21H25NO4), must not be less than 0.2mg containing corydalis tuber.
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; With 0.1% phosphoric acid solution (diethylamine adjust pH to 6.0)-methyl alcohol (42: 58) for mobile phase; Determined wavelength is 280nm.Number of theoretical plate calculates should be not less than 3000 by tetrahydropalmatine.
The preparation of reference substance solution: it is appropriate that precision takes tetrahydropalmatine reference substance, adds methyl alcohol and makes the solution of every 1ml containing 50ug, to obtain final product.
The preparation of need testing solution: this product of getting under content uniformity is about 1g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, precision adds strong ammonia solution-methyl alcohol (1: 20) mixed solution 50ml, weighed weight, ultrasonic process 30 minutes (power 250W, frequency 50KHz), let cool, weighed weight again, supplies the weight of less loss, shakes up with strong ammonia solution-methyl alcohol (1: 20) mixed solution.Filter.Precision measures subsequent filtrate 25ml, evaporate to dryness, and residue adds methyl alcohol and dissolves, and is transferred in 10ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product.
(2) determation of tanshinone ⅡA.
A, chromatographic condition and system suitability test: take octadecylsilane chemically bonded silica as filling agent; With methanol-water (82:18) for mobile phase; Determined wavelength is 270nm.Number of theoretical plate calculates should be not less than 2000 by tanshinone IIA peak.
The preparation of b, reference substance solution: get tanshinone IIA reference substance appropriate, accurately weighed, put in brown measuring bottle, add methyl alcohol and make the solution of every 1ml containing tanshinone IIA 16 μ g, to obtain final product
The preparation of c, need testing solution: get this product and be about 1g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, precision adds solvent methanol 50ml, weighed weight, and ultrasonic process 30 minutes, lets cool, more weighed weight, supplies the weight of less loss, shake up with methyl alcohol.Filter.Precision measures subsequent filtrate 25ml, evaporate to dryness, and residue adds methyl alcohol and dissolves, and is transferred in 10ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product.
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product.
This product every g sample in single tanshinone IIA, must not be less than 0.3mg containing corydalis tuber.
(3) protocatechuic acid assay.
A, chromatographic condition and system suitability test: be filling agent with octadecylsilane chemically bonded silica, with 0.1% phosphoric acid solution-methyl alcohol (90: 10) for mobile phase; Determined wavelength is 260nm.Number of theoretical plate calculates should be not less than 3000 by protocatechuic acid.
The preparation of b, reference substance solution: it is appropriate that precision takes protocatechuic acid reference substance, adds methyl alcohol and makes the solution of every 1ml containing 50ug, to obtain final product.
The preparation of c, need testing solution: get this product and be about 3g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, precision adds 80% methanol solution 100ml, weighed weight, ultrasonic process 30 minutes (power 250W, frequency 50KHz), let cool, weighed weight again, supplies the weight of less loss, shakes up with 80% methanol solution.Filter.Precision measures subsequent filtrate 50ml, evaporate to dryness, and residue adds methyl alcohol and dissolves, and is transferred in 5ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product.
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product.
The every g sample of this product in protocatechuic acid (C7H6O4), must not be less than 0.01mg containing excrementum pteropi.
Claims (8)
1. a quality determining method for dysmenorrhes particle, is characterized in that, the method is made up of following steps:
(1) Content determination of dl-tetrahydropalmatine measures:
A, chromatographic condition and system suitability test: be filling agent with octadecylsilane chemically bonded silica, with 0.1% phosphoric acid solution-methyl alcohol for mobile phase, phosphoric acid solution diethylamine adjust pH to 6.0, the volume ratio of phosphoric acid solution and methyl alcohol is 35 ~ 45: 65 ~ 55; Determined wavelength is 280nm; Number of theoretical plate calculates should be not less than 3000 by tetrahydropalmatine;
The preparation of b, reference substance solution: it is appropriate that precision takes tetrahydropalmatine reference substance, adds methyl alcohol and makes the solution of every 1ml containing 50ug, to obtain final product;
The preparation of c, need testing solution: get this product and be about 1g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, enriching ammonia 2ml soaks, place 15 minutes, precision adds solvent 50ml, weighed weight, extracts, let cool, more weighed weight, supply the weight of less loss with solvent, shake up, filter; Precision measures subsequent filtrate 25ml, and low temperature volatilizes solvent, and residue adds methyl alcohol and dissolves, and is transferred in 10ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product; Described molten Ji is ether or chloroform;
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product;
The every g sample of this product in tetrahydropalmatine, must not be less than 0.2mg containing corydalis tuber;
(2) determation of tanshinone ⅡA:
A, chromatographic condition and system suitability test: take octadecylsilane chemically bonded silica as filling agent; Take methanol-water as mobile phase, the volume ratio of methyl alcohol and water is 65 ~ 85: 35 ~ 15; Determined wavelength is 270nm; Number of theoretical plate calculates should be not less than 2000 by tanshinone IIA peak;
The preparation of b, reference substance solution: get tanshinone IIA reference substance appropriate, accurately weighed, put in brown measuring bottle, add methyl alcohol and make the solution of every 1ml containing tanshinone IIA 16 μ g, to obtain final product;
The preparation of c, need testing solution: get this product and be about 1g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, precision adds solvent 50ml, weighed weight, and ultrasonic process 30 minutes, lets cool, more weighed weight, supplies the weight of less loss, shake up with solvent, filters; Precision measures subsequent filtrate 25ml, evaporate to dryness, and residue adds methyl alcohol and dissolves, and is transferred in 10ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product; Described solvent is the one in methyl alcohol, ethanol or normal butyl alcohol;
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product;
The every g sample of this product in tanshinone IIA, must not be less than 0.3mg containing the red sage root;
(3) protocatechuic acid assay:
A, chromatographic condition and system suitability test: be filling agent with octadecylsilane chemically bonded silica, with 0.1% phosphoric acid solution-methyl alcohol for mobile phase, the volume ratio of phosphoric acid solution and methyl alcohol is 95 ~ 85: 5 ~ 15; Determined wavelength is 260nm, and number of theoretical plate calculates should be not less than 3000 by protocatechuic acid;
The preparation of b, reference substance solution: it is appropriate that precision takes protocatechuic acid reference substance, adds methyl alcohol and makes the solution of every 1ml containing 50ug, to obtain final product;
The preparation of c, need testing solution: get this product and be about 3g, porphyrize, accurately weighed, put in 100ml tool plug conical flask, precision adds solvent 100ml, weighed weight, and ultrasonic process 30 minutes, lets cool, more weighed weight, supplies the weight of less loss, shake up with solvent, filters; Precision measures subsequent filtrate 50ml, evaporate to dryness, and residue adds methyl alcohol and dissolves, and is transferred in 5ml measuring bottle, and is diluted to scale, shake up, and filters, gets subsequent filtrate, to obtain final product; Described solvent is 60% ~ 90% methyl alcohol or 60% ~ 90% ethanol;
D, mensuration: accurate absorption reference substance solution and each 10ul of need testing solution, respectively injection liquid chromatography, measure, to obtain final product;
The every g sample of this product in protocatechuic acid, must not be less than 0.01mg containing excrementum pteropi.
2. the quality determining method of a kind of dysmenorrhes particle according to claim 1, is characterized in that: described in step (1), the volume ratio of phosphoric acid solution and methyl alcohol is 40 ~ 45: 60 ~ 55.
3. the quality determining method of a kind of dysmenorrhes particle according to claim 1, is characterized in that: the extracting method described in step (1) is that ultrasonic extraction, refluxing extraction or apparatus,Soxhlet's extract 60 minutes.
4. the quality determining method of a kind of dysmenorrhes particle according to claim 3, is characterized in that: the extracting method described in step (1) is ultrasonic extraction.
5. the quality determining method of a kind of dysmenorrhes particle according to claim 1, is characterized in that: described in step (2), the volume ratio of methyl alcohol and water is 75 ~ 85: 25 ~ 15.
6. the quality determining method of a kind of dysmenorrhes particle according to claim 1, is characterized in that: described in step (2), solvent is methyl alcohol.
7. the quality determining method of a kind of dysmenorrhes particle according to claim 1, is characterized in that: described in step (3), the volume ratio of phosphoric acid solution and methyl alcohol is 92 ~ 87: 8 ~ 13.
8. the quality determining method of a kind of dysmenorrhes particle according to claim 1, it is characterized in that: described in step (3), the power of ultrasonic process is 250W, frequency is 50KHz.
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Cited By (4)
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CN106404974A (en) * | 2016-08-30 | 2017-02-15 | 普正药业股份有限公司 | Woman dysmenorrheal granule fingerprint establishing method |
CN106680395A (en) * | 2017-01-09 | 2017-05-17 | 广东省第二中医院(广东省中医药工程技术研究院) | Method for measuring content of protocatechuic acid in jasminum elongatum |
CN107102093A (en) * | 2017-05-23 | 2017-08-29 | 四川逢春制药有限公司 | The detection method of compound pellet mattress cream |
CN110333303A (en) * | 2019-06-30 | 2019-10-15 | 广西万寿堂药业有限公司 | The content assaying method of the antipruritic Chinese materia medica preparation of gynaecology |
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106404974A (en) * | 2016-08-30 | 2017-02-15 | 普正药业股份有限公司 | Woman dysmenorrheal granule fingerprint establishing method |
CN106680395A (en) * | 2017-01-09 | 2017-05-17 | 广东省第二中医院(广东省中医药工程技术研究院) | Method for measuring content of protocatechuic acid in jasminum elongatum |
CN107102093A (en) * | 2017-05-23 | 2017-08-29 | 四川逢春制药有限公司 | The detection method of compound pellet mattress cream |
CN107102093B (en) * | 2017-05-23 | 2018-05-22 | 四川逢春制药有限公司 | The detection method of compound pellet mattress cream |
CN110333303A (en) * | 2019-06-30 | 2019-10-15 | 广西万寿堂药业有限公司 | The content assaying method of the antipruritic Chinese materia medica preparation of gynaecology |
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