CN100420463C - Preparation for treating hysteritis and its quality control method - Google Patents

Abstract

The present invention relates to a gynecology Chinese medicine preparation which is prepared by five traditional Chinese medicine: shing bugleweed herb, shinyleaf pricklyash roots, Chinese angelica, chuanposhi, five-finger fig, etc. The preparation simultaneously comprises gallic acid, chlorinate nitidine and ferulic acid; a quality control method of the present invention adopts an analysis method of high efficiency liquid chromatography or an ultraviolet spectroscopy method to quantitatively determine tannin or the gallic acid which are contained in the formulation, or the quality control method of the present invention can simultaneously adopt a thin-layer chromatography method to identify the Gallic acid in the preparation is from one or more than two kinds of the chuanposhi, the shing bugleweed herb, the shinyleaf pricklyash roots and constituent contained by the five-finger figure.

Description

A kind of Gongyanping preparation and method of quality control thereof

Invention field

The present invention relates to a kind of method of quality control of gynecological Chinese medicine preparation, particularly a kind of Gongyanping preparation of making by Chinese medicine of the five flavours such as Herba Melastomatis dodecandri, Radix Zanthoxyli, Radix Angelicae Sinensis, Radix Cudraniae, Herba Fici Simplicissimae (as pharmaceutical dosage forms such as tablet, capsule, dispersible tablet, granule, soft capsule, solution, microcapsule, injection) and method of quality control thereof.

Background technology:

China Patent No.: 00103132.5 " coated Gongyanping tablet in special shape and manufacture method thereof " discloses a kind of preparation method of GONGYANPING tablet.In recent years, further investigation along with Gongyanping preparation, the preparation method of GONGYANPING novel form has obtained concern, patent application as application number 03118640.8 relates to the capsular preparation of a kind of GONGYANPING, relate to a kind of preparation of GONGYANPING dispersible tablet as the application of application number 200410045022.6, relate to the preparation of several novel forms of GONGYANPING as the application of application number 200410058353.3.

In the GONGYANPING PIAN standard,, only there is the thin layer of Herba Melastomatis dodecandri and Radix Zanthoxyli control medicinal material to differentiate the quality of the restive Gongyanping preparation of this quality standard in " Chinese traditional patent formulation preparation " the 17 for the quality control of Gongyanping preparation.For guaranteeing the safe and effective of Gongyanping preparation, the quality standard that has carried out said preparation improves research.

2004 4 phases of Guiyang Medical College journal disclose the method for a kind of " content of gallic acid in the high effective liquid chromatography for measuring GONGYANPING capsule ", and are significantly different with the present invention.

Summary of the invention:

The invention discloses the detection method of quantitative control Gongyanping preparation quality, this method specificity is strong, and sensitivity, precision, repeatability all satisfy the requirement of drug quality control:

Technical scheme of the present invention is:

A kind of Gongyanping preparation, said preparation contain Herba Melastomatis dodecandri, Radix Zanthoxyli, Radix Angelicae Sinensis, Radix Cudraniae, Herba Fici Simplicissimae Chinese medicine of the five flavours, make oral formulations or external preparation or injection by suitable method, as tablet, capsule, drop pill, suppository, injection.Said preparation contains three kinds of compositions such as gallic acid, nitidine chloride, ferulic acid simultaneously.

Above-mentioned Gongyanping preparation contains tannin or gallic acid (C ₇H ₆O ₅) be no less than 0.01mg; Ferulic acid (C ₁₀H ₁₀O ₄) be no less than 0.01mg, contain the chlorination nitidine and be no less than 0.01mg.Gallic acid detects with method disclosed by the invention, and ferulic acid, nitidine chloride are used the inventor: 200510035226.6 application disclosed methods or other known methods detect.

Adopt the gallic acid that contains in the HPLC (high performance liquid chromatography) quantitative assay said preparation; Can adopt simultaneously thin layer chromatography differentiate in the preparation gallic acid or from one or more of the contained composition of Radix Cudraniae or Herba Melastomatis dodecandri or Radix Zanthoxyli or Herba Fici Simplicissimae.

Wherein the content assaying method of gallic acid is in the Gongyanping preparation:

Detection wavelength when high-efficient liquid phase technique is measured gallic acid content is: 220nm to 280nm; , or the optimum detection wavelength is 272 ± 2nm; Or the optimum detection wavelength is 272nm;

The chromatographic column condition is:

1. chromatographic column is filler with the octadecylsilane chemically bonded silica,

2. mobile phase is: the acetonitrile of different proportion and the mixed liquor of phosphoric acid, or the acetonitrile of different proportion, triethylamine, phosphoric acid; Or condition optimization is:

1. chromatographic column: octadecylsilane chemically bonded silica is a filler;

2. mobile phase: the methanol of different proportion combination, the mixed liquor of water; Or the mixed liquor of acetonitrile, triethylamine, phosphoric acid, water; Or the mixed liquor of acetonitrile, phosphoric acid, water, or optimum selection is the mixed liquor (3: 97) of acetonitrile-0.05% phosphoric acid;

When measuring gallic acid content, make test sample by following steps:

Get Gongyanping preparation, porphyrize, the accurate title, decide, and adds 2%～10% hydrochloric acid solution, puts in the flask, claim decide weight, put in the boiling water bath back hydrolysis 1 to 6 hour, put coldly, weight decided in title again, supply the weight that subtracts mistake with 2%～10% hydrochloric acid solution, shake up, microporous filter membrane filters, promptly.

Or get Gongyanping preparation, and porphyrize accurate claims surely, adds 2%～10% hydrochloric acid solution, puts in the volumetric flask, puts into the boiling water bath back hydrolysis 1 to 6 hour, puts coldly, adds 2%～10% hydrochloric acid solution to scale, shakes up, microporous filter membrane filters, promptly.

Or preferably by following reagent dosage operation:

Get and be equivalent to crude drug 10g Gongyanping preparation, should remove coating if any coating, solid preparation is answered porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, and promptly gets need testing solution.

The high-efficient liquid phase technique of said determination gallic acid can integral body be preferably:

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.05% phosphoric acid (3: 97) is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 272nm; Column temperature: room temperature; Theoretical cam curve should be not less than 5000 by the gallic acid peak;

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding methanol, to make concentration be the solution that every 1ml contains 40 μ g, promptly;

10 of this product are got in the preparation of need testing solution, remove coating, porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, promptly;

Accurate respectively reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

This product is equivalent to contain the gallic acid (C of tannin with hydrolysis in the Gongyanping preparation of 1g crude drug ₇H ₆O ₅) meter, must not be less than 0.01mg.

The present invention can differentiate the thin layer discriminating combination in any use of Radix Cudraniae or Herba Melastomatis dodecandri or Radix Zanthoxyli or Herba Fici Simplicissimae medical material or gallic acid reference substance with qualitative thin layer about the assay of Gongyanping preparation.

The thin layer discrimination method of Radix Cudraniae is in the Gongyanping preparation:

Get Gongyanping preparation, porphyrize adds dissolve with methanol or Different concentrations of alcohol and extracts, extracting solution filters, the filtrate evaporate to dryness, residue is dissolved in water, and adds acid, heating hydrolysis, cooling is used the organic solvent not miscible with water (ether or ethyl acetate or chloroform or butanols etc.) extraction, the extract evaporate to dryness, residue adds chloroform makes dissolving, as need testing solution; Other gets the Radix Cudraniae control medicinal material, shines medical material solution in pairs with legal system; According to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, draw need testing solution, control medicinal material solution, put respectively on same silica gel g thin-layer plate, with petroleum ether-Ethyl formate-formic acid is developing solvent, launches, and takes out, dry, under ultra-violet lamp, inspect; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the fluorescence speckle of same color is arranged;

Or be preferably: get Gongyanping preparation an amount of (being equivalent to crude drug 8g approximately), should remove coating if any coating, solid preparation is answered porphyrize, add methanol 20ml, ultrasonic 30 minutes, filter the filtrate evaporate to dryness, residue adds water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, cooling immediately, with extracted with diethyl ether 2 times, each 20ml merges the ether solution evaporate to dryness, and residue adds chloroform 1ml makes dissolving, as need testing solution, other gets Radix Cudraniae control medicinal material 1g, shines medical material solution in pairs with legal system, according to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, draw need testing solution, each 5 μ l of control medicinal material solution, putting respectively on same silica gel g thin-layer plate, is developing solvent with petroleum ether (30-60 ℃)-Ethyl formate-formic acid (4: 1: 0.1), launches, take out, dry, under ultra-violet lamp (365nm wavelength), inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, the fluorescence speckle of same color is arranged.

The thin layer discrimination method of Herba Melastomatis dodecandri, gallic acid is in the Gongyanping preparation:

Get Gongyanping preparation, add methanol or Different concentrations of alcohol and extract, extracting solution filters, the filtrate evaporate to dryness, residue is dissolved in water, add acid, reflux certain hour, cooling, the organic solvent extraction not miscible of cooled aqueous solution with water, organic solvent extract evaporate to dryness, residue adds dissolve with methanol, as need testing solution; Get the Herba Melastomatis dodecandri control medicinal material, shine medical material solution in pairs with legal system; According to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, draw above-mentioned need testing solution, control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid is developing solvent, launches, and takes out, dry, spray is with the ferric chloride alcoholic solution; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the speckle of same color is arranged;

Or: get Gongyanping preparation, adding methanol or Different concentrations of alcohol extracts, extracting solution filters, and residue is dissolved in water, and adds hydrochloric acid, the reflux certain hour, cooling, the cooled aqueous solution organic solvent extraction not miscible with water, organic solvent extract evaporate to dryness, residue adds dissolve with methanol, as need testing solution; Get the gallic acid reference substance, add methanol and make reference substance solution; According to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, draw above-mentioned need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid is developing solvent, launches, and takes out, dry, spray is with the ferric chloride alcoholic solution; In the test sample chromatograph, with reference substance chromatograph relevant position on, the speckle of same color is arranged;

Or be preferably: get Gongyanping preparation an amount of (being equivalent to crude drug 10g approximately), should remove coating if any coating, solid preparation is answered porphyrize, add methanol 30ml, ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, cooling immediately, ethyl acetate extraction 2 times of cooled aqueous solution, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Get Herba Melastomatis dodecandri control medicinal material 1g, shine medical material solution in pairs with legal system; Other gets the gallic acid reference substance, add methanol and make the solution that every 1ml contains 1mg, product solution according to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, is drawn need testing solution, control medicinal material solution each 10 μ l and reference substance solution 5 μ l in contrast, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid (6: 3: 1) is developing solvent, launches, and takes out, dry, spray is with 1% ferric chloride alcoholic solution; In the test sample chromatograph, with reference substance and control medicinal material chromatograph relevant position on, the speckle of same color is arranged.

The thin layer discrimination method of Radix Zanthoxyli is in the Gongyanping preparation:

It is an amount of to get Gongyanping preparation, adds Different concentrations of alcohol or methanol extraction and filters, and filtrate evaporate to dryness, residue add ethanol makes dissolving, as need testing solution; Other gets the Radix Zanthoxyli control medicinal material, shines medical material solution in pairs with legal system; According to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, draw need testing solution, control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate is developing solvent, and ammonia is saturated, launches, take out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the fluorescence speckle of same color is arranged;

Or be preferably: get Gongyanping preparation an amount of (being equivalent to crude drug 6g approximately), should remove coating if any coating, solid preparation is answered porphyrize, adds ethanol 15ml, warm macerating (40-50 ℃) 30 minutes ultrasonic 30 minutes, filters, filtrate evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets Radix Zanthoxyli control medicinal material 1g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia), draw need testing solution, each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (4: 1) is developing solvent, saturated 15 minutes of ammonia launches, and takes out, dry, put under the ultra-violet lamp (365nm wavelength) and inspect; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the fluorescence speckle of same color is arranged.

Or adopt following method, also can differentiate the Radix Zanthoxyli medical material:

It is an amount of to get Gongyanping preparation, add alkaline solution, and adding Different concentrations of alcohol or methanol extraction, extracting solution filters, the filtrate evaporate to dryness, residue adds water makes dissolving, with water saturated n-butyl alcohol or other and the immiscible organic solvent extraction of water, merge n-butyl alcohol or other organic solvent liquid, evaporate to dryness, residue adds methanol or other organic solvent makes dissolving, as need testing solution; Other gets the Radix Zanthoxyli control medicinal material, shines medical material solution in pairs with legal system, according to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, draw need testing solution and control medicinal material solution, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-acetone-formic acid, launches, take out, dry, put under the ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, the fluorescence speckle of same color is arranged;

Or be preferably: get Gongyanping preparation an amount of (being equivalent to crude drug 6g approximately), should remove coating if any coating, solid preparation is answered porphyrize, put in the conical flask, add an amount of moistening of ammonia after, add methanol 20ml supersound process 30 minutes, put coldly, filter the filtrate evaporate to dryness, residue adds water 15ml makes dissolving, puts in the separatory funnel, with twice of water saturated n-butanol extraction, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 1mL makes dissolving, as need testing solution; Other gets Radix Zanthoxyli control medicinal material 0.2g, shine medical material solution in pairs with legal system, according to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, draw need testing solution 10 μ l, control medicinal material solution 5ul, put respectively in same be on the silica gel g thin-layer plate of binding agent with 0.5% sodium carboxymethyl cellulose, with chloroform-acetone-formic acid (15: 4: 1) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (254nm wavelength) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, the fluorescence speckle of same color is arranged.

The thin layer discrimination method of Herba Fici Simplicissimae is in the Gongyanping preparation:

It is an amount of to get Gongyanping preparation, adds Different concentrations of alcohol or methanol extraction, and extracting solution filters, the filtrate evaporate to dryness, and residue is dissolved in water, and uses the organic solvent extraction not miscible with water, organic solvent extract evaporate to dryness, residue adds chloroform makes dissolving, as need testing solution; Get the Herba Fici Simplicissimae control medicinal material, shine medical material solution in pairs with legal system; According to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia) test, draw need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate-formic acid is developing solvent, launches, and takes out, dry, spray is heated to colour developing with ethanol solution of sulfuric acid; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the speckle of same color is arranged;

Or be preferably: get Gongyanping preparation an amount of (being equivalent to crude drug 4g approximately), should remove coating if any coating, solid preparation is answered porphyrize, adds methanol 20ml ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 15ml dissolving, use ethyl acetate extraction 2 times, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution; Get Herba Fici Simplicissimae control medicinal material 1g, shine medical material solution in pairs with legal system; Test according to thin layer chromatography (appendix thin layer chromatography of Chinese Pharmacopoeia), draw need testing solution, each 8 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate-formic acid (4: 1: 0.1) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and hot blast blows to colour developing; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the speckle of same color is arranged.

The assay method of content of tannin in the Gongyanping preparation may further comprise the steps:

1. the preparation of reference substance solution: it is an amount of that precision takes by weighing the gallic acid reference substance, is dissolved in water, and makes the solution of certain content, promptly;

Or be preferably: precision takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, puts in the brown volumetric flask, is dissolved in water, and makes the solution that every 1ml contains 105 μ g, promptly;

2. the preparation of need testing solution: the weighting profit requires 1 described gynecological Chinese medicine preparation an amount of, is dissolved in water, extracts, and filters, and discards filtrate just, and it is an amount of that precision is measured subsequent filtrate, behind the dilute with water, shakes up, promptly;

Or be preferably: the weighting profit requires 1 described gynecological Chinese medicine preparation an amount of, is equivalent to crude drug 10g approximately, should remove coating, porphyrize if any coating, precision takes by weighing in right amount (being equivalent to crude drug 2g approximately), puts in the brown volumetric flask of 50ml, adds water 50ml, placement is spent the night, and supersound process 10min is put cold, shake up, leave standstill and make precipitation of solid material, filter, discard filtrate just, precision is measured subsequent filtrate 20ml, puts in the brown volumetric flask of 50ml, be diluted with water to scale, shake up, promptly;

3. algoscopy: measure according to an appendix content of tannin of Chinese Pharmacopoeia assay method, calculate: the described gynecological Chinese medicine preparation of claim 1 per unit preparation (being equivalent to crude drug 1g) contains tannin more than 0.01mg.

Effect of the present invention:

Though GONGYANPING PIAN is recorded in " Chinese traditional patent formulation preparation " the 17, the description and the assay of relevant said preparation effective ingredient are rarely put down in writing.

The present invention is by research, confirmation is made GONGYANPING oral formulations or external preparation or injection by specific extracting method with Herba Melastomatis dodecandri, Radix Zanthoxyli, Radix Angelicae Sinensis, Radix Cudraniae, Herba Fici Simplicissimae, as tablet, capsule, drop pill, suppository, injection etc., be equivalent to contain tannin or gallic acid (C in the per unit preparation of crude drug 1g ₇H ₆O ₅) be no less than 0.01mg; Ferulic acid is no less than 0.01mg, contain the chlorination nitidine is no less than 0.01mg.

The contained effective ingredient of this Gongyanping preparation is clear relatively, and quality controllability is good.

Modern pharmacological research shows, the flat preparation of cervicitis product has multiple effects such as antiinflammatory, analgesia, spasmolytic, has close dependency with contained gallic acid, psoralen etc. in the preparation, by gallic acid quantitatively being reached the foundation of other qualitative indexes, the inherent quality that can reflect preparation, the correct capacity of control raw medicinal material is used, and helps preventing the appearance of counterfeit drug, substandard drug.

Capsules of Gongyanping for cervicitis causes difficulty to Radix Zanthoxyli is differentiated because greasing base is extracting in the separation, and method of the present invention is stablized feasible, has overcome the interference of substrate.

Through the methodology checking, content control method specificity disclosed by the invention is strong, and sensitivity, precision, repeatability all satisfy the requirement of drug quality control.

The present invention has set up Gongyanping preparation gallic acid content assay method, effectively in comprehensive control GONGYANPING quality.

Specific embodiment:

Embodiment 1: a kind of gallic acid, nitidine chloride, ferulic acid GONGYANPING Film coated tablets preparation of containing simultaneously

Herba Melastomatis dodecandri 450g Radix Zanthoxyli 170g Radix Angelicae Sinensis 140g

Herba Fici Simplicissimae 100g Radix Cudraniae 140g

The above five tastes decoct with water secondary, each 2 hours, filter, merging filtrate, being concentrated into relative density is 1.25 (55～60 ℃), add ethanol and reach 50%, left standstill 24 hours, filter to containing the alcohol amount, filtrate recycling ethanol is concentrated into the thick paste shape, and cold drying is ground into fine powder, add appropriate amount of auxiliary materials, mixing is granulated, be pressed into 1000, coating, promptly.

Said preparation is after measured: contain gallic acid＞0.01mg, nitidine chloride＞0.01mg, ferulic acid＞0.01mg.

The detection method of gallic acid is:

Measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005);

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.05% phosphoric acid (3: 97) is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 272nm; Column temperature: room temperature; Theoretical cam curve should be not less than 5000 by the gallic acid peak;

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding methanol, to make concentration be the solution that every 1ml contains 40 μ g, promptly;

10 of this product are got in the preparation of need testing solution, remove coating, porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, promptly;

Accurate respectively reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly;

Every of this product contains Herba Melastomatis dodecandri with gallic acid (C ₇H ₆O ₅) meter, be no less than 0.01mg.

Embodiment 2: a kind of gallic acid, nitidine chloride, ferulic acid GONGYANPING Film coated tablets preparation of containing simultaneously

Herba Melastomatis dodecandri 450g Radix Zanthoxyli 170g Radix Angelicae Sinensis 140g

Herba Fici Simplicissimae 100g Radix Cudraniae 140g

Herba Melastomatis dodecandri, Radix Angelicae Sinensis be with 75% alcohol reflux 2.0 hours, and extracting solution reclaims behind the ethanol standby, and medicinal residues and its excess-three after Herba Melastomatis dodecandri refluxes with Radix Angelicae Sinensis are distinguished the flavor of, decoct with water secondary, each 2 hours, filter merging filtrate, being concentrated into relative density is 1.25 (55～60 ℃), add ethanol and reach 50%, left standstill 24 hours, filter to containing the alcohol amount, filtrate recycling ethanol, be concentrated into the thick paste shape, merge with above-mentioned ethanol extraction, cold drying is ground into fine powder, add appropriate amount of auxiliary materials, mixing is granulated, and is pressed into 1000, coating, promptly.

Said preparation is after measured: contain gallic acid＞0.01mg, nitidine chloride＞0.01mg, ferulic acid＞0.01mg.

Embodiment 3: a kind of gallic acid, nitidine chloride, ferulic acid GONGYANPING Film coated tablets preparation of containing simultaneously

Herba Melastomatis dodecandri 450g Radix Zanthoxyli 170g Radix Angelicae Sinensis 140g

Herba Fici Simplicissimae 100g Radix Cudraniae 140g

Herba Melastomatis dodecandri, Radix Angelicae Sinensis, Radix Zanthoxyli be with 65% alcohol reflux 2.0 hours, and extracting solution reclaims behind the ethanol standby, and the medicinal residues after Herba Melastomatis dodecandri and Radix Angelicae Sinensis, Radix Zanthoxyli are refluxed and all the other two are distinguished the flavor of, decoct with water secondary, each 2 hours, filter merging filtrate, being concentrated into relative density is 1.25 (55～60 ℃), add ethanol and reach 50%, left standstill 24 hours, filter to containing the alcohol amount, filtrate recycling ethanol, be concentrated into the thick paste shape, merge with above-mentioned ethanol extraction, cold drying is ground into fine powder, add appropriate amount of auxiliary materials, mixing is granulated, and is pressed into 1000, coating, promptly.

Said preparation is after measured: contain gallic acid＞0.01mg, nitidine chloride＞0.01mg, ferulic acid＞0.01mg.

Embodiment 4: a kind of gallic acid, nitidine chloride, ferulic acid GONGYANPING Film coated tablets preparation of containing simultaneously

Herba Melastomatis dodecandri 450g Radix Zanthoxyli 170g Radix Angelicae Sinensis 140g

Herba Fici Simplicissimae 100g Radix Cudraniae 140g

Herba Melastomatis dodecandri, Radix Angelicae Sinensis, Radix Zanthoxyli be with 95% alcohol reflux 2.0 hours, and extracting solution reclaims behind the ethanol standby, and the medicinal residues after Herba Melastomatis dodecandri and Radix Angelicae Sinensis, Radix Zanthoxyli are refluxed and all the other two are distinguished the flavor of, decoct with water secondary, each 2 hours, filter merging filtrate, being concentrated into relative density is 1.25 (55～60 ℃), add ethanol and reach 50%, left standstill 24 hours, filter to containing the alcohol amount, filtrate recycling ethanol, be concentrated into the thick paste shape, merge with above-mentioned ethanol extraction, cold drying is ground into fine powder, add appropriate amount of auxiliary materials, mixing is granulated, and is pressed into 1000, coating, promptly.

Said preparation is after measured: contain gallic acid＞0.01mg, nitidine chloride＞0.01mg, ferulic acid＞0.01mg.

Embodiment 5: get commercially available GONGYANPING Film coated tablets assay.

Measure according to high performance liquid chromatography (appendix of Chinese Pharmacopoeia version in 2005).

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.05% phosphoric acid (3: 97) is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 272nm; Column temperature: room temperature.Theoretical cam curve should be not less than 5000 by the gallic acid peak.

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding methanol, to make concentration be the solution that every 1ml contains 40 μ g, promptly.

10 of this product are got in the preparation of need testing solution, remove coating, porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, promptly.

Accurate respectively reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

Every of this product contains Herba Melastomatis dodecandri with gallic acid (C ₇H ₆O ₅) meter, be 0.1mg.

Embodiment 6: get commercially available GONGYANPING PIAN, (" appendix of Chinese pharmacopoeia year version is measured according to high performance liquid chromatography.

Measure according to high performance liquid chromatography (appendix of Chinese Pharmacopoeia version in 2005).

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.05% phosphoric acid (3: 97) is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 272nm; Column temperature: room temperature.Theoretical cam curve should be not less than 5000 by the gallic acid peak.

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding methanol, to make concentration be the solution that every 1ml contains 40 μ g, promptly.

10 of this product are got in the preparation of need testing solution, remove coating, porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, promptly.

Accurate respectively reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

The GONGYANPING dispersible tablet that is equivalent to crude drug in whole 1g contains gallic acid (C ₇H ₆O ₅) meter, be 0.01mg.

Embodiment 7: get commercially available GONGYANPING dispersible tablet gallic acid content and measure.

Measure according to high performance liquid chromatography (appendix of Chinese Pharmacopoeia version in 2005).

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.05% phosphoric acid (5: 95) is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 220nm; Column temperature: room temperature.Theoretical cam curve should be not less than 5000 by the gallic acid peak.

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding 50% methanol, to make concentration be the solution that every 1ml contains 60 μ g, promptly.

10 of this product are got in the preparation of need testing solution, remove coating, porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, promptly.

Accurate respectively reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

The GONGYANPING dispersible tablet that is equivalent to crude drug in whole 1g contains gallic acid (C ₇H ₆O ₅) meter, be 0.05mg.

Embodiment 8: the linear dependence of getting gallic acid content in the commercially available capsules of Gongyanping for cervicitis is measured:

Measure according to high performance liquid chromatography (appendix of Chinese Pharmacopoeia version in 2005).

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.05% phosphoric acid (3: 97) is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 272nm; Column temperature: room temperature.Theoretical cam curve should be not less than 5000 by the gallic acid peak.

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding methanol, to make concentration be the solution that every 1ml contains 80 μ g, promptly.

Accurate reference substance solution 1,2,4,6, the 8 μ l that draw, injecting chromatograph, detect by above-mentioned chromatographic condition, with the reference substance sample size is abscissa, peak area is a vertical coordinate, the drawing standard curve, and calculating recurrence (adding initial point) equation is Y=3524.7X-4.1189, correlation coefficient r=0.9999, gallic acid is good in 0.08-0.64 μ g scope internal linear relation.

Embodiment 9: get commercially available GONGYANPING drop pill assay.

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.05% phosphoric acid (5: 95) is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 220nm; Column temperature: room temperature.Theoretical cam curve should be not less than 5000 by the gallic acid peak.

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding methanol, to make concentration be the solution that every 1ml contains 60 μ g, promptly.

30 of GONGYANPING drop pill are got in the preparation of need testing solution, porphyrize, and precision takes by weighing 0.5g, add 5% hydrochloric acid solution 50ml, put in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with 5% hydrochloric acid solution, microporous filter membrane filters, promptly.

Accurate respectively reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

The GONGYANPING drop pill that is equivalent to crude drug in whole 1g contains gallic acid (C ₇H ₆O ₅) meter, be 0.05mg.

The thin layer of Radix Cudraniae is differentiated in embodiment 10, the GONGYANPING PIAN:

Get 8 of GONGYANPING PIAN, remove coating, porphyrize, add methanol 20ml, ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, extracted with diethyl ether 2 times are used in cooling immediately, each 20ml, merge the ether solution evaporate to dryness, residue adds chloroform 1ml makes dissolving, as need testing solution.Other gets Radix Cudraniae control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-Ethyl formate-formic acid (4: 1: 0.1) is developing solvent, launch, take out, dry, under the 365nm wavelength, inspect.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the fluorescence speckle of same color is arranged.

The thin layer of Radix Cudraniae is differentiated in embodiment 11, the capsules of Gongyanping for cervicitis:

Get 4 of capsules of Gongyanping for cervicitis, get content, porphyrize, add methanol 20ml, ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, extracted with diethyl ether 2 times are used in cooling immediately, each 20ml, merge the ether solution evaporate to dryness, residue adds chloroform 1ml makes dissolving, as need testing solution.Other gets Radix Cudraniae control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-Ethyl formate-formic acid (4: 1: 0.1) is developing solvent, launch, take out, dry, under the 365nm wavelength, inspect.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the fluorescence speckle of same color is arranged.

The thin layer of embodiment 12, GONGYANPING PIAN gallic acid and Herba Melastomatis dodecandri is differentiated:

Get 10 of GONGYANPING PIAN, remove coating, porphyrize, add methanol 30ml, ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, cooling immediately, ethyl acetate extraction 2 times of cooled aqueous solution, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Get Herba Melastomatis dodecandri control medicinal material 1g, shine medical material solution in pairs with legal system.Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, control medicinal material solution each 10 μ l and reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid (6: 3: 1) is developing solvent, launch, take out, dry, spray is with 1%Fecl ₃Alcoholic solution.In the test sample chromatograph, with reference substance and control medicinal material chromatograph relevant position on, the speckle of same color is arranged.

The thin layer of gallic acid and Herba Melastomatis dodecandri is differentiated in embodiment 13, the GONGYANPING drop pill:

Get 30 of GONGYANPING drop pill, porphyrize adds methanol 30ml, ultrasonic 30 minutes, filter the filtrate evaporate to dryness, residue adds water 10ml dissolving, adds hydrochloric acid 2ml, reflux 30 minutes, cooling immediately, ethyl acetate extraction 2 times of cooled aqueous solution, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution.Get Herba Melastomatis dodecandri control medicinal material 1g, shine medical material solution in pairs with legal system.Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, control medicinal material solution each 10 μ l and reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid (6: 3: 1) is developing solvent, launch, take out, dry, spray is with 1%Fecl ₃Alcoholic solution.In the test sample chromatograph, with reference substance and control medicinal material chromatograph relevant position on, the speckle of same color is arranged.

Embodiment 14: the thin layer of Radix Zanthoxyli is differentiated in the GONGYANPING PIAN:

Get 6 of GONGYANPING PIAN, remove coating, porphyrize adds ethanol 15ml, warm macerating (40-50 ℃) 30 minutes, and ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Other gets Radix Zanthoxyli control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (4: 1) is developing solvent, saturated 15 minutes of ammonia launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the fluorescence speckle of same color is arranged.

Embodiment 15: the thin layer of Radix Zanthoxyli is differentiated in the capsules of Gongyanping for cervicitis:

Get the content of 3 of capsules of Gongyanping for cervicitis, put in the conical flask, after adding 2ml ammonia and soaking into, add methanol 20ml supersound process 30 minutes, put coldly, filter, filtrate evaporate to dryness, residue add water 20ml makes dissolving, places separatory funnel, with water saturated n-butanol extraction twice, each 15ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 1ml makes dissolving as test sample. and Radix Zanthoxyli control medicinal material 1.0g in addition, shine medical material solution, product in contrast in pairs with legal system; Draw control medicinal material solution 5 μ l, each 10 μ l of need testing solution, putting respectively on same 0.5%CMC-Na silica gel g thin-layer plate, is developing solvent with chloroform-acetone-formic acid (15: 4: 1), launches, take out, dry, place under the uviol lamp (365nm), in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, the fluorescence speckle of same color is arranged.

Embodiment 16: the thin layer of the Herba Fici Simplicissimae in the GONGYANPING PIAN is differentiated:

Get 4 of GONGYANPING PIAN, remove coating, porphyrize added methanol 20ml ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 15ml dissolving, use ethyl acetate extraction 2 times, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution.Get Herba Fici Simplicissimae control medicinal material 1g, shine medical material solution in pairs with legal system.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw need testing solution, each 8 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate-formic acid (4: 1: 0.1) is developing solvent, launches, and takes out, dry, spray is with 10%H ₂SO ₄Alcoholic solution, hot blast blows to colour developing.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the speckle of same color is arranged.

Embodiment 17: the thin layer of the Herba Fici Simplicissimae in the GONGYANPING dispersible tablet is differentiated:

Get 4 of GONGYANPING dispersible tablets, remove coating, porphyrize added methanol 20ml ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 15ml dissolving, use ethyl acetate extraction 2 times, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution.Get Herba Fici Simplicissimae control medicinal material 1g, shine medical material solution in pairs with legal system.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw need testing solution, each 8 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate-formic acid (4: 1: 0.1) is developing solvent, launches, and takes out, dry, spray is with 10%H ₂SO ₄Alcoholic solution, hot blast blows to colour developing.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, the speckle of same color is arranged.

Embodiment 18: the thin layer of psoralen is differentiated in the GONGYANPING PIAN:

Get said preparation 5g, porphyrize adds methanol 50ml merceration 2 hours, filters evaporate to dryness, and residue adds methanol 2ml makes dissolving. as need testing solution; Get psoralen and make the 0.4mg/ml reference substance solution;

Draw each 5 μ l of Herba Fici Simplicissimae control medicinal material solution and need testing solution, psoralen reference substance solution 2 μ l, point is on same silica gel G (containing 0.7%CMC-Na) lamellae, launch with normal hexane-ethyl acetate (7: 3), exhibition is taken out apart from 10cm, dry, observe down at uviol lamp (365nm), in the need testing solution chromatograph, with psoralen reference substance and control medicinal material chromatograph relevant position on each shows faint yellow fluorescence speckle; With 10% potassium hydroxide ethanol solution spraying colour developing, visible corresponding yellow is entangled the hot spot point on former corresponding faint yellow fluorescence speckle displacement again.

Embodiment 19: the mensuration of GONGYANPING PIAN content of tannin:

1. the preparation of reference substance solution: precision takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, puts in the brown volumetric flask, is dissolved in water, and makes the solution that every 1ml contains 105 μ g, promptly.

2. the preparation of need testing solution: the weighting profit requires 1 described gynecological Chinese medicine preparation an amount of, is equivalent to crude drug 10g approximately, should remove coating, porphyrize if any coating, precision takes by weighing 0.5g, puts in the brown volumetric flask of 50ml, adds water 50ml, placement is spent the night, and supersound process 10 minutes is put cold, shake up, leave standstill and make precipitation of solid material, filter, discard filtrate just, precision is measured subsequent filtrate 20ml, puts in the brown volumetric flask of 50ml, be diluted with water to scale, shake up, promptly.

3. algoscopy: measure according to an appendix content of tannin of Chinese Pharmacopoeia assay method, calculate, GONGYANPING per unit preparation (being equivalent to crude drug 1 gram) contains tannin 0.05mg.

Embodiment 20: the mensuration of capsules of Gongyanping for cervicitis content of tannin:

1. the preparation of reference substance solution: precision takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, puts in the brown volumetric flask, is dissolved in water, and makes the solution that every 1ml contains 85 μ g, promptly.

2. the preparation of need testing solution: get 5 of capsules of Gongyanping for cervicitis, content, porphyrize, precision takes by weighing 0.5g, puts in the brown volumetric flask of 50ml, adds water 50ml, placement is spent the night, and supersound process 10min is put cold, shake up, leave standstill and make precipitation of solid material, filter, discard filtrate just, precision is measured subsequent filtrate 20ml, puts in the brown volumetric flask of 50ml, be diluted with water to scale, shake up, promptly.

3. algoscopy: measure according to an appendix content of tannin of Chinese Pharmacopoeia assay method, calculate, the capsules of Gongyanping for cervicitis that must be equivalent to crude drug in whole 1g contains tannin 0.01mg.

Embodiment 21: the mensuration of GONGYANPING granule content:

1. the preparation of reference substance solution: precision takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, puts in the brown volumetric flask, is dissolved in water, and makes the solution that every 1ml contains 105 μ g, promptly.

2. the preparation of need testing solution: get 3 bags of GONGYANPING granules, be equivalent to crude drug 8g, be equivalent to crude drug 12g approximately, porphyrize, precision takes by weighing 0.5g, puts in the brown volumetric flask of 50ml, adds water 50ml, placement is spent the night, and supersound process 10min is put cold, shake up, leave standstill and make precipitation of solid material, filter, discard filtrate just, precision is measured subsequent filtrate 20ml, puts in the brown volumetric flask of 50ml, be diluted with water to scale, shake up, promptly.

3. algoscopy: measure according to an appendix content of tannin of Chinese Pharmacopoeia assay method, calculate, GONGYANPING per unit preparation (being equivalent to crude drug 1 gram) contains tannin 0.01mg.

Embodiment 22: the method for quality control of a kind of GONGYANPING sugar-coat or Film coated tablets

Herba Melastomatis dodecandri 450g Radix Zanthoxyli 170g Radix Angelicae Sinensis 140g

Herba Fici Simplicissimae 100g Radix Cudraniae 140g

The above five tastes decoct with water secondary, each 2 hours, filter, merging filtrate, being concentrated into relative density is 1.25 (55～60 ℃), add ethanol and reach 50%, left standstill 24 hours, filter to containing the alcohol amount, filtrate recycling ethanol is concentrated into the thick paste shape, and cold drying is ground into fine powder, add appropriate amount of auxiliary materials, mixing is granulated, be pressed into 1000, coating, promptly.

This product is sugar-coat or Film coated tablets, removes to show shallow sepia behind the coating to brownish black; Feeble QI, bitter in the mouth, acid, little puckery.

Differentiate:

(1) gets 8 of this product, remove coating, porphyrize, add methanol 20ml, ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, extracted with diethyl ether 2 times are used in cooling immediately, each 20ml, merge the ether solution evaporate to dryness, residue adds chloroform 1ml makes dissolving, as need testing solution.Other gets Radix Cudraniae control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-Ethyl formate-formic acid (4: 1: 0.1) is developing solvent, launch, take out, dry, under the 365nm wavelength, inspect.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color.

(2) get 10 of this product, remove coating, porphyrize, add methanol 30ml, ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, cooling immediately, ethyl acetate extraction 2 times of cooled aqueous solution, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Get Herba Melastomatis dodecandri control medicinal material 1g, shine medical material solution in pairs with legal system.Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, control medicinal material solution each 10 μ l and reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid (6: 3: 1) is developing solvent, launch, take out, dry, spray is with 1%Fecl ₃Alcoholic solution.In the test sample chromatograph, with reference substance and control medicinal material chromatograph relevant position on, show the speckle of same color.

(3) get 6 of this product, remove coating, porphyrize adds ethanol 15ml, warm macerating (40-50 ℃) 30 minutes, and ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Other gets Radix Zanthoxyli control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (4: 1) is developing solvent, saturated 15 minutes of ammonia launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color.

(4) get 4 of this product, remove coating, porphyrize added methanol 20ml ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 15ml dissolving, use ethyl acetate extraction 2 times, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution.Get Herba Fici Simplicissimae control medicinal material 1g, shine medical material solution in pairs with legal system.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw need testing solution, each 8 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate-formic acid (4: 1: 0.1) is developing solvent, launches, and takes out, dry, spray is with 10%H ₂SO ₄Alcoholic solution, hot blast blows to colour developing.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

Assay:

Measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005).

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.05% phosphoric acid (3: 97) is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 272nm; Column temperature: room temperature.Theoretical cam curve should be not less than 5000 by the gallic acid peak.

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding methanol, to make concentration be the solution that every 1ml contains 40 μ g, promptly.

10 of this product are got in the preparation of need testing solution, remove coating, porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, promptly.

Accurate respectively reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

Every of this product contains Herba Melastomatis dodecandri with gallic acid (C ₇H ₆O ₅) meter, be 0.05mg.

Claims (5)

1. the method for quality control of a Gongyanping preparation: the content assaying method that it is characterized in that this method is:

Adopt Determination of Gallic content:

1. adopt the detection wavelength of Determination of Gallic content to be: 220nm to 280nm,

2. adopt the filler of the chromatographic column of Determination of Gallic content to be: octadecylsilane chemically bonded silica;

3. adopt the mobile phase of Determination of Gallic content to be: the acetonitrile of different proportion combination, the mixed liquor of phosphoric acid.

2. according to the content control method of right 1 described Gongyanping preparation, it is characterized in that the content assaying method of this method is:

Adopt Determination of Gallic content:

1. adopt the detection wavelength of Determination of Gallic content to be: 272nm;

2. adopt the filler of the chromatographic column of Determination of Gallic content to be: octadecylsilane chemically bonded silica;

3. adopt the mobile phase of Determination of Gallic content to be: 3 parts of acetonitriles, 97 parts of 0.05% phosphoric acid solutions.

3. according to the content control method of right 1 described Gongyanping preparation, when adopting Determination of Gallic content, it is characterized in that getting Gongyanping preparation and make need testing solution by following steps:

Get Gongyanping preparation, porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claims decide weight, puts in the boiling water bath back hydrolysis 4 hours, puts coldly, claims to decide weight again, supplies the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, the microporous filter membrane filtration, promptly.

4. the method for quality control of Gongyanping preparation according to claim 1 when adopting Determination of Gallic content, is characterized in that:

According to high performance liquid chromatography, i.e. appendix high effective liquid chromatography for measuring of Chinese Pharmacopoeia:

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; Acetonitrile-0.05% phosphoric acid is mobile phase at 3: 97; Detect wavelength 272nm; Theoretical cam curve should be not less than 5000 by the gallic acid peak;

The preparation of reference substance solution: precision takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, adds methanol and makes concentration and be the solution that every 1ml contains 40 μ g, and is standby;

The preparation of need testing solution: it is an amount of to get the Gongyanping preparation that is equivalent to crude drug 10g, should remove coating if any coating, and solid preparation is answered porphyrize, precision takes by weighing in right amount, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, and is standby;

Algoscopy: respectively accurate reference substance solution, each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, calculate: it is C with the molecular formula that the Gongyanping preparation per unit preparation that is equivalent to crude drug 1g contains Herba Melastomatis dodecandri ₇H ₆O ₅The gallic acid meter, must not be less than 0.01mg.

5. the method for quality control of a Gongyanping preparation is characterized in that its discrimination method and content assaying method implement according to the following steps:

(1) get the Gongyanping preparation that is equivalent to crude drug 8g, should remove coating if any coating, solid preparation is answered porphyrize, add methanol 20ml, ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, extracted with diethyl ether 2 times are used in cooling immediately, each 20ml, merge the ether solution evaporate to dryness, residue adds chloroform 1ml makes dissolving, as need testing solution; Other gets Radix Cudraniae control medicinal material 1g, shines medical material solution in pairs with legal system; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw need testing solution, each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with volume ratio be: petroleum ether-Ethyl formate of 4: 1: 0.1-formic acid mixed liquor is developing solvent, launch, take out, dry, under the 365nm wavelength, inspect; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color;

(2) get the Gongyanping preparation that is equivalent to crude drug 10g, should remove coating if any coating, solid preparation is answered porphyrize, add methanol 30ml, ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 10ml dissolving, add hydrochloric acid 2ml, reflux 30 minutes, cooling immediately, ethyl acetate extraction 2 times of cooled aqueous solution, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Get Herba Melastomatis dodecandri control medicinal material 1g, shine medical material solution in pairs with legal system; Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw need testing solution, control medicinal material solution each 10 μ l and reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with volume ratio is that chloroform-ethyl acetate-formic acid of 6: 3: 1 is developing solvent, launch, take out, dry, spray is with 1%FeCl ₃Alcoholic solution; In the test sample chromatograph, with reference substance and control medicinal material chromatograph relevant position on, show the speckle of same color;

(3) get the Gongyanping preparation that is equivalent to crude drug 6g, should remove coating if any coating, solid preparation is answered porphyrize, adds ethanol 15ml, 40-50 ℃ of warm macerating 30 minutes, and ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets Radix Zanthoxyli control medicinal material 1g, shines medical material solution in pairs with legal system; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw need testing solution, each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that normal hexane-ethyl acetate of 4: 1 is developing solvent, saturated 15 minutes of ammonia launches, and takes out, dry, put under the ultra-violet lamp 365nm and inspect; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color; (4) get the Gongyanping preparation that is equivalent to crude drug 4g, should remove coating if any coating, solid preparation is answered porphyrize, adds methanol 20ml ultrasonic 30 minutes, filter, filtrate evaporate to dryness, residue add water 15ml dissolving, use ethyl acetate extraction 2 times, each 20ml, combined ethyl acetate liquid evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution; Get Herba Fici Simplicissimae control medicinal material 1g, shine medical material solution in pairs with legal system; According to thin layer chromatography test of Chinese Pharmacopoeia, draw need testing solution, each 8 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that normal hexane-ethyl acetate-formic acid of 4: 1: 0.1 is developing solvent, launches, and takes out, dry, spray is with 10%H ₂SO ₄Alcoholic solution, hot blast blow to colour developing; In the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color;

(5) assay:

According to appendix high effective liquid chromatography for measuring of Chinese Pharmacopoeia;

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; Volume ratio is that 3: 97 acetonitrile-0.05% phosphoric acid is mobile phase; Flow velocity 1.0ml/ minute; Detect wavelength 272nm; Column temperature: room temperature; Theoretical cam curve should be not less than 5000 by the gallic acid peak;

The preparation precision of reference substance solution takes by weighing that to be dried to the gallic acid reference substance of constant weight through phosphorus pentoxide an amount of, and adding methanol, to make concentration be the solution that every 1ml contains 40 μ g, promptly;

The Gongyanping preparation that is equivalent to crude drug 10g is got in the preparation of need testing solution, should remove coating if any coating, and solid preparation is answered porphyrize, precision takes by weighing 0.5g, adds 5% hydrochloric acid solution 50ml, puts in the flask, claim to decide weight, put in the boiling water bath back hydrolysis 4 hours, and put coldly, claim to decide weight again, supply the weight that subtracts mistake with 5% hydrochloric acid solution, shake up, microporous filter membrane filters, promptly;

Accurate respectively reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly;

The Gongyanping preparation per unit preparation that is equivalent to crude drug 1g contains Herba Melastomatis dodecandri in gallic acid, must not be less than 0.01mg.