CN1785268A

CN1785268A - Quality control method of sinew soothing and pain relieving preparation

Info

Publication number: CN1785268A
Application number: CNA2005100033129A
Authority: CN
Inventors: 叶湘武; 张梅
Original assignee: Guizhou Yibai Pharmaceutical Co Ltd
Current assignee: Guizhou Yibai Pharmaceutical Co Ltd
Priority date: 2005-12-09
Filing date: 2005-12-09
Publication date: 2006-06-14
Anticipated expiration: 2025-12-09
Also published as: CN100402059C

Abstract

A quality control method for the blood-nourishing Chinese medicine containing Chinese angelica root features that a higher quality standard is disclosed for higher curative effect.

Description

The method of quality control of sinew soothing and pain relieving preparation

Technical field: the present invention relates to a kind of method of quality control of sinew soothing and pain relieving preparation, belong to the field of medicine technology.

Technical background: traumatic injury, chronic lumbago and skelalgia, rheumatic arthralgia are the commonly encountered diseases in our daily life, sinew soothing and pain relieving preparation by Eupolyphaga Seu Steleophaga, Olibanum, Myrrha, Pyritum, Flos Carthami, Rhizoma Drynariae, Radix Et Rhizoma Rhei, Borax and Radix Angelicae Sinensis totally nine the flavor medicines form; Have promoting blood circulation to remove blood stasis, the effect of reducing swelling and alleviating pain, to traumatic injury, chronic lumbago and skelalgia, rheumatism numbness is painful etc., and disease has definite curative effect.Wherein " SHUJIN DINGTONG PIAN " publication is the 15 of Drug Standard of Ministry of Public Health of the Peoples Republic of China, this medicine is used for many years clinically, in traumatic injury, chronic lumbago and skelalgia, aspects such as rheumatic arthralgia obtain satisfied therapeutic effect, but discover that through us existing sinew soothing and pain relieving preparation exists method of quality control to fall behind the uppity shortcoming of product quality.Because Rhizoma Drynariae has the kidney invigorating bone strengthening, the continuous analgesic effect of hindering, in preparation of the present invention, play an important role, in addition, Flos Carthami, Radix Et Rhizoma Rhei and Radix Angelicae Sinensis also are the materials of performance main pharmacodynamics in the preparation of the present invention, and effective ingredient in the Rhizoma Drynariae are not carried out assay in the existing method of quality control or Rhizoma Drynariae is differentiated.Other drug is not differentiated yet,, so existing method of quality control can not effectively be controlled the quality of sinew soothing and pain relieving preparation, thereby will influence the production of product and ensure the quality of products

Summary of the invention: the objective of the invention is to: a kind of method of quality control of sinew soothing and pain relieving preparation is provided, and said preparation is an oral solid formulation, comprises capsule, tablet and granule.

Sinew soothing and pain relieving preparation of the present invention is to constitute like this: calculate according to composition by weight: it mainly is prepared from by Eupolyphaga Seu Steleophaga 50-200g, Olibanum (vinegar system) 30-100g, Myrrha (vinegar system) 30-100g, Pyritum (vinegar is forged) 30-100g, Flos Carthami 30-100g, Rhizoma Drynariae 30-100g, Radix Et Rhizoma Rhei 30-100g, Borax (forging) 60g, Radix Angelicae Sinensis 30-100g.

Preferred prescription is formed: calculate according to composition by weight: it mainly is prepared from by Eupolyphaga Seu Steleophaga 100g, Olibanum (vinegar system) 60g, Myrrha (vinegar system) 60g, Pyritum (vinegar is forged) 60g, Flos Carthami 60g, Rhizoma Drynariae 60g, Radix Et Rhizoma Rhei 60g, Borax (forging) 60g, Radix Angelicae Sinensis 60g.

Preparation of the present invention prepares like this: Radix Et Rhizoma Rhei, Borax, Pyritum, Olibanum, Myrrha are ground into the above fine powder of 80 orders, and be standby; Flos Carthami, Rhizoma Drynariae, Eupolyph aga sinesis Walker are broken into coarse powder, make solvent with 40-85% ethanol, carry out percolation according to the percolation under Chinese Pharmacopoeia fluid extract and the extractum item and extract, and collect percolate, and be standby; Radix Angelicae Sinensis powder is broken into coarse powder, makes solvent with 40-85% ethanol, carries out percolation according to the percolation under Chinese Pharmacopoeia fluid extract and the extractum item and extracts, and collects percolate, and is standby; Merge above percolate, reclaim ethanol, be condensed into cream, the ethanol condensed cream is added powder such as Radix Et Rhizoma Rhei, mixing again according to conventional method, adds different auxiliary material, makes capsule, tablet or granule.

Eupolyphaga Seu Steleophaga is principal agent in the preparation of the present invention, but Eupolyphaga Seu Steleophaga is an animal drugs, can't carry out assay to its ingredient.By analysis,, continuously hinder the analgesic effect, in preparation of the present invention, play an important role, and the Rhizoma Drynariae medical material extracts through percolation,, more can reflect product quality, guarantee curative effect as the contained naringin of Rhizoma Drynariae is carried out assay because Rhizoma Drynariae has the kidney invigorating bone strengthening.If how to select chromatographic condition and preparation need testing solution to become difficult point of the present invention in addition.Through a large amount of experiments, it is C18 or C4 or C8 post that the present invention selects chromatographic column, with methanol: acetic acid: water=20-50: 1-10: 40-80 is a mobile phase, the detection wavelength is 200-500nm, and number of theoretical plate must not calculate the content that is lower than naringin in 2000 the high effective liquid chromatography for measuring Rhizoma Drynariae with the naringin peak.As a result, extracts active ingredients is complete in the medicine, and under this condition, component to be measured is separated fully (separating degree＞1.5) with other components, and precision, stability etc. all can meet the requirements.In addition, the present invention also carries out the thin layer Study on Identification to Flos Carthami, Rhizoma Drynariae, Radix Et Rhizoma Rhei and four medical materials of Radix Angelicae Sinensis, and to select with the Flos Carthami control medicinal material be contrast, and with chloroform: ethyl acetate: methanol: water=5-20: 20-60: 10-30: 5-20 is a Flos Carthami in the developing solvent discriminating side in the solution of stratified lower floor below 10 ℃; With the naringin reference substance is contrast, and with ethyl acetate: the mixed solution that the upper solution 10-30ml of acetone: water=10-30: 5-20: 5-25 adds acetone 1-5ml and formic acid 0.05-3ml is a Rhizoma Drynariae in the developing solvent discriminating side; With the Radix Et Rhizoma Rhei control medicinal material is contrast, and with petroleum ether (30～60 ℃): the upper solution of Ethyl formate: formic acid=5-30: 1-10: 0.5-5 is a Radix Et Rhizoma Rhei in the developing solvent discriminating side; With the Radix Angelicae Sinensis control medicinal material is contrast, is Radix Angelicae Sinensis in the developing solvent discriminating side with normal hexane: ethyl acetate=1-9: 9-1.Its separating degree is good as a result, and the speckle colour developing is clear, and negative control is noiseless, the method favorable reproducibility.So adopt method of quality control of the present invention can effectively control the quality of sinew soothing and pain relieving preparation, thereby guarantee the clinical efficacy of said preparation.

Method of quality control of the present invention may further comprise the steps:

Character is observed, content is differentiated, official method is checked content, and the naringin that contains is carried out assay.

Wherein character is observed and comprises:

Character observed comprises:

For capsule, character is: the product content thing is that yellowish-brown is to filemot fine powder; Feeble QI perfume (or spice), bitter in the mouth;

For tablet, character is: medicine shows yellowish-brown to yellowish-brown; Feeble QI perfume (or spice), bitter in the mouth;

For granule, character is: product is that yellowish-brown is to filemot granule;

Content is differentiated, be may further comprise the steps:

(1) gets capsule, tablet or granule respectively, add the water supersound process, filter, filtrate is transferred in the separatory funnel, uses ether extraction 1-8 time, abandon ether solution, water liquid merges n-butyl alcohol liquid, water bath method with the n-butanol extraction of water saturation 1-8 time, residue adds methanol makes dissolving, as need testing solution; Other gets the Flos Carthami control medicinal material, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform: ethyl acetate: methanol: water=5-20: 20-60: 10-30: 5-20 is developing solvent in the solution of stratified lower floor below 10 ℃, launches, and takes out, dry, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

(2) get capsule, tablet or granule respectively, add the water supersound process, filter, filtrate is transferred in the separatory funnel, ether extraction 1-8 time is abandoned ether solution, and water liquid merges n-butyl alcohol liquid with the n-butanol extraction of water saturation 1-8 time, water bath method, residue add methanol makes dissolving, as need testing solution; Other gets the naringin reference substance, adds dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: the mixed solution that the upper solution 10-30ml of acetone: water=10-30: 5-20: 5-25 adds acetone 1-5ml and formic acid 0.05-3ml is developing solvent, launch, take out, dry, spray is with the aluminum chloride test solution, put under the ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

(3) get capsule, tablet or granule respectively, add the methanol supersound process, filter, the filtrate evaporate to dryness, residue is dissolved in water, add hydrochloric acid again, heating and refluxing extraction, ether extraction 1-5 time used in cooling immediately, merge ether solution, evaporate to dryness, residue add the chloroform dissolving, as need testing solution; Other gets the Radix Et Rhizoma Rhei control medicinal material, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose, with petroleum ether (30～60 ℃): the upper solution of Ethyl formate: formic acid=5-30: 1-10: 0.5-5 is developing solvent, 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, show the fluorescence speckle of same color.

(4) get capsule, tablet or granule respectively, add the ethanol supersound process, filter, filtrate volatilizes, and residue adds water makes dissolving, filters, filtrate is transferred in the separatory funnel, and ethyl acetate extraction 1-5 time merges ethyl acetate liquid, add active carbon, after water-bath volatilizes, peroxidating aluminum post, use the ethyl acetate eluting, collect eluent, water bath method, residue adds anhydrous alcohol solution, as need testing solution; Other gets the Radix Angelicae Sinensis control medicinal material, adds ethyl acetate, and supersound process filters, and filtrate volatilizes, and residue adds anhydrous alcohol solution, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be developing solvent with normal hexane: ethyl acetate=1-9: 9-1, launch, take out, dry, put under the ultra-violet lamp and inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color.

Official method comprises content inspection:

Capsule of the present invention, tablet or granule should meet Chinese Pharmacopoeia about the pertinent regulations under capsule, tablet or the granule item.

The naringin that contains is carried out assay be may further comprise the steps:

Naringin adopts high performance liquid chromatography, and chromatographic column is C18 or C4 or C8 post, and with methanol: acetic acid: water=20-50: 1-10: 40-80 is a mobile phase, and the detection wavelength is 200-500nm; Precision takes by weighing the naringin reference substance that is dried to constant weight at 110 ℃, uses dissolve with methanol, promptly gets reference substance solution; Get capsule, tablet or granule under the content uniformity item, accurate claim surely, add the methanol supersound process, put coldly, supply the weight that subtracts mistake, filter, get filtrate, promptly get need testing solution with methanol; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; In this oral formulations, contain Rhizoma Drynariae in the capsule, must not be less than in 0.3mg/g, the tablet and contain Rhizoma Drynariae, must not be less than in 0.3mg/g, the granule and contain Rhizoma Drynariae, must not be less than 0.03mg/g in naringin in naringin in naringin.

Concrete method of quality control is:

Character observed comprises:

Content is differentiated, comprised

(1) gets capsule, each 2g of tablet respectively, or get granule 5g, add water 10-100ml, supersound process 10-60 minute, filter, filtrate is transferred in the separatory funnel, ether extraction 1-8 time, each 10-100ml abandons ether solution, water liquid is with the n-butanol extraction of water saturation 1-8 time, each 10-100ml merges n-butyl alcohol liquid, water bath method, residue adds methanol 0.5-2ml makes dissolving, as need testing solution; Other gets Flos Carthami control medicinal material 0.5g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-25 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform: ethyl acetate: methanol: water=5-20: 20-60: 10-30: 5-20 is developing solvent in the solution of stratified lower floor below 10 ℃, launches, and takes out, dry, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

(2) get capsule, each 2g of tablet respectively, or get granule 5g, add water 10-100ml, supersound process 10-60 minute, filter, filtrate is transferred in the separatory funnel, ether extraction 1-8 time, each 10-100ml abandons ether solution, water liquid is with the n-butanol extraction of water saturation 1-8 time, each 10-100ml merges n-butyl alcohol liquid, water bath method, residue adds methanol 0.5-2ml makes dissolving, as need testing solution; Other gets the naringin reference substance, adds methanol and makes the solution that every 1ml contains 0.5-1mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-25 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: the mixed solution that the upper solution 10-30ml of acetone: water=10-30: 5-20: 5-25 adds acetone 1-5ml and formic acid 0.05-3ml is developing solvent, launch, take out, dry, spray is with the aluminum chloride test solution, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

(3) get capsule, each 1g of tablet respectively, or get granule 5g, add methanol 10-100ml, supersound process 10-100 minute, filter the filtrate evaporate to dryness, residue adds water 5-50ml makes dissolving, adds hydrochloric acid 0.5-5ml again, reflux 10-60 minute, ether extraction 1-5 time used in cooling immediately, each 10-50ml, merge ether solution, evaporate to dryness, residue add chloroform 0.5-2ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.1g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-25 μ l of above-mentioned solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose, with petroleum ether (30～60 ℃): the upper solution of Ethyl formate: formic acid=5-30: 1-10: 0.5-5 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

(4) get capsule, each 2g of tablet respectively, or get granule 10g, add ethanol 10-100ml, supersound process 10-100 minute, filter, filtrate volatilizes, and residue adds water 10-50ml makes dissolving, filter, filtrate is transferred in the separatory funnel, ethyl acetate extraction 1-5 time, each 5-50ml, merge ethyl acetate liquid, add active carbon 0.5-5g, after water-bath volatilizes, peroxidating aluminum post (1.5 * 20cm, 3g), with ethyl acetate 10-50ml eluting, collect eluent, water bath method, residue adds dehydrated alcohol 0.5-2ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 0.2g, adds ethyl acetate 10-50ml, and supersound process 5-60 minute, filter, filtrate volatilizes, and residue adds dehydrated alcohol 0.5-2ml makes dissolving, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-20 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be developing solvent with normal hexane: ethyl acetate=1-9: 9-1, launch, take out, dry, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color.

Official method to content inspection is:

Contained naringin is carried out assay, may further comprise the steps:

Naringin adopts high performance liquid chromatography, and chromatographic column is C18 or C4 or C8 post, and with methanol: acetic acid: water=20-50: 1-10: 40-80 is a mobile phase, and the detection wavelength is 200-500nm, and number of theoretical plate must not calculate with the naringin peak and is lower than 2000; Precision takes by weighing the naringin reference substance that is dried to constant weight at 110 ℃, with dissolve with methanol and be diluted to every 1ml and contain naringin 0.01-0.1mg, promptly gets reference substance solution; Get capsule, tablet or granule 1-5g under the content uniformity item, the accurate title, decide, and added methanol 20-30ml supersound process 30-60 minute, put coldly, supply the weight that subtracts mistake, shake up with methanol, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and gets filtrate, promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; In this oral formulations, contain Rhizoma Drynariae in the capsule, must not be less than in 0.3mg/g, the tablet and contain Rhizoma Drynariae, must not be less than in 0.3mg/g, the granule and contain Rhizoma Drynariae, must not be less than 0.03mg/g in naringin in naringin in naringin.

Method of quality control is more specifically:

Character observed comprises:

Content is differentiated, being comprised:

(1) gets capsule, each 2g of tablet respectively, or get granule 5g, add water 30ml, supersound process 30 minutes filters, and filtrate is transferred in the separatory funnel, ether extraction 3 times, each 20ml abandons ether solution, water liquid is with the n-butanol extraction of water saturation 3 times, be respectively 20ml, 15ml, 15ml, merge n-butyl alcohol liquid, water bath method, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Flos Carthami control medicinal material 0.5g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform: ethyl acetate: methanol: water=15: 40: 22: 10 is developing solvent in the solution of stratified lower floor below 10 ℃, launches, and takes out, dry, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

(2) get capsule, each 2g of tablet respectively, or get granule 5g, add water 30ml, supersound process 30 minutes filters, and filtrate is transferred in the separatory funnel, ether extraction 3 times, each 20ml abandons ether solution, water liquid is with the n-butanol extraction of water saturation 3 times, be respectively 20ml, 15ml, 15ml, merge n-butyl alcohol liquid, water bath method, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the naringin reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: the mixed solution that the upper solution 20ml of water=20: 10: 16 adds acetone 2ml and formic acid 0.1ml is developing solvent, launch, take out, dry, spray is with the aluminum chloride test solution, put under the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

(3) get capsule, each 1g of tablet respectively, or get granule 5g, add methanol 25ml, supersound process 20 minutes filters the filtrate evaporate to dryness, residue adds water 20ml makes dissolving, adds hydrochloric acid 1ml again, reflux 30 minutes, ether extraction 2 times are used in cooling immediately, each 20ml, merge ether solution, evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.1g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose, with petroleum ether (30～60 ℃): Ethyl formate: the upper solution of formic acid=15: 5: 1 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

(4) get capsule, each 2g of tablet respectively, or get granule 10g, add ethanol 25ml, supersound process 30 minutes, filter, filtrate volatilizes, and residue adds water 20ml makes dissolving, filter, filtrate is transferred in the separatory funnel, and ethyl acetate extraction 2 times is respectively 20ml, 10ml, merge ethyl acetate liquid, add active carbon 1g, after water-bath volatilizes, peroxidating aluminum post (1.5 * 20cm, 3g), with ethyl acetate 25ml eluting, collect eluent, water bath method, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 0.2g, adds ethyl acetate 20ml, and supersound process 10 minutes filters, and filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane: ethyl acetate=8: 7 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 254nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color.

Official method to content inspection is:

Should meet Chinese Pharmacopoeia about the pertinent regulations under capsule, tablet or the granule item.

Contained naringin is carried out assay, may further comprise the steps:

Naringin adopts high performance liquid chromatography, and chromatographic column is the C18 post, and with methanol: acetic acid: water=35: 4: 65 is mobile phase, and the detection wavelength is 283nm, and number of theoretical plate must not calculate with the naringin peak and is lower than 3000; Precision takes by weighing the naringin reference substance that is dried to constant weight at 110 ℃, with dissolve with methanol and be diluted to every 1ml and contain naringin 0.06mg, promptly gets reference substance solution; Get capsule, tablet or granule 1g under the content uniformity item, accurate claim surely, added methanol 25ml supersound process 45 minutes, put coldly, supply the weight that subtracts mistake, shake up,, get filtrate, promptly get need testing solution with the microporous filter membrane filtration of 0.45 μ m with methanol; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; In this oral formulations, contain Rhizoma Drynariae in the capsule, must not be less than in 0.3mg/g, the tablet and contain Rhizoma Drynariae, must not be less than in 0.3mg/g, the granule and contain Rhizoma Drynariae, must not be less than 0.03mg/g in naringin in naringin in naringin.

Method of quality control of the present invention is the preferred plan that obtains through a large amount of screenings, and following experimentation is a preferred process of the present invention.

One, method for measuring naringin content research

1, the selection of mobile phase:

Mobile phase 1: the mixed solution with methanol, water and acetic acid is a mobile phase

Mobile phase 2: the mixed solution with acetonitrile and glacial acetic acid aqueous solution is a mobile phase;

Mobile phase 3: the mixed solution with acetonitrile and water is a mobile phase.

Result: with methanol: acetic acid: water=20-50: 1-10: 40-80 is a mobile phase, the negative sample chromatogram is at non-false positive peak, naringin position, naringin separates fully (separating degree＞1.5) with close impurity peaks, promptly naringin separates with other components fully under this condition.Optimal flow is mutually: methanol: acetic acid: water=35: 4: 65.

2, need testing solution preparation method research:

2.1 the selection of extracting method

Get preparation 1.0g, respectively add methanol 50ml with methods such as immersion, water-bath reflux, extract,, ultrasonic extraction respectively, claim decide weight, extract one hour respectively after, put coldly, methanol is supplied the weight that subtracts mistake, its content is measured in filtration, the results are shown in following table:

Extracting method	Soak	The water-bath reflux, extract,	Supersound extraction
Extracting method	Soak	The water-bath reflux, extract,	Supersound extraction	Content (mg/g) RSD (%)	0.9538 2.05	1.3992 2.53	1.3968 1.19

Result of the test shows, soaks not extract naringin in its preparation is extracted fully, and supersound extraction, water-bath reflux, extract, can be extracted naringin in the preparation fully, and the two differs minimum, for easy and simple to handle, selects ultrasonic extracting method for use.

2.2 extraction choice of Solvent

Get preparation 1.0g, add methanol, 50% ethanol, ethanol 50ml respectively, claim decide weight, supersound extraction after 1 hour is respectively put coldly, supplies the weight that subtracts mistake, and its content is measured in filtration, the results are shown in following table:

Extract solvent	Methanol	50% ethanol	Ethanol
Extract solvent	Methanol	50% ethanol	Ethanol	Content (mg/g) RSD (%)	1.4005 1.32	1.3321 0.79	0.9832 1.48

As seen, methanol more can leach the naringin in the preparation as extracting solvent, so the selective extraction solvent is a methanol.

3, replica test

Get preparation, the preparation method by above-mentioned test liquid prepares 5 parts of test liquids respectively, and sample introduction is measured peak area (Fig. 9), and the naringin average content is 1.4092mg/g, and RSD is 2.48%.Show that repeatability is good, the results are shown in following table

The sample introduction number of times	1	2	3	4	5	Meansigma methods	RSD(％)
The sample introduction number of times	1	2	3	4	5	Meansigma methods	RSD(％)	Content (mg/g)	1.4082	1.3538	1.4486	1.4096	1.4256	1.4092	2.48

4, naringin stability test in the need testing solution

Get preparation, by the preparation method of above-mentioned test liquid, measure its naringin peak area (Figure 10) respectively at 0,2,4,6,8 hour, average peak area is 344072, and RSD is 1.16%.Show that naringin is stable in 8 hours in the need testing solution, the results are shown in following table.

Time (hour)	0	2	4	6	8	Meansigma methods	RSD(％)
Time (hour)	0	2	4	6	8	Meansigma methods	RSD(％)	Peak area	340885	342341	340412	348107	348617	344072	1.16

Two, the Flos Carthami thin layer chromatography is signed and Yan Jiu not

With the Flos Carthami control medicinal material is contrast.

Need testing solution preparation method one: get preparation 2g, add 80% acetone soln 20ml, supersound process 20 minutes filters, and filtrate volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution.Other gets Flos Carthami control medicinal material 0.5g and lacks the negative test sample of Flos Carthami, shines medical material solution and negative test liquid in pairs with legal system.

Need testing solution preparation method two: get preparation 2g, add water 30ml, supersound process 30 minutes, filter, filtrate is transferred in the separatory funnel, ether extraction 3 times, each 20ml, abandon ether solution, water liquid merges n-butyl alcohol liquid, water bath method with the n-butanol extraction 3 times (20ml, 15ml, 15ml) of water saturation, residue adds methanol 1ml makes dissolving, as need testing solution.

Need testing solution preparation method three: get preparation 2g, add 5 of ethanol 30ml, chloroform 10ml, concentrated sulphuric acids, the water-bath reflux, extract, is after 1 hour, filter, filtrate volatilizes, and residue adds water 20ml makes dissolving, filters, filtrate is used chloroform extraction 2 times, each 20ml, the combined chloroform extracting solution volatilizes, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution.

Developing solvent is selected: respectively with the mixed solution of ethyl acetate formic acid water and the various ratios of methanol; The mixed solution of chloroform, ethyl acetate, the various ratios of first alcohol and water is in the solution of stratified lower floor below 10 ℃; The mixed solution of normal hexane, the various ratios of ethyl acetate is developing solvent.

Result: adopt method two to prepare need testing solution, with chloroform: ethyl acetate: methanol: water=5-20: 20-60: 10-30: 5-20 is developing solvent in the solution of stratified lower floor below 10 ℃, and its separating degree is good, and the speckle colour developing is clear, negative control is noiseless, the method favorable reproducibility.Best developing solvent is: chloroform: ethyl acetate: methanol: water=15: 40: 22: 10 in the solution of stratified lower floor below 10 ℃.

Three, the Rhizoma Drynariae thin layer chromatography is signed and Yan Jiu not

With the naringin reference substance is contrast.

Need testing solution preparation method one: get preparation 1.5g, add methanol 25ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets the naringin reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution.Make the negative test liquid that lacks Rhizoma Drynariae with method.

Need testing solution preparation method two: get preparation 2g, add water 30ml, supersound process 30 minutes, filter, filtrate is transferred in the separatory funnel, ether extraction 3 times, each 20ml, abandon ether solution, water liquid merges n-butyl alcohol liquid, water bath method with the n-butanol extraction 3 times (20ml, 15ml, 15ml) of water saturation, residue adds methanol 1ml makes dissolving, as need testing solution.Make the negative test liquid that lacks Rhizoma Drynariae with method.

Developing solvent is selected: respectively with the upper strata liquid with the various mixed solution of benzene, ethyl acetate, formic acid and water; The upper strata liquid of ethyl acetate, acetone, the various mixed solution of water and the mixed solution of acetone and formic acid are developing solvent.

Result: adopt method two to prepare need testing solution, with ethyl acetate: the mixed solution that the upper solution 10-30ml of acetone: water=10-30: 5-20: 5-25 adds acetone 1-5ml and formic acid 0.05-3ml is developing solvent, its separating degree is good, speckle is more obvious, negative noiseless, the method favorable reproducibility.Best developing solvent is: with ethyl acetate: acetone: the mixed solution that the upper solution 20ml of water=20: 10: 16 adds acetone 2ml and formic acid 0.1ml is developing solvent.

Four, the Radix Et Rhizoma Rhei thin layer chromatography is signed and Yan Jiu not

With the Radix Et Rhizoma Rhei control medicinal material is contrast.

Need testing solution preparation method one: get preparation 1g, add methanol 25ml, supersound process 20 minutes, filter, the filtrate evaporate to dryness adds water 20ml and makes dissolving, add hydrochloric acid 1ml again, put in the water-bath and heated 30 minutes, immediately cooling, divide 2 extractions with ether, each 20ml merges ether solution, evaporate to dryness, residue adds chloroform 1ml makes dissolving, as need testing solution; Make the negative test liquid of scarce Radix Et Rhizoma Rhei with method.

Need testing solution preparation method two: get preparation 1g, porphyrize, the 20ml that adds diethyl ether, supersound process 2 times, each 20min filters, and merging filtrate, water bath method, residue add methanol 2ml makes dissolving, as need testing solution; Make the negative test liquid of scarce Radix Et Rhizoma Rhei with method.

Developing solvent is selected: respectively with the mixed solution of petroleum ether (30～60 ℃), Ethyl formate and the various ratios of formic acid; The upper solution of petroleum ether (30～60 ℃), formic acid second fat and the various ratios of methyl ether is developing solvent.

The result: employing method one preparation need testing solution, with petroleum ether (30～60 ℃): the upper solution of Ethyl formate: formic acid=5-30: 1-10: 0.5-5 is developing solvent, and its separating degree is good, and the speckle colour developing is clear, and negative control is noiseless, the method favorable reproducibility.Best developing solvent is for petroleum ether (30～60 ℃): Ethyl formate: the upper solution of formic acid=15: 5: 1 is developing solvent.

Five, the Radix Angelicae Sinensis thin layer chromatography is signed and Yan Jiu not

With the Radix Angelicae Sinensis control medicinal material is contrast.

Need testing solution preparation method one: get preparation 2g, add ethyl acetate 25ml, supersound process 30 minutes filters, and filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution, makes the negative controls that lacks Radix Angelicae Sinensis with method.

Need testing solution preparation method two: get preparation 2g, add ethanol 25ml, supersound process 30 minutes filters, filtrate volatilizes, residue adds water 20ml makes dissolving, filters and is transferred in the separatory funnel, and ethyl acetate extraction 2 times (20,10ml) merges ethyl acetate liquid, water-bath volatilizes, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution, makes the negative controls that lacks Radix Angelicae Sinensis with method.

Need testing solution preparation method three: get preparation 2 product g, add ethanol 25ml, supersound process 30 minutes filters, and filtrate volatilizes, residue adds water 20ml makes dissolving, filter also and be transferred in the separatory funnel, ethyl acetate extraction 2 times (20,10ml) merges ethyl acetate liquid, add active carbon 1g, after water-bath volatilizes, and peroxidating aluminum post (1.5 * 20cm, 3g), with ethyl acetate 25ml eluting, collect eluent, water bath method, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution, make the negative controls that lacks Radix Angelicae Sinensis with method.

Developing solvent is selected: respectively with normal hexane: the mixed solution of the various ratios of ethyl acetate; Ethyl acetate: formic acid: the mixed solution of glacial acetic acid and the various ratios of water; The mixed solution of petroleum ether (30～60 ℃), chloroform and the various ratios of glacial acetic acid; The mixed solution of cyclohexane extraction, ethyl acetate and the various ratios of glacial acetic acid; The mixed solution of cyclohexane extraction, ethyl acetate, benzene and the various ratios of formic acid is developing solvent.

The result: employing method three preparation need testing solutions are developing solvent with normal hexane: ethyl acetate=1-9: 9-1, and its separating degree is good, and the speckle colour developing is clear, and negative control is noiseless, the method favorable reproducibility.Best developing solvent is a normal hexane: ethyl acetate=9: 1.The invention has the advantages that: method of quality control of the present invention is on former statutory standards basis, increases the content of high effective liquid chromatography for measuring naringin; Utilize thin layer chromatography to differentiate each flavour of a drug in this compound preparation, formed new method of quality control (standard).This method has guaranteed that the quality inspection standard of preparation of the present invention can have accuracy and advance than the qualitative character of effectively controlling preparation comprehensively, can be used as the effective technology means of the stability of quality control and investigation technology.Be of great importance to improving the quality of products.

Compared with the prior art, it is simple to the present invention is directed to original preparation SHUJIN DINGTONG PIAN quality control standard, shortcomings such as product quality is wayward, method of quality control to this preparation is studied, improved the quality standard of this sinew soothing and pain relieving preparation, guarantee the clinical efficacy of this preparation, reached goal of the invention.

The specific embodiment: further specify the present invention by the following examples, but not as limitation of the present invention.

The quality control of embodiment 1 capsule

For the capsule character be: the product content thing is that yellowish-brown is to filemot fine powder; Feeble QI perfume (or spice), bitter in the mouth;

Content is differentiated, comprised

(1) get capsule 2g and add water 100ml, supersound process 60 minutes filters, filtrate is transferred in the separatory funnel, ether extraction 8 times, each 100ml abandons ether solution, water liquid is with the n-butanol extraction of water saturation 8 times, each 100ml merges n-butyl alcohol liquid, water bath method, residue adds methanol 0.5-2ml makes dissolving, as need testing solution; Other gets Flos Carthami control medicinal material 0.5g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 25 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform: ethyl acetate: methanol: water=20: 20: 10: 5 is developing solvent in the solution of stratified lower floor below 10 ℃, launches, and takes out, dry, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

(2) get capsule 2g, add water 100ml, supersound process 60 minutes, filter, filtrate is transferred in the separatory funnel, ether extraction 8 times, each 100ml, abandon ether solution, water liquid is with the n-butanol extraction of water saturation 8 times, each 100ml, merge n-butyl alcohol liquid, water bath method, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the naringin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 25 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: the mixed solution that the upper solution 30ml of water=10: 5: 25 adds acetone 5ml and formic acid 3ml is developing solvent, launch, take out, dry, spray is with the aluminum chloride test solution, put under the ultra-violet lamp 500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

(3) get capsule 1g, add methanol 100ml, supersound process 100 minutes, filter, filtrate evaporate to dryness, residue add water 50ml makes dissolving, add hydrochloric acid 5ml again, reflux 60 minutes, cooling immediately, with ether extraction 5 times, each 50ml merges ether solution, evaporate to dryness, residue adds chloroform 2ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.1g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 25 μ l of above-mentioned solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose, with petroleum ether (60 ℃): Ethyl formate: the upper solution of formic acid=5: 10: 0.5 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 500nm and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

(4) get capsule 2g, add ethanol 100ml, supersound process 100 minutes filters, filtrate volatilizes, and residue adds water 50ml makes dissolving, filters, and filtrate is transferred in the separatory funnel, ethyl acetate extraction 5 times, each 50ml merges ethyl acetate liquid, add active carbon 5g, after water-bath volatilizes, peroxidating aluminum post (1.5 * 20cm, 3g), with ethyl acetate 50ml eluting, collect eluent, water bath method, residue add dehydrated alcohol 2ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 0.2g, adds ethyl acetate 50ml, and supersound process 60 minutes filters, and filtrate volatilizes, and residue adds dehydrated alcohol 2ml makes dissolving, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 20 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane: ethyl acetate=9: 1 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 500nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color.

Official method to content inspection is

Capsule of the present invention should meet Chinese Pharmacopoeia about the pertinent regulations under the capsule item.

Contained naringin is carried out assay, may further comprise the steps:

Naringin adopts high performance liquid chromatography, and chromatographic column is the C8 post, and with methanol: acetic acid: water=20: 1: 80 is mobile phase, and the detection wavelength is 500nm, and number of theoretical plate must not calculate with the naringin peak and is lower than 2000; Precision takes by weighing the naringin reference substance that is dried to constant weight at 110 ℃, with dissolve with methanol and be diluted to every 1ml and contain naringin 0.1mg, promptly gets reference substance solution; Get capsule, tablet or granule 5g under the content uniformity item, the accurate title, decide, and added methanol 30ml supersound process 60 minutes, put coldly, supply the weight that subtracts mistake, shake up with methanol, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and gets filtrate, promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; In this oral formulations, contain Rhizoma Drynariae in the capsule, must not be less than 0.3mg/g in naringin.

Embodiment 2: the quality control of tablet

For the tablet character be: medicine shows yellowish-brown to yellowish-brown; Feeble QI perfume (or spice), bitter in the mouth;

Content is differentiated, comprised

(1) gets tablet 2g, add water 10ml, supersound process 10 minutes, filter, filtrate is transferred in the separatory funnel, ether extraction 1 time, each 10ml, abandon ether solution, water liquid is with the n-butanol extraction of water saturation 1 time, each 10ml, merge n-butyl alcohol liquid, water bath method, residue add methanol 0.5ml makes dissolving, as need testing solution; Other gets Flos Carthami control medicinal material 0.5g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform: ethyl acetate: methanol: water=5: 60: 30: 20 is developing solvent in the solution of stratified lower floor below 10 ℃, launches, and takes out, dry, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

(2) get tablet 2g, add water 10ml, supersound process 10 minutes, filter, filtrate is transferred in the separatory funnel, ether extraction 1 time, each 10ml, abandon ether solution, water liquid is with the n-butanol extraction of water saturation 1 time, each 10ml, merge n-butyl alcohol liquid, water bath method, residue add methanol 0.5ml makes dissolving, as need testing solution; Other gets the naringin reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: the mixed solution that the upper solution 10ml of water=30: 20: 5 adds acetone 1ml and formic acid 0.05ml is developing solvent, launch, take out, dry, spray is with the aluminum chloride test solution, put under the ultra-violet lamp 200nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

(3) get tablet 1g, add methanol 10ml, supersound process 10 minutes, filter, filtrate evaporate to dryness, residue add water 5ml makes dissolving, add hydrochloric acid 0.5ml again, reflux 10 minutes, cooling immediately, with ether extraction 1 time, each 10ml merges ether solution, evaporate to dryness, residue adds chloroform 0.5ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.1g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose, with petroleum ether (30 ℃): Ethyl formate: the upper solution of formic acid=30: 1: 5 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200nm and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

(4) get tablet 2g, add ethanol 10ml, supersound process 10 minutes filters, filtrate volatilizes, and residue adds water 10ml makes dissolving, filters, and filtrate is transferred in the separatory funnel, ethyl acetate extraction 1 time, each 5ml merges ethyl acetate liquid, add active carbon 0.5g, after water-bath volatilizes, peroxidating aluminum post (1.5 * 20cm, 3g), with ethyl acetate 10ml eluting, collect eluent, water bath method, residue add dehydrated alcohol 0.5ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 0.2g, adds ethyl acetate 10ml, and supersound process 5 minutes filters, and filtrate volatilizes, and residue adds dehydrated alcohol 0.5ml makes dissolving, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane: ethyl acetate=1: 9 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color.

Official method to content inspection is

Tablet of the present invention should meet Chinese Pharmacopoeia about the pertinent regulations under the tablet item.

Contained naringin is carried out assay, may further comprise the steps:

Naringin adopts high performance liquid chromatography, and chromatographic column is the C18 post, and with methanol: acetic acid: water=50: 10: 40 is mobile phase, and the detection wavelength is 200nm, and number of theoretical plate must not calculate with the naringin peak and is lower than 2000; Precision takes by weighing the naringin reference substance that is dried to constant weight at 110 ℃, with dissolve with methanol and be diluted to every 1ml and contain naringin 0.01mg, promptly gets reference substance solution; Get capsule, tablet or granule 1g under the content uniformity item, the accurate title, decide, and added methanol 20ml supersound process 30 minutes, put coldly, supply the weight that subtracts mistake, shake up with methanol, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and gets filtrate, promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; In this oral formulations, contain Rhizoma Drynariae in the tablet, must not be less than 0.3mg/g in naringin.

The quality control of embodiment 3 granules

Character observed comprises:

Content is differentiated, being comprised:

(1) gets granule 5g, add water 30ml, supersound process 30 minutes, filter, filtrate is transferred in the separatory funnel, ether extraction 3 times, each 20ml, abandon ether solution, water liquid is respectively 20ml, 15ml, 15ml with the n-butanol extraction of water saturation 3 times, merge n-butyl alcohol liquid, water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Flos Carthami control medicinal material 0.5g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform: ethyl acetate: methanol: water=15: 40: 22: 10 is developing solvent in the solution of stratified lower floor below 10 ℃, launches, and takes out, dry, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

(2) get granule 5g, add water 30ml, supersound process 30 minutes, filter, filtrate is transferred in the separatory funnel, ether extraction 3 times, each 20ml, abandon ether solution, water liquid is respectively 20ml, 15ml, 15ml with the n-butanol extraction of water saturation 3 times, merge n-butyl alcohol liquid, water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the naringin reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: the mixed solution that the upper solution 20ml of water=20: 10: 16 adds acetone 2ml and formic acid 0.1ml is developing solvent, launch, take out, dry, spray is with the aluminum chloride test solution, put under the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

(3) get granule 5g, add methanol 25ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue add water 20ml makes dissolving, add hydrochloric acid 1ml again, reflux 30 minutes, cooling immediately, with ether extraction 2 times, each 20ml merges ether solution, evaporate to dryness, residue adds chloroform 1ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.1g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose, with petroleum ether (30～60 ℃): Ethyl formate: the upper solution of formic acid=15: 5: 1 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

(4) get granule 10g, add ethanol 25ml, supersound process 30 minutes filters, filtrate volatilizes, and residue adds water 20ml makes dissolving, filters, and filtrate is transferred in the separatory funnel, ethyl acetate extraction 2 times is respectively 20ml, 10ml, merges ethyl acetate liquid, add active carbon 1g, after water-bath volatilizes, peroxidating aluminum post (1.5 * 20cm, 3g), with ethyl acetate 25ml eluting, collect eluent, water bath method, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 0.2g, adds ethyl acetate 20ml, and supersound process 10 minutes filters, and filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane: ethyl acetate=8: 7 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 254nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the fluorescence speckle of same color.

Official method to content inspection is:

Should meet Chinese Pharmacopoeia about the pertinent regulations under the granule item.

Contained naringin is carried out assay, may further comprise the steps:

Naringin adopts high performance liquid chromatography, and chromatographic column is the C18 post, and with methanol: acetic acid: water=35: 4: 65 is mobile phase, and the detection wavelength is 283nm, and number of theoretical plate must not calculate with the naringin peak and is lower than 3000; Precision takes by weighing the naringin reference substance that is dried to constant weight at 110 ℃, with dissolve with methanol and be diluted to every 1ml and contain naringin 0.06mg, promptly gets reference substance solution; Get the granule 1g under the content uniformity item, accurate claim surely, added methanol 25ml supersound process 45 minutes, put coldly, supply the weight that subtracts mistake, shake up,, get filtrate, promptly get need testing solution with the microporous filter membrane filtration of 0.45 μ m with methanol; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content;

Contain Rhizoma Drynariae in naringin in this granule, must not be less than 0.03mg/g.

Claims

1, a kind of method of quality control of sinew soothing and pain relieving preparation is characterized in that, comprises character is observed, and content is differentiated, official method is checked content, the naringin that contains is carried out the step of assay.

2, the method for claim 1 is characterized in that, described sinew soothing and pain relieving preparation is an oral solid formulation.

3, the method for claim 2 is characterized in that, described oral solid formulation is granule, tablet or capsule.

4, the method for claim 1 is characterized in that, described discrimination method is selected from one or more in the following method:

The step of described assay comprises:

5, method according to claim 4 is characterized in that, described discrimination method is selected from one or more in the following method:

6, method according to claim 4 is characterized in that, the step of described assay comprises:

According to the method for claim 1, it is characterized in that 7, described sinew soothing and pain relieving preparation is made by following Chinese medicine raw materials by weight proportion:

Eupolyphaga Seu Steleophaga 50-200g, Olibanum (vinegar system) 30-100g, Myrrha (vinegar system) 30-100g, Pyritum (vinegar is forged) 30-100g, Flos Carthami 30-100g, Rhizoma Drynariae 30-100g, Radix Et Rhizoma Rhei 30-100g, Borax (forging) 60g, Radix Angelicae Sinensis 30-100g.

8, according to the method for claim 1, it is characterized in that, wherein character is observed and comprise: for the capsule character be: the product content thing is that yellowish-brown is to filemot fine powder; Feeble QI perfume (or spice), bitter in the mouth; For the tablet character be: medicine shows yellowish-brown to yellowish-brown; Feeble QI perfume (or spice), bitter in the mouth; For the granule character be: product is that yellowish-brown is to filemot granule.

According to the method for claim 1, it is characterized in that 9, official method comprises content inspection: capsule, tablet or granule should meet Chinese Pharmacopoeia about the pertinent regulations under capsule, tablet or the granule item.

10, according to the method for claim 1, it is characterized in that:

Character observed comprises:

Content is differentiated, being comprised:

Official method to content inspection is:

Contained naringin is carried out assay, may further comprise the steps: