CN1785220A

CN1785220A - Method for quality control of blood-bonifying Chinese angelica prepn.

Info

Publication number: CN1785220A
Application number: CN 200510003311
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: CN100402063C

Abstract

A quality control method for the blood-mourishing Chinese angelica root extract is disclosed, which features that the quality standard of said medicine is increased for ensuring its curative effect.

Description

The method of quality control of 'Buxuedanggui '

Technical field: the present invention relates to a kind of method of quality control of 'Buxuedanggui ', belong to the field of medicine technology.

Technical background: weak, the anemia of menoxenia, puerperal blood deficiency is the commonly encountered diseases in our daily life, 'Buxuedanggui ' by Radix Angelicae Sinensis, Radix Rehmanniae Preparata, the Radix Paeoniae Alba, Rhizoma Chuanxiong, Radix Codonopsis, Radix Glycyrrhizae and the Radix Astragali altogether seven flavor medicine form; Effect with nourishing qi and blood, to dizziness, anemia, weak, the menoxenia of health, blood deficiency in puerperal disease such as weak has definite curative effect.Wherein " blood-bonifying Chinese angelica essence " publication is the tenth of Drug Standard of Ministry of Public Health of the Peoples Republic of China, this medicine is used for many years clinically, obtain satisfied therapeutic effect at treatment dizziness, anemia, health aspect weak, but discover through us, existing 'Buxuedanggui ' exists method of quality control to fall behind the uppity shortcoming of product quality.Since Radix Angelicae Sinensis have enrich blood invigorate blood circulation, effects such as menstruction regulating and pain relieving, loosening bowel to relieve constipation, it is main effective ingredient in the preparation of the present invention, in addition, the Radix Paeoniae Alba, the Radix Astragali and Radix Glycyrrhizae also are the materials of performance main pharmacodynamics in the preparation of the present invention, and Radix Angelicae Sinensis is not differentiated in the existing method of quality control, or effective ingredient in the Radix Angelicae Sinensis is carried out assay.Other drug is not differentiated yet,, thereby will be influenced the production of product and ensure the quality of products so existing method of quality control can not effectively be controlled the quality of 'Buxuedanggui '.

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

'Buxuedanggui ' of the present invention is to constitute like this: calculate according to composition by weight: it mainly is prepared from by Radix Angelicae Sinensis 300-500g, Radix Rehmanniae Preparata (wine steams) 10-50g, the Radix Paeoniae Alba (wine stir-fry) 10-50g, Rhizoma Chuanxiong (wine stir-fry) 5-20g, the Radix Astragali (processed with honey) 10-50g, Radix Codonopsis 10-50g, Radix Glycyrrhizae 5-20g.

Preferred prescription is formed: calculate according to composition by weight: it mainly is prepared from by Radix Angelicae Sinensis 400g, Radix Rehmanniae Preparata (wine steams) 24g, the Radix Paeoniae Alba (wine stir-fry) 24g, Rhizoma Chuanxiong (wine stir-fry) 12g, the Radix Astragali (processed with honey) 24g, Radix Codonopsis 24g, Radix Glycyrrhizae 12g.

Preparation of the present invention prepares like this: above seven flavors, pulverize separately becomes coarse powder, Radix Angelicae Sinensis, the Radix Paeoniae Alba, Rhizoma Chuanxiong, the Radix Astragali are made solvent with 20-85% ethanol respectively, carrying out percolation according to the percolation under Chinese Pharmacopoeia fluid extract and the extractum item extracts, collect 8-15 respectively and doubly measure percolate, the liquid decompression recycling ethanol of filtering; Radix Rehmanniae Preparata, Radix Glycyrrhizae, Radix Codonopsis decoct with water 1-5 time, and collecting decoction filters, and filtrate concentrates, and adds ethanol 1-10 and doubly measures, stir evenly, leave standstill, filter filtrate recycling ethanol, merge with above-mentioned medicinal liquid,, add different auxiliary material, make capsule, tablet or granule again according to conventional method.

In the prescription Radix Angelicae Sinensis have enrich blood invigorate blood circulation, effect such as menstruction regulating and pain relieving, loosening bowel to relieve constipation, it is main effective ingredient in the preparation of the present invention, so in the 'Buxuedanggui ' in the reply Radix Angelicae Sinensis main effective ingredient set up content assaying method, can control the quality of said preparation well, if but how to select chromatographic condition and preparation need testing solution to become difficult point of the present invention.Through a large amount of experiments, the present invention selects chromatographic column to be C18 or C4 or C8 post, and fine with methanol or second: 0.05-0.5% phosphoric acid solution=1-9: 9-1 is a mobile phase; The detection wavelength is 200-500nm, and number of theoretical plate must not calculate with the ferulic acid peak and is lower than content of ferulic acid in 4000 the high effective liquid chromatography for measuring Radix Angelicae Sinensis.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 Radix Angelicae Sinensis, the Radix Paeoniae Alba, the Radix Astragali and four medical materials of Radix Glycyrrhizae, and to select with the Radix Angelicae Sinensis control medicinal material be contrast, is Radix Angelicae Sinensis in the developing solvent discriminating side with normal hexane: ethyl acetate=1-9: 9-1; With the Radix Paeoniae reference substance is contrast, and with ethyl acetate: formic acid: glacial acetic acid: water=5-30: 0.5-5: 0.5-5: 0.5-5 is the Radix Paeoniae Alba in the developing solvent discriminating side; With the astragaloside reference substance is contrast, and with chloroform: lower floor's solution of methanol: water=5-30: 3-15: 1-10 is the Radix Astragali in the developing solvent discriminating side; With the Radix Glycyrrhizae control medicinal material is contrast, and with ethyl acetate: formic acid: glacial acetic acid: water=5-30: 0.5-5: 0.5-5: 0.5-5 is a Radix Glycyrrhizae in the developing solvent discriminating side; 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 'Buxuedanggui ', 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 ferulic acid that contains is carried out assay.

Wherein character is observed and comprises:

For capsule, character is: the product content thing is brown xanchromatic fine powder or granule, acrid in the mouth, little hardship;

For the tablet character be: medicine shows brown xanchromatic fine powder or granule; Acrid in the mouth, little hardship;

For the granule character be: product is the xanchromatic granule of palm fibre;

Content is differentiated, be may further comprise the steps:

(1) get capsule, tablet or granule respectively, add petroleum ether, supersound process, extracting solution filters, the filtrate evaporate to dryness, residue adds anhydrous alcohol solution, as need testing solution; Other gets the Radix Angelicae Sinensis 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, putting respectively on same silica gel g thin-layer plate, is developing solvent with normal hexane: ethyl acetate=1-9: 9-1, 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, show the fluorescence speckle of same color.

(2) get capsule, tablet or granule respectively, add ethyl acetate, supersound extraction, extracting solution filters, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol of water saturation again, and supersound extraction filters, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 1-8 time is abandoned water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution is transferred to the macroporous resin column of having handled well, the water eluting, abandon water liquid, reuse 20-85% ethanol elution is collected eluent, evaporate to dryness, residue add methanol makes dissolving, as need testing solution; Other gets the Radix Paeoniae 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 on the silica gel g thin-layer plate of same sodium hydroxide solution preparation with 0.5-5%, with ethyl acetate: formic acid: glacial acetic acid: water=5-30: 0.5-5: 0.5-5: 0.5-5 is developing solvent, launch, take out, dry, spray is with ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) get capsule, tablet or granule respectively, add methanol, reflux, extract,, extracting solution filters, the filtrate evaporate to dryness, residue adds water makes dissolving, with the n-butanol extraction of water saturation 1-8 time, merges n-butyl alcohol liquid, with ammonia solution washing 1-8 time, abandon the ammonia washing liquid, water lotion is abandoned in the water washing of the saturated mistake of n-butyl alcohol liquid reuse n-butyl alcohol 1-5 time, n-butyl alcohol liquid is put evaporate to dryness in the water-bath, residue adds dissolve with methanol, crosses the neutral alumina post, with 20-85% methanol solution eluting, collect eluent, put water bath method, residue adds methanol makes dissolving, as need testing solution; Other gets the astragaloside 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 on same silica gel g thin-layer plate, with chloroform: lower floor's solution of methanol: water=5-30: 3-15: 1-10 is developing solvent, launch, take out, dry, spray is with ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with contrast chromatograph corresponding position on, daylight shows identical sepia speckle down; Ultra-violet lamp shows identical orange-yellow fluorescence speckle down.

(4) get capsule, tablet or granule respectively, add ethyl acetate, supersound extraction, extracting solution filters, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol of water saturation again, and supersound extraction filters, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 1-8 time, abandon water lotion, n-butyl alcohol liquid water bath method, residue water dissolution, be transferred to the D101 type macroporous resin column of having handled well, the water eluting is abandoned water liquid, reuse 20-85% ethanol elution, collect eluent, evaporate to dryness, residue add methanol makes dissolving, as need testing solution; The extracting liquorice control medicinal material adds diethyl ether reflux, extract, in addition, filter, abandon filtrate, medicinal residues add methanol, the water-bath reflux, extract, filters the filtrate evaporate to dryness, residue is dissolved in water, and is transferred in the separatory funnel, with the n-butanol extraction of water saturation 1-8 time, merge n-butyl alcohol liquid, water lotion is abandoned in the washing of the saturated mistake of reuse n-butyl alcohol 1-5 time, n-butyl alcohol liquid water bath method, residue adds methanol makes dissolving, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned solution, put respectively on the silica gel g thin-layer plate of same sodium hydroxide solution preparation with 0.5-5%, with ethyl acetate: formic acid: glacial acetic acid: water=5-30: 0.5-5: 0.5-5: 0.5-5 is developing solvent, launches, take out, dry, under daylight, inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the 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 ferulic acid that contains is carried out assay be may further comprise the steps:

Ferulic acid adopts high performance liquid chromatography, and chromatographic column is C18 or C4 or C8 post, and is fine with methanol or second: 0.05-0.5% phosphoric acid solution=1-9: 9-1 is a mobile phase; The detection wavelength is 200-500nm; Precision takes by weighing the ferulic acid reference substance, uses dissolve with methanol, promptly gets reference substance solution; Get capsule, tablet or granule respectively,

The accurate methanol solution water-bath reflux, extract, that adds is put coldly, supplies the weight that subtracts mistake with methanol, shakes up, and leaves standstill, and with 0.65 μ m and less than the microporous filter membrane filtration of 0.65 μ m, 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 Radix Angelicae Sinensis in the capsule, must not be less than in 0.05mg/g, the tablet and contain Radix Angelicae Sinensis, must not be less than in 0.05mg/g, the granule and contain Radix Angelicae Sinensis, must not be less than 0.005mg/g in ferulic acid in ferulic acid in ferulic acid.

Concrete method of quality control comprises:

Character observed comprises:

For the capsule character be: the product content thing is brown xanchromatic fine powder or granule, acrid in the mouth, little hardship;

Content is differentiated, comprised

(1) get capsule, each 1g of tablet respectively, or get granule 5g, add petroleum ether 20-150ml, supersound process 10-60 minute, filter, filtrate evaporate to dryness, residue add dehydrated alcohol 0.5-2ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 0.1-0.5g, 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, putting respectively on same silica gel g thin-layer plate, is developing solvent with normal hexane: ethyl acetate=1-9: 9-1, launches, 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.

(2) get capsule respectively, each 5g of tablet, or get granule 20g, add ethyl acetate 20-200ml, supersound extraction 10-60 minute, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 20-100ml of water saturation again, supersound extraction 0.5-3 hour, filter the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 1-8 time, each 10-50ml, abandon water lotion, n-butyl alcohol liquid water bath method, residue water dissolution, be transferred to the D101 type macroporous resin column of having handled well (1.5 * 15cm), use the 20-100ml water elution, abandon water liquid, reuse 20-85% ethanol 20-100ml eluting, collect eluent, evaporate to dryness, residue add methanol 0.5-2ml makes dissolving, as need testing solution; Other gets the Radix Paeoniae reference substance, adds methanol and makes the solution that every 1ml contains 0.2-1mg, 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 on the silica gel g thin-layer plate of same sodium hydroxide solution preparation with 0.5-5%, with ethyl acetate: formic acid: glacial acetic acid: water=5-30: 0.5-5: 0.5-5: 0.5-5 is developing solvent, launch, take out, dry, spray is with the 5-20% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) get capsule respectively, each 5g of tablet, or get granule 20g, add methanol 20-200ml, reflux, extract, 10-100 minute, filter, filtrate evaporate to dryness, residue add water 10-100ml makes dissolving, with the n-butanol extraction of water saturation 1-8 time, each 10-100ml, merge n-butyl alcohol liquid, with ammonia solution washing 1-8 time, each 10-100ml, abandon the ammonia washing liquid, water lotion is abandoned in the water 10-100ml washing of the saturated mistake of n-butyl alcohol liquid reuse n-butyl alcohol 1-5 time, and n-butyl alcohol liquid is put evaporate to dryness in the water-bath, residue adds methanol 2-10ml makes dissolving, (1.5 * 20cm 5g), uses 20-85% methanol solution 50-200ml eluting to cross the neutral alumina post, collect eluent, put water bath method, residue adds methanol 0.2-1ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.2-1mg, 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 on same silica gel g thin-layer plate, with chloroform: lower floor's solution of methanol: water=5-30: 3-15: 1-10 is developing solvent, launch, take out, dry, spray is with the 5-20% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with contrast chromatograph corresponding position on, daylight shows identical sepia speckle down; Ultra-violet lamp 200-500nm shows identical orange-yellow fluorescence speckle down.

(4) get capsule respectively, each 5g of tablet, or get granule 20g, add ethyl acetate 20-200ml, supersound extraction 10-60 minute, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 20-100ml of water saturation again, supersound extraction 0.5-3 hour, filter the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 1-8 time, each 10-50ml, abandon water lotion, n-butyl alcohol liquid water bath method, residue water dissolution, be transferred to the D101 type macroporous resin column of having handled well (1.5 * 15cm), use the 20-100ml water elution, abandon water liquid, reuse 20-85% ethanol 20-100ml eluting, collect eluent, evaporate to dryness, residue add methanol 0.5-2ml makes dissolving, as need testing solution; Other is extracting liquorice control medicinal material 0.5g, the 10-100ml that adds diethyl ether after reflux, extract, 0.5-5 hour, filters, abandon filtrate, medicinal residues add methanol 10-100ml, and water-bath reflux, extract, 0.5-5 hour filters, the filtrate evaporate to dryness, residue adds water 10-50ml makes dissolving, and is transferred in the separatory funnel, with the n-butanol extraction of water saturation 1-8 time, each 10-100ml, merge n-butyl alcohol liquid, the washing of the saturated mistake of reuse n-butyl alcohol 1-5 time, each 10-50ml, abandon water lotion, n-butyl alcohol liquid water bath method, residue add methanol 0.5-2ml makes dissolving, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-25 μ l of above-mentioned solution, put respectively on the silica gel g thin-layer plate of same sodium hydroxide solution preparation with 0.5-5%, with ethyl acetate: formic acid: glacial acetic acid: water=5-30: 0.5-5: 0.5-5: 0.5-5 is developing solvent, launches, take out, dry, under daylight, inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

Official method to content inspection is

To the contained assay that carries out, may further comprise the steps:

Ferulic acid adopts high performance liquid chromatography, and chromatographic column is C18 or C4 or C8 post, and is fine with methanol or second: 0.05-0.5% phosphoric acid solution=1-9: 9-1 is a mobile phase; The detection wavelength is 200-500nm, and number of theoretical plate must not calculate with the ferulic acid peak and is lower than 4000; Precision takes by weighing the ferulic acid reference substance, with dissolve with methanol and make every 1ml and contain ferulic acid 5-20 μ g, promptly gets reference substance solution; Get capsule, each 1.0g of tablet or granule 5g, the accurate title, decide, and puts in the Backflow bottle, the accurate methanol 10-100ml that adds claims to decide weight, water-bath backflow 20-80 minute, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, leave standstill, 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 each 5-25 μ l of need testing solution of drawing injects hplc determination content; In this oral formulations, contain Radix Angelicae Sinensis in the capsule, must not be less than in 0.05mg/g, the tablet and contain Radix Angelicae Sinensis, must not be less than in 0.05mg/g, the granule and contain Radix Angelicae Sinensis, must not be less than 0.005mg/g in ferulic acid in ferulic acid in ferulic acid.

Method of quality control comprises more specifically:

Character observed comprises:

Content is differentiated, be may further comprise the steps:

(1) get capsule, each 1g of tablet respectively, or get granule 5g, add petroleum ether 30ml, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 0.3g, 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 on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=4: 1 is developing solvent, launches, take 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.

(2) get capsule respectively, each 5g of tablet, or get granule 20g, add ethyl acetate 40ml, supersound extraction 30 minutes filters, and abandons acetic acid ethyl fluid, residue adds the n-butyl alcohol 60ml of water saturation again, supersound extraction 1 hour filters, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 3 times, each 20ml, abandon water lotion, n-butyl alcohol liquid water bath method, residue water dissolution, be transferred to the D101 type macroporous resin column of having handled well (1.5 * 15cm), use the 50ml water elution, abandon water liquid, reuse 70% ethanol 50ml eluting, collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the Radix Paeoniae 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 on same silica gel g thin-layer plate with 1% sodium hydroxide solution preparation, with ethyl acetate: formic acid: glacial acetic acid: water=15: 1: 1: 2 is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) get capsule respectively, each 5g of tablet, or get granule 20g, add methanol 50ml, reflux, extract, 40 minutes filters the filtrate evaporate to dryness, residue adds water 30ml makes dissolving, with the n-butanol extraction of water saturation 2 times, and each 25ml, merge n-butyl alcohol liquid, with ammonia solution washing 2 times, each 25ml, abandon the ammonia washing liquid, water lotion is abandoned in the water 30ml washing of the saturated mistake of n-butyl alcohol liquid reuse n-butyl alcohol 1 time, and n-butyl alcohol liquid is put evaporate to dryness in the water-bath, residue adds methanol 5ml makes dissolving, cross the neutral alumina post (1.5 * 20cm, 5g), with 40% methanol solution 100ml eluting, collect eluent, put water bath method, residue adds methanol 0.5ml makes dissolving, as need testing solution; Other gets the astragaloside 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 on same silica gel g thin-layer plate, with chloroform: methanol: lower floor's solution of water=13: 7: 2 is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with contrast chromatograph corresponding position on, daylight shows identical sepia speckle down; Ultra-violet lamp 365nm shows identical orange-yellow fluorescence speckle down.

(4) get capsule respectively, each 5g of tablet, or get granule 20g, add ethyl acetate 40ml, supersound extraction 30 minutes filters, and abandons acetic acid ethyl fluid, residue adds the n-butyl alcohol 60ml of water saturation again, supersound extraction 1 hour filters, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 3 times, each 20ml, abandon water lotion, n-butyl alcohol liquid water bath method, residue water dissolution, be transferred to the D101 type macroporous resin column of having handled well (1.5 * 15cm), use the 50ml water elution, abandon water liquid, reuse 70% ethanol 50ml eluting, collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g in addition, the 20ml that adds diethyl ether, reflux, extract, filtered after 1 hour, abandon filtrate, medicinal residues add methanol 30ml, and water-bath reflux, extract, 1 hour filters, filtrate evaporate to dryness, residue add water 20ml makes dissolving, and is transferred in the separatory funnel, with the n-butanol extraction of water saturation 3 times, each 30ml merges n-butyl alcohol liquid, the washing of the saturated mistake of reuse n-butyl alcohol 3 times, each 30ml abandons water lotion, n-butyl alcohol liquid water bath method, residue add methanol 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 on same silica gel g thin-layer plate with the preparation of 1% sodium hydroxide solution, with ethyl acetate: formic acid: glacial acetic acid: water=15: 1: 1: 2 be developing solvent, expansion, take out, dry, under daylight, inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

Official method comprises content inspection:

Ferulic acid adopts high performance liquid chromatography, and chromatographic column is the C18 post, and is fine with methanol or second: 0.085% phosphoric acid solution=1.6: 8.4 is a mobile phase; The detection wavelength is 324nm, and number of theoretical plate must not calculate with the ferulic acid peak and is lower than 5000; Precision takes by weighing the ferulic acid reference substance, with dissolve with methanol and make every 1ml and contain ferulic acid 10 μ g, promptly gets reference substance solution; Get capsule, each 1.0g of tablet or granule 5g, the accurate title, decide, and puts in the Backflow bottle, the accurate methanol 25ml that adds claims to decide weight, and water-bath refluxed 45 minutes, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, leave standstill, microporous filter membrane with 0.45 μ m filters, and gets filtrate, promptly gets need testing solution; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject hplc determination content; In this oral formulations, contain Radix Angelicae Sinensis in the capsule, must not be less than in 0.05mg/g, the tablet and contain Radix Angelicae Sinensis, must not be less than in 0.05mg/g, the granule and contain Radix Angelicae Sinensis, must not be less than 0.005mg/g in ferulic acid in ferulic acid in ferulic acid.

Compared with the prior art, it is simple to the present invention is directed to original preparation blood-bonifying Chinese angelica extract Iuality control criterion, shortcomings such as product quality is wayward, method of quality control to this preparation is studied, improved the quality standard of 'Buxuedanggui ', guarantee the clinical efficacy of this preparation, reached the purpose of invention.

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, ferulaic acid content study on determination method

1, the selection of mobile phase:

Mobile phase 1: mixed solution fine with methanol or second, phosphoric acid solution is a mobile phase;

Mobile phase 2: the mixed solution with methanol, water and glacial acetic acid is a mobile phase;

Mobile phase 3: the mixed solution with methanol and glacial acetic acid is a mobile phase.

Result: fine with methanol or second: 0.05-0.5% phosphoric acid solution=1-9: 9-1 is a mobile phase, the negative sample chromatogram is at non-false positive peak, ferulic acid position, ferulic acid separates fully (separating degree＞1.5) with close impurity peaks, promptly ferulic acid separates with other components fully under this condition.Optimal flow is mutually: second is fine: 0.085% phosphoric acid solution=16: 84.

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 1 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 (%)	0.1538 2.75	0.2392 2.82	0.2268 1.89

Result of the test shows that reflux, extract, more can be extracted the ferulic acid in the preparation fully, so the selective extraction method is the water-bath reflux, extract.

2.2 extraction choice of Solvent

Get preparation 1.0g, add methanol, 50% ethanol, ethanol 50ml respectively, claim decide weight, water-bath reflux, extract, 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 (%)	0.2405 1.32	0.2381 0.79	0.1932 1.48

Result of the test shows that methanol more can leach the ferulic acid 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, and the ferulic acid average content is 0.2407mg/g, and RSD is 1.88%.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)	0.2412	0.2438	0.2386	0.2456	0.2341	0.2407	1.88

4, ferulic acid stability test in the need testing solution

Get preparation, by the preparation method of above-mentioned test liquid, measure its ferulic acid peak area respectively at 0,2,4,6,8 hour, average peak area is 404106, and RSD is 1.19%.Show that ferulic acid is good at 8 hours internal stabilities 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	406474	409737	406178	399316	398826	404106	1.19

Two, Radix Angelicae Sinensis thin layer Study on Identification

With the Radix Angelicae Sinensis control medicinal material is contrast.

Need testing solution preparation method one: get preparation 1g, add diethyl ether, reflux, extract, 30min is put coldly, filters, and the low temperature evaporate to dryness, residue adds anhydrous alcohol solution, as need testing solution; Lack the Radix Angelicae Sinensis negative controls with the method preparation.

Need testing solution preparation method two: get preparation 1g, add petroleum ether, supersound process 30 minutes filters, the filtrate evaporate to dryness, and residue adds anhydrous alcohol solution, as need testing solution; Lack the Radix Angelicae Sinensis negative controls with the method preparation.

Developing solvent is selected: the mixed solution of normal hexane and the various ratios of ethyl acetate is developing solvent respectively.

The result: adopting method two to prepare need testing solution, is 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: normal hexane: ethyl acetate=4: 1.

Three, Radix Paeoniae Alba Study on Identification

With the Radix Paeoniae reference substance is contrast.

Need testing solution preparation method one: get preparation 5g respectively, add ethyl acetate, supersound extraction 30 minutes, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 50ml of water saturation again, supersound extraction 1 hour, filter, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 3 times, each 20ml abandons water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution, be transferred to the D101 type macroporous resin column handled well (1.5 * 15cm), use the 50ml water elution, abandon water liquid, reuse 70% ethanol 50ml eluting is collected eluent, evaporate to dryness, residue adds methanol 1mL makes dissolving, as need testing solution; Lack the Radix Paeoniae negative controls with the method preparation.

Need testing solution preparation method two: get preparation 5g, add methanol 3, supersound process 30min, put coldly, filter evaporate to dryness, residue adds water 10mL, and dissolving adds water-saturated n-butanol and extracts 2 times, each 20mL merges water bath method, residue adds ethanol 20mL, puts on the neutral alumina post of having handled well (＜115cm, neutral alumina 5g), water bath method, residue add methanol 1mL dissolving, as need testing solution; Lack the Radix Paeoniae negative controls with the method preparation.

Developing solvent is selected: respectively with the mixed solution of ethyl acetate, formic acid, glacial acetic acid and the various ratios of water; The mixed solution of chloroform, ethyl acetate and the various ratios of methanol is developing solvent.

The result: employing method one preparation need testing solution, with ethyl acetate: formic acid: glacial acetic acid: water=5-30: 0.5-5: 0.5-5: 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: ethyl acetate: formic acid: glacial acetic acid: water=15: 2: 2: 1.5.

Four, Radix Astragali Study on Identification

With the astragaloside reference substance is contrast.

Need testing solution preparation method one: get preparation 5g, add methanol 50ml, reflux, extract, 40 minutes, filter, filtrate evaporate to dryness, residue add water 30ml makes dissolving, with the n-butanol extraction of water saturation 2 times, each 25ml merges n-butyl alcohol liquid, with ammonia solution washing 2 times, each 25ml abandons the ammonia washing liquid, the water 30ml washing of the saturated mistake of n-butyl alcohol liquid reuse n-butyl alcohol 1 time, abandon water lotion, n-butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methanol 5ml makes dissolving, crosses neutral alumina post (1.5 * 20cm, 5g), with 40% methanol solution 100ml eluting, collect eluent, put water bath method, residue adds methanol 0.5ml makes dissolving, as need testing solution; Lack Radix Astragali negative controls with the method preparation.

Need testing solution preparation method two: get preparation 5g, porphyrize adds methanol 50mL supersound process 20min, filter, evaporate to dryness, residue add water 20mL dissolving, be transferred in the separatory funnel, the 20mL that adds diethyl ether extraction 2 times discards ether solution, adds water-saturated n-butanol 20mL, extract 2 times, merge n-butyl alcohol liquid, water bath method, residue add methanol 2mL dissolving as test liquid; Lack Radix Astragali negative controls with the method preparation.

Developing solvent is selected: the mixed solution with chloroform, the various ratios of first alcohol and water is developing solvent respectively.

The result: employing method one preparation need testing solution, with chloroform: lower floor's solution of methanol: water=5-30: 3-15: 1-10 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: chloroform: methanol: lower floor's solution of water=15: 10: 4.

Five, Radix Glycyrrhizae Study on Identification

With the Radix Glycyrrhizae control medicinal material is contrast.

Need testing solution preparation method one: get preparation 5g, add dilute hydrochloric acid 1ml, water 40ml shakes up, adding water-saturated n-butanol extracts 5 times, each 30ml merges n-butyl alcohol liquid, water bath method, residue dissolves with methanol 5ml gradation, be transferred in the 100ml round-bottomed flask, volatilization goes methanol solution, residue to add hydrochloric acid 5ml, chloroform 50ml, reflux 3h.Put coldly, divide and to get chloroform layer, filter, with 10ml chloroform gradation washing filter paper, merging filtrate, water bath method, residue add dissolve with methanol and are settled to 2ml, as test sample liquid; Lack the Radix Glycyrrhizae negative controls with the method preparation.

Need testing solution preparation method two: get preparation 5g respectively, add ethyl acetate, supersound extraction 30 minutes, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 50ml of water saturation again, supersound extraction 1 hour, filter, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 3 times, each 20ml abandons water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution, be transferred to the D101 type macroporous resin column handled well (1.5 * 15cm), use the 50ml water elution, abandon water liquid, reuse 70% ethanol 50ml eluting is collected eluent, evaporate to dryness, residue adds methanol 1mL makes dissolving, as need testing solution; Lack the Radix Glycyrrhizae negative controls with the method preparation.

Need testing solution preparation method three: get preparation 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, and reflux 1 hour is put coldly, filters, and filtrate evaporate to dryness, residue add ethanol or methanol 2ml makes dissolving, as need testing solution;

Developing solvent is selected: respectively with 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 petroleum ether (60～90 ℃), benzene, ethyl acetate 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: adopt method two to prepare need testing solution, with ethyl acetate: formic acid: glacial acetic acid: water=5-30: 0.5-5: 0.5-5: 0.5-5 is that its separating degree of developing solvent is good, and the speckle colour developing is clear, and negative control is noiseless, the method favorable reproducibility.Best developing solvent is an ethyl acetate: formic acid: glacial acetic acid: water=20: 3: 2: 1.

The invention has the advantages that: method of quality control of the present invention is on former statutory standards basis, increases the high effective liquid chromatography for measuring content of ferulic acid; 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.

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

Content is differentiated, comprised

(1) get capsule 1g, add petroleum ether 20ml, supersound process 10 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 0.5ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis 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 on same silica gel g thin-layer plate, 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.

(2) get capsule 5g, add ethyl acetate 20ml, supersound extraction 10 minutes, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 20ml of water saturation again, supersound extraction 0.5 hour, filter, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 1 time, each 10ml abandons water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution, be transferred to the D101 type macroporous resin column handled well (1.5 * 15cm), use the 20ml water elution, abandon water liquid, reuse 20% ethanol 20ml eluting is collected eluent, evaporate to dryness, residue adds methanol 0.5ml makes dissolving, as need testing solution; Other gets the Radix Paeoniae reference substance, adds methanol and makes the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate with 0.5% sodium hydroxide solution preparation, with ethyl acetate: formic acid: glacial acetic acid: water=5: 0.5: 5: 5 is developing solvent, launch, take out, dry, spray is with 5% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) get capsule 5g, add methanol 20ml, reflux, extract, 10 minutes, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, with the n-butanol extraction of water saturation 1 time, each 10ml merges n-butyl alcohol liquid, with ammonia solution washing 1 time, each 10ml abandons the ammonia washing liquid, the water 10ml washing of the saturated mistake of n-butyl alcohol liquid reuse n-butyl alcohol 1 time, abandon water lotion, n-butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methanol 2ml makes dissolving, crosses neutral alumina post (1.5 * 20cm, 5g), with 20% methanol solution 50ml eluting, collect eluent, put water bath method, residue adds methanol 0.2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: lower floor's solution of water=5: 15: 1 is developing solvent, launch, take out, dry, spray is with 5% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with contrast chromatograph corresponding position on, daylight shows identical sepia speckle down; Ultra-violet lamp 200nm shows identical orange-yellow fluorescence speckle down.

(4) get capsule 5g, add ethyl acetate 20ml, supersound extraction 10 minutes, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 20ml of water saturation again, supersound extraction 0.5 hour, filter, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 1 time, each 10ml abandons water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution, be transferred to the D101 type macroporous resin column handled well (1.5 * 15cm), use the 20ml water elution, abandon water liquid, reuse 20% ethanol 20ml eluting is collected eluent, evaporate to dryness, residue adds methanol 0.5ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g in addition, the 10ml that adds diethyl ether, reflux, extract, filtered after 0.5 hour, abandon filtrate, medicinal residues add methanol 10ml, and water-bath reflux, extract, 0.5 hour filters, filtrate evaporate to dryness, residue add water 10ml makes dissolving, and is transferred in the separatory funnel, with the n-butanol extraction of water saturation 1 time, each 10ml merges n-butyl alcohol liquid, the washing of the saturated mistake of reuse n-butyl alcohol 1 time, each 10ml abandons water lotion, n-butyl alcohol liquid water bath method, residue add methanol 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 on same silica gel g thin-layer plate with the preparation of 0.5% sodium hydroxide solution, with ethyl acetate: formic acid: glacial acetic acid: water=5: 5: 0.5: 0.5 be developing solvent, expansion, take out, dry, under daylight, inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the 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 ferulic acid is carried out assay, may further comprise the steps:

Ferulic acid adopts high performance liquid chromatography, and chromatographic column is C4, and with methanol: 0.05% phosphoric acid solution=1: 9 is a mobile phase; The detection wavelength is 200nm, and number of theoretical plate must not calculate with the ferulic acid peak and is lower than 4000; Precision takes by weighing the ferulic acid reference substance, with dissolve with methanol and make every 1ml and contain ferulic acid 5 μ g, promptly gets reference substance solution; Get capsule 1.0g, the accurate title, decide, and puts in the Backflow bottle, the accurate methanol 10ml that adds claims to decide weight, and water-bath refluxed 20 minutes, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, leave standstill, 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 each 5 μ l of need testing solution of drawing inject hplc determination content; In this oral formulations, contain Radix Angelicae Sinensis in the capsule, must not be less than 0.05mg/g in ferulic acid.

The quality control of embodiment 2 tablets

Content is differentiated, comprised

(1) get tablet 1g, add petroleum ether 150ml, supersound process 60 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 0.5-2ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis 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 on same silica gel g thin-layer plate, 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.

(2) get tablet 5g, add ethyl acetate 200ml, supersound extraction 60 minutes, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 100ml of water saturation again, supersound extraction 3 hours, filter, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 8 times, each 50ml abandons water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution, be transferred to the D101 type macroporous resin column handled well (1.5 * 15cm), use the 100ml water elution, abandon water liquid, reuse 85% ethanol 100ml eluting is collected eluent, evaporate to dryness, residue adds methanol 2ml makes dissolving, as need testing solution; Other gets the Radix Paeoniae 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 on same silica gel g thin-layer plate with 5% sodium hydroxide solution preparation, with ethyl acetate: formic acid: glacial acetic acid: water=30: 5: 0.5: 0.5 is developing solvent, launch, take out, dry, spray is with 20% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) get tablet 5g, add methanol 200ml, reflux, extract, 100 minutes, filter, filtrate evaporate to dryness, residue add water 100ml makes dissolving, with the n-butanol extraction of water saturation 8 times, each 100ml merges n-butyl alcohol liquid, with ammonia solution washing 8 times, each 100ml abandons the ammonia washing liquid, the water 100ml washing of the saturated mistake of n-butyl alcohol liquid reuse n-butyl alcohol 5 times, abandon water lotion, n-butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methanol 10ml makes dissolving, crosses neutral alumina post (1.5 * 20cm, 5g), with 85% methanol solution 200ml eluting, collect eluent, put water bath method, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the astragaloside 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 on same silica gel g thin-layer plate, with chloroform: methanol: lower floor's solution of water=30: 3: 10 is developing solvent, launch, take out, dry, spray is with 20% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with contrast chromatograph corresponding position on, daylight shows identical sepia speckle down; Ultra-violet lamp 500nm shows identical orange-yellow fluorescence speckle down.

(4) get tablet 5g, add ethyl acetate 200ml, supersound extraction 60 minutes, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 100ml of water saturation again, supersound extraction 3 hours, filter, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 8 times, each 50ml abandons water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution, be transferred to the D101 type macroporous resin column handled well (1.5 * 15cm), use the 100ml water elution, abandon water liquid, reuse 85% ethanol 100ml eluting is collected eluent, evaporate to dryness, residue adds methanol 2ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g in addition, the 100ml that adds diethyl ether, reflux, extract, filtered after 5 hours, abandon filtrate, medicinal residues add methanol 100ml, and water-bath reflux, extract, 5 hours filters, filtrate evaporate to dryness, residue add water 50ml makes dissolving, and is transferred in the separatory funnel, with the n-butanol extraction of water saturation 8 times, each 100ml merges n-butyl alcohol liquid, the washing of the saturated mistake of reuse n-butyl alcohol 5 times, each 50ml abandons water lotion, n-butyl alcohol liquid water bath method, residue add methanol 2ml makes dissolving, in contrast medical material solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 25 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate with the preparation of 5% sodium hydroxide solution, with ethyl acetate: formic acid: glacial acetic acid: water=30: 0.5: 5: 5 be developing solvent, expansion, take out, dry, under daylight, inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the 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 ferulic acid is carried out assay, may further comprise the steps:

Ferulic acid adopts high performance liquid chromatography, and chromatographic column is the C8 post, is mobile phase with fine 0.5% phosphoric acid solution of second=9: 1; The detection wavelength is 500nm, and number of theoretical plate must not calculate with the ferulic acid peak and is lower than 4000; Precision takes by weighing the ferulic acid reference substance, with dissolve with methanol and make every 1ml and contain ferulic acid 20 μ g, promptly gets reference substance solution; It is fixed to get the accurate title of tablet 1.0g, puts in the Backflow bottle, and the accurate methanol 100ml that adds claims to decide weight, water-bath backflow 80 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, leave standstill, 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 each 25 μ l of need testing solution of drawing inject hplc determination content; In this oral formulations, contain Radix Angelicae Sinensis in the tablet, must not be less than 0.05mg/g in ferulic acid.

The quality control of embodiment 3 granules:

Discriminating to granule comprises:

(1) get granule 5g, add petroleum ether 30ml, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 0.3g, 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 on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=4: 1 is developing solvent, launches, take 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.

(2) get granule 20g, add ethyl acetate 40ml, supersound extraction 30 minutes, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 60ml of water saturation again, supersound extraction 1 hour, filter, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 3 times, each 20ml abandons water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution, be transferred to the D101 type macroporous resin column handled well (1.5 * 15cm), use the 50ml water elution, abandon water liquid, reuse 70% ethanol 50ml eluting is collected eluent, evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the Radix Paeoniae 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 on same silica gel g thin-layer plate with 1% sodium hydroxide solution preparation, with ethyl acetate: formic acid: glacial acetic acid: water=15: 1: 1: 2 is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) get granule 20g, add methanol 50ml, reflux, extract, 40 minutes, filter, filtrate evaporate to dryness, residue add water 30ml makes dissolving, with the n-butanol extraction of water saturation 2 times, each 25ml merges n-butyl alcohol liquid, with ammonia solution washing 2 times, each 25ml abandons the ammonia washing liquid, the water 30ml washing of the saturated mistake of n-butyl alcohol liquid reuse n-butyl alcohol 1 time, abandon water lotion, n-butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methanol 5ml makes dissolving, crosses neutral alumina post (1.5 * 20cm, 5g), with 40% methanol solution 100ml eluting, collect eluent, put water bath method, residue adds methanol 0.5ml makes dissolving, as need testing solution; Other gets the astragaloside 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 on same silica gel g thin-layer plate, with chloroform: methanol: lower floor's solution of water=13: 7: 2 is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with contrast chromatograph corresponding position on, daylight shows identical sepia speckle down; Ultra-violet lamp 365nm shows identical orange-yellow fluorescence speckle down.

(4) get granule 20g, add ethyl acetate 40ml, supersound extraction 30 minutes, filter, abandon acetic acid ethyl fluid, residue adds the n-butyl alcohol 60ml of water saturation again, supersound extraction 1 hour, filter, the washing of the saturated mistake of filtrate usefulness n-butyl alcohol 3 times, each 20ml abandons water lotion, n-butyl alcohol liquid water bath method, the residue water dissolution, be transferred to the D101 type macroporous resin column handled well (1.5 * 15cm), use the 50ml water elution, abandon water liquid, reuse 70% ethanol 50ml eluting is collected eluent, evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g in addition, the 20ml that adds diethyl ether, reflux, extract, filtered after 1 hour, abandon filtrate, medicinal residues add methanol 30ml, and water-bath reflux, extract, 1 hour filters, filtrate evaporate to dryness, residue add water 20ml makes dissolving, and is transferred in the separatory funnel, with the n-butanol extraction of water saturation 3 times, each 30ml merges n-butyl alcohol liquid, the washing of the saturated mistake of reuse n-butyl alcohol 3 times, each 30ml abandons water lotion, n-butyl alcohol liquid water bath method, residue add methanol 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 on same silica gel g thin-layer plate with the preparation of 1% sodium hydroxide solution, with ethyl acetate: formic acid: glacial acetic acid: water=15: 1: 1: 2 be developing solvent, expansion, take out, dry, under daylight, inspect, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.

Official method to content inspection is

Granule of the present invention should meet Chinese Pharmacopoeia about the pertinent regulations under the granule item.

To the contained assay that carries out, may further comprise the steps:

Ferulic acid adopts high performance liquid chromatography, and chromatographic column is the C18 post, and is fine with methanol or second: 0.085% phosphoric acid solution=1.6: 8.4 is a mobile phase; The detection wavelength is 324nm, and number of theoretical plate must not calculate with the ferulic acid peak and is lower than 5000; Precision takes by weighing the ferulic acid reference substance, with dissolve with methanol and make every 1ml and contain ferulic acid 10 μ g, promptly gets reference substance solution; Get granule 5g, the accurate title, decide, and puts in the Backflow bottle, and the accurate methanol 25ml that adds claims to decide weight, water-bath refluxed 45 minutes, put coldly, claimed to decide weight again, supplied the weight that subtracts mistake with methanol, shook up, leave standstill,, get filtrate, promptly get need testing solution with the microporous filter membrane filtration of 0.45 μ m; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject hplc determination content;

Contain Radix Angelicae Sinensis in the granule in ferulic acid, must not be less than 0.005mg/g.

Claims

1, a kind of method of quality control of 'Buxuedanggui ' is characterized in that, comprises character is observed, and content is differentiated, official method is checked content, the ferulic acid that contains is carried out the step of assay.

2, the method for claim 1 is characterized in that, described 'Buxuedanggui ' 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:

(3) get capsule respectively, each 5g of tablet, or get granule 20g, add methanol 20-200ml, reflux, extract, 10-100 minute, filter, filtrate evaporate to dryness, residue add water 10-100ml makes dissolving, with the n-butanol extraction of water saturation 1-8 time, each 10-100ml, merge n-butyl alcohol liquid, with ammonia solution washing 1-8 time, each 10-100ml, abandon the ammonia washing liquid, water lotion is abandoned in the water 10-100ml washing of the saturated mistake of n-butyl alcohol liquid reuse n-butyl alcohol 1-5 time, and n-butyl alcohol liquid is put evaporate to dryness in the water-bath, residue adds methanol 2-10ml makes dissolving, (1.5 * 20cm 5g), uses 20-85% methanol solution 50-200ml eluting to cross the neutral alumina post, collect eluent, put water bath method, residue adds methanol 0.2-1ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.2-1mg, 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 on-silica gel g thin-layer plate, with chloroform: lower floor's solution of methanol: water=5-30: 3-15: 1-10 is developing solvent, launch, take out, dry, spray is with the 5-20% ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with contrast chromatograph corresponding position on, daylight shows identical sepia speckle down; Ultra-violet lamp 200-500nm shows identical orange-yellow fluorescence speckle down.

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 'Buxuedanggui ' is made by following Chinese medicine raw materials by weight proportion:

Radix Angelicae Sinensis 300-500g, Radix Rehmanniae Preparata (wine steams) 10-50g, the Radix Paeoniae Alba (wine stir-fry) 10-50g, Rhizoma Chuanxiong (wine stir-fry) 5-20g, the Radix Astragali (processed with honey) 10-50g, Radix Codonopsis 10-50g, Radix Glycyrrhizae 5-20g.

8, according to the method for claim 1, it is characterized in that, wherein character is observed and comprise: for capsule, character is: the product content thing is brown xanchromatic fine powder or granule, acrid in the mouth, little hardship; For the tablet character be: medicine shows brown xanchromatic fine powder or granule; Acrid in the mouth, little hardship; For the granule character be: product is the xanchromatic granule of palm fibre.

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, be may further comprise the steps:

Official method comprises content inspection: