CN101187651B

CN101187651B - Detection method of tongsaimai formualtion

Info

Publication number: CN101187651B
Application number: CN2006100978429A
Authority: CN
Inventors: 夏月; 陈荣明; 李吉峰; 殷书梅; 秦小荣
Original assignee: NANXING PHARMACEUTICAL CO Ltd JIANGSU PROVINCE
Current assignee: JIANGSU KANION YANGGUANG PHARMACEUTICAL CO., LTD.; Jiangsu Kanion Pharmaceutical Co Ltd
Priority date: 2006-11-15
Filing date: 2006-11-15
Publication date: 2012-06-13
Anticipated expiration: 2026-11-15
Also published as: CN101187651A

Abstract

The invention discloses a method for controlling quality of a traditional Chinese medicine preparation, which is applied to test a passing plug vein preparation. The passing plug vein preparation is composed of the traditional Chinese medicines as astragalus root, campanumaea pilosula, dendrobium nobile, honeysuckle flower, achyranthes and the like. The quality control method of the invention deletes an identification of anoelica sinensis, and TLC qualitative identifications of the astragal us root, scrophularia root, licorice root, scoparone and the astragalus root are increased further. An HPLC determination method and a content limiting regulation of glycyrrhizin acid, galuteolin and astragaloside are also increased further. Preparation quality of the passing plug vein preparation can be further controlled thoroughly through the invention. The method of controlling quality of the traditional Chinese medicine preparation of the invention is mainly applicable to passing plug vein oralpreparations as pills, granules, capsules, powder medicines, soft capsules and dropping pills or oral liquid.

Description

A kind of detection method of logical plug arteries and veins preparation

Technical field

The present invention relates to a kind of detection method of Chinese medicine preparation, particularly a kind of detection method of logical plug arteries and veins preparation.

Background technology

TONGSAIMAI PIAN State Food and Drug Administration standard medicine, its standard is following:

[prescription] Radix Angelicae Sinensis, the root of bidentate achyranthes, the Radix Astragali, Radix Codonopsis, the stem of noble dendrobium, radix scrophulariae, honeysuckle, Radix Glycyrrhizae;

[proterties] these article are sugar coated tablet or Film coated tablets, remove to show sepia behind the dressing; Sweet, the little hardship, puckery of distinguishing the flavor of.

3 of these article are got in [discriminating] (1), remove dressing, and porphyrize adds ethanol 40ml, reflux 2 hours; Filter, the filtrating evaporate to dryness, residue adds water 15ml makes dissolving, extracts 2 times with the ethyl acetate jolting, each 15ml; Merge ethyl acetate liquid, evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution.Other gets the forulic acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution.Test according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000); Drawing need testing solution 10～20 μ l, reference substance solution 10 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with chloroform-ethyl acetate-formic acid (5: 4: 0.5); Launch; Take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

(2) get 10 of these article, remove dressing, porphyrize adds ethanol 40ml, reflux 30 minutes; Filter, filtrating is steamed near and is done, and residue adds water 10ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 10ml; Reflux 3 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 2 times with the chloroform jolting; Each 15ml, combined chloroform liquid is concentrated into 1ml, as need testing solution.Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution.Test according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000); Drawing each 10 μ l of above-mentioned two kinds of solution, put on same silica gel g thin-layer plate respectively, is developping agent with thiacyclohexane-acetone-ethyl acetate (4: 2: 1), launches; Take out; Dry, spray is with 10% phosphomolybdic acid ethanol solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(3) in the chromatogram of these article gained under (assay) item, should present the chromatographic peak identical with the reference substance chromatographic retention.

[inspection] should meet each item regulation (appendix ID of Chinese Pharmacopoeia version in 2000) relevant under the tablet item.

[assay] measured according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000).

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent; Methanol-water-glacial acetic acid (9: 88: 3) is a moving phase; The detection wavelength is 330nm.Number of theoretical plate calculates by the chlorogenic acid peak should be not less than 3000.

The preparation precision of reference substance solution takes by weighing chlorogenic acid reference substance 8mg, puts in the brown measuring bottle of 50ml, adds 90% methyl alcohol and makes dissolving in right amount and be diluted to scale; Shake up, precision is measured 5ml, puts in the brown measuring bottle of 50ml; Add 90% methyl alcohol to scale, shake up, promptly get (containing chlorogenic acid 16 μ g among every 1ml).

10 of these article are got in the preparation of need testing solution, remove dressing, and porphyrize is got 1g, and accurate the title decides; Put in the tool plug conical flask, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight, reflux 1 hour; Take out, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methyl alcohol; Shake up, filter, get continuous solution, promptly get.

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

Every of these article contain honeysuckle in chlorogenic acid (C16H18O9), must not be less than 0.11mg.

[function with cure mainly] promoting blood circulation and removing obstruction in channels, supplementing qi and nourishing yin.Be used for light moderate atherosclerotic snow bolt property cerebral infarction (ishemic stroke apoplex involving the channels and collaterals) convalescence blood stagnancy due to deficiency of QI disease, symptom shows as hemiplegia, hemianesthesia, facial paralysis, speech is unfavorable, limbs are felt to go down or disappearance etc.; The scorchingly hot disease that is used for thromboangiitis (gangrene of finger or toe).

Summary of the invention

Technical matters to be solved by this invention is the deficiency to prior art, and a kind of new method of quality control that can further comprehensively control the Chinese medicine preparation of the logical plug arteries and veins quality of the pharmaceutical preparations is provided.

Technical matters to be solved by this invention is to realize through following technical scheme.The present invention is a kind of method of quality control of Chinese medicine preparation, is characterized in, it is applicable to detects logical plug arteries and veins preparation, and its discrimination method is following,

(1) get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 1～5g, porphyrize is with methyl alcohol ultrasonic Extraction 1-3 time; Add methyl alcohol 40-60ml at every turn and extracted 10-30 minute, filter behind the merging extract, the filtrating evaporate to dryness adds water 20-40ml and makes dissolving; Extract 2-4 time with water-saturated n-butanol, each 20-40ml merges normal butyl alcohol liquid; With ammonia solution washing 1-3 time, each 20-40ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 3-8ml makes dissolving, last D-101 macroporous adsorptive resins; Water 40-60ml wash-out discards water liquid, with 30-50% ethanol 20-40ml wash-out respectively; Discard eluent, continue to collect eluent with 60-80% ethanol 40-60ml wash-out; Evaporate to dryness, residue add methyl alcohol 1-3ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with the 5-15% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 1～5g, porphyrize adds ethanol 30-50ml, reflux 20-40 minute; Filter, filtrating is steamed near and is done, and residue adds water 5-15ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 5-15ml; Reflux 2-4 hour, take out, put and steam in the water-bath to there not being the alcohol flavor, extract 1-3 time with the chloroform jolting; Each 10-20ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 5-15% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent, and methanol-water-glacial acetic acid (17: 80: 3) is a moving phase, and the detection wavelength is 330nm, and number of theoretical plate is pressed chlorogenic acid and calculated, and should be lower than 1500;

It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, puts in the brown bottle, adds 50% methyl alcohol and process the solution that every 1ml contains 0.01mg～0.05mg, as reference substance solution;

This product is got in the preparation of need testing solution, perhaps gets the dry fine powder of this product or this product dry extract fine powder, takes by weighing 0.5g～1g; The accurate title, decide, and places the 50ml volumetric flask, adds the nearly scale of 50% methyl alcohol; Extracted 15～20 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and in the gained chromatogram, should present the chromatographic peak identical with the reference substance chromatographic retention.

Technical matters to be solved by this invention can also further realize through following technical scheme.The method of quality control of above-described a kind of Chinese medicine preparation is characterized in, wherein the discrimination method of Radix Codonopsis is following: get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder; Add 5～15 times of amount methyl alcohol, reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color.

Technical matters to be solved by this invention can also further realize through following technical scheme.The method of quality control of above-described a kind of Chinese medicine preparation is characterized in, wherein the discrimination method of radix scrophulariae is following, gets radix scrophulariae control medicinal material 0.2g, shines medicinal material solution according to need testing solution preparation method described in the discriminating of above-mentioned Radix Codonopsis in pairs with legal system; According to thin-layered chromatography test, draw need testing solution and each 10 μ l of above-mentioned control medicinal material solution in the discriminating of above-mentioned Radix Codonopsis, put respectively on same silica gel g thin-layer plate; With 7: 1: 2 normal butyl alcohol-glacial acetic acid-water was developping agent; Launch 14cm, take out, dry; Spray is with the vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color.

Technical matters to be solved by this invention can also further realize through following technical scheme.The method of quality control of above-described a kind of Chinese medicine preparation is characterized in, wherein the discrimination method of Radix Glycyrrhizae is following, gets these article, perhaps gets the dry fine powder of these article or these article dry extract fine powder; Add 5～15 times of amount methyl alcohol, reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds water 30ml makes dissolving; Extract 3 times with water saturated normal butyl alcohol jolting, each 20ml merges normal butyl alcohol liquid, extracts 3 times with the 1%NaOH jolting; Each 20ml merges alkaline solution, with watery hydrochloric acid adjust pH to 3, extracts 3 times with water saturated normal butyl alcohol jolting; Each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

Technical matters to be solved by this invention can also further realize through following technical scheme.The method of quality control of above-described a kind of Chinese medicine preparation is characterized in, gets these article, perhaps gets the dry fine powder of these article or these article dry extract fine powder; Add 5～15 times of amount methyl alcohol, reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with the chloroform jolting, each 30ml, combined chloroform extract; Put and be concentrated into driedly in the water-bath, precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

Technical matters to be solved by this invention can also further realize through following technical scheme.The method of quality control of above-described a kind of Chinese medicine preparation is characterized in, wherein the chlorogenic acid contents assay method is following, according to high effective liquid chromatography for measuring,

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent, and methanol-water-glacial acetic acid (17: 80: 3) is a moving phase, and the detection wavelength is 330nm; Number of theoretical plate is pressed chlorogenic acid and is calculated, and should be lower than 1500;

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder 0.5g～1g, and accurate the title decides; Place 50～100ml volumetric flask; Add the nearly scale of 50% methyl alcohol, extracted 15～20 minutes, put cold; Add 50% methyl alcohol to scale, get need testing solution with filtering with microporous membrane;

Technical matters to be solved by this invention can also further realize through following technical scheme.The method of quality control of above-described a kind of Chinese medicine preparation is characterized in, wherein the content assaying method of ammonium glycyrrhetate is following,

According to high effective liquid chromatography for measuring,

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent; Methyl alcohol-0.1% phosphoric acid solution was a moving phase in 64: 36; The detection wavelength is 250nm; Number of theoretical plate should be not less than 2000 by the glycyrrhizic acid peak;

It is an amount of that the preparation precision of reference substance solution takes by weighing the ammonium glycyrrhetate reference substance, adds methyl alcohol and process the solution that every 1ml contains 0.2mg, amounts to glycyrrhizic acid 0.1959mg, promptly gets;

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, and accurate the title decides, and puts in the flask; 10～30 times of amounts of accurate adding, 70% ethanol claim to decide weight, and circumfluence method was extracted 45 minutes, put to room temperature; Claim again to decide weight, supply the weight that subtracts mistake, shake up with 70% ethanol; Filter, get subsequent filtrate, promptly get;

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and calculate, and promptly get.

Technical matters to be solved by this invention can also further realize through following technical scheme.The method of quality control of above-described a kind of Chinese medicine preparation is characterized in, wherein the content assaying method of galuteolin is following,

According to high effective liquid chromatography for measuring,

Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica, are mobile phase A with the acetonitrile, are Mobile phase B with 0.5% glacial acetic acid solution, and according to the form below carries out gradient elution; Detect wavelength 350nm, number of theoretical plate calculates by the galuteolin peak should be not less than 2000;

It is an amount of that the preparation precision of reference substance solution takes by weighing the galuteolin reference substance, adds 70% ethanol and process the solution that every ml contains 10 μ g;

These article are got in the preparation of need testing solution, perhaps get these article through the dry thing fine powder after the pre-treatment, and accurate the title decides; Place volumetric flask (embodiment can be listed as: 25ml or 50ml or 100ml); Add an amount of ultrasonic dissolving that makes of 70% ethanol, be settled to scale, filter with miillpore filter; Get subsequent filtrate, promptly get;

Accurate respectively reference substance solution and the need testing solution 10 μ l of drawing of determination method inject liquid chromatograph, measure, and calculate, and promptly get.

Technical matters to be solved by this invention can also further realize through following technical scheme.The method of quality control of above-described a kind of Chinese medicine preparation is characterized in, wherein the content assaying method of Astragaloside IV is following,

According to high effective liquid chromatography for measuring,

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent; 77: 23 methanol-waters are moving phase; Detect with EISD.Number of theoretical plate should be not less than 2000 by the Astragaloside IV peak;

It is an amount of that the preparation precision of reference substance solution takes by weighing the Astragaloside IV reference substance, adds methyl alcohol and process the solution that every 1ml contains 0.2mg, promptly gets;

This product is got in the preparation of need testing solution, perhaps gets the dry fine powder of this product or this product dry extract fine powder, and accurate the title decides, and puts in the flask, and precision adds 10～20 times of amount methyl alcohol ultrasonic Extraction 20min; Filter, residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount; Merging filtrate and washing lotion are concentrated into driedly, and residue adds water 10ml, and low-grade fever makes dissolving, extract 3 times with water saturated n-butanol jolting; Each 20ml merges n-butanol extracting liquid, extracts 2 times with ammonia solution, and each 20ml discards ammoniacal liquor; N-butanol liquid evaporate to dryness, residue dissolves with methyl alcohol and is transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;

Accurate respectively reference substance solution 5 μ l, 20 μ l and the need testing solution 10 μ l of drawing of determination method inject liquid chromatograph, calculate with external standard two-point method logarithmic equation, promptly get.

The formulation of the logical plug arteries and veins preparation described in the present invention can be any formulation on the pharmacy, and the preferred oral preparation comprises pill, granule, tablet, capsule, powder, soft capsule, pill and oral liquid; Preferred tablet in the oral formulations.Refer generally in " get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder " described in the technical scheme of the present invention:, can directly be ground into the fine powder sampling when preparation is pill or granule; When preparation was tablet, plain sheet directly was ground into the fine powder sampling, and Film coated tablets or sugar coated tablet are ground into the fine powder sampling after removing dressing; After preparation removes photoresist capsule during for capsule, be ground into the fine powder sampling, as be soft capsule, after removing the capsule skin; Add 3～5 times of water gagings, mixing is again to hang down extractions such as polar organic solvent such as ether, chloroform, methylene chloride, sherwood oil 2～3 times; Aqueous suspension after the de-oiling is dry, is ground into the fine powder sampling again; If other formulation can be handled according to this area conventional treatment method.

Its prescription of medicine involved in the present invention " logical plug arteries and veins preparation " is identical with the prescription described in the background technology, is made up of Radix Angelicae Sinensis, honeysuckle, Radix Codonopsis, radix scrophulariae, the Radix Astragali, the root of bidentate achyranthes, the stem of noble dendrobium and Radix Glycyrrhizae.Its function with cure mainly: the training fill blood, replenishing the vital essence and removing heat, promoting blood circulation and removing blood stasis, clearing and activating the channels and collaterals.The scorchingly hot card that is used for Buerger's disease (gangrene of finger or toe).

Below be example with the TONGSAIMAI PIAN agent, [drafting of (quality standard (draft)) describes to the method for quality control of the logical plug of the present invention arteries and veins preparation.

TONGSAIMAI PIAN is made up of Chinese medicines such as the Radix Astragali, Radix Codonopsis, the stem of noble dendrobium, honeysuckle, the roots of bidentate achyranthes.Since kind protection in 1994; The annual production of this product increases year by year; Supply falls short of demand for product, for preventing fake products, guarantees product quality; Tentative TONGSAIMAI PIAN standard [YBZ08372004] detection method of former country studied, increased the TLC qualitative identification method of Radix Codonopsis, radix scrophulariae, Radix Glycyrrhizae, the Radix Astragali, the stem of noble dendrobium newly; The HPLC content assaying method of glycyrrhizic acid, galuteolin, Astragaloside IV and content limit regulation.

[discriminating] left out the discriminating of Radix Angelicae Sinensis; Differentiate in the tentative standard that (1) item is down the thin layer discriminating of Radix Angelicae Sinensis; But we find the forulic acid less stable as reference substance in the study on the stability of product, and the meeting natural degradation is unfavorable for the control of the quality stability of TONGSAIMAI PIAN finished product.So consider it is left out.Increased the TLC qualitative identification of Radix Codonopsis, radix scrophulariae, Radix Glycyrrhizae, escoparone, the Radix Astragali.

[assay]

One, chlorogenic acid: owing to strengthened raw material control aborning, use authentic medicinal herbs, and each workshop section is tightened control, reduce the loss of effective ingredient, make that the chlorogenic acid detection level increases in the dry extract.Detection method Central Plains test sample disposal route is comparatively loaded down with trivial details, chlorogenic acid peak shape broad, use 50% methyl alcohol sonicated instead after, prepare easier; Number of theoretical plate is higher, and peak shape is better, investigates good stability through methodology; The recovery is higher, and method is simple and feasible, can better control its content.Assay method is following:

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

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent, and methanol-water-glacial acetic acid (17: 80: 3) is a moving phase, and the detection wavelength is 330nm.Number of theoretical plate is pressed chlorogenic acid and is calculated, and should be lower than 1500.

It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, puts in the brown bottle, adds 50% methyl alcohol and process the solution that every 1ml contains 0.01mg, as reference substance solution.

10 of these article are got in the preparation of need testing solution, remove dressing, take by weighing 0.5g, and accurate the title decides; Place the 50ml volumetric flask, add the nearly scale of 50% methyl alcohol, ultrasonic Extraction 20 minutes (40KHz 250W); Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale.

Every of these article contain honeysuckle with chlorogenic acid (C ₁₆H ₁₈O ₉) meter, must not be less than 0.35mg.

Learn the investigation checking through efficient liquid-phase chromatography method, above-mentioned determination of chlorogenic acid method can detect chlorogenic acid contents in logical plug arteries and veins preparation and the intermedium accurately.

Two, glycyrrhizic acid: Radix Glycyrrhizae is one of flavour of a drug in the TONGSAIMAI PIAN prescription, have invigorate the spleen and benefit qi, the effect of clearing heat and detoxicating, expelling phlegm and arresting coughing, relieving spasm to stop pain, coordinating the drug actions of a prescription.In order to control the quality of TONGSAIMAI PIAN better, we select for use the principal ingredient glycyrrhizic acid of Radix Glycyrrhizae as the assay index, adopt efficient liquid-phase chromatography method to make an experiment, and method is easy, and favorable reproducibility is easy to operate, and accuracy is high.

(1) instrument and reagent and reagent

Instrument: Agilent 1100 liquid chromatographs; Mettler AE240 electronic balance (100,000/); H66025 supersonic wave cleaning machine (Wuxi Ultrasonic Electronic Equipment Factory); PTHW type electric jacket (Ying Yu of Gongyi City gives magnificent instrument plant).

Reagent: methyl alcohol (chromatographically pure, TEDIA, USA); Water is ultrapure water; All the other reagent are analyzed pure all available from Shanghai chemical reagent company limited.

Reagent: the ammonium glycyrrhetate reference substance (available from Nat'l Pharmaceutical & Biological Products Control Institute, supplies assay usefulness, lot number: 110731-200408); TONGSAIMAI PIAN is provided by the Southern Star medicine company; The TONGSAIMAI PIAN blank sample (lacking the Radix Glycyrrhizae preparation) that lacks Radix Glycyrrhizae by full prescription.

(2) chromatographic condition

Chromatographic column: ZORBAX SB-C ₁₈(5 μ m, 4.6mm * 250mm);

Use octadecylsilane chemically bonded silica to be filling agent; Methyl alcohol-0.1% phosphoric acid solution (64: 36) is a moving phase; The detection wavelength is 250nm.Number of theoretical plate should be not less than 2000 by the glycyrrhizic acid peak.

The preparation of reference substance solution: it is an amount of that precision takes by weighing the ammonium glycyrrhetate reference substance, adds methyl alcohol and process the solution that every 1ml contains 0.2mg, promptly gets.

Flow velocity: 1.0ml/min; Column temperature: 30 ℃.

Measure the selection of wavelength: 250nm is as the detection wavelength of ammonium glycyrrhetate.

The system suitability test: under these conditions, greater than 2000, degree of separation reaches baseline separation greater than 1.5 with glycyrrhizic acid peak theory of computation plate number.

(3) preparation of need testing solution

1. method for distilling is investigated

These article of getting (lot number: 060208) an amount of, remove film-coating after, porphyrize is got 2g, parallel 2 parts; The accurate title, decide, and puts in the flask, and the accurate 70% methyl alcohol 50ml that adds claims to decide weight, uses ultrasonic (power 300W respectively; Frequency 20kHz) and circumfluence method extracted 30 minutes, put to room temperature, claim again to decide weight, supply the weight that subtracts mistake with 70% methyl alcohol, shake up; Filter with miillpore filter (0.45 μ m), get subsequent filtrate, measure its content, the result sees the following form.

Different Extraction Method is extracted the result relatively

Result of the test shows, this product uses circumfluence method to extract, and institute's glycyrrhizic acid content of surveying is higher, so definite use circumfluence method is as the test sample method for distilling.

2. extracting solvent investigates

(1) extracting solvent investigates

These article of getting (lot number: 060208) an amount of, remove film-coating after, porphyrize is got 2g, parallel 2 parts; The accurate title, decide, and puts in the flask, adds different solvents 50ml respectively, claims to decide weight, and circumfluence method was extracted 30 minutes; Put to room temperature, claim again to decide weight, supply the weight that subtracts mistake, shake up with same solvent; Filter with miillpore filter (0.45 μ m), get subsequent filtrate, measure its content, the result sees the following form.

Different solvents extracts the result relatively

Test findings shows, with ethanol is to extract solvent, and institute's glycyrrhizic acid content of surveying is higher, so definite ethanol is test sample extraction solvent.

(2) extracting solvent strength investigates

These article of getting (lot number: 060208) an amount of, remove film-coating after, porphyrize is got 2g, parallel five parts; The accurate title, decide, and puts in the flask, adds different concentration ethanol 50ml respectively, claims to decide weight, and circumfluence method was extracted 30 minutes; Put to room temperature, claim again to decide weight, supply the weight that subtracts mistake, shake up with same solvent; Filter with miillpore filter (0.45 μ m), get subsequent filtrate, measure its content, the result sees the following form.

Different concentration ethanol is extracted the result relatively

Test findings shows, serves as to extract solvent with 70% ethanol, and institute's glycyrrhizic acid content of surveying is higher, so definite 70% ethanol is test sample extraction solvent.

3. extraction time is investigated

These article of getting (lot number: 060207) an amount of, remove film-coating after, porphyrize is got 2g, parallel four parts; The accurate title, decide, and puts in the flask, adds 70% ethanol 50ml, claims to decide weight, and circumfluence method was extracted 15 minutes, 30 minutes, 45 minutes, 60 minutes respectively; Put to room temperature, claim again to decide weight, supply the weight that subtracts mistake, shake up with 70% ethanol; Filter with miillpore filter (0.45 μ m), get subsequent filtrate, measure its content, the result sees the following form.

Different extraction times are extracted the result relatively

Test findings shows, with 70% alcohol reflux 45min, the glycyrrhizic acid content of surveying higher, confirm that reflux extracting time is 45min.

According to above test findings, confirm that the test sample preparation method is: it is an amount of to get these article, remove film-coating after, porphyrize is got 2g; The accurate title, decide, and puts in the flask, adds 70% ethanol 50ml, claims to decide weight; Circumfluence method was extracted 45 minutes, put to room temperature, claimed to decide weight again, supplied the weight that subtracts mistake with 70% ethanol; Shake up, filter, get subsequent filtrate, measure its content with miillpore filter (0.45 μ m).

Learn the investigation checking through efficient liquid-phase chromatography method, the content assaying method of glycyrrhizic acid can detect the content of glycyrrhizic acid in logical plug arteries and veins preparation and the intermedium accurately.

According to the assay test findings of many batches of TONGSAIMAI PIANs, glycyrrhizic acid content should be not less than the 0.4mg/ sheet in tentative these article.

Three, galuteolin: honeysuckle is one of main flavour of a drug in the TONGSAIMAI PIAN prescription, has effect clearing heat and detoxicating, wind-heat dissipating, because its principal ingredient chlorogenic acid ubiquity in Chinese crude drug; In order to control the quality of TONGSAIMAI PIAN better, we select for use characteristic effective constituent galuteolin in the honeysuckle as the assay index, adopt efficient liquid-phase chromatography method to make an experiment; Method is easy; Favorable reproducibility, easy to operate, accuracy is high.

(1) instrument and reagent and reagent:

Instrument: Agilent 1100 liquid chromatographs; Mettler AE240 electronic balance (100,000/); H66025 supersonic wave cleaning machine (Wuxi Ultrasonic Electronic Equipment Factory).

Reagent: acetonitrile (chromatographically pure, TEDIA, USA); Water is ultrapure water; All the other reagent are analyzed pure all available from Shanghai chemical reagent company limited.

Reagent: the galuteolin reference substance (available from Nat'l Pharmaceutical & Biological Products Control Institute, supplies assay usefulness, lot number: 111520-200201); TONGSAIMAI PIAN is provided by the Southern Star medicine company; The TONGSAIMAI PIAN blank sample (lacking the honeysuckle preparation) that lacks honeysuckle by full prescription.

(2) chromatographic condition

Chromatographic condition: ZORBAX SB-C ₁₈(5 μ m, 4.6mm * 150mm), be mobile phase A with the acetonitrile, be Mobile phase B with 0.5% glacial acetic acid solution, according to the form below carries out gradient elution; Detect wavelength 350nm, flow velocity: 1.0ml/min; Column temperature: 30 ℃.

(3) sample preparation methods

The preparation of reference substance solution: it is an amount of that precision takes by weighing the galuteolin reference substance, adds 70% ethanol and process the solution that every ml contains 10 μ g.

The preparation of need testing solution: get 10 of these article, remove film-coating after, porphyrize is got 1.5g; The accurate title, decide, and places the 25ml volumetric flask, adds 70% ethanol ultrasonic in right amount (power 300W, frequency 20kHz) and make dissolving; Be settled to scale, filter, get subsequent filtrate, promptly get with miillpore filter.

The preparation of negative need testing solution: get the TONGSAIMAI PIAN dry extract that does not contain honeysuckle, press the sample solution preparation, promptly get.

Learn the investigation checking through efficient liquid-phase chromatography method, the content assaying method of galuteolin can detect the content of galuteolin in logical plug arteries and veins preparation and the intermedium accurately.

According to the content data of many batches of galuteolins, in conjunction with big production actual conditions, the content limit of ordering galuteolin in the TONGSAIMAI PIAN temporarily is: every contains honeysuckle with galuteolin (C ₂₁H ₂₀O ₁₁) meter, must not be less than 0.10mg.

Four, Astragaloside IV: the Radix Astragali is one of main flavour of a drug in the TONGSAIMAI PIAN prescription; In order to control the quality of TONGSAIMAI PIAN better, we select for use characteristic effective constituent Astragaloside IV in the Radix Astragali as the assay index, adopt efficient liquid-phase chromatography method to make an experiment; Method is easy; Favorable reproducibility, easy to operate, accuracy is high.

(1) instrument and reagent and reagent: instrument: Waters 600 liquid chromatographs, Alltech 500 type EISDs; Mettler AE240 electronic balance (100,000/); H66025 supersonic wave cleaning machine (Wuxi Ultrasonic Electronic Equipment Factory).

Reagent: the Astragaloside IV reference substance (available from Nat'l Pharmaceutical & Biological Products Control Institute, supplies assay usefulness, lot number: 0781-200109); TONGSAIMAI PIAN is provided by the Southern Star medicine company; The TONGSAIMAI PIAN blank sample (lacking Radix Astragali preparation) that lacks the Radix Astragali by full prescription.

(2) chromatographic condition.Chromatographic condition: Lichrospher 5-C ₁₈(5 μ m, 4.6mm * 250mm), be moving phase with methanol-water (75: 25), flow velocity: 1.0ml/min; Column temperature: 30 ℃; Detect with EISD.

(3) sample preparation methods

The preparation of reference substance solution: it is an amount of that precision takes by weighing the Astragaloside IV reference substance, adds methyl alcohol and process the solution that every 1ml contains 0.2mg, promptly gets.

The preparation of need testing solution: get 10 of these article, remove film-coating after, porphyrize is got 1.5g, accurate claims surely, adds methyl alcohol 20ml ultrasonic Extraction (power 300W; Frequency 20kHz) 20min filters, and residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount; Merging filtrate and washing lotion are concentrated into driedly, and residue adds water 10ml, and low-grade fever makes dissolving, extract 3 times with water saturated normal butyl alcohol jolting; Each 20ml merges n-butanol extracting liquid, extracts 2 times with ammonia solution, and each 20ml discards ammoniacal liquor; Normal butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets.

The preparation of negative need testing solution: get the TONGSAIMAI PIAN dry extract that does not contain the Radix Astragali, press the sample solution preparation and promptly get.

Learn the investigation checking through efficient liquid-phase chromatography method, the content assaying method of Astragaloside IV can detect the content of Astragaloside IV in logical plug arteries and veins preparation and the intermedium accurately.

According to the content data of many batches of Astragaloside IVs, in conjunction with big production actual conditions, the content limit of ordering Astragaloside IV in the TONGSAIMAI PIAN temporarily is every and contains the Radix Astragali with Astragaloside IV (C ₄₁H ₆₈O ₁₄) meter, must not be less than 0.04mg.

Embodiment

Embodiment 1.A kind of method of quality control of Chinese medicine preparation, it is applicable to detects logical plug arteries and veins preparation, and its discrimination method is following,

(1) get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 1g, porphyrize is with methyl alcohol ultrasonic Extraction 1 time; Add methyl alcohol 60ml at every turn and extracted 30 minutes, filter behind the merging extract, the filtrating evaporate to dryness adds water 20ml and makes dissolving; Extract 2 times with water-saturated n-butanol, each 40ml merges normal butyl alcohol liquid; With ammonia solution washing 1 time, each 40ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 3ml makes dissolving, last D-101 macroporous adsorptive resins; Water 40ml wash-out discards water liquid, with 30% ethanol 40ml wash-out respectively; Discard eluent, continue to collect eluent with 60% ethanol 60ml wash-out; Evaporate to dryness, residue add methyl alcohol 1ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with 5% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 1g, porphyrize adds ethanol 30ml, reflux 40 minutes; Filter, filtrating is steamed near and is done, and residue adds water 5ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 5ml; Reflux 2 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 1 time with the chloroform jolting; Each 20ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 5% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, take by weighing 0.5g, and accurate the title decides; Place the 50ml volumetric flask, add the nearly scale of 50% methyl alcohol, extracted 15 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

Embodiment 2.A kind of method of quality control of Chinese medicine preparation, it is applicable to detects logical plug arteries and veins preparation, and its discrimination method is following,

(1) get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 5g, porphyrize is with methyl alcohol ultrasonic Extraction 3 times; Add methyl alcohol 40ml at every turn and extracted 10 minutes, filter behind the merging extract, the filtrating evaporate to dryness adds water 40ml and makes dissolving; Extract 4 times with water-saturated n-butanol, each 20ml merges normal butyl alcohol liquid; With ammonia solution washing 3 times, each 20ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 8ml makes dissolving, last D-101 macroporous adsorptive resins; Water 60ml wash-out discards water liquid, with 50% ethanol 20ml wash-out respectively; Discard eluent, continue to collect eluent with 80% ethanol 40ml wash-out; Evaporate to dryness, residue add methyl alcohol 3ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with 15% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 5g, porphyrize adds ethanol 50ml, reflux 20 minutes; Filter, filtrating is steamed near and is done, and residue adds water 15ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 15ml; Reflux 4 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 3 times with the chloroform jolting; Each 10ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 15% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, take by weighing 1g, and accurate the title decides; Place the 50ml volumetric flask, add the nearly scale of 50% methyl alcohol, extracted 20 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

Embodiment 3.A kind of method of quality control of Chinese medicine preparation, it is applicable to detects logical plug arteries and veins preparation, and its discrimination method is following,

(1) get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 2g, porphyrize is with methyl alcohol ultrasonic Extraction 2 times; Add methyl alcohol 50ml at every turn and extracted 20 minutes, filter behind the merging extract, the filtrating evaporate to dryness adds water 30ml and makes dissolving; Extract 3 times with water-saturated n-butanol, each 30ml merges normal butyl alcohol liquid; With ammonia solution washing 2 times, each 30ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 5ml makes dissolving, last D-101 macroporous adsorptive resins; Water 50ml wash-out discards water liquid, with 40% ethanol 30ml wash-out respectively; Discard eluent, continue to collect eluent with 70% ethanol 50ml wash-out; Evaporate to dryness, residue add methyl alcohol 2ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 2g, porphyrize adds ethanol 40ml, reflux 30 minutes; Filter, filtrating is steamed near and is done, and residue adds water 10ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 10ml; Reflux 3 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 2 times with the chloroform jolting; Each 15ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 10% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, take by weighing 0.8g, and accurate the title decides; Place the 50ml volumetric flask, add the nearly scale of 50% methyl alcohol, extracted 18 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

Embodiment 4.Like the method for quality control of embodiment 1 described a kind of Chinese medicine preparation, wherein the discrimination method of Radix Codonopsis is following: get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder, add 5 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color.

Embodiment 5.Like the method for quality control of embodiment 2 described a kind of Chinese medicine preparations, wherein the discrimination method of Radix Codonopsis is following: get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder, add 15 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color.

Embodiment 6.Like the method for quality control of embodiment 1 described a kind of Chinese medicine preparation, wherein the discrimination method of radix scrophulariae is following, gets radix scrophulariae control medicinal material 0.2g, shines medicinal material solution according to need testing solution preparation method described in the discriminating of Radix Codonopsis among the embodiment 4 or 5 in pairs with legal system; According to thin-layered chromatography test, draw need testing solution and each 10 μ l of above-mentioned control medicinal material solution in the discriminating of Radix Codonopsis among the embodiment 4 or 5, put respectively on same silica gel g thin-layer plate; With 7: 1: 2 normal butyl alcohol-glacial acetic acid-water was developping agent; Launch 14cm, take out, dry; Spray is with the vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color.

Embodiment 7.Like the method for quality control of embodiment 5 described a kind of Chinese medicine preparations, wherein the discrimination method of radix scrophulariae is following, gets radix scrophulariae control medicinal material 0.2g, shines medicinal material solution according to need testing solution preparation method described in the discriminating of Radix Codonopsis in pairs with legal system; According to the thin-layered chromatography test, need testing solution and each 10 μ l of above-mentioned control medicinal material solution put respectively on same silica gel g thin-layer plate in the discriminating of absorption Radix Codonopsis; With 7: 1: 2 normal butyl alcohol-glacial acetic acid-water was developping agent; Launch 14cm, take out, dry; Spray is with the vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color.

Embodiment 8.Like the method for quality control of embodiment 1 described a kind of Chinese medicine preparation, wherein the discrimination method of Radix Glycyrrhizae is following, gets these article, perhaps gets the dry fine powder of these article or these article dry extract fine powder, adds 5 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds water 30ml makes dissolving; Extract 3 times with water saturated normal butyl alcohol jolting, each 20ml merges normal butyl alcohol liquid, extracts 3 times with the 1%NaOH jolting; Each 20ml merges alkaline solution, with watery hydrochloric acid adjust pH to 3, extracts 3 times with water saturated normal butyl alcohol jolting; Each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

Embodiment 9.Like the method for quality control of embodiment 7 described a kind of Chinese medicine preparations, wherein the discrimination method of Radix Glycyrrhizae is following, gets these article, perhaps gets the dry fine powder of these article or these article dry extract fine powder, adds 15 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds water 30ml makes dissolving; Extract 3 times with water saturated normal butyl alcohol jolting, each 20ml merges normal butyl alcohol liquid, extracts 3 times with the 1%NaOH jolting; Each 20ml merges alkaline solution, with watery hydrochloric acid adjust pH to 3, extracts 3 times with water saturated normal butyl alcohol jolting; Each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

Embodiment 10.Like the method for quality control of embodiment 1 described a kind of Chinese medicine preparation, wherein the discrimination method of escoparone is following, gets these article, perhaps gets the dry fine powder of these article or these article dry extract fine powder; Add 5 times of amount methyl alcohol, reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with the chloroform jolting, each 30ml, combined chloroform extract; Put and be concentrated into driedly in the water-bath, precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

Embodiment 11.Like the method for quality control of embodiment 9 described a kind of Chinese medicine preparations, wherein the discrimination method of escoparone is following, gets these article, perhaps gets the dry fine powder of these article or these article dry extract fine powder; Add 15 times of amount methyl alcohol, reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with the chloroform jolting, each 30ml, combined chloroform extract; Put and be concentrated into driedly in the water-bath, precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

Embodiment 12.Method of quality control like embodiment 1 described a kind of Chinese medicine preparation: wherein the chlorogenic acid contents assay method is following, according to high effective liquid chromatography for measuring,

It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, puts in the brown bottle, adds 50% methyl alcohol and process the solution that every 1ml contains 0.01mg, as reference substance solution;

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder 0.5g, and accurate the title decides; Place the 50ml volumetric flask, add the nearly scale of 50% methyl alcohol, extracted 15 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

Embodiment 13.Method of quality control like embodiment 11 described a kind of Chinese medicine preparations: wherein the chlorogenic acid contents assay method is following, according to high effective liquid chromatography for measuring,

It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, puts in the brown bottle, adds 50% methyl alcohol and process the solution that every 1ml contains 0.05mg, as reference substance solution;

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder 1g, and accurate the title decides; Place the 100ml volumetric flask, add the nearly scale of 50% methyl alcohol, extracted 20 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

Embodiment 14.Like the method for quality control of embodiment 1 described a kind of Chinese medicine preparation, wherein the content assaying method of ammonium glycyrrhetate is following, according to high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, and accurate the title decides, and puts in the flask; 10 times of amounts of accurate adding, 70% ethanol claim to decide weight, and circumfluence method was extracted 45 minutes, put to room temperature; Claim again to decide weight, supply the weight that subtracts mistake, shake up with 70% ethanol; Filter, get subsequent filtrate, promptly get;

Embodiment 15.Like the method for quality control of embodiment 13 described a kind of Chinese medicine preparations, wherein the content assaying method of ammonium glycyrrhetate is following, according to high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, and accurate the title decides, and puts in the flask; 30 times of amounts of accurate adding, 70% ethanol claim to decide weight, and circumfluence method was extracted 45 minutes, put to room temperature; Claim again to decide weight, supply the weight that subtracts mistake, shake up with 70% ethanol; Filter, get subsequent filtrate, promptly get;

Embodiment 16.Like the method for quality control of embodiment 1 described a kind of Chinese medicine preparation, wherein the content assaying method of galuteolin is following,

According to high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, perhaps get these article through the dry thing fine powder after the pre-treatment, and accurate the title decides, and places the 25ml volumetric flask, add an amount of ultrasonic dissolving that makes of 70% ethanol, are settled to scale, filter with miillpore filter, get subsequent filtrate, promptly get;

Embodiment 17.Like the method for quality control of embodiment 15 described a kind of Chinese medicine preparations, wherein the content assaying method of galuteolin is following,

According to high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, perhaps get these article through the dry thing fine powder after the pre-treatment, and accurate the title decides, and places the 50ml volumetric flask, add an amount of ultrasonic dissolving that makes of 70% ethanol, are settled to scale, filter with miillpore filter, get subsequent filtrate, promptly get;

Embodiment 18.Like the method for quality control of embodiment 1 described a kind of Chinese medicine preparation, wherein the content assaying method of Astragaloside IV is following, according to high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, and accurate the title decides, and put in the flask, and precision adds 10 times of amount methyl alcohol ultrasonic Extraction 20min; Filter, residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount; Merging filtrate and washing lotion are concentrated into driedly, and residue adds water 10ml, and low-grade fever makes dissolving, extract 3 times with water saturated normal butyl alcohol jolting; Each 20ml merges n-butanol extracting liquid, extracts 2 times with ammonia solution, and each 20ml discards ammoniacal liquor; Normal butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;

Embodiment 19.Like the method for quality control of embodiment 17 described a kind of Chinese medicine preparations, wherein the content assaying method of Astragaloside IV is following, according to high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, and accurate the title decides, and put in the flask, and precision adds 20 times of amount methyl alcohol ultrasonic Extraction 20min; Filter, residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount; Merging filtrate and washing lotion are concentrated into driedly, and residue adds water 10ml, and low-grade fever makes dissolving, extract 3 times with water saturated normal butyl alcohol jolting; Each 20ml merges n-butanol extracting liquid, extracts 2 times with ammonia solution, and each 20ml discards ammoniacal liquor; Normal butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;

Embodiment 20.The method of quality control of a kind of TONGSAIMAI PIAN agent or granule or pill or powder.

[discriminating]

(1) get these article, 1g, porphyrize is with methyl alcohol ultrasonic Extraction 2 times; Add methyl alcohol 50ml at every turn and extracted 20 minutes, filter behind the merging extract, the filtrating evaporate to dryness adds water 30ml and makes dissolving; Extract 3 times with water-saturated n-butanol, each 30ml merges normal butyl alcohol liquid; With ammonia solution washing 2 times, each 30ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 5ml makes dissolving, last D-101 macroporous adsorptive resins; Water 50ml wash-out discards water liquid, with 40% ethanol 30ml wash-out respectively; Discard eluent, continue to collect eluent with 70% ethanol 50ml wash-out; Evaporate to dryness, residue add methyl alcohol 2ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get these article 1g, porphyrize adds ethanol 40ml, reflux 30 minutes; Filter, filtrating is steamed near and is done, and residue adds water 10ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 10ml; Reflux 3 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 2 times with the chloroform jolting; Each 15ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 10% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

These article fine powder is got in the preparation of need testing solution, takes by weighing 0.5g, accurate claims surely, places the 50ml volumetric flask, adds the nearly scale of 50% methyl alcohol, extracts 15 minutes, puts coldly, adds 50% methyl alcohol to scale, gets need testing solution with filtering with microporous membrane;

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and in the gained chromatogram, should present the chromatographic peak identical with the reference substance chromatographic retention;

(4) get these article fine powder, add 5 times of amount methyl alcohol, reflux 1 hour filters; The filtrating evaporate to dryness, residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color;

(5) get radix scrophulariae control medicinal material 0.2g, shine medicinal material solution in pairs with legal system according to need testing solution preparation method described in the discriminating of Radix Codonopsis under (4) item; According to thin-layered chromatography test, draw (4) down need testing solution and each 10 μ l of above-mentioned control medicinal material solution in the discriminating of Radix Codonopsis, put respectively on same silica gel g thin-layer plate; With 7: 1: 2 normal butyl alcohol-glacial acetic acid-water was developping agent; Launch 14cm, take out, dry; Spray is with the vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color;

(6) get these article fine powder, add 10 times of amount methyl alcohol, reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, and each 20ml merges normal butyl alcohol liquid, extracts 3 times with the 1%NaOH jolting; Each 20ml merges alkaline solution, with watery hydrochloric acid adjust pH to 3, extracts 3 times with water saturated normal butyl alcohol jolting; Each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(7) get these article fine powder, add 10 times of amount methyl alcohol, reflux 1 hour filters; The filtrating evaporate to dryness, residue adds water 30ml makes dissolving, extracts 3 times with the chloroform jolting, each 30ml; The combined chloroform extract is put and is concentrated into driedly in the water-bath, and precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

[the content side is fixed]

Chlorogenic acid shines high effective liquid chromatography for measuring,

These article fine powder 0.5g is got in the preparation of need testing solution, accurate claims surely, places the 50ml volumetric flask, adds the nearly scale of 50% methyl alcohol, extracts 15 minutes, puts coldly, adds 50% methyl alcohol to scale, gets need testing solution with filtering with microporous membrane;

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

Ammonium glycyrrhetate shines high effective liquid chromatography for measuring,

This product fine powder is got in the preparation of need testing solution, and accurate the title decides, and puts in the flask, and 20 times of amounts of accurate adding, 70% ethanol claim to decide weight; Circumfluence method was extracted 45 minutes, put to room temperature, claimed to decide weight again, supplied the weight that subtracts mistake with 70% ethanol; Shake up, filter, get subsequent filtrate, promptly get;

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and calculate, and promptly get;

Galuteolin shines high effective liquid chromatography for measuring,

These article fine powder is got in the preparation of need testing solution, and accurate the title decides, and places the 100ml volumetric flask, adds an amount of ultrasonic dissolving that makes of 70% ethanol, is settled to scale, filters with miillpore filter, gets subsequent filtrate, promptly gets;

Astragaloside IV shines high effective liquid chromatography for measuring,

This product fine powder is got in the preparation of need testing solution, and accurate the title decides, and puts in the flask, and 15 times of amounts of accurate adding methyl alcohol ultrasonic Extraction 20min filter; Residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount, merging filtrate and washing lotion; Be concentrated into driedly, residue adds water 10ml, and low-grade fever makes dissolving, extracts 3 times with water saturated n-butanol jolting, each 20ml; Merge n-butanol extracting liquid, extract 2 times with ammonia solution, each 20ml discards ammoniacal liquor, n-butanol liquid evaporate to dryness; Residue dissolves with methyl alcohol and is transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;

Embodiment 21.The method of quality control of a kind of logical plug arteries and veins Film coated tablets or sugar coated tablet.

[discriminating]

(1) gets these article, remove dressing, get fine powder 2g, porphyrize; With methyl alcohol ultrasonic Extraction 2 times, add methyl alcohol 50ml at every turn and extracted 20 minutes, filter the filtrating evaporate to dryness after merging extract; Add water 30ml and make dissolving, extract 3 times with water-saturated n-butanol, each 30ml merges normal butyl alcohol liquid; With ammonia solution washing 2 times, each 30ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 5ml makes dissolving, last D-101 macroporous adsorptive resins; Water 50ml wash-out discards water liquid, with 40% ethanol 30ml wash-out respectively; Discard eluent, continue to collect eluent with 70% ethanol 50ml wash-out; Evaporate to dryness, residue add methyl alcohol 2ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get these article, remove dressing, get fine powder 2g, porphyrize adds ethanol 40ml; Reflux 30 minutes filters, and filtrating is steamed near and done, and residue adds water 10ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 10ml; Reflux 3 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 2 times with the chloroform jolting; Each 15ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 10% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

These article are got in the preparation of need testing solution, remove dressing, get fine powder, take by weighing 1g, accurately claim surely, place the 50ml volumetric flask, add the nearly scale of 50% methyl alcohol, extract 20 minutes, put coldly, add 50% methyl alcohol to scale, get need testing solution with filtering with microporous membrane;

(4) get these article, remove dressing, get fine powder, add 10 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color;

(6) get these article, remove dressing, get fine powder, add 10 times of amount methyl alcohol, reflux 1 hour; Filter, the filtrating evaporate to dryness, residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Merge normal butyl alcohol liquid, extract 3 times with the 1%NaOH jolting, each 20ml merges alkaline solution, with watery hydrochloric acid adjust pH to 3; Extract 3 times with water saturated normal butyl alcohol jolting, each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(7) get these article, remove dressing, get fine powder, add 10 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds water 30ml makes dissolving; Extract 3 times each 30ml, combined chloroform extract with the chloroform jolting; Put and be concentrated into driedly in the water-bath, precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

[the content side is fixed]

Chlorogenic acid shines high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, remove dressing, get fine powder 1g, accurately claim surely, place the 100ml volumetric flask, add the nearly scale of 50% methyl alcohol, extract 20 minutes, put coldly, add 50% methyl alcohol to scale, get need testing solution with filtering with microporous membrane;

This product is got in the preparation of need testing solution, removes dressing, gets fine powder, and accurate the title decides, and puts in the flask; 20 times of amounts of accurate adding, 70% ethanol claim to decide weight, and circumfluence method was extracted 45 minutes, put to room temperature, claimed to decide weight again; Supply the weight that subtracts mistake with 70% ethanol, shake up, filter, get subsequent filtrate, promptly get;

Galuteolin shines high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, remove dressing, get fine powder, and accurate the title decides, and places the 50ml volumetric flask, add an amount of ultrasonic dissolving that makes of 70% ethanol, are settled to scale, filter with miillpore filter, get subsequent filtrate, promptly get;

Astragaloside IV shines high effective liquid chromatography for measuring,

This product is got in the preparation of need testing solution, removes dressing, gets fine powder, and accurate the title decides, and puts in the flask, and precision adds 15 times of amount methyl alcohol ultrasonic Extraction 20min; Filter, residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount; Merging filtrate and washing lotion are concentrated into driedly, and residue adds water 10ml, and low-grade fever makes dissolving, extract 3 times with water saturated n-butanol jolting; Each 20ml merges n-butanol extracting liquid, extracts 2 times with ammonia solution, and each 20ml discards ammoniacal liquor; N-butanol liquid evaporate to dryness, residue dissolves with methyl alcohol and is transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;

Embodiment 22.A kind of method of quality control of TONGSAIMAI JIAONANG agent.

[discriminating]

(1) gets these article, behind the capsule that removes photoresist, get fine powder 3g, porphyrize; With methyl alcohol ultrasonic Extraction 2 times, add methyl alcohol 50ml at every turn and extracted 20 minutes, filter the filtrating evaporate to dryness after merging extract; Add water 30ml and make dissolving, extract 3 times with water-saturated n-butanol, each 30ml merges normal butyl alcohol liquid; With ammonia solution washing 2 times, each 30ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 3-8ml makes dissolving, last D-101 macroporous adsorptive resins; Water 30ml wash-out discards water liquid, with 40% ethanol 30ml wash-out respectively; Discard eluent, continue to collect eluent with 70% ethanol 50ml wash-out; Evaporate to dryness, residue add methyl alcohol 2ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get these article, behind the capsule that removes photoresist, get fine powder 3g, porphyrize adds ethanol 40ml; Reflux 30 minutes filters, and filtrating is steamed near and done, and residue adds water 10ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 10ml; Reflux 3 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 2 times with the chloroform jolting; Each 15ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 10% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, puts in the brown bottle, adds 50% methyl alcohol and process the solution that every 1ml contains 0.02mg, as reference substance solution;

These article are got in the preparation of need testing solution, behind the capsule that removes photoresist, get fine powder 1g, accurate claim surely, place the 50ml volumetric flask, add the nearly scale of 50% methyl alcohol, extract 15 minutes, put coldly, add 50% methyl alcohol to scale, get need testing solution with filtering with microporous membrane;

(4) get these article, behind the capsule that removes photoresist, get fine powder, add 10 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color;

(6) get these article, behind the capsule that removes photoresist, get fine powder, add 10 times of amount methyl alcohol, reflux 1 hour; Filter, the filtrating evaporate to dryness, residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Merge normal butyl alcohol liquid, extract 3 times with the 1%NaOH jolting, each 20ml merges alkaline solution, with watery hydrochloric acid adjust pH to 3; Extract 3 times with water saturated normal butyl alcohol jolting, each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(7) get these article, behind the capsule that removes photoresist, get fine powder, add 10 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds water 30ml makes dissolving; Extract 3 times each 30ml, combined chloroform extract with the chloroform jolting; Put and be concentrated into driedly in the water-bath, precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

[the content side is fixed]

Chlorogenic acid shines high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder 0.5g, and accurate the title decides; Place the 5ml volumetric flask, add the nearly scale of 50% methyl alcohol, extracted 15 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

This product is got in the preparation of need testing solution, behind the capsule that removes photoresist, gets fine powder, and accurate the title decides, and puts in the flask; 20 times of amounts of accurate adding, 70% ethanol claim to decide weight, and circumfluence method was extracted 45 minutes, put to room temperature, claimed to decide weight again; Supply the weight that subtracts mistake with 70% ethanol, shake up, filter, get subsequent filtrate, promptly get;

Galuteolin shines high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, behind the capsule that removes photoresist, get fine powder, and accurate the title decides, and places the 25m volumetric flask, add an amount of ultrasonic dissolving that makes of 70% ethanol, are settled to scale, filter with miillpore filter, get subsequent filtrate, promptly get;

Astragaloside IV shines high effective liquid chromatography for measuring,

This product is got in the preparation of need testing solution, behind the capsule that removes photoresist, gets fine powder, and accurate the title decides, and puts in the flask, 20 times of amounts of accurate adding methyl alcohol ultrasonic Extraction 20min; Filter, residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount; Merging filtrate and washing lotion are concentrated into driedly, and residue adds water 10ml, and low-grade fever makes dissolving, extract 3 times with water saturated n-butanol jolting; Each 20ml merges n-butanol extracting liquid, extracts 2 times with ammonia solution, and each 20ml discards ammoniacal liquor; N-butanol liquid evaporate to dryness, residue dissolves with methyl alcohol and is transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;

Embodiment 23.A kind of method of quality control of logical plug arteries and veins soft capsule.

[discriminating]

(1) get these article, remove the capsule skin after, the dry thing fine powder 3g after the pre-treatment of learning from else's experience, porphyrize; With methyl alcohol ultrasonic Extraction 2 times, add methyl alcohol 50ml at every turn and extracted 20 minutes, filter the filtrating evaporate to dryness after merging extract; Add water 30ml and make dissolving, extract 3 times with water-saturated n-butanol, each 30ml merges normal butyl alcohol liquid; With ammonia solution washing 2 times, each 30ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 3-8ml makes dissolving, last D-101 macroporous adsorptive resins; Water 30ml wash-out discards water liquid, with 40% ethanol 30ml wash-out respectively; Discard eluent, continue to collect eluent with 70% ethanol 50ml wash-out; Evaporate to dryness, residue add methyl alcohol 2ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get these article, remove the capsule skin after, the dry thing fine powder 3g after the pre-treatment of learning from else's experience, porphyrize adds ethanol 40ml; Reflux 30 minutes filters, and filtrating is steamed near and done, and residue adds water 10ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 10ml; Reflux 3 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 2 times with the chloroform jolting; Each 15ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 10% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

This product is got in the preparation of need testing solution, remove the capsule skin after, the dry thing fine powder 1g after the pre-treatment of learning from else's experience, accurate claim fixed; Place the 50ml volumetric flask, add the nearly scale of 50% methyl alcohol, extracted 15 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

(4) get these article, remove the capsule skin after, the dry thing fine powder after the pre-treatment of learning from else's experience, add 10 times the amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color;

(6) get these article, remove the capsule skin after, the dry thing fine powder after the pre-treatment of learning from else's experience, add 10 times the amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds water 30ml makes dissolving; Extract 3 times with water saturated normal butyl alcohol jolting, each 20ml merges normal butyl alcohol liquid, extracts 3 times with the 1%NaOH jolting; Each 20ml merges alkaline solution, with watery hydrochloric acid adjust pH to 3, extracts 3 times with water saturated normal butyl alcohol jolting; Each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(7) get these article, remove the capsule skin after, the dry thing fine powder after the pre-treatment of learning from else's experience, add 10 times the amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds water 30ml makes dissolving; Extract 3 times each 30ml, combined chloroform extract with the chloroform jolting; Put and be concentrated into driedly in the water-bath, precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

[the content side is fixed]

Chlorogenic acid shines high effective liquid chromatography for measuring,

This product is got in the preparation of need testing solution, remove the capsule skin after, the dry thing fine powder 1g after the pre-treatment of learning from else's experience, accurate claim fixed; Place the 100ml volumetric flask, add the nearly scale of 50% methyl alcohol, extracted 20 minutes; Put coldly, add 50% methyl alcohol, get need testing solution with filtering with microporous membrane to scale;

This product is got in the preparation of need testing solution, remove the capsule skin after, the dry thing fine powder after the pre-treatment of learning from else's experience, accurate claim fixed; Put in the flask, 20 times of amounts of accurate adding, 70% ethanol claim to decide weight, and circumfluence method was extracted 45 minutes; Put to room temperature, claim again to decide weight, supply the weight that subtracts mistake, shake up with 70% ethanol; Filter, get subsequent filtrate, promptly get;

Galuteolin shines high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, remove the capsule skin after, the dry thing fine powder after the pre-treatment of learning from else's experience accurate is claimed surely, places the 25m volumetric flask, adds an amount of ultrasonic dissolving that makes of 70% ethanol, is settled to scale, filters with miillpore filter, gets subsequent filtrate, promptly gets;

Astragaloside IV shines high effective liquid chromatography for measuring,

This product is got in the preparation of need testing solution, remove the capsule skin after, the dry thing fine powder after the pre-treatment of learning from else's experience accurate is claimed surely, puts in the flask, accurately adds 20 times of amount methyl alcohol ultrasonic Extraction 20min; Filter, residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount; Merging filtrate and washing lotion are concentrated into driedly, and residue adds water 10ml, and low-grade fever makes dissolving, extract 3 times with water saturated n-butanol jolting; Each 20ml merges n-butanol extracting liquid, extracts 2 times with ammonia solution, and each 20ml discards ammoniacal liquor; N-butanol liquid evaporate to dryness, residue dissolves with methyl alcohol and is transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;

Embodiment 24.The method of quality control of a kind of logical plug arteries and veins pill or oral liquid.

[discriminating]

(1) get the dry fine powder of these article or these article dry extract fine powder 3g, porphyrize with methyl alcohol ultrasonic Extraction 2 times, adds methyl alcohol 50ml extraction 20 minutes at every turn; Filter after merging extract, the filtrating evaporate to dryness adds water 30ml and makes dissolving; Extract 3 times with water-saturated n-butanol, each 30ml merges normal butyl alcohol liquid; With ammonia solution washing 2 times, each 30ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 3-8ml makes dissolving, last D-101 macroporous adsorptive resins; Water 30ml wash-out discards water liquid, with 40% ethanol 30ml wash-out respectively; Discard eluent, continue to collect eluent with 70% ethanol 50ml wash-out; Evaporate to dryness, residue add methyl alcohol 2ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of normal butyl alcohol-ethyl acetate-water (5: 1: 4); Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) get the dry fine powder of these article or these article dry extract fine powder 3g, porphyrize adds ethanol 40ml, reflux 30 minutes; Filter, filtrating is steamed near and is done, and residue adds water 10ml makes dissolving, adds 5% sulfuric acid-ethanol (1: 1) mixed solution 10ml; Reflux 3 hours is taken out, and puts and steams in the water-bath to there not being the alcohol flavor, extracts 2 times with the chloroform jolting; Each 15ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With cyclohexane-acetone-ethyl acetate (4: 2: 1) is developping agent, launches, and takes out; Dry, spray is with 10% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) according to high effective liquid chromatography for measuring;

The dry fine powder of these article or these article dry extract fine powder 1g are got in the preparation of need testing solution, and accurate the title places the 50ml volumetric flask calmly, adds the nearly scale of 50% methyl alcohol, extracts 15 minutes, put coldly, add 50% methyl alcohol to scale, get need testing solution with filtering with microporous membrane;

(4) get the dry fine powder of these article or these article dry extract fine powder, add 10 times of amount methyl alcohol, reflux 1 hour; Filter, the filtrating evaporate to dryness, residue adds water 30ml makes dissolving; Extract 3 times with water saturated normal butyl alcohol jolting, each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath; Residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color;

(6) get the dry fine powder of these article or these article dry extract fine powder, add 10 times of amount methyl alcohol, reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, and each 20ml merges normal butyl alcohol liquid, extracts 3 times with the 1%NaOH jolting; Each 20ml merges alkaline solution, with watery hydrochloric acid adjust pH to 3, extracts 3 times with water saturated normal butyl alcohol jolting; Each 20ml, normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(7) get the dry fine powder of these article or these article dry extract fine powder, add 10 times of amount methyl alcohol, reflux 1 hour; Filter, the filtrating evaporate to dryness, residue adds water 30ml makes dissolving; Extract 3 times each 30ml, combined chloroform extract with the chloroform jolting; Put and be concentrated into driedly in the water-bath, precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

[the content side is fixed]

Chlorogenic acid shines high effective liquid chromatography for measuring,

The dry fine powder of these article or these article dry extract fine powder 1g are got in the preparation of need testing solution, and accurate the title places the 100ml volumetric flask calmly, adds the nearly scale of 50% methyl alcohol, extracts 20 minutes, put coldly, add 50% methyl alcohol to scale, get need testing solution with filtering with microporous membrane;

The dry fine powder of this product or this product dry extract fine powder are got in the preparation of need testing solution, and accurate the title decides, and put in the flask, and precision adds 20 times of amount 70% ethanol; Claim to decide weight, circumfluence method was extracted 45 minutes, put to room temperature; Claim again to decide weight, supply the weight that subtracts mistake, shake up with 70% ethanol; Filter, get subsequent filtrate, promptly get;

Galuteolin shines high effective liquid chromatography for measuring,

The dry fine powder of these article or these article dry extract fine powder are got in the preparation of need testing solution, and accurate the title decides, and places the 25m volumetric flask, add an amount of ultrasonic dissolving that makes of 70% ethanol, are settled to scale, filter with miillpore filter, get subsequent filtrate, promptly get;

Astragaloside IV shines high effective liquid chromatography for measuring,

The dry fine powder of this product or this product dry extract fine powder are got in the preparation of need testing solution, and accurate the title decides, and put in the flask, and precision adds 20 times of amount methyl alcohol ultrasonic Extraction 20min, filters; Residue adds methyl alcohol 20ml together with filter paper, and ultrasonic Extraction 20min filters again, and residue washs with methyl alcohol in right amount, merging filtrate and washing lotion; Be concentrated into driedly, residue adds water 10ml, and low-grade fever makes dissolving, extracts 3 times with water saturated n-butanol jolting, each 20ml; Merge n-butanol extracting liquid, extract 2 times with ammonia solution, each 20ml discards ammoniacal liquor, n-butanol liquid evaporate to dryness; Residue dissolves with methyl alcohol and is transferred in the 2ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;

Claims

1. the detection method of a logical plug arteries and veins preparation is characterized in that its discrimination method is following,

(1) discrimination method of Astragaloside IV: get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 1～5g, porphyrize is with methyl alcohol ultrasonic Extraction 1-3 time; Add methyl alcohol 40-60ml at every turn and extracted 10-30 minute, filter behind the merging extract, the filtrating evaporate to dryness adds water 20-40ml and makes dissolving; Extract 2-4 time with water-saturated n-butanol, each 20-40ml merges normal butyl alcohol liquid; With ammonia solution washing 1-3 time, each 20-40ml discards ammoniacal liquor; With normal butyl alcohol liquid evaporate to dryness, residue adds water 3-8ml makes dissolving, last D-101 macroporous adsorptive resins; Water 40-60ml wash-out discards water liquid, with 30-50% ethanol 20-40ml wash-out respectively; Discard eluent, continue to collect eluent with 60-80% ethanol 40-60ml wash-out; Evaporate to dryness, residue add methyl alcohol 1-3ml dissolving as need testing solution; Other gets the Astragaloside IV reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution put respectively on same silica gel g thin-layer plate, be developping agent with the upper solution of 5: 1: 4 normal butyl alcohol-ethyl acetate-water; Launch, take out, dry; Spray is with the 5-15% ethanol solution of sulfuric acid, and hot blast blows to clear spot, in the test sample chromatogram; With the corresponding position of reference substance chromatogram on, show the spot of same color;

(2) discrimination method of oleanolic acid: get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder 1～5g, porphyrize adds ethanol 30-50ml, reflux 20-40 minute; Filter, filtrating is steamed near and is done, and residue adds water 5-15ml makes dissolving, adds 5% sulfuric acid-alcohol mixed solution 5-15ml of 1: 1; Reflux 2-4 hour, take out, put and steam in the water-bath to there not being the alcohol flavor, extract 1-3 time with the chloroform jolting; Each 10-20ml, combined chloroform liquid is concentrated into 1ml, as need testing solution; Other evens up pier tartaric acid reference substance, adds ethanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 4: 2: 1 cyclohexane-acetone-ethyl acetates was developping agent, launched, and took out; Dry, spray is with 5-15% phosphomolybdic acid ethanol test solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) high performance liquid chromatography of chlorogenic acid:

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent, and 17: 80: 3 methanol-water-glacial acetic acid are moving phase, and the detection wavelength is 330nm, and number of theoretical plate is pressed chlorogenic acid and calculated, and should be lower than 1500;

(4) discrimination method of Radix Codonopsis: get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder, add 5～15 times of amount methyl alcohol; Reflux 1 hour filters, the filtrating evaporate to dryness; Residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting, each 20ml; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets Radix Codonopsis control medicinal material 1g, shines medicinal material solution in pairs with legal system; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 7: 1: 0.5 normal butyl alcohol-glacial acetic acid-water was developping agent, launched 16cm, took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the main fluorescence spot of same color;

(5) discrimination method of radix scrophulariae: get radix scrophulariae control medicinal material 0.2g, shine medicinal material solution in pairs with legal system according to need testing solution preparation method described in the discriminating of Radix Codonopsis; According to the thin-layered chromatography test, need testing solution and each 10 μ l of above-mentioned control medicinal material solution put respectively on same silica gel g thin-layer plate in the discriminating of absorption Radix Codonopsis; With 7: 1: 2 normal butyl alcohol-glacial acetic acid-water was developping agent; Launch 14cm, take out, dry; Spray is with the vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color;

(6) discrimination method of Radix Glycyrrhizae: get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder, add 5～15 times of amount methyl alcohol, reflux 1 hour; Filter, the filtrating evaporate to dryness, residue adds water 30ml makes dissolving, extracts 3 times with water saturated normal butyl alcohol jolting; Each 20ml merges normal butyl alcohol liquid, extracts 3 times with the 1%NaOH jolting, each 20ml; Merge alkaline solution,, extract 3 times, each 20ml with water saturated normal butyl alcohol jolting with watery hydrochloric acid adjust pH to 3; Normal butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With 15: 1: 1: 2 ethyl acetate-formic acid-glacial acetic acid-water was developping agent, launched, and took out; Dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear, puts under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(7) discrimination method of escoparone: get these article, perhaps get the dry fine powder of these article or these article dry extract fine powder, add 5～15 times of amount methyl alcohol, reflux 1 hour; Filter, the filtrating evaporate to dryness, residue adds water 30ml makes dissolving; Extract 3 times each 30ml, combined chloroform extract with the chloroform jolting; Put and be concentrated into driedly in the water-bath, precision adds chloroform and makes dissolving to 1ml in the residue, as need testing solution; Other gets the escoparone reference substance, and chlorination is copied into the solution that every 1ml contains 0.3mg, as reference substance solution; According to thin-layered chromatography test, draw reference substance solution, each 10 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, be developping agent with lower floor's solution of 1: 1: 1 ethyl acetate-chloroform-liquor ammoniae fortis; Launch, exhibition is taken out apart from 8cm, dries up; Be developping agent with 2: 3: 3 cyclohexane-chloroform-ethyl acetates again, launch that exhibition is apart from 18cm; Take out, dry, under the 365nm ultraviolet lamp, inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

(8) content assaying method of ammonium glycyrrhetate:

According to high effective liquid chromatography for measuring,

(9) content assaying method of galuteolin:

According to high effective liquid chromatography for measuring,

These article are got in the preparation of need testing solution, perhaps get the dry fine powder of these article or these article dry extract fine powder, and accurate the title decides; Place 25ml or 50ml or 100ml volumetric flask, add an amount of ultrasonic dissolving that makes of 70% ethanol, be settled to scale; Filter with miillpore filter, get subsequent filtrate, promptly get;

Accurate respectively reference substance solution and the need testing solution 10 μ l of drawing of determination method inject liquid chromatograph, measure, and calculate, and promptly get;

(10) content assaying method of Astragaloside IV:

According to high effective liquid chromatography for measuring,

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent; 77: 23 methanol-waters are moving phase; Detect with EISD; Number of theoretical plate should be not less than 2000 by the Astragaloside IV peak;

Accurate respectively reference substance solution 5 μ l, 20 μ l and the need testing solution 10 μ l of drawing of determination method inject liquid chromatograph, calculate with external standard two-point method logarithmic equation, promptly get;

2. the detection method of a kind of logical plug arteries and veins preparation according to claim 1 is characterized in that: the formulation of said preparation is an oral formulations, is selected from pill, granule, tablet, capsule, powder and oral liquid.