CN1935199A - Quality control method for Chinese medicine compound preparation - Google Patents

Abstract

The present invention relates to a quality control method of Chinese medicine compound preparation made up by using the Chinese medicinal material of fleabane and red peony (or white peony). Said quality control method includes fingerprint chromatogram test method and/or identification test method and/or content determination method of fleabane and red peony (or white peony).

Description

A kind of method of quality control of compound Chinese medicinal preparation

Technical field

The present invention is a kind of method of quality control of compound Chinese medicinal preparation, belongs to the technical field of medicine.

Background technology

This compound Chinese medicinal preparation is that raw material is made with Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba, it is characterized in that: calculate according to ratio of weight and number, it is through extracting refining forming by 1～10 part of 1～10 part of Herba Erigerontis and Radix Paeoniae Rubra or the Radix Paeoniae Alba; Or the medicinal substances extract that is obtained after extracting by corresponding weight portion Herba Erigerontis and corresponding weight portion Radix Paeoniae Rubra or white Peony Root is made; This compound Chinese medicinal preparation has activating blood circulation to dissipate blood stasis, TONGMAI SHULUO, improves blood circulation and metabolism effect, be applicable to the hemiplegia of apoplexy syndrome of static blood blocking collaterals, facial hemiparalysis, speech is stuttering puckery, white and thin fur or thin white greasy, deep-thready pulse etc., and cerebral infarction is plugged with the above-mentioned symptom of opinion, except that being used for the treatment of cardiovascular and cerebrovascular diseases such as coronary heart disease, angina pectoris, arrhythmia, cerebral thrombosis, alzheimer disease, also be used for diseases such as hepatorenal syndrome, heart and lung diseases, diabetes and complication thereof clinically.Cardiovascular and cerebrovascular disease such as coronary heart disease, cerebral thrombosis, alzheimer disease etc. all are one of diseases that the world today is the most common and harm is maximum, have become human mortality's one of the main reasons in many countries.According to investigations, the sickness rate of such disease has and increases trend year by year in recent years, and in, young patient constantly increases, and become commonly encountered diseases, the frequently-occurring disease of harm China people ' s health.The applicant is doing a lot of work aspect the medicine of research control cardiovascular and cerebrovascular disease, and has once applied for to Patent Office of the People's Republic of China that it was 200410040776.2 patent of invention that name is called " compound recipe fleabane preparation of treatment cardiovascular and cerebrovascular disease and preparation method thereof ", number of patent application.But finding in the research that further pharmaceutical preparation must be on the basis that guarantees the constant product quality controllable safety, constantly more new development.In order better to control the quality of said preparation, guarantee the safety of medication, better instruct and produce, make technology controlling and process rationally strict more, make consumer's energy full appreciation product quality, need research, control this composing prescription preparation method for quality, the applicant has furtherd investigate its method of quality control.At present, this prescription product does not have other any related datas, in other the pharmaceutical preparation that contains Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba, serves as to detect index with scutellarin, peoniflorin only generally.But the chemical constituent of Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba has tens kinds, if its inherent quality is described with one, two kinds of active component, have certain one-sidedness, said nothing of the index components of no efficacy, and the phase mutual interference of composition does not have correlational study especially between the two.Therefore, the applicant has not only studied with the active component of Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba or characteristic component and the discriminating and the content assaying method that carry out, more set up the technical scheme of finger printing, its material group integral body has been controlled, effectively characterized its quality on the whole.Chinese medicine fingerprint is meant chromatograph or spectrographic collection of illustrative plates common, that have distinctive certain class or number constituents in certain Chinese crude drug or the Chinese patent medicine.In present stage, the most effective ingredient of Chinese medicine does not have under the clear and definite situation, and finger printing has useful effect for the quality of effective control product of the present invention.Discriminating, assay, finger printing use in conjunction, broad covered area, high specificity is outstanding substantive distinguishing features and an obvious improvement of this method of quality control.

Summary of the invention

The objective of the invention is to: a kind of method of quality control of compound Chinese medicinal preparation is provided, and this method provides means, technical method of the index that detects, detection or the like to relevant production, testing agency; So that better control the quality of said preparation, guarantee the safety of medication, can better instruct production, make controlling of production process rationally strict more, make consumer's energy full appreciation product quality.

The present invention constitutes like this: a kind of method of quality control of the compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba, and it is characterized in that: this method comprises following all or part of content:

(1) based on the fingerprint test method of all or part of composition characteristics in Herba Erigerontis, the Radix Paeoniae Alba or the Radix Paeoniae Rubra;

(2) the differential test method of all or part of composition in Herba Erigerontis medical material, Radix Paeoniae Rubra or white Peony Root, scutellarin, peoniflorin, the gallic acid;

(3) content test method of all or part of composition in scutellarin, peoniflorin, total flavones, Radix Paeoniae Rubra total glycosides or the Radix Paeoniae Alba total glucosides.

The method of quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba is characterized in that this method comprises all or part of content of following finger printing test:

(1) preparation of need testing solution: it is an amount of to get compound Chinese medicinal preparation to be measured, is solvent with in water or methanol or the ethanol one or more, extracts or dilution or dissolving the preparation need testing solution;

(2) preparation of object of reference solution: get scutellarin or peoniflorin, one or more in water or methanol or the ethanol are solvent, are dissolved to suitable concn, as object of reference solution;

(3) chromatographic condition: chromatographic column adopting alkyl silane bonded silica gel is a filler: mobile phase A is 50～100% acetonitriles or 50～100% methanol, Mobile phase B is 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate or 0.1%～3% glacial acetic acid or 0.1%～3% formic acid or 0.03%～1% phosphoric acid solution, gradient elution, mobile phase A ratio excursion is 10～90%, flow velocity is that 0.5～1.5ml/min, detection wavelength are one or several in the 190-400nm scope, and column temperature is at 20～60 ℃;

(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the standard finger-print of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulating standard finger-print, is benchmark with the retention time of the object of reference chromatographic peak determined, calculates the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 4～20;

(5) with the described method in (1)～(3) as in the compound Chinese medicinal preparation to be measured based on the means of testing of the finger printing of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics, the finger printing of preparation testing sample;

(6) finger printing of compound Chinese medicinal preparation to be measured, in should meeting the following requirements partly or entirely:

I. the similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.80～1.00;

II. non-total peak area must not surpass 10% of total peak area;

III. the ratio of the relative retention time of total chromatographic peak of other except that the object of reference chromatographic peak and object of reference chromatographic peak and standard finger-print compare, and relative deviation must not surpass ± 20%;

IV. unimodal area accounts for total peak area than greater than 20% peak in the total peak, must not surpass ± 30% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 10% less than 20% peak in the total peak, must not surpass ± 35% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 5% less than 10% peak in the total peak, must not surpass ± 40% with the ratio relative deviation of standard finger-print respective peaks peak area;

The method of quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba is characterized in that this method comprises all or part of content of following finger printing test:

(1) preparation of need testing solution: it is an amount of that precision takes by weighing compound Chinese medicinal preparation to be measured, and the solution that every 1ml contains 1mg is made in thin up or dissolving, filters, promptly;

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methanol and make the solution that every 1ml contains 0.15mg, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is 80% acetonitrile, and Mobile phase B is 0.1% phosphoric acid solution, gradient elution, and solvent ratios was from 0 minute to 80 minutes, and the ratio of mobile phase A rises to 70% by 15%, and the ratio of Mobile phase B reduces to 30% by 85%; Flow velocity is 1.0ml/min, and the detection wavelength is 230 ± 2nm, and column temperature is 40 ℃;

(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the standard finger-print of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics; Accurate respectively object of reference solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid respectively, according to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, retention time with definite object of reference chromatographic peak is a benchmark, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 5～12;

(5) with the described method in (1)～(3) as in the compound Chinese medicinal preparation to be measured based on the means of testing of the finger printing of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics, the finger printing of preparation testing sample;

(6) finger printing of compound Chinese medicinal preparation to be measured, in should meeting the following requirements partly or entirely:

I. the similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.90～1.00;

II. non-total peak area must not surpass 5% of total peak area;

III. the ratio of the relative retention time of total chromatographic peak of other except that the object of reference chromatographic peak and object of reference chromatographic peak and standard finger-print compare, and relative deviation must not surpass ± 10%;

IV. unimodal area accounts for total peak area than greater than 20% peak in the total peak, must not surpass ± 20% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 10% less than 20% peak in the total peak, must not surpass ± 25% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 5% less than 10% peak in the total peak, must not surpass ± 30% with the ratio relative deviation of standard finger-print respective peaks peak area;

The method of quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba is characterized in that this method comprises all or part of content in the following differential test method:

One or both thin layer chromatography discrimination method in scutellarin, the Herba Erigerontis in a, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds ethanol or methanol or n-butanol extraction, and filtration or centrifugal is got filtrate or supernatant as need testing solution; Get the Herba Erigerontis control medicinal material, first weeding of grease solubility impurity refers again to the need testing solution method for making, makes control medicinal material solution; Other gets the scutellarin reference substance, adds methanol or ethanol and makes the reference substance solution that every 1ml contains 1mg; The test of employing thin layer chromatography, one or both and the test solution drawn respectively in the above-mentioned contrast solution are an amount of, point is on same silica gel thin-layer plate or polyamide film, with methanol or ethanol-ethyl acetate or Ethyl formate or butyl acetate-formic acid or acetic acid=5～15: be developing solvent at 0.5～5: 0.01～3, expansion, take out, dry, spray is containing aluminum chloride or ferric chloride developer, in the test sample chromatograph, with contrast chromatograph corresponding position on, should show the same color speckle;

The liquid chromatograph discrimination method of scutellarin in b, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, is solvent with in methanol or ethanol or the water one or more, extracts or dilution or dissolving, is made as need testing solution; Prepare reference substance solution with the scutellarin reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, with methanol or acetonitrile-water or 0.2%～3% glacial acetic acid aqueous solution or 0.2%～3% aqueous formic acid or 0.01%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=15～70: 85～30 is mobile phase, detects wavelength and be among 200～410nm one or more; Employing liquid chromatography test is drawn need testing solution respectively with reference substance solution is an amount of, injects chromatograph of liquid, tests, and in the test sample chromatograph, answers tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end;

One or more thin layer chromatography discrimination method in Radix Paeoniae Rubra or white Peony Root, peoniflorin, the gallic acid in c, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds in ethanol or methanol or n-butyl alcohol or the water one or more for solvent extraction, the preparation need testing solution; Get Radix Paeoniae Rubra or Radix Paeoniae Alba control medicinal material,, make control medicinal material solution with reference to the need testing solution preparation method; Get one or both of peoniflorin reference substance, gallic acid reference substance, make reference substance solution; The test of employing thin layer chromatography, one or both and the test solution drawn in the above-mentioned contrast solution are an amount of, put respectively on same silica gel thin-layer plate, with chloroform or dichloromethane-ethyl acetate or Ethyl formate or butyl acetate-methanol or ethanol-formic acid or acetic acid=2～10: be developing solvent at 0.5～5: 0.5～5: 0.01～5, launch, take out, dry, successively or spray with the developer that contains ferric chloride or developer that contains aluminum chloride or vitriolated developer respectively or contain in the developer of vanillin one or more, in the test sample chromatograph, with contrast chromatograph corresponding position on, should show the same color speckle;

The liquid chromatograph discrimination method of peoniflorin in d, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds one or more extractions or the dissolving in methanol or ethanol or the water or is diluted to suitable concn, as need testing solution; Prepare reference substance solution with the peoniflorin reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, methanol or acetonitrile-water or 0.05%～3% glacial acetic acid aqueous solution or 0.05%～3% aqueous formic acid or 0.001%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=15～60: 85～40 mobile phases, detection wavelength are one or several among 200～410nm; Employing liquid chromatography test is drawn need testing solution respectively with reference substance solution is an amount of, injects chromatograph of liquid, tests, and in the test sample chromatograph, answers tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end;

The method of quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba is characterized in that this method comprises all or part of content in the following differential test method:

One or both thin layer chromatography discrimination method in Herba Erigerontis medical material, the scutellarin in a, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds methanol extraction, the preparation need testing solution; Other gets the scutellarin reference substance, adds methanol and makes the solution that every 1ml contains 1mg; Get the Herba Erigerontis control medicinal material, first weeding of grease solubility impurity refers again to the need testing solution method for making, makes control medicinal material solution; The test of employing thin layer chromatography, one or both and the test solution drawn respectively in the above-mentioned contrast solution are an amount of, point be developing solvent with methanol-ethyl acetate-formic acid=7: 2: 1 on same polyamide membrane, expansion, take out, dry, spray is with 2% ferric chloride alcoholic solution, in the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;

The liquid chromatograph discrimination method of scutellarin in b, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds dissolve with methanol or is diluted to suitable concn, shakes up, and filters, and gets subsequent filtrate as need testing solution; Methanol solution with the scutellarin reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% phosphate aqueous solution=50: 50 is a mobile phase, and the detection wavelength is 335nm; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end;

The thin layer chromatography discrimination method of Radix Paeoniae Rubra medical material, peoniflorin, gallic acid in c, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds methanol extraction, centrifugal, gets supernatant as need testing solution; The methanol solution that other gets peoniflorin, gallic acid reference substance is product solution in contrast; Get the Radix Paeoniae Rubra medical material, make control medicinal material solution with reference to the preparation method of need testing solution; The test of employing thin layer chromatography, draw respectively in control medicinal material or the reference substance solution one or both, and test solution an amount of, point is on same silica gel g thin-layer plate, with chloroform-ethyl acetate-methanol-formic acid=6: 1.5: 1.5: 0.5 be developing solvent, expansion, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph earlier, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the same color speckle; Spray with the vanillin sulfuric acid solution, it is clear to be heated to speckle colour developing again, in the test sample chromatograph, with the corresponding position of contrast chromatograph on, show the same color speckle;

The liquid chromatograph discrimination method of peoniflorin in d, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds methanol extraction or dilution, filters, and gets subsequent filtrate as need testing solution; Methanol solution with the peoniflorin reference substance is a reference substance solution, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% formic acid solution=40: 60 is a mobile phase, and the detection wavelength is 230nm; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end.

The method of quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba is characterized in that this method comprises all or part of content of following assay:

The assay of scutellarin in a, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, is solvent with in methanol or ethanol or the water one or more, extracts or dilution or dissolving, is made as need testing solution; Prepare reference substance solution with the scutellarin reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, with methanol or acetonitrile-water or 0.2%～3% glacial acetic acid aqueous solution or 0.2%～3% aqueous formic acid or 0.01%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=15～70: 85～30 is mobile phase, detects wavelength and be among 200～410nm one or more; The test of employing liquid chromatography, accurate respectively absorption need testing solution and reference substance solution are an amount of, inject chromatograph of liquid, test, calculate with one point external standard method or standard curve method, must not be less than 15mg with what dosage contained scutellarin every day in the compound Chinese medicinal preparation;

Content of paeoniflorin is measured in b, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds one or more extractions or the dissolving in methanol or ethanol or the water or is diluted to suitable concn, filters, as need testing solution; Prepare reference substance solution with the peoniflorin reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, methanol or acetonitrile-water or 0.05%～3% glacial acetic acid aqueous solution or 0.05%～3% aqueous formic acid or 0.001%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=15～60: 85～40 mobile phases, detection wavelength are one or several among 200～410nm; Employing liquid chromatography test is drawn need testing solution respectively with reference substance solution is an amount of, and the injection chromatograph of liquid calculates with one point external standard method or standard curve method, and must not be less than 15mg with what dosage contained peoniflorin every day in the compound Chinese medicinal preparation;

Content of total flavone is measured in c, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, with one or more extractions in methanol or water or the ethanol or be diluted to suitable concn, shakes up, as need testing solution; With scutellarin product in contrast, get reference substance solution with legal system; With the retinue solvent is blank, adopts spectrophotography, and place in 300～600nm wavelength or two places measure trap, calculate with one point external standard method or standard curve method; Contain total flavones with dosage and must not be less than 20mg every day in the compound Chinese medicinal preparation;

The assay of Radix Paeoniae Rubra total glycosides or Radix Paeoniae Alba total glucosides in d, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, puts in the measuring bottle, adds strong base solution and makes dilution or dissolving and fixed to scale in right amount, shake up, precision is measured in right amount, puts in the tool plug test tube, put the boiling water bath hydrolysis 0.5 hour～5 hours, and took out, put to room temperature, quantitatively be transferred in the measuring bottle, swing repeatedly with methanol or ethanol or mobile phase and to wash test tube wall, fixed to scale, shake up, filter with microporous filter membrane, get subsequent filtrate as need testing solution; Preparation reference substance solution with the benzoic acid reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, methanol or acetonitrile-water or 0.05%～3% glacial acetic acid aqueous solution or 0.05%～3% aqueous formic acid or 0.001%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/ sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=10～60: 90～40 mobile phases, the detection wavelength is one or several in 200～410nm wave-length coverage, and column temperature is in 20～60 ℃ of scopes; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, calculate behind the content again by formula Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides content %=benzoic acid content % * 480.48/122.13 with one point external standard method or standard curve method, calculate the content of Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides must not be less than 20mg.

The method of quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba is characterized in that this method comprises all or part of content of following assay:

The assay of scutellarin in a, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, and accurate the title decides or measure, and adds methanol extraction or is diluted to suitable concn, shakes up, and filters, and gets subsequent filtrate as need testing solution; Methanol solution with the scutellarin reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% phosphate aqueous solution=50: 50 is a mobile phase, and the detection wavelength is 335nm; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, calculate with one point external standard method, must not be less than 30mg with what dosage contained scutellarin every day in the compound Chinese medicinal preparation;

Content of paeoniflorin is measured in b, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, and accurate the title decides or measure, and adds dissolve with methanol or is diluted to suitable concn, shakes up, and filters, and gets subsequent filtrate as need testing solution; Methanol solution with the peoniflorin reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% formic acid solution=40: 60 is a mobile phase, and the detection wavelength is 230nm; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, calculate with one point external standard method, must not be less than 30mg with what dosage contained peoniflorin every day in the compound Chinese medicinal preparation;

Content of total flavone is measured in c, the compound Chinese medicinal preparation:

Get in the compound Chinese medicinal preparation to be measured in right amount, put in the measuring bottle, add methanol and make dissolving in right amount and shake up to scale surely, precision is measured 1ml, puts in the 50ml measuring bottle, adds methanol to scale, shakes up, as need testing solution; With scutellarin product in contrast, get reference substance solution with legal system.With the retinue solvent is blank, according to spectrophotography, measures trap at the wavelength place of 335nm, calculate with one point external standard method, each preparation is unit quantity to be equivalent to every day with output, and the per unit amount contains the limit of total flavones in scutellarin, must not be less than 40mg;

The assay of Radix Paeoniae Rubra total glycosides or Radix Paeoniae Alba total glucosides in d, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, and accurate the title decides or measure, and puts in the measuring bottle, add 5% sodium hydroxide solution and make dissolving and fixed to scale in right amount, shake up, precision is measured in right amount, put in the tool plug test tube, put the boiling water bath hydrolysis 2 hours, take out, put to room temperature, quantitatively be transferred in the 10ml measuring bottle, swing repeatedly with methanol or mobile phase and wash test tube wall, fixed to scale, shake up, filter, get subsequent filtrate as need testing solution with microporous filter membrane; Methanol solution with the benzoic acid reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-0.02mol/L sodium dihydrogen phosphate=30: 70 is a mobile phase, and the detection wavelength is 230nm, 35 ℃ of column temperatures; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, calculate with one point external standard method, Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides content=benzoic acid content * 480.48/122.13, calculate the content of Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides, compound Chinese medicinal preparation must not be less than 40mg with the limit that contains Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides in the dosage every day.

The method of quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba, it is characterized in that: the assay result of described compound Chinese medicinal preparation, calculate according to percentage by weight, the total amount that can measure composition accounts for deduction adjuvant and outer more than 25% of total solid of moisture in the preparation.

Compared with prior art, the present invention's quality of the compound Chinese medicinal preparation made with Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba of perfect control more.The Chinese medicine ingredients complexity, if only with wherein one, two kind of composition illustrate its inherent quality, has certain one-sidedness, more can't judge the index components of its drug effect.Therefore the applicant has formulated the quality that finger printing, discriminating and assay are controlled injection comprehensively.The chemical constituent contained owing to every kind of medical material in the compound Chinese medicinal preparation is all very complicated, and phase mutual interference behind the preparations shaping, all can affect greatly discriminating, assay, finger printing.Originally once be applicable to discriminating, assay or the fingerprint test method of single medical material, when being used to test the compound preparation that contains this medical material, often can't implement.Therefore, the method for quality control of compound preparation must be done with bigger improvement on the basis of former single medical material correlation method, or abandons former method and set up diverse new method.That is to say that quality control method of the present invention is not to be the simple superposition of single medicinal material material control method in the prescription, but after a large amount of experimental studies, invent.

Proof by experiment, method of quality control of the present invention is more effective to the quality control of the compound Chinese medicinal preparation product made with Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba, and method precision, stability are all higher.

In experimental example 1 injectable sterile powder based on the research of the finger printing of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics.

The selection of 1 chromatographic column:

In the research process, having selected conventional octadecylsilane chemically bonded silica for use is the liquid-phase chromatographic column of filler, has tried out Inertsil ODS-3 (250mm * 4.6mm, 5 μ m) Kromasil C respectively ₁₈(200mm * 4.6mm, 5 μ m), Diamonsil C ₁₈The chromatographic column of (250mm * 4.6mm, 5 μ m) three kinds of trades mark, the result shows that the chromatographic column of three kinds of trades mark all can reach separating effect preferably, wherein DiamonsilC ₁₈The chromatographic column separating effect is best, and post is imitated the highest.So select Diamonsil C for use ₁₈(250mm * 4.6mm, 5 μ m) are the experimentation post.

The selection of 2 mobile phases:

Investigated (1) methanol-water (25: 75) in the research process respectively, (2) acetonitrile-water (50: 50); (3) methanol-0.05 potassium dihydrogen phosphate (10: 90); (4) acetonitrile solution-0.5% phosphoric acid (gradient elution); (5) methanol solution-0.005moL/L sodium dihydrogen phosphate (gradient elution); (6) acetonitrile solution-0.1% phosphoric acid 6 kinds of flow phase system such as (gradient elutions); The result shows that under mobile phase (1) condition, minority peak-to-peak shape is bad and have overlapping; Under mobile phase (2) condition, the peak shape hangover, peak and peak overlapping are more; Under mobile phase (3) condition, it is less to go out the peak in 1 hour, goes out the peak after still having many components to be trapped in; Under mobile phase (4), (5), (6) condition, 50～100% acetonitriles or 50～100% methanol: 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate or 0.1%～3% glacial acetic acid or 0.1%～3% formic acid or 0.03%～1% phosphoric acid solution=10～90: 90～10 scope inside gradient eluting, effect is preferable; Optimum condition is: mobile phase A is 80% acetonitrile, and Mobile phase B is 0.1% phosphoric acid, gradient elution, from 0 minute to 80 minutes, the ratio of mobile phase A rose to 70% by 15%, and flow velocity is 1.0ml/min, under this condition, peak shape is good, and it is fast and fully and be evenly distributed to go out the peak.

3 detect wavelength determination:

Under above-mentioned optimal flow phase condition, investigated the chromatographic peak situation under 190～410nm respectively, the result shows that 230nm can take into account kurtosis, separating degree and the baseline of each composition chromatographic peak when detecting, best results is so select 230nm to detect wavelength for this product finger printing.

4 instrument parameters: selected liquid-phase chromatographic analysis field mainstream configuration and well behaved LC-10Avp high performance liquid chromatograph for use, WML-2010 chromatographic work station, 40 ℃ of column temperatures, flow velocity 1.0ml/min.

The preparation of 5 need testing solutions:

It is an amount of that precision takes by weighing this product, adds water and make the solution that every 1ml contains 1mg, promptly.

The preparation of 6 object of reference solution:

It is an amount of to take by weighing the scutellarin reference substance, adds methanol and makes the solution that every 1ml contains 0.15mg approximately, shakes up, promptly.

7 formulate standard finger-print according to 10 batch samples, and it is as follows to formulate the fingerprint image standard of compound Chinese medicinal preparation to be measured:

I. the similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.90～1.00;

II. non-total peak area must not surpass 5% of total peak area;

III. the ratio of the relative retention time of total chromatographic peak of other except that the object of reference chromatographic peak and object of reference chromatographic peak and standard finger-print compare, and relative deviation must not surpass ± 10%;

IV. unimodal area accounts for total peak area than greater than 20% peak in the total peak, must not surpass ± 20% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 10% less than 20% peak in the total peak, must not surpass ± 25% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 5% less than 10% peak in the total peak, must not surpass ± 30% with the ratio relative deviation of standard finger-print respective peaks peak area;

The thin layer chromatography discrimination method of Herba Erigerontis medical material, scutellarin in experimental example 2 injection:

For the feature of outstanding Herba Erigerontis, selected Herba Erigerontis control medicinal material, scutellarin as its feature, but owing to had composition, for example peoniflorin in the Radix Paeoniae Rubra, peonin like more, the polar phase close in the medical material with the scutellarin structure.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of thin layer chromatography effect quality is the composition of unfolding condition, particularly developing solvent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:

The thin layer chromatography discrimination method of Herba Erigerontis medical material, scutellarin

Conditional outcome

(5: 5: 10: 4) the polyamide film reference substance was expanded to the forward position to normal hexane-benzene-Ethyl formate-formic acid

Ethyl acetate-benzene-acetic acid (5: 4: 3) silica gel H lamellae feminine gender has interference

Ethyl acetate-benzene-formic acid (5: 4: 3) silica gel g thin-layer plate separates unintelligible

Ethanol-acetone-formic acid (5: 5: 2) polyamide film feminine gender has interference

Toluene-ethyl acetate (8: 1) silica gel H lamellae does not fully launch

Benzene-ethyl acetate-glacial acetic acid (10: 5: 0.5) silica gel g thin-layer plate separates unintelligible

It is clear to separate, and Rf value is moderate, and negative do not have

Methanol-ethyl acetate-formic acid (7: 2: 1) polyamide film

Disturb

Through screening, determined best unfolding condition: being immobile phase with the polyamide membrane, is developing solvent with methanol-ethyl acetate-formic acid (7: 2: 1), and with this understanding, the Rf value of scutellarin is moderate, and it is clear to separate with other speckle, negative noiseless.

The liquid chromatograph discrimination method of scutellarin in experimental example 3 concentrated solution for injection:

Feature for outstanding Herba Erigerontis, selected scutellarin as its characteristic component, but owing to there is composition like more, the polar phase close in the medical material with the scutellarin structure, the for example peoniflorin in the Radix Paeoniae Rubra, peonin, have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of liquid chromatography effect quality is the composition of elution requirement, particularly mobile phase.Therefore, experiment sieving multiple elution requirement, part elution requirement and result are as follows:

The liquid chromatograph discrimination method of scutellarin in the concentrated solution for injection

Conditional outcome

Methanol-0.05mol/L sodium hydrogen phosphate (80: 20) separates unintelligible

Eight alkyl silane bonded silica gels

Acetonitrile-0.05mol/L sodium hydrogen phosphate (90: 10) separates unintelligible

Eight alkyl silane bonded silica gels

Methanol-oxolane-0.05mol/L sodium hydrogen phosphate (20: 10: 70) feminine gender has interference

Eight alkyl silane bonded silica gels

Acetonitrile-oxolane-0.05mol/L sodium hydrogen phosphate (20: 10: 70) peak shape is slightly asymmetric

Eight alkyl silane bonded silica gels

Methanol-oxolane-1% glacial acetic acid aqueous solution (20: 10: 70) feminine gender has interference

Octadecylsilane chemically bonded silica

Acetonitrile-oxolane-1% glacial acetic acid aqueous solution (20: 10: 70) peak shape is slightly asymmetric

Octadecylsilane chemically bonded silica

It is clear that methanol-0.1% phosphate aqueous solution (50: 50) separates, and the peak is capable sharp-pointed, symmetry, the moon

Octadecylsilane chemically bonded silica is noiseless

Through screening, determined optimum condition: be immobile phase with the octadecylsilane chemically bonded silica, methanol-0.1% phosphate aqueous solution (50: 50) is a mobile phase, and with this understanding, the scutellarin retention time is moderate, and the peak is capable sharp-pointed, and symmetry is negative noiseless.

The thin layer chromatography discrimination method of peoniflorin, gallic acid in experimental example 4 injectable sterile powders:

Feature for outstanding Radix Paeoniae Rubra or white Peony Root, selected Radix Paeoniae Rubra or white Peony Root, peoniflorin, gallic acid as its feature speckle, but because have in the medical material that more and peoniflorin, gallic acid structure are close, composition, for example scutellarin in the Herba Erigerontis like the polar phase.The key factor of thin layer chromatography effect quality is the composition of unfolding condition, particularly developing solvent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:

The thin layer chromatography discrimination method of peoniflorin, gallic acid in the injectable sterile powder

Conditional outcome

Benzene-ethyl acetate (5: 1) silica gel g thin-layer plate reference substance is expanded to the forward position

Methylene chloride-methanol-formic acid (7: 6: 5) silica gel H lamellae separates unintelligible

Chloroform-ethanol-glacial acetic acid (9: 5: 2) silica gel H lamellae feminine gender has interference

Methanol-ethyl acetate-formic acid (10: 1: 0.2) silica gel g thin-layer plate feminine gender has interference

Chloroform-ethyl acetate-glacial acetic acid (7: 1: 2) silica gel H lamellae separates unintelligible

Petroleum ether-ethyl acetate-ethanol-glacial acetic acid (10: 5: 1.5: 0.01) separated unintelligible by silica gel G

Lamellae

(6: 1.5: 1.5: 0.5) the silica gel G separation was clear, and Rf value is moderate, and negative do not have for chloroform-ethyl acetate-methanol-formic acid

Lamellae disturbs

Through screening, determined with the silica gel g thin-layer plate to be immobile phase, with chloroform-ethyl acetate-methanol-formic acid (6: 1.5: 1.5: 0.5) be developing solvent, with this understanding, the Rf value of peoniflorin, gallic acid is moderate, and it is clear to separate with other speckle, and is negative noiseless.Chloroform or dichloromethane-ethyl acetate or Ethyl formate or butyl acetate-methanol or ethanol-formic acid or acetic acid=2～10: 0.5～5: 0.5～5: 0.01～5

The assay of scutellarin in experimental example 5 injectable sterile powders

1 instrument and Tianjin, reagent island HPLC system comprise the LC-10ATvp pump, and the SPD-10Avp ultraviolet-visible light detects; Rheodyne 7725i hand sampling valve (U.S. Rheodyne company); WML chromatographic work station (Guangxi Weil-McLain dragon chromatograph scientific ﹠ technical corporation); TCQ-250 ultrasonic cleaner (Beijing armarium two factories, power 250W; Frequency 19～33kHz).

2 chromatographic conditions select test to select Hypersil C for use ₁₈(5 μ m, 200mm * 5.0mm, Dalian Yi Lite); Kromasil C ₁₈(5 μ m, 200mm * 5.0mm, Dalian Yi Lite) and Diamonsil (diamond) C ₁₈Chromatographic column (5 μ m, 250mm * 4.6mm, Di Ma company); Guard column is SecurityGuard C ₁₈4mm * 3mm (U.S. Phenomenex inc.); Mobile phase is mobile phase with methanol-0.1% phosphoric acid solution, methanol-0.1% formic acid solution, acetonitrile-0.1% phosphoric acid solution, the result is mobile phase the best to use methanol-0.1% phosphoric acid solution (50: 50), above chromatographic column all can be separated scutellarin in the preparation preferably, take all factors into consideration factors such as post pressure, disengaging time and separation case again, determine that column temperature is 40 ℃.

3 detect the testing result of the selection of wavelength according to scutellarin reference substance ultraviolet spectrogram.Scutellarin has absorption maximum at the 335nm place, so select 335nm as detecting wavelength.

The negative control solution that 4 negative interference tests are got scutellarin reference substance solution, preparation need testing solution, scarce Herba Erigerontis medical material respectively injects chromatograph of liquid respectively, the record chromatogram.The result lacks Herba Erigerontis medical material negative control collection of illustrative plates at the no chromatographic peak in place, scutellarin peak position.

Preparation this product of 5 need testing solutions is a powder, easily is dissolved in the water, so it is an amount of to get this product, accurately claims surely, with water dissolution and be diluted to finite concentration, shakes up, promptly.

6 linear relationships are investigated and working curve is drawn

6.1 the preparation of reference substance storing solution takes by weighing scutellarin reference substance 15.50mg (content: 97.12%) in the 50ml volumetric flask, add dissolve with methanol respectively and be diluted to scale, shake up, make the reference substance storing solution that every 1ml contains 0.3010mg.

6.2 linear relationship is investigated respectively precision and is sucted and state reference substance storing solution 1,1.5,2,2.5,3,3.5ml in the 10ml volumetric flask, add methanol and be diluted to scale, shake up, get every 1ml and contain scutellarin 0.03010,0.04515,0.06020,0.07525,0.09030,0.10535mg the reference substance working solution, the above-mentioned working solution 10 μ l of accurate respectively absorption, inject liquid chromatograph, the record chromatograph, with peak area (Y) is vertical coordinate, sample size (X) is an abscissa, draws out working curve, calculates with method of least square, get regression equation Y=182202944X-13956, r=0.9998.The linear equation that fitted to initial point is Y=18022825X, to calculate respectively in two equations in the peak area substitution of minimum sample introduction concentration and maximum sample introduction concentration gained respectively, the relative deviation of low concentration sample introduction measurement result is 0.33% as a result, the relative deviation of high concentration sample introduction measurement result is 0.12%, illustrate that the working curve intercept is approximately zero, so adopt one point external standard method to calculate content.Under this experiment condition, scutellarin sample introduction concentration is in linear relation in 0.0301～0.105mg/ml scope.

The scutellarin linear relationship is investigated

Sample introduction concentration 0.03010 0.04515 0.06020 0.07525 0.09030 0.10535

(mg/ml)

Peak area 540,002 805,785 1,083,704 1,349,363 1,615,394 1918785

7 replica tests are got a test sample, press 9 parts of test liquids of preparation method preparation of need testing solution, and sample introduction is measured respectively, and the result is as follows:

Scutellarin assay replica test in the preparation

Numbering	Weighing (mg)	Scutellarin content (%)	Meansigma methods (%)	RSD(％)
					1 2	76.27 74.52	14.23 14.34	14.33	0.73

3 4 5 6 7 8 9

75.66 103.2 101.5 101.9 126.3 123.4 125.6

14.37 14.17 14.31 14.27 14.35 14.40 14.53

The result shows that method repeatability is good.

8 recovery tests adopt the average recovery test, and precision takes by weighing the sample of measuring scutellarin content (14.33%), and the accurate respectively scutellarin that adds is an amount of, prepares test liquid by the need testing solution preparation method, measures, and the result is as follows:

Scutellarin recovery test result

Numbering	Weighing (mg)	Contain scutellarin (mg)	Add scutellarin (mg)	The amount of recording (mg)	The response rate (%)	Meansigma methods (%)	RSD(％)
								1 2 3 4 5 6 7 8 9	52.63 48.76 51.21 50.94 53.27 49.12 54.11 51.96 47.81	7.542 6.987 7.338 7.300 7.634 7.039 7.754 7.444 6.851	3.512 3.512 3.512 7.640 7.640 7.640 11.25 11.25 11.25	11.14 10.45 10.95 15.01 15.11 14.58 19.43 18.61 17.86	102.4 98.60 102.8 100.9 97.85 98.70 103.8 99.25 97.86	100.2	2.3

The result shows that method is good to the response rate of scutellarin.

9 need testing solution study on the stability are got same test sample, prepare test liquid by the need testing solution preparation method, and respectively at 0,1,2,4,8 hour sample introduction, the result was as follows:

The need testing solution study on the stability

Times (h) 01248 meansigma methods RSD (%)

Peak area 1,104,601 1,098,676 1,082,647 1,100,740 1,083,609 1,094,054 0.93

The result shows that need testing solution is stable in 8 hours.

10 sample determinations are got finished product preparation, measure in accordance with the law, and the result is as follows.

Scutellarin assay result (mg/ props up) in the test agent in three crowdes

Lot number 12 meansigma methodss

1 batch 32.06 30.62 31.34

2 batches 29.81 29.55 29.68

3 batches 28.14 28.84 28.49

Paeoniflorin content is measured in experimental example 6 injectable sterile powders

1 instrument and reagent

Peoniflorin reference substance Chinese biological goods calibrating institute uses for assay.

Hypersil C has been selected in the selection test of 2 chromatographic column conditions for use ₁₈(5 μ m, 200mm * 5.0mm, Dalian Yi Lite); Kromasil C ₁₈(5 μ m, 200mm * 5.0mm, Dalian Yi Lite) and Diamonsil (diamond) C ₁₈Chromatographic column (5 μ m, 250mm * 4.6mm, Di Ma company); Guard column is SecurityGuard C ₁₈4mm * 3mm (U.S. Phenomenex inc.); Mobile phase is mobile phase with methanol-0.1% phosphoric acid solution, methanol-0.1% formic acid solution, acetonitrile-0.1% phosphoric acid solution.The result is a mobile phase with methanol-0.1% formic acid solution (40: 60), chromatographic column adopting Diamonsil (diamond) C ₁₈Chromatographic column the best is taken all factors into consideration factors such as post pressure, disengaging time and separation case again, determines that column temperature is 40 ℃.

3 detect the testing result of the selection of wavelength according to the ultraviolet spectrogram of peoniflorin reference substance.Peoniflorin has absorption maximum at the 230nm place, so select 230nm for detecting wavelength.

The negative control solution that 4 negative interference tests are got peoniflorin reference substance solution, preparation need testing solution, scarce Radix Paeoniae Rubra medical material respectively injects chromatograph of liquid respectively, the record chromatogram.As a result, lack Radix Paeoniae Rubra medical material negative control collection of illustrative plates at the no absworption peak in place, peoniflorin peak position.

This product 0.1g is got in the preparation of 5 need testing solutions, accurate claims surely, puts in the 50ml measuring bottle, with water dissolution and be diluted to scale, shakes up, and filters, and gets subsequent filtrate promptly.

6 linear relationships are investigated and working curve is drawn

6.1 the preparation of reference substance storing solution takes by weighing peoniflorin 12.12mg in the 25ml volumetric flask, adds dissolve with methanol respectively and is diluted to scale, shakes up, and makes the reference substance storing solution that every 1ml contains 0.4848mg.

6.2 linear relationship is investigated respectively precision and is sucted and state reference substance storing solution 0.5,1,2,3,4,5ml is in the 10ml volumetric flask, add methanol and be diluted to scale, shake up, get every 1ml and contain peoniflorin 0.02424,0.04848,0.09696,0.14544,0.19392,0.2424mg the reference substance working solution, the above-mentioned working solution 10 μ l of accurate respectively absorption, inject liquid chromatograph, the record chromatograph, with peak area (Y) is vertical coordinate, sample size (X) is an abscissa, and the drawing curve calculates with method of least square, get regression equation Y=11283302X-2530, r=0.9999.The linear equation that fitted to initial point is Y=11268659X, the peak area of minimum sample introduction concentration and maximum sample introduction concentration gained is calculated in two equations in the substitution respectively, the relative deviation of low concentration sample introduction measurement result is 0.4% as a result, the relative deviation of high concentration sample introduction measurement result is 0.1%, under this experiment condition, reference substance solution sample introduction concentration is in linear relation with peak area in 0.024～0.24mg/ml scope, and intercept is approximately zero, so adopt one point external standard method to calculate content.

The peoniflorin linear relationship is investigated

Sample introduction concentration

0.02424 0.04848 0.09696 0.14544 0.19392 0.2424

(mg/ml)

Peak area 273,620 536,873 1,113,552 1,614,915 2,185,113 2739469

7 replica tests are got a test sample, press 9 parts of test liquids of preparation method preparation of need testing solution, the difference sample introduction, and the result is as follows for the record chromatograph:

Replica test

Numbering	Weighing (mg)	Paeoniflorin content (%)	Meansigma methods (%)	RSD(％)
					1 2 3 4 5 6 7 8 9	73.61 77.28 76.36 102.9 106.2 101.7 128.5 123.2 125.8	22.98 23.09 22.89 23.07 22.66 23.10 22.43 23.50 23.16	22.99	1.30

Result of the test shows that the repeatability of method is good.

8 recovery tests adopt the average recovery test, and it is an amount of that precision takes by weighing the finished product of measuring paeoniflorin content, and the accurate peoniflorin that adds prepares test liquid by the need testing solution preparation method, measures, and the result is as follows:

Peoniflorin recovery test result

Numbering	Weighing (mg)	Content (mg)	Addition (mg)	The amount of recording (mg)	The response rate (%)	Meansigma methods (%)	RSD(％)
								1 2 3 4 5	55.61 50.27 49.34 51.32 53.11	12.78 11.56 11.34 11.80 12.21	6.252 6.252 6.252 11.96 11.96	18.91 19.05 17.56 23.54 25.74	97.97 103.8 99.44 98.17 97.97	100.2	2.4

6 7 8 9

52.48 50.9l 48.89 54.03

12.07 11.70 11.24 12.42

11.96 13.81 13.81 13.81

24.78 30.40 28.75 32.06

101.1 102.1 98.15 103.5

The result shows that method is good to the peoniflorin response rate.

9 need testing solution study on the stability are got same test sample, prepare test liquid by the need testing solution preparation method, and respectively at 0,1,2,4,8 hour sample introduction, the result was as follows:

Need testing solution peoniflorin study on the stability

Times (h) 01248 meansigma methods RSD (%)

Peak area 1,041,206 1,028,914 1,074,929 1,025,477 1,047,162 1,043,538 1.9

The result shows that peoniflorin is stable in 8 hours in the need testing solution.

10 sample determinations

Get finished product preparation, measure in accordance with the law and calculate paeoniflorin content.

Paeoniflorin content measurement result (mg/ props up) in three batches of pilot scale formulation samples

Lot number 12 meansigma methodss

1 batch 48.69 48.27 48.48

2 batches 46.05 46.59 46.32

3 batches 48.73 47.85 48.29

The specific embodiment

Embodiment 1 adopts in the liquid chromatography for measuring injectable sterile powder finger printing based on Herba Erigerontis, Radix Paeoniae Rubra composition characteristics.

(1) preparation of need testing solution: it is an amount of that precision takes by weighing sample powder, adds methanol and make the solution that every 1ml contains 1mg, shakes up, and uses filtering with microporous membrane, gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methanol and make every 1ml and contain O.15mg solution, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is 80% acetonitrile, and Mobile phase B is 0.1% phosphoric acid solution, gradient elution, and solvent ratios was from 0 minute to 80 minutes, and the ratio of mobile phase A rises to 70% by 15%, and flow velocity is 1.0ml/min, and the detection wavelength is 230 ± 2nm, column temperature is 40 ℃;

(4) based on the formulation of the standard finger-print of Herba Erigerontis and Radix Paeoniae Rubra composition characteristics: accurate respectively each the 10 μ l of object of reference solution and need testing solution that draw, inject chromatograph of liquid respectively, according to 10 batches of collection of illustrative plates that test sample is measured, the formulation standard finger-print; Retention time with definite object of reference chromatographic peak is a benchmark, calculates the relative retention time of other total chromatographic peak, and in the described standard finger-print, total peak has 8;

(5) with said method as in the compound Chinese medicinal preparation based on the means of testing of the finger printing of Herba Erigerontis, Radix Paeoniae Rubra composition characteristics, the finger printing of preparation testing sample;

(6) finger printing of compound Chinese medicinal preparation to be measured should meet following requirement:

I. the similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.90～1.00;

II. non-total peak area must not surpass 5% of total peak area;

III. the ratio of the relative retention time of total chromatographic peak of other except that the object of reference chromatographic peak and object of reference chromatographic peak and standard finger-print compare, and relative deviation must not surpass ± 10%;

IV. unimodal area accounts for total peak area than greater than 20% peak in the total peak, must not surpass ± 20% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 10% less than 20% peak in the total peak, must not surpass ± 25% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 5% less than 10% peak in the total peak, must not surpass ± 30% with the ratio relative deviation of standard finger-print respective peaks peak area;

Embodiment 2 adopts in the liquid chromatography for measuring concentrated solution for injection finger printing based on Herba Erigerontis, Radix Paeoniae Alba composition characteristics.

(1) preparation of need testing solution: it is an amount of that precision is measured sample, adds 10 times of methanol dilutions, shakes up, and uses filtering with microporous membrane, gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methanol and make the solution that every 1ml contains 0.15mg, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is 50% acetonitrile, and Mobile phase B is the 0.3mol/L potassium dihydrogen phosphate, gradient elution, and solvent ratios was from 0 minute to 80 minutes, and the ratio of mobile phase A rises to 90% by 20%, and flow velocity is 1.5ml/min, and the detection wavelength is 337 ± 2nm, column temperature is 60 ℃;

(4) based on the formulation of the standard finger-print of Herba Erigerontis and Radix Paeoniae Alba composition characteristics: accurate respectively each the 5 μ l of object of reference solution and need testing solution that draw, inject chromatograph of liquid respectively, according to 10 batches of collection of illustrative plates that test sample is measured, the formulation standard finger-print; Retention time with definite object of reference chromatographic peak is a benchmark, calculates the relative retention time of other total chromatographic peak, and in the described standard finger-print, total peak has 7;

(5) with said method as in the compound Chinese medicinal preparation based on the means of testing of the finger printing of Herba Erigerontis, Radix Paeoniae Alba composition characteristics, the finger printing of preparation testing sample;

(finger printing of (6) compound Chinese medicinal preparation to be measured should meet following requirement:

I. the similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.90～1.00;

II. non-total peak area must not surpass 5% of total peak area;

Embodiment 3 adopts in the liquid chromatography for measuring injection finger printing based on Herba Erigerontis, Radix Paeoniae Rubra composition characteristics.

(1) preparation of need testing solution: sample thief, with the filtering with microporous membrane of 0.45 μ m, get subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing peoniflorin, adds 50% ethanol and make the solution that every 1ml contains 0.05mg, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is a methanol, Mobile phase B is 0.1% formic acid solution, gradient elution, and solvent ratios was from 0 minute to 40 minutes, the ratio of mobile phase A rises to 60% by 10%, from 40 minutes to 60 minutes, the ratio of mobile phase A rose to 80% by 60%, from 60 minutes to 65 minutes, the ratio of mobile phase A reduces to 10% by 80%, flow velocity is 0.5ml/min, and the detection wavelength is 190 ± 2nm, and column temperature is 20 ℃;

(4) based on the formulation of the standard finger-print of Herba Erigerontis and Radix Paeoniae Rubra composition characteristics: accurate respectively each the 10 μ l of object of reference solution and need testing solution that draw, inject chromatograph of liquid respectively, according to 10 batches of collection of illustrative plates that test sample is measured, the formulation standard finger-print; Retention time with definite object of reference chromatographic peak is a benchmark, calculates the relative retention time of other total chromatographic peak, and in the described standard finger-print, total peak has 4;

(5) with said method as in the compound Chinese medicinal preparation based on the means of testing of the finger printing of Herba Erigerontis, Radix Paeoniae Rubra composition characteristics, the finger printing of preparation testing sample;

(6) with the finger printing and the contrast of above-mentioned standard finger-print of compound Chinese medicinal preparation to be measured, similarity should be 0.90～1.00.

Embodiment 4 adopts in the liquid chromatography for measuring injectable sterile powders finger printing based on Herba Erigerontis, Radix Paeoniae Rubra composition characteristics.

(1) preparation of need testing solution: it is an amount of that precision takes by weighing sample powder, adds 80% methanol and make the solution that every 1ml contains 1mg, shakes up, and with the filtering with microporous membrane of 0.45 μ m, gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds 80% methanol and make the solution that every 1ml contains 0.1mg, as object of reference solution;

(3) chromatographic condition: chromatographic column adopting eight alkyl silane bonded silica gels are filler; Mobile phase A is an acetonitrile, and Mobile phase B is the 0.005mol/L sodium dihydrogen phosphate, gradient elution, and solvent ratios was from 0 minute to 60 minutes, and the ratio of mobile phase A rises to 70% by 10%, and flow velocity is 1.2ml/min, and the detection wavelength is 303 ± 2nm, column temperature is 40 ℃;

(4) based on the formulation of the standard finger-print of Herba Erigerontis and Radix Paeoniae Rubra composition characteristics: accurate respectively each the 10 μ l of object of reference solution and need testing solution that draw, inject chromatograph of liquid respectively, according to 10 batches of collection of illustrative plates that test sample is measured, the formulation standard finger-print; Retention time with definite object of reference chromatographic peak is a benchmark, calculates the relative retention time of other total chromatographic peak, and in the described standard finger-print, total peak has 20;

(5) with said method as in the compound Chinese medicinal preparation based on the means of testing of the finger printing of Herba Erigerontis, Radix Paeoniae Rubra composition characteristics, the finger printing of preparation testing sample;

(6) finger printing of compound Chinese medicinal preparation to be measured should meet following requirement:

I. the similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.80～1.00;

II. non-total peak area must not surpass 10% of total peak area;

III. the ratio of the relative retention time of total chromatographic peak of other except that the object of reference chromatographic peak and object of reference chromatographic peak and standard finger-print compare, and relative deviation must not surpass ± 20%;

IV. unimodal area accounts for total peak area than greater than 20% peak in the total peak, must not surpass ± 30% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 10% less than 20% peak in the total peak, must not surpass ± 35% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 5% less than 10% peak in the total peak, must not surpass ± 40% with the ratio relative deviation of standard finger-print respective peaks peak area;

The thin layer chromatography discrimination method of Herba Erigerontis, scutellarin in embodiment 5 injectable sterile powders:

Get this product 0.1g, add methanol 20ml and extract, the preparation need testing solution; Other gets the scutellarin reference substance, adds methanol and makes the solution that every 1ml contains 1mg; Get Herba Erigerontis control medicinal material 1g, with chloroform weeding of grease solubility impurity, refer again to the need testing solution method for making earlier, make control medicinal material solution; The test of employing thin layer chromatography, draw each 3 μ l of above-mentioned solution, putting on same polyamide membrane with strip tape respectively, is developing solvent with methanol-ethyl acetate-formic acid=7: 2: 1, launches, take out, dry, spray is with 2% ferric chloride alcoholic solution, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the streak of same color.

The thin layer chromatography discrimination method of Herba Erigerontis, scutellarin in embodiment 6 injectable sterile powders:

Get this product 0.1g, add 50% ethanol 20ml and extract, the preparation need testing solution; Other gets the scutellarin reference substance, adds 50% ethanol and makes the solution that every 1ml contains 1mg; Get Herba Erigerontis control medicinal material 0.5g, with chloroform weeding of grease solubility impurity, refer again to the need testing solution method for making earlier, make control medicinal material solution; The test of employing thin layer chromatography, drawing each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, is developing solvent with methanol-ethyl acetate-formic acid=10: 4: 1, launch, take out, dry, spray is with 2% aluminum chloride alcoholic solution, 105 ℃ were dried by the fire 5 minutes, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the speckle of same color.

The thin layer chromatography discrimination method of scutellarin in embodiment 7 injection:

Get this product 5ml, evaporate to dryness, residue add methanol 10ml and extract, the preparation need testing solution; Other gets the scutellarin reference substance, adds methanol and makes the solution that every 1ml contains 1mg; The test of employing thin layer chromatography, draw each 2 μ l of above-mentioned solution, putting on same polyamide membrane with strip tape respectively, is developing solvent with ethanol-butyl acetate-formic acid=5: 0.5: 3, launches, take out, dry, spray is with 4% ferric chloride alcoholic solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the streak of same color.

The thin layer chromatography discrimination method of scutellarin in the embodiment 8 injection dopes:

Get this product 2ml, with 5 times of amount ethanol dilutions, preparation need testing solution; Other gets the scutellarin reference substance, adds methanol and makes the solution that every 1ml contains 1mg; The test of employing thin layer chromatography, draw each 10 μ l of above-mentioned solution, putting respectively on same polyamide membrane, is developing solvent with ethanol-butyl acetate-formic acid=5: 5: 0.01, launches, take out, dry, spray is with 5% aluminum chloride alcoholic solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The liquid chromatograph discrimination method of scutellarin in embodiment 9 injectable sterile powders:

Sample thief 10mg adds methanol 20ml and extracts, and uses filtering with microporous membrane, gets subsequent filtrate as need testing solution; Get the scutellarin reference substance, add methanol and make the contrast solution that every 1ml contains 0.1mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% phosphate aqueous solution=50: 50 is a mobile phase, and the detection wavelength is 335nm; Draw each 5 μ l of need testing solution and reference substance solution respectively, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end;

The liquid chromatograph discrimination method of scutellarin in embodiment 10 concentrated solution for injection:

Sample thief 0.2ml adds ethanol 20ml dilution, the preparation need testing solution; Get the scutellarin reference substance, add methanol and make the contrast solution that every 1ml contains 0.05mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.01% phosphate aqueous solution=15: 85 is a mobile phase, and the detection wavelength is 200nm; Draw each 15 μ l of need testing solution and reference substance solution respectively, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end;

The liquid chromatograph discrimination method of scutellarin in embodiment 11 injection:

Sample thief 2ml adds ethanol 20ml dilution, the preparation need testing solution; Get the scutellarin reference substance, add ethanol and make the contrast solution that every 1ml contains 0.05mg, adopt liquid chromatography, chromatographic column is a filler with eight alkyl silane bonded silica gels, acetonitrile-0.3mol/L sodium dihydrogen phosphate=70: 30 is a mobile phase, and the detection wavelength is 410nm; Draw each 10 μ l of need testing solution and reference substance solution respectively, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end;

The thin layer chromatography discrimination method of peoniflorin, gallic acid in embodiment 12 injectable sterile powders:

Sample thief 0.2g adds methanol 20ml and extracts, the preparation need testing solution; Other gets peoniflorin, gallic acid reference substance, adds methanol and makes the reference substance solution that every ml contains peoniflorin 2mg, gallic acid 0.5mg; The test of employing thin layer chromatography, draw test solution and contrast solution 1 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-methanol-formic acid=6: 1.5: 1.5: 0.5 was developing solvent, launches, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph earlier, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the same color speckle; Spray again with 5% vanillin sulfuric acid solution, 105 ℃ dry by the fire to speckle colour developing clear, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the same color speckle;

The thin layer chromatography discrimination method of white Peony Root, peoniflorin, gallic acid in embodiment 13 concentrated solution for injection:

Sample thief 0.2ml adds methanol 20ml dilution, the preparation need testing solution; Other gets peoniflorin, gallic acid reference substance, adds methanol and makes the reference substance solution that every ml contains peoniflorin 2mg, gallic acid 0.5mg; Get white Peony Root 0.5g, add methanol 20ml and extract, preparation control medicinal material solution; The test of employing thin layer chromatography, it is an amount of to draw test solution and contrast solution, put respectively on same silica gel g thin-layer plate, with dichloromethane-ethyl acetate-methanol-formic acid=10: 0.5: 5: 0.01 was developing solvent, launches, take out, dry, spray is with 10% aluminum chloride alcoholic solution, in the test sample chromatograph earlier, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the same color speckle; Spray with 10% vanillin sulfuric acid solution, heating dot colour developing is clear again, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the same color speckle;

The thin layer chromatography discrimination method of Radix Paeoniae Rubra medical material, peoniflorin in embodiment 14 injection:

Sample thief 2ml adds ethanol 20ml dilution, the preparation need testing solution; Other gets the peoniflorin reference substance, adds ethanol and makes the reference substance solution that every ml contains peoniflorin 1mg; Get Radix Paeoniae Rubra medical material 0.5g, add methanol 20ml and extract, preparation control medicinal material solution; The test of employing thin layer chromatography, draw test solution and contrast solution 3 μ l, put respectively on same silica gel g thin-layer plate, with dichloromethane-ethyl acetate-methanol-formic acid=2: 0.5: 5: 5 was developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, the heating dot colour developing is clear, in the test sample chromatograph, with reference substance and the corresponding position of control medicinal material chromatograph on, show the same color speckle;

The liquid chromatograph discrimination method of peoniflorin in embodiment 15 injectable sterile powders:

Sample thief 20mg adds methanol 20ml and extracts, and uses filtering with microporous membrane, gets subsequent filtrate as need testing solution; Get the peoniflorin reference substance and add methanol and make the reference substance solution that every ml contains 0.1mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% formic acid solution=40: 60 is a mobile phase, and the detection wavelength is 230nm; Draw each 10 μ l of need testing solution and reference substance solution respectively, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end.

The liquid chromatograph discrimination method of peoniflorin in embodiment 16 concentrated solution for injection:

Sample thief 0.1ml adds methanol 20ml dilution, the preparation need testing solution; Get the peoniflorin reference substance and add methanol and make the reference substance solution that every ml contains 0.1mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-1% phosphoric acid=15: 85 is mobile phase, and the detection wavelength is 200nm; Draw each 5 μ l of need testing solution and reference substance solution respectively, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end.

The liquid chromatograph discrimination method of peoniflorin in embodiment 17 injection:

Sample thief 1ml adds ethanol 20ml dilution, the preparation need testing solution; Getting the peoniflorin reference substance adds 95% ethanol and makes the reference substance solution that every ml contains 0.05mg, adopt liquid chromatography, chromatographic column is a filler with the dialkyl silane bonded silica gel, and acetonitrile-0.005mol/L potassium dihydrogen phosphate=60: 40 is a mobile phase, and the detection wavelength is 410nm; Draw each 1～10 μ l of need testing solution and reference substance solution respectively, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end.

The assay of scutellarin in embodiment 18 injectable sterile powders:

Sample thief 20mg, the accurate title, decide, and adds methanol 50ml and extract, the preparation need testing solution; Get the scutellarin reference substance, add methanol and make the reference substance solution that every ml contains 0.1mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% phosphate aqueous solution=50: 50 is a mobile phase, and the detection wavelength is 335nm; Respectively accurate each the 10 μ l of need testing solution and reference substance solution that draw, the injection chromatograph of liquid calculates with one point external standard method, and must not be less than 40mg with what dosage contained scutellarin every day in the preparation;

The assay of scutellarin in embodiment 19 concentrated solution for injection:

Precision is measured sample 0.2ml, adds ethanol 50ml dilution, uses filtering with microporous membrane, gets subsequent filtrate as need testing solution; Get the scutellarin reference substance, add ethanol and make the reference substance solution that every ml contains 0.05mg, adopt liquid chromatography, chromatographic column is a filler with the dialkyl silane bonded silica gel, acetonitrile-0.3mol/L potassium dihydrogen phosphate=15: 85 is a mobile phase, and the detection wavelength is 410nm; Respectively accurate each the 10 μ l of need testing solution and reference substance solution that draw, the injection chromatograph of liquid calculates with one point external standard method, and must not be less than 15mg with what dosage contained scutellarin every day in the preparation;

The assay of scutellarin in embodiment 20 injection:

Precision is measured sample 0.5ml, adds methanol 50ml dilution, uses filtering with microporous membrane, gets subsequent filtrate as need testing solution; Get the scutellarin reference substance, add methanol and make the reference substance solution that every ml contains 0.05mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-0.2% aqueous formic acid=70: 30 is a mobile phase, and the detection wavelength is 200nm; Respectively accurate each the 5 μ l of need testing solution and reference substance solution that draw, the injection chromatograph of liquid calculates with standard curve method, and must not be less than 30mg with what dosage contained scutellarin every day in the preparation;

Content of paeoniflorin is measured in embodiment 21 injectable sterile powders:

Sample thief 20mg, the accurate title, decide, and adds methanol 50ml and extract, and uses filtering with microporous membrane, gets subsequent filtrate as need testing solution; Get the peoniflorin reference substance, add methanol and make the reference substance solution that every ml contains 0.1mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% formic acid solution=40: 60 is a mobile phase, and the detection wavelength is 230nm; Respectively accurate each the 10 μ l of need testing solution and reference substance solution that draw, the injection chromatograph of liquid calculates with one point external standard method, and must not be less than 60mg with what dosage contained peoniflorin every day in the preparation;

Content of paeoniflorin is measured in embodiment 22 injectable sterile powders:

Sample thief 100mg, the accurate title, decide, and adds methanol 100ml and extract, and uses filtering with microporous membrane, gets subsequent filtrate as need testing solution; Get the peoniflorin reference substance, add methanol and make the reference substance solution that every ml contains 0.02mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-3% formic acid solution=15: 85 is a mobile phase, and the detection wavelength is 200nm; Respectively accurate each the 10 μ l of need testing solution and reference substance solution that draw, the injection chromatograph of liquid calculates with standard curve method, and must not be less than 15mg with what dosage contained peoniflorin every day in the preparation;

Content of paeoniflorin is measured in embodiment 23 injection:

Precision is measured sample 1ml, adds methanol and is diluted to 10ml, the preparation need testing solution; Get the peoniflorin reference substance, add methanol and make the reference substance solution that every ml contains 0.05mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, acetonitrile-0.005mol/L sodium dihydrogen phosphate=60: 40 is a mobile phase, and the detection wavelength is 410nm; Respectively accurate each the 20 μ l of need testing solution and reference substance solution that draw, the injection chromatograph of liquid calculates with standard curve method, and must not be less than 20mg with what dosage contained peoniflorin every day in the preparation;

Content of paeoniflorin is measured in embodiment 24 concentrated solution for injection:

Precision is measured sample 0.3ml, adds ethanol dilution to 10ml, uses filtering with microporous membrane, gets subsequent filtrate as need testing solution; Get the peoniflorin reference substance, add 50% ethanol and make the reference substance solution that every ml contains 0.05mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, methanol-3% glacial acetic acid aqueous solution=30: 70 is a mobile phase, and the detection wavelength is 360nm; Respectively accurate each the 10 μ l of need testing solution and reference substance solution that draw, the injection chromatograph of liquid calculates with standard curve method, and must not be less than 30mg with what dosage contained peoniflorin every day in the preparation;

Content of total flavone is measured in embodiment 25 concentrated solution for injection:

Precision is measured sample 1ml, adds methanol and is diluted to 100ml, the preparation need testing solution; Get the scutellarin reference substance, add methanol and make the reference substance solution that every ml contains scutellarin 0.1mg.With the retinue solvent is blank, according to spectrophotography, measures trap at the wavelength place of 335nm; Accurate respectively need testing solution and each 10 μ l of reference substance solution of drawing, inject chromatograph of liquid, calculate with one point external standard method, each preparation is unit quantity to be equivalent to every day with output, the per unit amount contains the limit of total flavones in scutellarin, must not be less than 20mg;

Content of total flavone is measured in embodiment 26 injectable sterile powders:

Precision is measured sample 1g, adds ethanol 100ml and extracts, the preparation need testing solution; Get the scutellarin reference substance, add ethanol and make the reference substance solution that every ml contains scutellarin 0.1mg.With the retinue solvent is blank, according to spectrophotography, measures trap at the wavelength place of 300nm, calculate with standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the per unit amount contains the limit of total flavones in scutellarin, must not be less than 40mg;

Content of total flavone is measured in embodiment 27 concentrated solution for injection:

Precision is measured sample 1ml, and thin up prepares need testing solution to 100ml; Get the scutellarin reference substance, add water alcohol and make the reference substance solution that every ml contains scutellarin 0.1mg.With the retinue solvent is blank, according to spectrophotography, measures trap at the wavelength place of 600nm, calculate with one point external standard method, each preparation is unit quantity to be equivalent to every day with output, and the per unit amount contains the limit of total flavones in scutellarin, must not be less than 30mg;

The assay of Radix Paeoniae Rubra total glycosides in embodiment 28 injection:

Precision is measured sample 1ml, puts in the 10ml measuring bottle, adds 5% sodium hydroxide solution and fixed to scale, shake up, precision is measured 5ml, puts in the tool plug test tube, put the boiling water bath hydrolysis 2 hours, and took out, put to room temperature, quantitatively be transferred in the 10ml measuring bottle, swing repeatedly with methanol and to wash test tube wall, fixed to scale, shake up, filter with microporous filter membrane, get subsequent filtrate as need testing solution; Get the benzoic acid reference substance, add methanol and make the reference substance solution that every ml contains benzoic acid 0.05mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, acetonitrile-0.02mol/L sodium dihydrogen phosphate=30: 70 is a mobile phase, and the detection wavelength is 230nm, 35 ℃ of column temperatures; Accurate respectively need testing solution and each 10 μ l of reference substance solution of drawing, inject chromatograph of liquid, calculate with one point external standard method, Radix Paeoniae Rubra total glycosides content=benzoic acid content * 480.48/122.13, calculate the content of Radix Paeoniae Rubra total glycosides, compound Chinese medicinal preparation must not be less than 20mg with the limit that contains Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides in the dosage every day.

The assay of Radix Paeoniae Alba total glucosides in embodiment 29 concentrated solution for injection:

Precision is measured sample 0.2ml, puts in the 10ml measuring bottle, adds 10% potassium hydroxide solution and fixed to scale, shake up, precision is measured 5ml, puts in the tool plug test tube, put the boiling water bath hydrolysis 5 hours, and took out, put to room temperature, quantitatively be transferred in the 10ml measuring bottle, swing repeatedly with ethanol and to wash test tube wall, fixed to scale, shake up, filter with microporous filter membrane, get subsequent filtrate as need testing solution; Get the benzoic acid reference substance, add methanol and make the reference substance solution that every ml contains benzoic acid 0.03mg, adopt liquid chromatography, chromatographic column is a filler with eight alkyl silane bonded silica gels, methanol-0.05% glacial acetic acid solution=10: 90 is a mobile phase, and the detection wavelength is 200nm, 20 ℃ of column temperatures; Accurate respectively need testing solution and each 15 μ l of reference substance solution of drawing, inject chromatograph of liquid, calculate with one point external standard method, Radix Paeoniae Alba total glucosides content=benzoic acid content * 480.48/122.13, calculate Radix Paeoniae Alba total glucosides content, preparation must not be less than 40mg with the limit that contains Radix Paeoniae Alba total glucosides in the dosage every day.

The assay of Radix Paeoniae Alba total glucosides in embodiment 30 injectable sterile powders:

Precision is measured sample 0.5g, puts in the 10ml measuring bottle, adds the dissolving of 5% potassium hydroxide solution and is settled to scale, shake up, precision is measured 5ml, puts in the tool plug test tube, put the boiling water bath hydrolysis 0.5 hour, and took out, put to room temperature, quantitatively be transferred in the 10ml measuring bottle, swing repeatedly with ethanol and to wash test tube wall, fixed to scale, shake up, filter with microporous filter membrane, get subsequent filtrate as need testing solution; Get the benzoic acid reference substance, add methanol and make the reference substance solution that every ml contains benzoic acid 0.08mg, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, methanol-0.3mol/L potassium dihydrogen phosphate=60: 40 is mobile phase, and the detection wavelength is 410nm, 60 ℃ of column temperatures; Accurate respectively need testing solution and each 20 μ 1 of reference substance solution of drawing, inject chromatograph of liquid, calculate with standard curve method, Radix Paeoniae Alba total glucosides content=benzoic acid content * 480.48/122.13, calculate Radix Paeoniae Alba total glucosides content, preparation must not be less than 30mg with the limit that contains Radix Paeoniae Alba total glucosides in the dosage every day.

Embodiment 31 adopts in the liquid chromatography for measuring injectable sterile powders finger printing based on Herba Erigerontis, Radix Paeoniae Rubra composition characteristics.

(1) preparation of need testing solution: it is an amount of that precision takes by weighing sample powder, adds methanol and make the solution that every 1ml contains 1mg, shakes up, and uses filtering with microporous membrane, gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methanol and make the solution that every 1ml contains 0.15mg, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is 80% acetonitrile, and Mobile phase B is 0.1% phosphoric acid solution, gradient elution, and solvent ratios was from 0 minute to 80 minutes, and the ratio of mobile phase A rises to 70% by 15%, and flow velocity is 1.0ml/min, and the detection wavelength is 230 ± 2nm, column temperature is 40 ℃;

(4) based on the formulation of the standard finger-print of Herba Erigerontis and Radix Paeoniae Rubra composition characteristics: accurate respectively each the 10 μ l of object of reference solution and need testing solution that draw, inject chromatograph of liquid respectively, according to 10 batches of collection of illustrative plates that test sample is measured, the formulation standard finger-print; Retention time with definite object of reference chromatographic peak is a benchmark, calculates the relative retention time of other total chromatographic peak, and in the described standard finger-print, total peak has 8;

(5) with said method as in the compound Chinese medicinal preparation based on the means of testing of the finger printing of Herba Erigerontis, Radix Paeoniae Rubra composition characteristics, the finger printing of preparation testing sample;

(6) similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.90～1.00.

Claims (8)

1, a kind of method of quality control of the compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba, it is characterized in that: this method comprises following all or part of content:

(1) based on the fingerprint test method of all or part of composition characteristics in Herba Erigerontis, the Radix Paeoniae Alba or the Radix Paeoniae Rubra;

(2) the differential test method of all or part of composition in Herba Erigerontis medical material, Radix Paeoniae Rubra or white Peony Root, scutellarin, peoniflorin, the gallic acid;

(3) content test method of all or part of composition in scutellarin, peoniflorin, total flavones, Radix Paeoniae Rubra total glycosides or the Radix Paeoniae Alba total glucosides.

2,, it is characterized in that this method comprises all or part of content of following finger printing test according to the method for quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba of claim 1:

(1) preparation of need testing solution: it is an amount of to get compound Chinese medicinal preparation to be measured, is solvent with in water or methanol or the ethanol one or more, extracts or dilution or dissolving the preparation need testing solution;

(2) preparation of object of reference solution: get scutellarin or peoniflorin, one or more in water or methanol or the ethanol are solvent, are dissolved to suitable concn, as object of reference solution;

(3) chromatographic condition: chromatographic column adopting alkyl silane bonded silica gel is a filler: mobile phase A is 50～100% acetonitriles or 50～100% methanol, Mobile phase B is 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate or 0.1%～3% glacial acetic acid or 0.1%～3% formic acid or 0.03%～1% phosphoric acid solution, gradient elution, mobile phase A ratio excursion is 10～90%, flow velocity is that 0.5～1.5ml/min, detection wavelength are one or several in the 190-400nm scope, and column temperature is at 20～60 ℃;

(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the standard finger-print of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulating standard finger-print, is benchmark with the retention time of the object of reference chromatographic peak determined, calculates the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 4～20;

(5) with the described method in (1)～(3) as in the compound Chinese medicinal preparation to be measured based on the means of testing of the finger printing of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics, the finger printing of preparation testing sample;

(6) finger printing of compound Chinese medicinal preparation to be measured, in should meeting the following requirements partly or entirely:

I. the similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.80～1.00;

II. non-total peak area must not surpass 10% of total peak area;

III. the ratio of the relative retention time of total chromatographic peak of other except that the object of reference chromatographic peak and object of reference chromatographic peak and standard finger-print compare, and relative deviation must not surpass ± 20%;

IV. unimodal area accounts for total peak area than greater than 20% peak in the total peak, must not surpass ± 30% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 10% less than 20% peak in the total peak, must not surpass ± 35% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 5% less than 10% peak in the total peak, must not surpass ± 40% with the ratio relative deviation of standard finger-print respective peaks peak area.

3,, it is characterized in that this method comprises all or part of content of following finger printing test according to the method for quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba of claim 2:

(1) preparation of need testing solution: it is an amount of that precision takes by weighing compound Chinese medicinal preparation to be measured, and the solution that every 1ml contains 1mg is made in thin up or dissolving, filters, promptly;

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methanol and make the solution that every 1ml contains 0.15mg, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is 80% acetonitrile, and Mobile phase B is 0.1% phosphoric acid solution, gradient elution, and solvent ratios was from 0 minute to 80 minutes, and the ratio of mobile phase A rises to 70% by 15%, and the ratio of Mobile phase B reduces to 30% by 85%; Flow velocity is 1.0ml/min, and the detection wavelength is 230 ± 2nm, and column temperature is 40 ℃;

(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the standard finger-print of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics; Accurate respectively object of reference solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid respectively, according to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, retention time with definite object of reference chromatographic peak is a benchmark, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 5～12;

(5) with the described method in (1)～(3) as in the compound Chinese medicinal preparation to be measured based on the means of testing of the finger printing of Herba Erigerontis, the Radix Paeoniae Alba or Radix Paeoniae Rubra composition characteristics, the finger printing of preparation testing sample;

(6) finger printing of compound Chinese medicinal preparation to be measured, in should meeting the following requirements partly or entirely:

I. the similarity of the finger printing of compound Chinese medicinal preparation to be measured and standard finger-print should be 0.90～1.00;

II. non-total peak area must not surpass 5% of total peak area;

III. the ratio of the relative retention time of total chromatographic peak of other except that the object of reference chromatographic peak and object of reference chromatographic peak and standard finger-print compare, and relative deviation must not surpass ± 10%;

IV. unimodal area accounts for total peak area than greater than 20% peak in the total peak, must not surpass ± 20% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 10% less than 20% peak in the total peak, must not surpass ± 25% with the ratio relative deviation of standard finger-print respective peaks peak area; Unimodal area accounts for total peak area than greater than 5% less than 10% peak in the total peak, must not surpass ± 30% with the ratio relative deviation of standard finger-print respective peaks peak area.

4, according to the method for quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba of claim 1, it is characterized in that this method comprises all or part of content in the following differential test method:

One or both thin layer chromatography discrimination method in scutellarin, the Herba Erigerontis in a, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds ethanol or methanol or n-butanol extraction, and filtration or centrifugal is got filtrate or supernatant as need testing solution; Get the Herba Erigerontis control medicinal material, first weeding of grease solubility impurity refers again to the need testing solution method for making, makes control medicinal material solution; Other gets the scutellarin reference substance, adds methanol or ethanol and makes the reference substance solution that every 1ml contains 1mg; The test of employing thin layer chromatography, one or both and the test solution drawn respectively in the above-mentioned contrast solution are an amount of, point is on same silica gel thin-layer plate or polyamide film, with methanol or ethanol-ethyl acetate or Ethyl formate or butyl acetate-formic acid or acetic acid=5～15: be developing solvent at 0.5～5: 0.01～3, expansion, take out, dry, spray is containing aluminum chloride or ferric chloride developer, in the test sample chromatograph, with contrast chromatograph corresponding position on, should show the same color speckle;

The liquid chromatograph discrimination method of scutellarin in b, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, is solvent with in methanol or ethanol or the water one or more, extracts or dilution or dissolving, is made as need testing solution; Prepare reference substance solution with the scutellarin reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, with methanol or acetonitrile-water or 0.2%～3% glacial acetic acid aqueous solution or 0.2%～3% aqueous formic acid or 0.01%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=15～70: 85～30 is mobile phase, detects wavelength and be among 200～410nm one or more; Employing liquid chromatography test is drawn need testing solution respectively with reference substance solution is an amount of, injects chromatograph of liquid, tests, and in the test sample chromatograph, answers tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end;

One or more thin layer chromatography discrimination method in Radix Paeoniae Rubra or white Peony Root, peoniflorin, the gallic acid in c, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds in ethanol or methanol or n-butyl alcohol or the water one or more for solvent extraction, the preparation need testing solution; Get Radix Paeoniae Rubra or Radix Paeoniae Alba control medicinal material,, make control medicinal material solution with reference to the need testing solution preparation method; Get one or both of peoniflorin reference substance, gallic acid reference substance, make reference substance solution; The test of employing thin layer chromatography, one or both and the test solution drawn in the above-mentioned contrast solution are an amount of, put respectively on same silica gel thin-layer plate, with chloroform or dichloromethane-ethyl acetate or Ethyl formate or butyl acetate-methanol or ethanol-formic acid or acetic acid=2～10: be developing solvent at 0.5～5: 0.5～5: 0.01～5, launch, take out, dry, successively or spray with the developer that contains ferric chloride or developer that contains aluminum chloride or vitriolated developer respectively or contain in the developer of vanillin one or more, in the test sample chromatograph, with contrast chromatograph corresponding position on, should show the same color speckle;

The liquid chromatograph discrimination method of peoniflorin in d, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds one or more extractions or the dissolving in methanol or ethanol or the water or is diluted to suitable concn, as need testing solution; Prepare reference substance solution with the peoniflorin reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, methanol or acetonitrile-water or 0.05%～3% glacial acetic acid aqueous solution or 0.05%～3% aqueous formic acid or 0.001%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=15～60: 85～40 mobile phases, detection wavelength are one or several among 200～410nm; Employing liquid chromatography test is drawn need testing solution respectively with reference substance solution is an amount of, injects chromatograph of liquid, tests, and in the test sample chromatograph, answers tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end.

5, according to the method for quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba of claim 4, it is characterized in that this method comprises all or part of content in the following differential test method:

One or both thin layer chromatography discrimination method in Herba Erigerontis medical material, the scutellarin in a, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds methanol extraction, the preparation need testing solution; Other gets the scutellarin reference substance, adds methanol and makes the solution that every 1ml contains 1mg; Get the Herba Erigerontis control medicinal material, first weeding of grease solubility impurity refers again to the need testing solution method for making, makes control medicinal material solution; The test of employing thin layer chromatography, one or both and the test solution drawn respectively in the above-mentioned contrast solution are an amount of, point be developing solvent with methanol-ethyl acetate-formic acid=7: 2: 1 on same polyamide membrane, expansion, take out, dry, spray is with 2% ferric chloride alcoholic solution, in the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;

The liquid chromatograph discrimination method of scutellarin in b, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds dissolve with methanol or is diluted to suitable concn, shakes up, and filters, and gets subsequent filtrate as need testing solution; Methanol solution with the scutellarin reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% phosphate aqueous solution=50: 50 is a mobile phase, and the detection wavelength is 335nm; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end;

The thin layer chromatography discrimination method of Radix Paeoniae Rubra medical material, peoniflorin, gallic acid in c, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds methanol extraction, centrifugal, gets supernatant as need testing solution; The methanol solution that other gets peoniflorin, gallic acid reference substance is product solution in contrast; Get the Radix Paeoniae Rubra medical material, make control medicinal material solution with reference to the preparation method of need testing solution; The test of employing thin layer chromatography, draw respectively in control medicinal material or the reference substance solution one or both, and test solution an amount of, point is on same silica gel g thin-layer plate, with chloroform-ethyl acetate-methanol-formic acid=6: 1.5: 1.5: 0.5 be developing solvent, expansion, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph earlier, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the same color speckle; Spray with the vanillin sulfuric acid solution, it is clear to be heated to speckle colour developing again, in the test sample chromatograph, with the corresponding position of contrast chromatograph on, show the same color speckle;

The liquid chromatograph discrimination method of peoniflorin in d, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds methanol extraction or dilution, filters, and gets subsequent filtrate as need testing solution; Methanol solution with the peoniflorin reference substance is a reference substance solution, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% formic acid solution=40: 60 is a mobile phase, and the detection wavelength is 230nm; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, in the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end.

6, according to the method for quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba of claim 1, it is characterized in that this method comprises all or part of content of following assay:

The assay of scutellarin in a, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, is solvent with in methanol or ethanol or the water one or more, extracts or dilution or dissolving, is made as need testing solution; Prepare reference substance solution with the scutellarin reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, with methanol or acetonitrile-water or 0.2%～3% glacial acetic acid aqueous solution or 0.2%～3% aqueous formic acid or 0.01%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=15～70: 85～30 is mobile phase, detects wavelength and be among 200～410nm one or more; The test of employing liquid chromatography, accurate respectively absorption need testing solution and reference substance solution are an amount of, inject chromatograph of liquid, test, calculate with one point external standard method or standard curve method, must not be less than 15mg with what dosage contained scutellarin every day in the compound Chinese medicinal preparation;

Content of paeoniflorin is measured in b, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, adds one or more extractions or the dissolving in methanol or ethanol or the water or is diluted to suitable concn, filters, as need testing solution; Prepare reference substance solution with the peoniflorin reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, methanol or acetonitrile-water or 0.05%～3% glacial acetic acid aqueous solution or 0.05%～3% aqueous formic acid or 0.001%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=15～60: 85～40 mobile phases, detection wavelength are one or several among 200～410nm; Employing liquid chromatography test is drawn need testing solution respectively with reference substance solution is an amount of, and the injection chromatograph of liquid calculates with one point external standard method or standard curve method, and must not be less than 15mg with what dosage contained peoniflorin every day in the compound Chinese medicinal preparation;

Content of total flavone is measured in c, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, with one or more extractions in methanol or water or the ethanol or be diluted to suitable concn, shakes up, as need testing solution; With scutellarin product in contrast, get reference substance solution with legal system; With the retinue solvent is blank, adopts spectrophotography, and place in 300～600nm wavelength or two places measure trap, calculate with one point external standard method or standard curve method; Contain total flavones with dosage and must not be less than 20mg every day in the compound Chinese medicinal preparation;

The assay of Radix Paeoniae Rubra total glycosides or Radix Paeoniae Alba total glucosides in d, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, puts in the measuring bottle, adds strong base solution and makes dilution or dissolving and fixed to scale in right amount, shake up, precision is measured in right amount, puts in the tool plug test tube, put the boiling water bath hydrolysis 0.5 hour～5 hours, and took out, put to room temperature, quantitatively be transferred in the measuring bottle, swing repeatedly with methanol or ethanol or mobile phase and to wash test tube wall, fixed to scale, shake up, filter with microporous filter membrane, get subsequent filtrate as need testing solution; Preparation reference substance solution with the benzoic acid reference substance, adopt liquid chromatography, chromatographic column is a filler with the alkyl silane bonded silica gel, methanol or acetonitrile-water or 0.05%～3% glacial acetic acid aqueous solution or 0.05%～3% aqueous formic acid or 0.001%～1% phosphate aqueous solution or 0.005mol/L～0.3mol/L sodium dihydrogen phosphate or 0.005mol/L～0.3mol/L potassium dihydrogen phosphate=10～60: 90～40 mobile phases, the detection wavelength is one or several in 200～410nm wave-length coverage, and column temperature is in 20～60 ℃ of scopes; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, calculate behind the content again by formula Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides content %=benzoic acid content % * 480.48/122.13 with one point external standard method or standard curve method, calculate the content of Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides must not be less than 20mg.

7, according to the method for quality control of the described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba of claim 6, it is characterized in that this method comprises all or part of content of following assay:

The assay of scutellarin in a, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, and accurate the title decides or measure, and adds methanol extraction or is diluted to suitable concn, shakes up, and filters, and gets subsequent filtrate as need testing solution; Methanol solution with the scutellarin reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% phosphate aqueous solution=50: 50 is a mobile phase, and the detection wavelength is 335nm; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, calculate with one point external standard method, must not be less than 30mg with what dosage contained scutellarin every day in the compound Chinese medicinal preparation;

Content of paeoniflorin is measured in b, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, and accurate the title decides or measure, and adds dissolve with methanol or is diluted to suitable concn, shakes up, and filters, and gets subsequent filtrate as need testing solution; Methanol solution with the peoniflorin reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-0.1% formic acid solution=40: 60 is a mobile phase, and the detection wavelength is 230nm; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, calculate with one point external standard method, must not be less than 30mg with what dosage contained peoniflorin every day in the compound Chinese medicinal preparation;

Content of total flavone is measured in c, the compound Chinese medicinal preparation:

Get in the compound Chinese medicinal preparation to be measured in right amount, put in the measuring bottle, add methanol and make dissolving in right amount and shake up to scale surely, precision is measured 1ml, puts in the 50ml measuring bottle, adds methanol to scale, shakes up, as need testing solution; With scutellarin product in contrast, get reference substance solution with legal system.With the retinue solvent is blank, according to spectrophotography, measures trap at the wavelength place of 335nm, calculate with one point external standard method, each preparation is unit quantity to be equivalent to every day with output, and the per unit amount contains the limit of total flavones in scutellarin, must not be less than 40mg;

The assay of Radix Paeoniae Rubra total glycosides or Radix Paeoniae Alba total glucosides in d, the compound Chinese medicinal preparation:

It is an amount of to get compound Chinese medicinal preparation to be measured, and accurate the title decides or measure, and puts in the measuring bottle, add 5% sodium hydroxide solution and make dissolving and fixed to scale in right amount, shake up, precision is measured in right amount, put in the tool plug test tube, put the boiling water bath hydrolysis 2 hours, take out, put to room temperature, quantitatively be transferred in the 10ml measuring bottle, swing repeatedly with methanol or mobile phase and wash test tube wall, fixed to scale, shake up, filter, get subsequent filtrate as need testing solution with microporous filter membrane; Methanol solution with the benzoic acid reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-0.02mol/L sodium dihydrogen phosphate=30: 70 is a mobile phase, and the detection wavelength is 230nm, 35 ℃ of column temperatures; Draw need testing solution respectively and reference substance solution is an amount of, inject chromatograph of liquid, calculate with one point external standard method, Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides content=benzoic acid content * 480.48/122.13, calculate the content of Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides, compound Chinese medicinal preparation must not be less than 40mg with the limit that contains Radix Paeoniae Alba total glucosides or Radix Paeoniae Rubra total glycosides in the dosage every day.

8, according to the method for quality control of claim 6 or the 7 described compound Chinese medicinal preparation made from Herba Erigerontis, Radix Paeoniae Rubra or the Radix Paeoniae Alba, it is characterized in that: the assay result of described compound Chinese medicinal preparation, calculate according to percentage by weight, the total amount that can measure composition accounts for deduction adjuvant and outer more than 25% of total solid of moisture in the preparation.