CN1919308A

CN1919308A - Quality controlling means of traditional medicine Injectio

Info

Publication number: CN1919308A
Application number: CN 200510092858
Authority: CN
Inventors: 于文风
Original assignee: Qiyuanyide Medicines Institute Beijing
Current assignee: Qiyuanyide Medicines Institute Beijing
Priority date: 2005-08-22
Filing date: 2005-08-22
Publication date: 2007-02-28

Abstract

The invention discloses a method for controlling quality of Chinese medicinal preparation for injection, which comprises herb of shortcape fleabane, ginseng (or red ginseng, Codonopsis pilosula) and ophiopogon root. The quality control method includes fingerprint pattern testing process and/or discrimination testing process and/or content determination process for each constituent in the injection.

Description

A kind of method of quality control of traditional medicine Injectio

Technical field

The present invention relates to a kind of method of quality control, belong to the field of medical technology with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis.

Background technology

Along with the development of human civilization, the raising of people's living standard, cardiovascular and cerebrovascular diseases such as coronary heart disease, angina pectoris have become human second largest killer.Exploitation heart disease class medicine has become instant challenge of pharmacy industry.The applicant is with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and carry out Radix Ophiopogonis having developed a kind of injection after the assembly, and it comprises the injection that is directly used in drug administration by injection, needs to be used for the concentrated solution for injection of intravenous drip after the dilution, directly for the venous transfusion of intravenous drip and injectable sterile powder and the aseptic block that makes with freeze-drying or spray drying method.Calculate according to components by weight percent, it is made through extracting refining and adding suitable adjuvant by 50～5000 parts of 10～1000 parts of 10～1000 parts of Radix Ophiopogonis, Radix Ginseng and Herba Erigerontiss, or the extract that is obtained after extracting by corresponding weight portion medical material is through refining and add suitable adjuvant and be made.For the effective quality of control product, the safety of medicine that satisfy the requirement of producing, guarantees to produce, reliable and curative effect is accurate, the applicant has set up its quality standard.The chemical constituent of known Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and Radix Ophiopogonis has tens kinds.If its inherent quality is described with one, two kinds of active component, has certain one-sidedness, said nothing of the index components of no efficacy, therefore, not only chosen Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or the active component of Radix Codonopsis and Radix Ophiopogonis and carried out assay,, more set up finger printing with the quality of how much judging of its content, its material group integral body is 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.Do not have under the clear and definite situation in the present stage Effective Components of Chinese Herb overwhelming majority, the quality for effective control Chinese crude drug or Chinese patent medicine has great importance.The Japan main manufacturing enterprise of Chinese prescription medicine just adopts the high-efficiency liquid-phase fingerprint control of quality in enterprises in the eighties in 20th century.Germany, France find that the medical function of Folium Ginkgo extract is extract gained material group's mass action result in the process that Folium Ginkgo extract is developed jointly, and to the quality control of such integral body, also adopt high performance liquid chromatography.In the post medical herbs guide that U.S. FDA is formulated clearly with the method for quality control of finger printing as the compounding substances group.Finger printing has become common recognition at present as Chinese herbal medicine and extraction of substance amount control method thereof.

Summary of the invention

The objective of the invention is by a kind of method of quality control with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis is provided, this method of utilizing the inventor to provide, can effectively control quality with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis, concrete guidance is carried out in production to medicine: this method that provides simultaneously is simple, the precision height, favorable reproducibility.

The present invention constitutes like this:

Method of quality control with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis comprises following all or part of content:

(1) finger printing test, comprise finger printing based on the Herba Erigerontis composition characteristics, based on the finger printing of Radix Codonopsis composition characteristics, based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics finger printing, based in the finger printing of composition characteristics Radix Ophiopogonis one or more;

(2) Herba Erigerontis medical material, Radix Ginseng Rubra or Radix Ginseng or codonopsis pilosula, Radix Ophiopogonis medical material, scutellarin, ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rf, ophiopogonin B, ophiopogonin D, ophiopogonin D ', the differential test method of all or part of composition in the Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, lobetyolin, atractylenoide;

(3) ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rf, ophiopogonin B, ophiopogonin D, ophiopogonin D ', the content test method of all or part of composition in the Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, total saponins, total flavones, lobetyolin, atractylenoide.

Described finger printing based on the Herba Erigerontis composition characteristics, based on the finger printing of Radix Codonopsis composition characteristics, based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics finger printing, based on Radix Ophiopogonis composition characteristics fingerprint test method be:

A, employing liquid chromatography test Herba Erigerontis composition characteristics are main finger printing:

(1) preparation of need testing solution: it is an amount of to get the Chinese medicine of making Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and Radix Ophiopogonis, adds dissolving of water or methanol equal solvent or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: get the main active reference substance scutellarin in an amount of Herba Erigerontis medical material, be diluted to suitable concn with methanol or dissolve with ethanol, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler: mobile phase is acetonitrile or methanol: 0.005mol/L～2mol/L sodium dihydrogen phosphate (phosphoric acid is regulated pH=2.0～5.0) or 0.2%～5% glacial acetic acid or 0.2%～5% formic acid or 0.2%～3% phosphoric acid solution, gradient elution, flow velocity is that 0.5～2.0ml/min, detection wavelength are one or several in the 190-400nm scope, and column temperature is in 20～60 ℃ of scopes;

(4) based on the formulation of the standard finger-print of Herba Erigerontis composition characteristics: accurate draw need testing solution and object of reference solution an amount of, inject chromatograph of liquid respectively, according to measured collection of illustrative plates, formulate standard finger-print;

(5) with the means of testing of said method as Herba Erigerontis ingredients fingerprint in the injection to be measured;

(6) with the finger printing and the contrast of above-mentioned standard finger-print of injection to be measured, calculate similarity, should be 0.80～1.00;

B, adopt liquid chromatography test Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics be main finger printing:

(1) preparation of need testing solution: it is an amount of to get the Chinese medicine of making Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and Radix Ophiopogonis, adds water or methanol or dissolve with ethanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: get the main active reference substance in an amount of Radix Ginseng Rubra or Radix Ginseng and the Radix Ophiopogonis medical material, comprise ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rf, ophiopogonin B, ophiopogonin D, Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, ophiopogonin D ' in one or more, water or methanol, dissolve with ethanol are diluted to suitable concn, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler: mobile phase is 20%～99% acetonitrile or 20%～99% methanol: 0.01mol/L～2mol/L sodium dihydrogen phosphate or 0.01mol/L～2mol/L potassium dihydrogen phosphate or 0.2%～3% glacial acetic acid or 0.2%～3% formic acid or 0.2%～3% phosphoric acid solution, gradient elution, flow velocity is that 0.5～2.0ml/min, detection wavelength are one or several in the 190-300nm scope, and column temperature is in 20～60 ℃ of scopes;

(4) based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics the formulation of standard finger-print: accurate draw need testing solution and object of reference solution an amount of, inject chromatograph of liquid respectively, according to measured collection of illustrative plates, formulate standard finger-print;

(5) with said method as Radix Ginseng Rubra or Radix Ginseng in the injection to be measured and Radix Ophiopogonis ingredients fingerprint means of testing;

C, employing liquid chromatography test Radix Codonopsis composition characteristics are main finger printing:

(2) preparation of object of reference solution: get the main active reference substance in an amount of codonopsis pilosula, comprise in lobetyolin, the atractylenoide one or more, water or methanol, dissolve with ethanol are diluted to suitable concn, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler: mobile phase is 5%～99%: 95%～5% acetonitrile or methanol-water or 0.01mol/L～2mol/L sodium dihydrogen phosphate or 0.2%～3% glacial acetic acid or 0.2%～3% formic acid or 0.2%～3% phosphoric acid solution, flow velocity is 0.5～2.0ml/min, detect wavelength is that one or several or evaporation photodetector in the 190-300nm scope detects, and column temperature is in 20～60 ℃ of scopes;

(4) based on the formulation of the standard finger-print of Radix Codonopsis composition characteristics: accurate draw need testing solution and object of reference solution an amount of, inject chromatograph of liquid respectively, according to measured collection of illustrative plates, formulate standard finger-print;

(5) with the means of testing of said method as Radix Codonopsis ingredients fingerprint in the injection to be measured;

D, employing liquid chromatography test composition characteristics Radix Ophiopogonis are main finger printing:

(2) preparation of object of reference solution: get the main active reference substance in an amount of Radix Ophiopogonis of the medical material, comprise ophiopogonin B, ophiopogonin D, Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, ophiopogonin D ' in one or more, water or methanol, dissolve with ethanol are diluted to suitable concn, as object of reference solution;

(4) based on the formulation of standard finger-print of composition characteristics Radix Ophiopogonis: accurate draw need testing solution and object of reference solution an amount of, inject chromatograph of liquid respectively, according to measured collection of illustrative plates, formulate standard finger-print;

(5) with said method as in the injection to be measured Radix Ophiopogonis ingredients fingerprint means of testing;

(6) with the finger printing and the contrast of above-mentioned standard finger-print of injection to be measured, calculate similarity, should be 0.80～1.00.

Described method of quality control with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis comprises one or more in the following finger printing:

(1) preparation of need testing solution: it is an amount of that precision takes by weighing this product, adds methanol and make the solution that every 1ml contains 50mg, filters with microporous filter 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 0.1mg, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is 0.02moL/L sodium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), gradient elution, solvent ratios was from 0 minute to 8 minutes, and the ratio of mobile phase A rises to 30% by 18%, from 8 minutes to 15 minutes, the ratio of acetonitrile rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of acetonitrile reduced to 18% by 50%, from 18 minutes to 60 minutes, the ratio of acetonitrile was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 40 ℃;

(4) based on the formulation of the standard finger-print of Herba Erigerontis 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 measured collection of illustrative plates, the formulation standard finger-print;

(6) with injection finger printing to be measured and the contrast of above-mentioned standard finger-print, calculate similarity, should be 0.90～1.00;

B, adopt liquid chromatography for measuring based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics finger printing:

(1) preparation of need testing solution: it is an amount of that precision takes by weighing this product, adds methanol and make the solution that every 1ml contains 50mg, filters, and gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing the ginsenoside Rg1, adds methanol and make the solution that every 1ml contains 0.3mg, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is the 0.05mol/L potassium dihydrogen phosphate, and Mobile phase B is acetonitrile-water 80: 20, gradient elution, solvent ratios was from 0 minute to 5 minutes, the ratio of Mobile phase B is 25%, and from 5 minutes to 40 minutes, the ratio of Mobile phase B rose to 64% by 25%, from 40 minutes to 50 minutes, the ratio of Mobile phase B is 64%, and from 50 minutes to 65 minutes, the ratio of Mobile phase B rose to 80% by 64%, from 65 minutes to 75 minutes, the ratio of Mobile phase B reduced to 25% by 80%; Flow velocity is 1.0ml/min, and the detection wavelength is 203 ± 2nm, and column temperature is 40 ℃;

(4) based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics the formulation of standard finger-print: 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 measured collection of illustrative plates, the formulation standard finger-print;

(1) preparation of need testing solution: it is an amount of to get the Chinese medicine of making Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and Radix Ophiopogonis, adds methanol and makes the solution that every 1ml contains 50mg, filters, and gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: get the main active atractylenoide in an amount of codonopsis pilosula, add methanol and make the solution that every 1ml contains 0.1mg, as object of reference solution;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler: mobile phase is methanol-water 67: 33, and flow velocity is that 1.0ml/min, evaporation photodetector detect, 30 ℃ of drift tube temperatures, and carrier gas flux 2.2L/min, column temperature are 30 ℃;

(6) with the finger printing and the contrast of above-mentioned standard finger-print of injection to be measured, calculate similarity, should be 0.90～1.00;

D, adopt the finger printing of liquid chromatography for measuring based on composition characteristics Radix Ophiopogonis:

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing ophiopogonin B, adds methanol and make the solution that every 1ml contains 0.3mg, as object of reference solution;

(4) based on the formulation of standard finger-print of composition characteristics Radix Ophiopogonis: 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 measured collection of illustrative plates, the formulation standard finger-print;

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

Described discrimination method with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis is following all or part of content:

The thin layer chromatography discrimination method of scutellarin in a, the injection:

It is an amount of to get each preparation, adds ethyl acetate or ethanol or methanol extraction, filters, and filtrate volatilizes, and residue is with methanol or dissolve with ethanol, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other gets the Herba Erigerontis control medicinal material, shines medical material solution in pairs with legal system; Get the scutellarin reference substance again, add methanol or ethanol and make the solution that every 1ml contains 1mg; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica GF254 lamellae, with ethyl acetate or Ethyl formate-formic acid or acetic acid-water 1～60: 0.2～10: 0.3～5 or benzene or toluene-ethyl acetate or Ethyl formate-formic acid or acetic acid-water 1～30: 0.5～15: 0.05～5: 0.01～3 is developing solvent, launch, take out, dry, spray is with 0.3～10% aluminum chloride ethanol or 0.3～10% aluminum chloride methanol solution, 90～130 ℃ were dried by the fire 1～15 minute, put under the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, should show the same color speckle;

The liquid chromatograph discrimination method of scutellarin in b, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol or mobile phase or ethanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol or alcoholic solution with the scutellarin reference substance are contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 5%～95%: 95%～5% methanol or acetonitrile-water or 0.05%～10% glacial acetic acid aqueous solution or 0.02%～2% phosphate aqueous solution or 2%～30%: 2%～30%: 96%～40% methanol or acetonitrile-oxolane-0.02%～2% phosphate aqueous solution or acetonitrile-0.005～0.3moL/L sodium dihydrogen phosphate (1～99% phosphoric acid is transferred pH=2.0～5.0) gradient elution system is a mobile phase, the detection wavelength is one or several in 200～410nm scope, in 20～50 ℃ of scopes of column temperature; In the test sample chromatograph, answer tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless;

In c, the injection Radix Ophiopogonis medical material the thin layer chromatography discrimination method:

It is an amount of to get each preparation, adds n-butyl alcohol or ethyl acetate or chloroform extraction, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other gets control medicinal material Radix Ophiopogonis, shines medical material solution in pairs with legal system; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate or silica gel H lamellae or silica GF254 lamellae, with benzene or toluene or chloroform-methanol or ethanol-glacial acetic acid or formic acid or water 8～300: 0.5～100: 0.01～30 or ethyl acetate or Ethyl formate-pyridine-water 0.2～10: 0.1～5: 0.2～10 is developing solvent, launch, take out, dry, put and inspect under uviol lamp 365nm or the 254nm or spray with 10% sulphuric acid or vanillin reagent or 50% sulphuric acid or 5% vanillin reagent or anisaldehyde reagent, 90 ℃～130 ℃ dry by the fire to speckle colour developing clear, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, the fluorescence speckle that should show same color, negative noiseless;

Ophiopogonin B in d, the injection, ophiopogonin D, ophiopogonin D ' the thin layer chromatography discrimination method:

It is an amount of to get each preparation, and with usefulness n-butyl alcohol and ether or ethyl acetate extraction behind the water extraction, extract evaporate to dryness, residue is with methanol or dissolve with ethanol, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other get ophiopogonin B, ophiopogonin D, ophiopogonin D ', add methanol or ethanol respectively and make the solution that every 1ml contains 1mg, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate or silica gel H lamellae or silica GF254 lamellae, with ethyl acetate or chloroform-methanol-water 3～50: 1～15: 0.1～5 is developing solvent, launch, take out, dry, spray is with 5～50% sulphuric acid ethanol reagent, 90 ℃～130 ℃ dry by the fire to speckle colour developing clear, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, the speckle that should show same color, negative noiseless;

Ophiopogonin B in e, the injection, ophiopogonin D, ophiopogonin D ' liquid chromatograph differentiate:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol or mobile phase or ethanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ophiopogonin B, ophiopogonin D, ophiopogonin D ' reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 1%～99%: 99%～1% methanol or acetonitrile-water or 0.05%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution are mobile phase, the detection wavelength is one or several in 200～410nm scope, in 20～50 ℃ of scopes of column temperature; In the test sample chromatograph, answer tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless;

The thin layer chromatography discrimination method of Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin in f, the injection:

It is an amount of to get each preparation, adds 0.5～38% sulphuric acid or hydrochloric acid hydrolysis, regulates pH to neutral, evaporate to dryness, and residue is with chloroform or ethyl acetate dissolving, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other gets Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, adds chloroform or ethyl acetate respectively and makes the solution that every 1ml contains 1mg, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate or silica gel H lamellae or silica GF254 lamellae, with normal hexane or methanol-ethyl acetate or chloroform or Ethyl formate-water 0.2～15: 0.2～40: 0.1～5 is developing solvent, launch, take out, dry, spray is with 5～50% sulphuric acid ethanol reagent, 90 ℃～130 ℃ dry by the fire to speckle colour developing clear, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, the speckle that should show same color, negative noiseless;

The liquid chromatograph of Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin is differentiated in g, the injection:

It is an amount of to get each preparation, puts in the round-bottomed flask, and take out with 0.5～38% sulphuric acid or hydrochloric acid hydrolysis the back that is dissolved in water, put to room temperature, transfer pH to neutral, with chloroform or ethyl acetate extraction, the extracting solution evaporate to dryness, residue filters with microporous filter membrane with methanol or dissolve with ethanol, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 5%～80%: 95%～20% methanol or acetonitrile-water or 0.05%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution are mobile phase, the detection wavelength is one or several in 200～410nm scope, in 20～50 ℃ of scopes of column temperature; In the test sample chromatograph, answer tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless;

Radix Ginseng Rubra or Radix Ginseng, ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rf's thin layer chromatography discrimination method in h, the injection:

It is an amount of to get each preparation, with n-butyl alcohol or ethanol or methanol extraction, filters, and filtrate is as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Get Radix Ginseng Rubra or Radix Ginseng control medicinal material, add the chloroform reflux, extract,, discard chloroform solution, residue adds water-saturated n-butanol and extracts, and extracting solution adds ammonia solution, and divide and get the upper strata, evaporate to dryness, residue is with methanol or dissolve with ethanol, medical material solution in contrast; Other gets ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rf's reference substance, adds methanol or ethanol respectively and makes the solution that every 1ml contains 2mg, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate or silica gel H lamellae or silica GF254 lamellae, lower floor's solution or n-butyl alcohol-ethyl acetate or the Ethyl formate-water 1～10: 0.2～2 placed below 5～40: 10～100: 5～50: 0.2～30 10 ℃ with chloroform-ethyl acetate or Ethyl formate-methanol-water: 1～15 upper solution is developing solvent, launch, take out, dry, spray is with 5～50% sulphuric acid ethanol reagent or 50% sulphate reagent, 90 ℃～130 ℃ dry by the fire to speckle colour developing clear, put respectively under daylight and the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, should show the speckle of same color, negative noiseless;

I. ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's liquid chromatograph is differentiated in the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds water or methanol or mobile phase to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels, methanol or acetonitrile-water or 0.5%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution or 0.5%～5% formic acid solution are mobile phase, gradient elution, flow velocity is 0.5～2.0ml/min, one or several or the evaporation photodetector that detect wavelength and be in 200～350nm scope detect, and column temperature is in 20～50 ℃ of scopes; In the test sample chromatograph, answer tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless;

The thin layer chromatography discrimination method of lobetyolin in j, the injection:

It is an amount of to get each preparation, be dissolved in water, with n-butyl alcohol or ethyl acetate or chloroform extraction, extract is concentrated into dried, and residue is with methanol or dissolve with ethanol, as need testing solution, or put in C18 or the C8 solid phase extraction column, use 5%～50% methanol, methanol-eluted fractions successively, collect meoh eluate, be concentrated into 1ml, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other gets the Radix Codonopsis control medicinal material, shines medical material solution in pairs with legal system; Get lobetyolin's reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica GF254 lamellae, n-butyl alcohol-glacial acetic acid or formic acid or ethanol or dehydrated alcohol-water 1～35: 0.1～10: 0.05～5 are developing solvent, launch, and take out, dry, spray is with 3%～30% ethanol solution of sulfuric acid, and 80～150 ℃ were heated 1～30 minute, and put under the ultra-violet lamp 365nm and inspect.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, should show the fluorescence speckle of same color, negative noiseless;

The liquid chromatograph discrimination method of lobetyolin in k, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol or ethanol or mobile phase to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with lobetyolin's reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 5%～80%: 95%～20% methanol or acetonitrile-water or 0.05%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution are mobile phase, the detection wavelength is one or several in 200～410nm scope, in 20～50 ℃ of scopes of column temperature; In the test sample chromatograph, answer tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless.

Described discrimination method with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis is following one or more:

It is an amount of to get each preparation, adds ethyl acetate extraction, filters, and filtrate volatilizes, and the residue dissolve with methanol is as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other gets the Herba Erigerontis control medicinal material, shines medical material solution in pairs with legal system; Get the scutellarin reference substance again, add methanol and make the solution that every 1ml contains 1mg; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, toluene-ethyl acetate-formic acid-water 10: 5: 1: 0.5 is developing solvent, launch, take out, dry, spray is with 3% aluminum chloride alcoholic solution, 105 ℃ were dried by the fire 5 minutes, and put under the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the same color speckle;

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with the scutellarin reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-0.1% phosphate aqueous solution is a mobile phase at 20: 80, and the detection wavelength is 335nm, 30 ℃ of column temperatures; In the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless;

The thin layer chromatography discrimination method of Radix Ophiopogonis in c, the injection:

It is an amount of to get each preparation, adds 7: 3 mixed solutions of chloroform-methanol and extracts, and filters, and filtrate volatilizes, and residue adds the chloroform dissolving, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other gets control medicinal material Radix Ophiopogonis, shines medical material solution in pairs with legal system; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 10 μ l of above-mentioned solution, put in same silica gel G F respectively ₂₅₄On the lamellae, be at 80: 5: 0.1 developing solvent, launch, put under the uviol lamp 254nm and inspect with toluene-methanol-glacial acetic acid, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, negative noiseless;

It is an amount of to get each preparation, with using n-butanol extraction behind the water dissolution, and the extract evaporate to dryness, the residue dissolve with methanol is as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other get ophiopogonin B, ophiopogonin D, ophiopogonin D ', add methanol respectively and make the solution that every 1ml contains 1mg, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-methanol-water is developing solvent at 15: 5: 1, launch, take out, dry, spray is with 10% sulphuric acid ethanol reagent, 105 ℃ dry by the fire to speckle colour developing clear, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, the speckle that shows same color, negative noiseless;

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ophiopogonin B, ophiopogonin D, ophiopogonin D ' reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-water is a mobile phase at 90: 10, and the detection wavelength is 203nm, 30 ℃ of column temperatures; In the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless;

It is an amount of to get each preparation, adds 3% sulphuric acid hydrolysis 4 hours, regulates pH to neutral, evaporate to dryness, and residue dissolves with chloroform, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other gets Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, adds chloroform respectively or the ethyl acetate equal solvent is made the solution that every 1ml contains 1mg, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate or-water is developing solvent at 1: 1: 1, launch, take out, dry, spray is with 10% sulphuric acid ethanol reagent, 90 ℃ dry by the fire to speckle colour developing clear, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, the speckle that shows same color, negative noiseless;

It is an amount of to get each preparation, puts in the round-bottomed flask, is dissolved in water, and refluxes 4 hours with 3% sulphuric acid, takes out, and puts to room temperature, transfers pH to neutral, uses chloroform extraction, the extracting solution evaporate to dryness, and the residue dissolve with methanol filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin reference substance is contrast, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-water is a mobile phase at 90: 10, the detection wavelength is 203nm, 30 ℃ of column temperatures; In the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless;

It is an amount of to get each preparation, uses n-butanol extraction, filters, and filtrate is as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Get Radix Ginseng Rubra or Radix Ginseng control medicinal material, add the chloroform reflux, extract,, discard chloroform solution, residue adds water-saturated n-butanol and extracts, and extracting solution adds ammonia solution, divides and gets the upper strata, evaporate to dryness, residue dissolve with methanol, medical material solution in contrast; Other gets ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rf's reference substance, adds methanol respectively and makes the solution that every 1ml contains 2mg, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 10 μ l of above-mentioned solution, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica gel G F ₂₅₄On the lamellae, with chloroform-ethyl acetate-methanol-water 15: 40: 22: lower floor's solution of placing below 1010 ℃ was developing solvent, launches, and takes out, dry, spray is with 10% sulphuric acid ethanol reagent, and 105 ℃ are dried by the fire to the speckle colour developing clearly, put respectively under daylight and the ultra-violet lamp 365nm and inspect, 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, negative noiseless;

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's reference substance is contrast, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, acetonitrile-water is a mobile phase, gradient elution, solvent ratios is from 0 minute to 35 minutes, the ratio of acetonitrile is 19%, from 35 minutes to 55 minutes, the ratio of acetonitrile rises to 29% by 19%, and from 55 minutes to 70 minutes, the ratio of acetonitrile was 29%, from 70 minutes to 100 minutes, the ratio of acetonitrile rose to 40% by 29%; Flow velocity is 1.0ml/min, detects wavelength 203nm, and column temperature is at 30 ℃; In the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless;

It is an amount of to get each preparation, add methanol 25ml, supersound process 30 minutes filters, the filtrate evaporate to dryness, residue adds water 2ml makes dissolving, puts in the C18 solid phase extraction column (500mg is with methanol, each the 10ml prewashing of 20% methanol), use 20% methanol, each 5ml eluting of methanol successively, collect meoh eluate, be concentrated into 1ml, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Other gets the Radix Codonopsis control medicinal material, shines medical material solution in pairs with legal system; It is an amount of to get lobetyolin's reference substance again, adds methanol and makes every 1ml and contain 1mg solution, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw need testing solution 6 μ l, reference substance solution 2 μ l, put respectively on same high-efficient silica gel G lamellae, with n-butyl alcohol-glacial acetic acid-water is developing solvent at 7: 1: 0.5, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ were heated 5 minutes, and put under the ultra-violet lamp 365nm and inspect.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color, negative noiseless;

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with lobetyolin's reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-water is a mobile phase at 22: 78, and the detection wavelength is 267nm, 30 ℃ of column temperatures; In the test sample chromatograph, tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless.

Described method of testing with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine injection agent content made Radix Ophiopogonis should comprise one or more in following:

The assay of scutellarin in a, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol or mobile phase or ethanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol or alcoholic solution with the scutellarin reference substance are contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 5%～95%: 95%～5% methanol or acetonitrile-water or 0.05%～10% glacial acetic acid aqueous solution or 0.02%～2% phosphate aqueous solution or 2%～30%: 2%～30%: 96%～40% methanol or acetonitrile-oxolane-0.02%～2% phosphate aqueous solution or acetonitrile-0.005～0.3moL/L sodium dihydrogen phosphate (1～99% phosphoric acid is transferred pH=2.0～5.0) gradient elution system is a mobile phase, the detection wavelength is one or several in 200～410nm scope, in 20～50 ℃ of scopes of column temperature; Calculate with external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains scutellarin must not be less than 6.0mg;

Content of total flavone is measured in b, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol or water is diluted 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, adopt the disclosed spectrophotography experiment of Chinese Pharmacopoeia appendix, measure trap at the wavelength place of 335 ± 10nm or get each preparation an amount of, put in the measuring bottle, thin up is to scale, shake up, precision is measured in right amount, puts in the 10ml measuring bottle, adds 50% methanol or water to 5ml, add 5% sodium nitrite solution, 0.3～1ml, shake up, placed 6 minutes, add 10% aluminum nitrate solution, 0.3～1ml, shake up, placed 6 minutes, hydro-oxidation sodium test solution 4～10ml adds 50% methanol or water again to scale, shake up, as need testing solution; With rutin or scutellarin product in contrast, get reference substance solution with legal system.With the retinue solvent is blank, adopt the Chinese Pharmacopoeia appendix disclosed according to the spectrophotography experiment, measure trap at 500 ± 10nm place, calculate with one point external standard method or standard curve 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 rutin or scutellarin, must not be less than 30.0mg;

C. ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's assay in the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol or mobile phase or ethanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels, methanol or acetonitrile-water or 0.5%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution or 0.5%～5% formic acid solution are mobile phase, gradient elution, flow velocity is 0.5～2.0ml/min, one or several or the evaporation photodetector that detect wavelength and be in 200～350nm scope detect, and column temperature is in 20～50 ℃ of scopes; Calculate with external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, the limit that the per unit amount contains the ginsenoside Rg1 must not be less than 1.0mg, the limit that contains the ginsenoside Re must not be less than 0.5mg, the limit that contains the ginsenoside Rb1 must not be less than 1.0mg, and the limit that contains ginsenoside Rg1, ginsenoside Re's summation must not be less than 2.5mg;

D. the assay of total saponins in the injection:

It is an amount of to get each preparation, puts in the measuring bottle, and adding distil water makes dissolving and fixed to scale in right amount, shake up, precision is measured in right amount, to measuring bottle, water bath method takes out immediately, and precision adds 5% vanillin-glacial acetic acid solution 0.2～4ml, perchloric acid 0.5～5ml shakes up, and heating is 15 minutes in 60 ℃ of water-baths, take out, with ice-water bath cooling 0.5～10 minute, precision added glacial acetic acid to scale immediately, shake up, as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; With the product of ginsenoside Rg1 or ginsenoside Rb1 or ginsenoside Re or ginsenoside Rf or ophiopogonin B or ophiopogonin D or ophiopogonin D ' in contrast, get reference substance solution with legal system.With the retinue solvent is blank, adopt the disclosed spectrophotography of Chinese Pharmacopoeia appendix, wavelength place at 547 ± 10nm measures trap, calculate with external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, the per unit amount contain total saponins in ginsenoside Rg1 or ginsenoside Rb1 or ginsenoside Re or ginsenoside Rf or ophiopogonin B or ophiopogonin D or ophiopogonin D ', must not be less than 18.0mg;

Ophiopogonin B, ophiopogonin D, ophiopogonin D ' assay in e, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ophiopogonin B, ophiopogonin D, ophiopogonin D ' reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 1%～99%: 99%～1% methanol or acetonitrile-water or 0.05%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution are mobile phase, one or several or the evaporation photodetector that detect wavelength and be in 200～410nm scope detect, in 20～50 ℃ of scopes of column temperature; Calculate with one point external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains ophiopogonin B must not be less than 0.05mg, and the limit that contains ophiopogonin D must not be less than 0.05mg; Contain ophiopogonin D ' limit must not be less than 0.01mg; Contain ophiopogonin B, ophiopogonin D, ophiopogonin D ' the limit of summation must not be less than 0.11mg;

Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin assay in f, the injection:

It is an amount of to get each preparation, puts in the round-bottomed flask, and take out with 0.5～38% sulphuric acid or hydrochloric acid hydrolysis the back that is dissolved in water, put to room temperature, transfer pH, use chloroform extraction, the extracting solution evaporate to dryness to neutral, the residue dissolve with methanol filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 5%～80%: 95%～20% methanol or acetonitrile-water or 0.05%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution are mobile phase, one or several or the evaporation photodetector that detect to detect wavelength and be in 200～410nm scope detect, in 20～50 ℃ of scopes of column temperature; Calculate with one point external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains Ruscus aculeatus L. sapogenin must not be less than 0.01mg, and the limit that contains diosgenin must not be less than 0.01mg; The limit that contains ruscogenin must not be less than 0.01mg; The limit that contains the summation of Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin must not be less than 0.03mg;

The assay of lobetyolin in g, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with lobetyolin's reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 5%～80%: 95%～20% methanol or acetonitrile-water or 0.05%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution are mobile phase, the detection wavelength is one or several in 200～410nm scope, in 20～50 ℃ of scopes of column temperature; Calculate with one point external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains lobetyolin must not be less than 0.1mg.

Described method of testing with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine injection agent content made Radix Ophiopogonis is following one or more methods:

The assay of scutellarin in a, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol or mobile phase equal solvent to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with the scutellarin reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-0.1% phosphate aqueous solution is a mobile phase at 20: 80, and the detection wavelength is 335nm, 30 ℃ of column temperatures; Calculate with external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains scutellarin must not be less than 6.0mg;

Content of total flavone is measured in b, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol and makes dissolving and fixed to scale in right amount, shakes up, and 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, adopt the disclosed spectrophotography of Chinese Pharmacopoeia appendix, wavelength place at 335nm measures trap, calculate with one point external standard method or standard curve 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 30.0mg;

C. ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's assay in the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's reference substance is contrast, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, acetonitrile-water is a mobile phase, gradient elution, solvent ratios is from 0 minute to 35 minutes, the ratio of acetonitrile is 19%, from 35 minutes to 55 minutes, the ratio of acetonitrile rises to 29% by 19%, and from 55 minutes to 70 minutes, the ratio of acetonitrile was 29%, from 70 minutes to 100 minutes, the ratio of acetonitrile rose to 40% by 29%; Flow velocity is 1.0ml/min, detects wavelength 203nm, and column temperature is in 10～50 ℃ of scopes; Calculate with external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, the limit that the per unit amount contains the ginsenoside Rg1 must not be less than 1.0mg, the limit that contains the ginsenoside Re must not be less than 0.5mg, the limit that contains the ginsenoside Rb1 must not be less than 1.0mg, and the limit that contains ginsenoside Rg1, ginsenoside Re's summation must not be less than 2.5mg;

D. the assay of total saponins in the injection:

It is an amount of to get each preparation, put in the 10ml measuring bottle, adding distil water makes dissolving in right amount and decides and shakes up to scale, and precision is measured 0.6ml, to the 25ml measuring bottle, water bath method takes out immediately, and precision adds 5% vanillin-glacial acetic acid solution 0.5ml, perchloric acid 2.0ml, shake up, heating is 15 minutes in 60 ℃ of water-baths, takes out, immediately with ice-water bath cooling 2 minutes, precision adds glacial acetic acid to 25ml, shakes up, and be blank with the retinue solvent, adopt the disclosed spectrophotography of Chinese Pharmacopoeia appendix, wavelength place at 547nm measures trap, calculates with external standard method or standard curve method, and each preparation is unit quantity to be equivalent to every day with output, the per unit amount contains total saponins in the ginsenoside Rg1, must not be less than 18.0mg;

Ophiopogonin B, ophiopogonin D, ophiopogonin D ' assay in e, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ophiopogonin B, ophiopogonin D, ophiopogonin D ' reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-water is a mobile phase at 90: 10, and the detection wavelength is 203nm, 30 ℃ of column temperatures; Calculate with one point external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains ophiopogonin B must not be less than 0.05mg, and the limit that contains ophiopogonin D must not be less than 0.05mg; Contain ophiopogonin D ' limit must not be less than 0.01mg; Contain ophiopogonin B, ophiopogonin D, ophiopogonin D ' the limit of summation must not be less than 0.11mg;

Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin assay in f, the injection:

It is an amount of to get each preparation, puts in the round-bottomed flask, and refluxing 4 hours with 3% sulphuric acid in the back that is dissolved in water, takes out, and puts to room temperature, transfers pH to neutral, uses chloroform extraction, the extracting solution evaporate to dryness, and the residue dissolve with methanol filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin reference substance is contrast, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-water is a mobile phase at 90: 10, the detection wavelength is 203nm, 30 ℃ of column temperatures; Calculate with one point external standard method or standard curve method, calculate with one point external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains Ruscus aculeatus L. sapogenin must not be less than 0.01mg, and the limit that contains diosgenin must not be less than 0.01mg; The limit that contains ruscogenin must not be less than 0.01mg; The limit that contains the summation of Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin must not be less than 0.03mg;

The assay of lobetyolin in g, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with lobetyolin's reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-water is a mobile phase at 22: 78, and the detection wavelength is 267nm, 30 ℃ of column temperatures; Calculate with one point external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains lobetyolin must not be less than 0.1mg.

Described with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, the method of quality control of the Chinese medicine of making Radix Ophiopogonis, it is characterized in that: calculate according to percentage by weight, all can be surveyed composition and comprise scutellarin in the described injection, the ginsenoside Rg1, the ginsenoside Rb1, the ginsenoside Re, total saponins, ophiopogonin B, ophiopogonin D, ophiopogonin D ', Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, lobetyolin, in the atractylenoide all or part of total content be not less than in the preparation deduction adjuvant amount and water quantities total solid 25%.

Compared with prior art, the present invention more perfect control with the quality of Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine injection products made Radix Ophiopogonis.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 no drug effect.Therefore the applicant formulated based on the finger printing of Herba Erigerontis composition characteristics based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics finger printing and control the quality of injection based on the finger printing of Radix Codonopsis composition characteristics comprehensively.But owing to contained complex chemical composition between each medical material that the present invention relates in the injection, formulation to finger printing causes interference, cause each several part finger printing feature instability,, just can obtain good finger printing so must control mobile phase isochromatic spectrum condition.That is to say, the finger printing that the present invention relates to preparation be not with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and Radix Ophiopogonis medical material or the finger printing simple superposition of preparation just getable, because four kinds of medical material ingredients interference effect each other in the prescription, cause the present invention relates to Herba Erigerontis part in the preparation, Radix Codonopsis part, Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis part the finger printing characteristic peak change, and have only the condition of the present invention of employing, just can obtain ideal finger printing.

Proof by experiment, method of quality control of the present invention is to more effective with the quality control of Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine injection products made Radix Ophiopogonis, and method precision, stability are all higher.

Experimental example 1 is based on the preparation of the finger printing of Herba Erigerontis composition characteristics

A, experimental apparatus, reagent and sample

Reference substance: scutellarin: Nat'l Pharmaceutical ﹠ Biological Products Control Institute;

B, chromatographic condition and system suitability experiment

1. the selection of chromatographic column

In the research process, having selected conventional octadecylsilane chemically bonded silica for use is the liquid-phase chromatographic column of filler, tried out Zorbax, Inertsil ODS-3 respectively, Diamonsil ODS (is C18,250mm * 4.6mm, 5 μ m) chromatographic column of three kinds of trades mark, result show that the chromatographic column of three kinds of trades mark all can reach separating effect preferably, and wherein Diamonsil ODS chromatographic column separating effect is best, post is imitated the highest, can reach 28000 (calculating with object of reference scutellarin peak).So finally selecting Diamonsil ODS chromatographic column (250 * 4.6mm, 5 μ m) for use is the experimentation post.

2. the selection of mobile phase

Investigated (1) methanol-water (10: 90) in the research process respectively, (2) methanol-0.1% glacial acetic acid (5: 95), (3) (the gradient elution volume proportion is from 0 minute to 8 minutes to methanol-0.5% glacial acetic acid (gradient elution) (4) acetonitrile-0.02moL/L sodium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), the ratio of acetonitrile rises to 30% by 18%, from 8 minutes to 15 minutes, the ratio of acetonitrile rises to 50% by 30%, from 15 minutes to 18 minutes, the ratio of acetonitrile reduces to 18% by 50%, from 18 minutes to 60 minutes, the ratio of acetonitrile was 18%) four kinds of flow phase system.The result shows that peak shape is relatively poor under mobile phase (1) condition, separates bad; Under mobile phase (2) condition, peak shape is better, but 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 (3) condition, peak shape is better, and it is incomplete to go out the peak; Under mobile phase (4) condition, peak shape is better, goes out the peak fully and be evenly distributed, so finally selected.

3. detection wavelength determination

In the research under acetonitrile-0.02moL/L sodium dihydrogen phosphate (25% phosphoric acid transfer pH=3.5) (gradient elution) mobile phase condition, investigated the chromatographic peak situation under different-waveband typical wavelengths 205,230,254,283,325nm respectively, the result shows, chromatographic peak is less 205,230, under the 325nm, 205, baseline drift is serious under the 230nm, 254, baseline is good under the 283nm, and wherein chromatographic peak is more under the 283nm, so finally select for use 283nm as detecting wavelength.

4. instrument, chromatographic column and integral parameter

4.1 instrument parameter: selected liquid-phase chromatographic analysis field mainstream configuration and well behaved Agilent1100 series of high efficiency chromatograph of liquid for use, the Chemstation chromatographic work station.Chromatographic column is Diamonsil ODS (250mm * 4.6mm, 5 μ m); 40 ℃ of column temperatures, flow velocity 1.0ml/min.

4.2 integral parameter: Slope Sensitivity:1, peak width:0.05, smallest peaks area are 5% of object of reference (S) peak-to-peak area, minimum peak height be the S peak-to-peak high 5%.So setting can be avoided some very calculating of the chromatographic peak of small size (unimodal area accounts for total peak area less than 0.5%), guarantees the dependency with object of reference simultaneously.

5. the preparation of need testing solution

It is an amount of that precision takes by weighing this product, adds water and make the solution that every 1ml contains 50mg, promptly.

6. the preparation of object of reference solution

Scutellarin is one of Herba Erigerontis main active, and its integral area proportion in finger printing more greatly and more stable is taken into account the research of intermediate and medical material simultaneously, and therefore selected scutellarin is as object of reference.

7. finger printing and technical parameter

Formulate standard finger-print according to 10 batch samples, test sample finger printing and standard finger-print are compared, similarity is all between 0.90～1.00.

Experimental example 2 based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics the preparation of finger printing

A, experimental apparatus, reagent and sample

Reference substance: ginsenoside Rg ₁: Nat'l Pharmaceutical ﹠ Biological Products Control Institute;

B, chromatographic condition and system suitability experiment

1. the selection of chromatographic column

In the research process, having selected conventional octadecylsilane chemically bonded silica for use is the liquid-phase chromatographic column of filler, tried out Zorbax, Inertsil ODS-3 respectively, Diamonsil ODS (is C18,4.6mm * 200mm, 5 μ m) chromatographic column of three kinds of trades mark, result show that the chromatographic column of three kinds of trades mark all can reach separating effect preferably, and wherein Diamonsil ODS chromatographic column separating effect is best, post is imitated the highest, can reach 5400 (calculating with the object of reference ginsenoside Rg1).So finally selecting Diamonsil ODS chromatographic column (4.6mm * 200mm, 5 μ m) for use is the experimentation post.

2. the selection of mobile phase

Investigated (1) methanol-water (20: 80) in the research process respectively, (2) acetonitrile-water (10: 80), (3) acetonitrile-water (gradient elution) (4) A is 50mmol.L ^-1KH ₂PO ₄Solution, B is that (the gradient elution volume proportion is from 0 minute to 5 minutes to acetonitrile-water (80: 20), the ratio of Mobile phase B is 25%, and from 5 minutes to 40 minutes, the ratio of Mobile phase B rose to 64% by 25%, from 40 minutes to 50 minutes, the ratio of Mobile phase B is 64%, and from 50 minutes to 65 minutes, the ratio of Mobile phase B rose to 80% by 64%, from 65 minutes to 75 minutes, the ratio of Mobile phase B reduced to 25% by 80%) four kinds of flow phase system.The result shows that peak shape is relatively poor under mobile phase (1) condition, and 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 (2) the mobile phase condition, peak shape is relatively poor, separates bad; (3) under the condition, the peak hangover is serious, and it is incomplete to go out the peak; Under mobile phase (4) condition, peak shape is better, goes out the peak fully and be evenly distributed, so finally selected.

3. detection wavelength determination

Be 50mmol.L at A in the research ^-1KH ₂PO ₄Solution, B are under acetonitrile-water (80: 20) (gradient elution) the mobile phase condition, have investigated the chromatographic peak situation under different-waveband typical wavelengths 203,210,230,254 respectively, the result shows, chromatographic peak is more under 203nm, and peak shape is better, so finally select for use 203nm as detecting wavelength.

4. instrument, chromatographic column and integral parameter

4.1 instrument parameter: selected liquid-phase chromatographic analysis field mainstream configuration and well behaved Agilent1100 series of high efficiency chromatograph of liquid for use, the Chemstation chromatographic work station.Chromatographic column is Diamonsil ODS (4.6mm * 200mm, 5 μ m); 40 ℃ of column temperatures, flow velocity 1.0ml/min.

5. the preparation of need testing solution

6. the preparation of object of reference solution

The ginsenoside Rg ₁Be one of Radix Ginseng Rubra or Radix Ginseng main active, its integral area proportion in finger printing more greatly and more stable is taken into account the research of intermediate and medical material simultaneously, therefore selected ginsenoside Rg ₁As object of reference.

7. finger printing and technical parameter

Experimental example 3 is differentiated

The present invention relates to the thin layer chromatography discrimination method of Herba Erigerontis in the injection

Feature for outstanding Herba Erigerontis, selected scutellarin as its feature speckle, but owing to there is composition like more, the polar phase close in the medical material with the scutellarin structure, usual terms is difficult to reach requirements for quality control, so we have screened following lamellae and unfolding condition launches scutellarin:

Condition question

Ethyl acetate-formic acid-water (15-5-1) silica gel g thin-layer plate reference substance is expanded to the forward position

Ethyl acetate-formic acid-water (15-3-1) silica gel H lamellae reference substance does not separate, and feminine gender has interference

Ethyl acetate-acetic acid-water (10-5-1) silica gel g thin-layer plate reference substance is expanded to the forward position

Benzene-ethyl acetate-formic acid-water (10-5-1-0.5) silica gel G reference substance does not separate, and feminine gender has interference

Lamellae

Toluene-ethyl acetate-acetic acid-water (10-5-1-0.5) silica gel reference substance does not separate, and feminine gender has interference

The G lamellae

Toluene-Ethyl formate-formic acid-water (13-3-1-0.5) silica gel reference substance does not separate, and feminine gender has interference

The G lamellae

Determine condition: toluene-ethyl acetate-acetic acid-separated form water is clear, and the moderate feminine gender of Rf value is noiseless

(10-5-1-0.5) silica gel g thin-layer plate

Through screening, determined with the silica gel g thin-layer plate to be immobile phase, be developing solvent with toluene-ethyl acetate-acetic acid-water (10-5-1-0.5), with this understanding, the Rf value of scutellarin is moderate, and it is clear to separate with other speckle, negative noiseless.

The present invention relates to the liquid chromatograph discrimination method of scutellarin in the 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, usual terms is difficult to reach requirements for quality control, so we have screened following chromatographic column and with mobile phase scutellarin are separated:

Condition question

Methanol-water (80: 20) appearance time is too fast

Octadecylsilane chemically bonded silica

Acetonitrile-water (40: 60) feminine gender has interference

Octadecylsilane chemically bonded silica

Methanol-0.2% glacial acetic acid aqueous solution (30: 70) eight alkyl silane feminine genders have interference

Bonded silica gel

Methanol-oxolane-0.2% phosphate aqueous solution (14: 14: 72) peak shape is slightly asymmetric

Octadecylsilane chemically bonded silica

There is bifurcated at acetonitrile-oxolane-0.2% phosphate aqueous solution (14: 14: 72) peak,

The dialkyl silane bonded silica gel

Methanol-water (40: 60) feminine gender has interference

Eight alkyl silane bonded silica gels

Acetonitrile-0.1% phosphate aqueous solution (20: 80) retention time is moderate, and the peak is capable sharp-pointed, symmetry,

The octadecylsilane chemically bonded silica feminine gender is noiseless

Through screening, determined with the octadecylsilane chemically bonded silica to be immobile phase, acetonitrile-0.1% phosphate aqueous solution (20: 80) 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 present invention relates to the thin layer chromatography discrimination method of Radix Ophiopogonis in the injection

Feature for outstanding Radix Ophiopogonis, selected the Radix Ophiopogonis control medicinal material as its feature speckle, but because have that more structure is close in the medical material, composition like the polar phase, usual terms is difficult to reach requirements for quality control, so we have screened following lamellae and unfolding condition to launching Radix Ophiopogonis:

Condition question

Benzene-methanol-glacial acetic acid (50-10-3) silica gel G F ₂₅₄The lamellae reference substance does not separate, and feminine gender has interference

Toluene-methanol-glacial acetic acid (50-10-3) silica gel H lamellae reference substance does not separate, and feminine gender has interference

Chloroform-ethanol-formic acid (50-20-5) silica gel g thin-layer plate reference substance is expanded to the forward position

Chloroform-methanol-formic acid (50-20-5) silica gel g thin-layer plate reference substance is expanded to the forward position

Ethyl acetate-pyridine-water (5-3-5) silica gel H lamellae reference substance does not separate, and feminine gender has interference

Benzene-ethanol-formic acid (60-10-2) silica gel g thin-layer plate reference substance does not separate, and feminine gender has interference

Determine condition: it is clear that toluene-methanol-glacial acetic acid (80: 5: 0.1) separates, and the moderate feminine gender of Rf value is noiseless

Silica gel G F ₂₅₄Lamellae

Through screening, determined with silica gel G F ₂₅₄Lamellae is an immobile phase, is developing solvent with toluene-methanol-glacial acetic acid (80: 5: 0.1), and with this understanding, the Rf value of principal spot is moderate, and it is clear to separate with other speckle, and is negative noiseless.

The present invention relates to ophiopogonin B in the injection, ophiopogonin D, ophiopogonin D ' the thin layer chromatography discrimination method

Feature for outstanding ophiopogonin, ophiopogonin B, ophiopogonin D, ophiopogonin D ' have been selected as its feature speckle, but because have in the medical material that more and ophiopogonin B, ophiopogonin D, ophiopogonin D ' structure are close, composition like the polar phase, usual terms is difficult to reach requirements for quality control, so we have screened following lamellae and unfolding condition to launching Radix Ophiopogonis:

Condition question

Ethyl acetate-methanol-water (10-10-0.5) silica gel G thin layer reference substance is expanded to the forward position

Plate

Chloroform-methanol-water (10-10-0.5) silica gel H lamellae reference substance is expanded to the forward position

Ethyl acetate-methanol-water (10-3-0.2) silica gel g thin-layer plate reference substance does not separate, and feminine gender has interference

Chloroform-methanol-water (15-10-2) silica gel G F ₂₅₄The lamellae reference substance is expanded to the forward position

Ethyl acetate-methanol-water (20-3-1) silica gel G F ₂₅₄The lamellae reference substance does not separate, and feminine gender has interference

Ethyl acetate-methanol-water (20-8-1) silica gel g thin-layer plate reference substance does not separate, and feminine gender has interference

Determine condition: it is clear to separate with ethyl acetate-methanol-water (15: 5: 1), and the moderate feminine gender of Rf value is noiseless

Silica gel g thin-layer plate

Through screening, determined with the silica gel g thin-layer plate to be immobile phase, be developing solvent with ethyl acetate-methanol-water (15: 5: 1), with this understanding, the Rf value of ophiopogonin B, ophiopogonin D, ophiopogonin D ' speckle is moderate, and it is clear to separate with other speckle, and is negative noiseless.

The present invention relates to ophiopogonin B in the injection, ophiopogonin D, ophiopogonin D ' the liquid chromatograph discrimination method

Feature for outstanding ophiopogonin, ophiopogonin B, ophiopogonin D, ophiopogonin D ' have been selected as its characteristic component, but because have in the medical material that more and ophiopogonin B, ophiopogonin D, ophiopogonin D ' structure are close, composition like the polar phase, usual terms is difficult to reach requirements for quality control, so we have screened following chromatographic column and mobile phase to ophiopogonin B, ophiopogonin D, ophiopogonin D ' separate:

Condition question

Methanol-water (99: 1) appearance time is too fast

Octadecylsilane chemically bonded silica

Acetonitrile-water (95: 5) appearance time is too fast

Octadecylsilane chemically bonded silica

Methanol-2% glacial acetic acid aqueous solution (95: 5) feminine gender has interference

Eight alkyl silane bonded silica gels

Methanol-0.2% phosphoric acid aqueous acid (95: 5) feminine gender has interference

Octadecylsilane chemically bonded silica

There is bifurcated at acetonitrile-1% glacial acetic acid aqueous solution (95: 5) peak,

Eight alkyl silane bonded silica gels

Acetonitrile-0.5% phosphoric acid aqueous acid (95: 5) feminine gender has interference

Octadecylsilane chemically bonded silica

Acetonitrile-water (90: 10) retention time is moderate, and the peak is capable sharp-pointed, and symmetry is negative noiseless

Octadecylsilane chemically bonded silica

Through screening, determined with the octadecylsilane chemically bonded silica to be immobile phase, acetonitrile-water (90: 10) be a mobile phase, and with this understanding, ophiopogonin B, ophiopogonin D, ophiopogonin D ' retention time are moderate, and the peak is capable sharp-pointed, symmetry, feminine gender is noiseless.

The present invention relates to the thin layer chromatography discrimination method of Ruscus aculeatus L. sapogenin in the injection, diosgenin, ruscogenin

Feature for outstanding ophiopogonin unit, selected Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin as its feature speckle, but because have in the medical material that more and Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin structure are close, composition like the polar phase, usual terms is difficult to reach requirements for quality control, so we have screened following lamellae and unfolding condition to launching Radix Ophiopogonis:

Condition question

Methanol-chloroform-water (5-3-2) silica gel g thin-layer plate reference substance is expanded to the forward position

Normal hexane-ethyl acetate-water (5-3-1) silica gel H lamellae reference substance is expanded to the forward position

Normal hexane-Ethyl formate-water (3-2-2) silica gel G F ₂₅₄The lamellae reference substance does not separate, and feminine gender has interference

Methanol-ethyl acetate-water (2-1-1) silica gel g thin-layer plate reference substance does not separate, and feminine gender has interference

Methanol-Ethyl formate-water (1-1-1) silica gel H lamellae reference substance does not separate, and feminine gender has interference

Methanol-Ethyl formate-water (2-1.5-0.5) silica gel H lamellae reference substance does not separate, and feminine gender has interference

Determine condition: it is clear that normal hexane-ethyl acetate-water (1: 1: 1) separates, the noiseless silica gel g thin-layer plate of the moderate feminine gender of Rf value

Through screening, determined with the silica gel g thin-layer plate to be immobile phase, be developing solvent with normal hexane-ethyl acetate-water (1: 1: 1), with this understanding, the Rf value of Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin is moderate, and it is clear to separate with other speckle, and is negative noiseless.

The present invention relates to the liquid chromatograph discrimination method of Ruscus aculeatus L. sapogenin in the injection, diosgenin, ruscogenin

Feature for outstanding ophiopogonin unit, selected Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin as its characteristic component, but because have in the medical material that more and Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin structure are close, composition like the polar phase, usual terms is difficult to reach requirements for quality control, so we have screened following chromatographic column and with mobile phase Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin are separated:

Condition question

Methanol-water (99: 1) octadecylsilane chemically bonded silica appearance time is too fast

Acetonitrile-water (95: 5) octadecylsilane chemically bonded silica appearance time is too fast

Eight alkyl silane bonded silica gels

Octadecylsilane chemically bonded silica

Eight alkyl silane bonded silica gels

Octadecylsilane chemically bonded silica

Acetonitrile-water (90: 10) retention time is moderate, and the peak is capable sharp-pointed, symmetry,

The octadecylsilane chemically bonded silica feminine gender is noiseless

Through screening, determined with the octadecylsilane chemically bonded silica to be immobile phase, acetonitrile-water (90: 10) is a mobile phase, with this understanding, wheat Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin retention time are moderate, and the peak is capable sharp-pointed, symmetry, negative noiseless.

The present invention relates to ginsenoside Rg in the injection ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf the thin layer chromatography discrimination method

For the feature of outstanding Radix Ginseng Rubra or Radix Ginseng, selected the ginsenoside Rg ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf be as its feature speckle, but owing to exist more and the ginsenoside Rg in the medical material ₁, ginsenoside Rb ₁, composition like close, the polar phase of ginsenoside Re, ginsenoside Rf's structure, usual terms is difficult to reach requirements for quality control, so we have screened following lamellae and unfolding condition to launching Radix Ophiopogonis:

Condition question

(20: 60: 40: 10) 10 reference substances were expanded to the forward position to chloroform-Ethyl formate-methanol-water

Lower floor's solution silica gel g thin-layer plate of placing below ℃

N-butyl alcohol-Ethyl formate-water (10-1-5) upper solution silica gel H reference substance is expanded to the forward position

Lamellae

(15: 40: 22: 10) 10 reference substances did not separate chloroform-Ethyl formate-methanol-water, and feminine gender has interference

Lower floor's solution silica gel g thin-layer plate of placing below ℃

(10: 40: 15: 5) 10 ℃ of reference substances did not separate chloroform-ethyl acetate-methanol-water, and feminine gender has interference

Following lower floor's solution silica gel g thin-layer plate of Fang Zhiing

N-butyl alcohol-Ethyl formate-water (5-1-5) upper solution reference substance does not separate, and feminine gender has interference

Silica gel g thin-layer plate

N-butyl alcohol-Ethyl formate-water (2-1.5-0.5) upper solution reference substance does not separate, and feminine gender has interference

The silica gel H lamellae

Determine condition: (15: 40: it was clear to separate, and the moderate negative nothing of Rf value is done for chloroform-ethyl acetate-methanol-water

22: 10) lower floor's solution silica gel g thin-layer plate of placing below 10 ℃ disturbs

Through screening, determined with the silica gel g thin-layer plate to be immobile phase, with chloroform-ethyl acetate-methanol-water (15: 40: 22: 10) lower floor's solution of placing below 10 ℃ was developing solvent, with this understanding, the ginsenoside Rg ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf Rf value moderate, it is clear to separate with other speckle, negative noiseless.

The present invention relates to ginsenoside Rg in the injection ₁, ginsenoside Rb ₁, the ginsenoside Re the liquid chromatograph discrimination method

For the feature of outstanding Radix Ginseng Rubra or Radix Ginseng, selected the ginsenoside Rg ₁, ginsenoside Rb ₁, the ginsenoside Re is as its characteristic component, but owing to exist more and the ginsenoside Rg in the medical material ₁, ginsenoside Rb ₁, composition like close, the polar phase of ginsenoside Re's structure, usual terms is difficult to reach requirements for quality control, so we have screened following chromatographic column and mobile phase to the ginsenoside Rg ₁, ginsenoside Rb ₁, the ginsenoside Re separates:

Condition question

Methanol-water (99: 5) appearance time is too fast

Octadecylsilane chemically bonded silica

Acetonitrile-water (95: 5) appearance time is too fast

Octadecylsilane chemically bonded silica

Methanol-2% glacial acetic acid aqueous solution (80: 20) feminine gender has interference

Eight alkyl silane bonded silica gels

Methanol-0.2% phosphoric acid aqueous acid (80: 20) feminine gender has interference

Octadecylsilane chemically bonded silica

Acetonitrile-1% glacial acetic acid aqueous solution (90: 10) feminine gender has interference

Eight alkyl silane bonded silica gels

Acetonitrile-0.5% phosphoric acid aqueous acid (85: 15) feminine gender has interference

Octadecylsilane chemically bonded silica

Acetonitrile-water is a mobile phase, gradient elution, and solvent ratios is that retention time is moderate, the peak is capable sharp-pointed, symmetry, negative do not have

From 0 minute to 35 minutes, the ratio of acetonitrile was 19%, disturbed

From 35 minutes to 55 minutes, the ratio of acetonitrile was by 19%

Rise to 29%, from 55 minutes to 70 minutes, acetonitrile

Ratio be 29%, from 70 minutes to 100 minutes, second

The ratio of nitrile rises to 40% by 29%

Octadecylsilane chemically bonded silica

Through screening, determined with the octadecylsilane chemically bonded silica to be immobile phase, acetonitrile-water is a mobile phase, gradient elution, solvent ratios are from 0 minute to 35 minutes, and the ratio of acetonitrile is 19%, from 35 minutes to 55 minutes, the ratio of acetonitrile rises to 29% by 19%, and from 55 minutes to 70 minutes, the ratio of acetonitrile was 29%, from 70 minutes to 100 minutes, the ratio of acetonitrile rises to 40% by 29%, with this understanding, and the ginsenoside Rg ₁, ginsenoside Rb ₁, ginsenoside Re's retention time is moderate, the peak is capable sharp-pointed, symmetry is negative noiseless.

Experimental example 4 assay project approaches are investigated

One, ginsenoside Rg ₁, the ginsenoside Re content

1. instrument and reagent

(1) instrument: Agilent1100 high performance liquid chromatograph, chemstationsys work station.The TU-1800SPC ultraviolet spectrophotometer.

(2) reagent: ginsenoside Rg ₁, the ginsenoside Re: Nat'l Pharmaceutical ﹠ Biological Products Control Institute;

2. detect the selection of wavelength: precision takes by weighing the ginsenoside Rg ₁, the ginsenoside Re, split in the 10ml measuring bottle, add methanol dilution and make the solution that every 1ml contains 0.5mg, scan in the 200-400nm wave-length coverage.The ginsenoside Rg ₁, the ginsenoside Re all has absorption maximum at the 203nm place, therefore selects the detection wavelength of 203nm as assay.

3. chromatographic condition: Dikma ODS (4.6mm * 250mm, 5um); Mobile phase: acetonitrile-water is a mobile phase, gradient elution, solvent ratios is from 0 minute to 35 minutes, the ratio of acetonitrile is 19%, and from 35 minutes to 55 minutes, the ratio of acetonitrile rose to 29% by 19%, from 55 minutes to 70 minutes, the ratio of acetonitrile is 29%, and from 70 minutes to 100 minutes, the ratio of acetonitrile rose to 40% by 29%; The detection wavelength is 203nm; Flow velocity: 1.0ml/min.

4. the preparation of reference substance solution: get the ginsenoside Rg1 and the ginsenoside Re is an amount of, accurately claim surely, add methanol respectively and be diluted to scale, shake up, every 1ml contain ginsenoside Rg1 0.30mg, the solution of ginsenoside Re 0.24mg.

5. the preparation of need testing solution: get the about 0.5g of this product, the accurate title, decide, and puts in the 5ml measuring bottle, adds to flow mutual-assistance dissolving and be diluted to scale, shakes up, promptly.

With this understanding, negative sample ginsenoside Rg in the disturbed specimen not ₁, the ginsenoside Re mensuration, and separating degree is good.

6. ginsenoside Rg ₁Take by weighing the ginsenoside Rg with ginsenoside Re's linear relationship precision ₁10.20mg, ginsenoside Re 10.72mg, put altogether in the 10ml measuring bottle, it is fixed to scale to add mobile phase, shakes up, product solution (contains the ginsenoside Rg among every 1ml in contrast ₁1.02mg, ginsenoside Rb ₁1.01mg, contain ginsenoside Re 1.072mg), precision is measured 2.5ml, puts in the 10ml measuring bottle, adds mobile phase to scale, shakes up, in contrast the product dilute solution.(contain the ginsenoside Rg among every 1ml ₁0.255mg, contain ginsenoside Re 0.268mg) and accurate reference substance solution 3 μ l, 6 μ l, the 10 μ l of drawing; Reference substance dilute solution 2 μ l, 5 μ l, 10 μ l; Injecting chromatograph of liquid, is vertical coordinate with the peak area, and ginsenoside Rg1 and ginsenoside Re's amount is figure for abscissa, the drawing standard curve.

Ginsenoside Rg1's linear relationship

Numbering	Ginsenoside Rg1 (μ g)	Peak area
Numbering	Ginsenoside Rg1 (μ g)	Peak area	1 2 3 4 5 6	0.510 1.275 2.550 3.060 6.120 10.200	150.04 324.27 856.87 1024.83 2098.79 3523.14

Regression equation: Y=351.97X-61.521

The coefficient of determination: r=0.9996

The ginsenoside Rg1 is good in 0.510～10.20 μ g scope internal linear relation.

Ginsenoside Re's linear relationship

Numbering	Ginsenoside Re (μ g)	Peak area
Numbering	Ginsenoside Re (μ g)	Peak area	1 2 3 4 5 6	0.536 1.340 2.680 3.216 6.432 10.720	129.65 283.75 759.26 902.30 1840.48 3075.59

Regression equation: Y=290.94.97X-43.21

The coefficient of determination: r=0.9995

The ginsenoside Re is good in 0.536～10.72 μ g scope internal linear relation.

7. ginsenoside Rg1 and ginsenoside Re's reference substance dilute solution (contain ginsenoside Rg1 0.255mg under the accurate absorption of the reference substance precision test standard curve item among every 1ml, contain ginsenoside Re 0.268mg) 10 μ l, inject chromatograph of liquid, continuous sample introduction is measured 5 times, investigates the precision of reference substance solution.

Ginsenoside Rg1 and the test of ginsenoside Re's reference substance precision

Test number (TN)	1	2	3	4	5	Meansigma methods	RSD(％)
Test number (TN)	1	2	3	4	5	Meansigma methods	RSD(％)	Ginsenoside Rg1's peak area ginsenoside Re peak area	859.78 746.19	847.26 774.10	846.53 740.15	856.40 736.24	845.30 730.54	851.054 740.044	0.772％ 0.135％

The result shows that ginsenoside Rg1 and ginsenoside Re's reference substance solution precision are good.

8. stability test

8.1. ginsenoside Rg1 and ginsenoside Re's reference substance dilute solution (contain ginsenoside Rg1 0.255mg under the accurate absorption of the reference substance stability test standard curve item among every 1ml, contain ginsenoside Re 0.268mg) 10 μ l, inject chromatograph of liquid, measure at 0,6,12,24,48 hour sample introduction.

Reference substance stability test result

Testing time (h)	0	6	12	24	48	On average	RSD(％)
Testing time (h)	0	6	12	24	48	On average	RSD(％)	Ginsenoside Rg1's peak area ginsenoside Re peak area	859.78 746.19	847.26 774.10	846.53 740.15	856.40 736.24	845.30 730.54	851.054 740.044	0.772％ 0.135％

The result shows that ginsenoside Rg1 and ginsenoside Re's reference substance solution have good stability.

8.2. the accurate test sample 10 μ l that draw of need testing solution stability test inject chromatograph of liquid, measure at 0,6,12,24,48 hour sample introduction respectively.

Need testing solution stability test result

Testing time (h)	0	6	12	24	48	On average	RSD(％)
Testing time (h)	0	6	12	24	48	On average	RSD(％)	Ginsenoside Rg1's peak area ginsenoside Re peak area	842.71 384.78	853.69 381.32	847.14 376.52	845.36 380.63	840.88 381.12	845.96 380.87	0.58 0.77

The result shows that need testing solution has good stability.

9. replica test

Get under this product content uniformity item 5 bottles on powder pin, get content, porphyrize is therefrom got about 0.5g, accurately claims surely, puts in the 5ml measuring bottle, adds the mutual-assistance dissolving and shake up to scale surely of flowing, and the accurate 10 μ l that draw inject chromatograph of liquid, measure, promptly.

Replica test

Test number	1	2	3	4	5	Meansigma methods	RSD(％)
Test number	1	2	3	4	5	Meansigma methods	RSD(％)	Ginsenoside Rg1's (mg/ bottle) ginsenoside Re (mg/ bottle)	1.32 0.73	1.35 0.74	1.33 0.72	1.34 0.75	1.31 0.74	1.33 0.74	1.19 1.55

10. average recovery test

Adopt the application of sample absorption method, precision takes by weighing the ginsenoside Rg ₁10.67mg ginsenoside Re 6.02mg puts in the 10ml measuring bottle altogether, adds mobile phase to scale, shakes up, precision is measured 1ml; Get this product under the content uniformity item, get content, mixing is therefrom got about 0.5g, and accurate the title decides, and puts altogether in the 5ml measuring bottle, adds mobile phase to scale, shakes up, and the accurate 10 μ l that draw inject chromatograph of liquid, measure, promptly.

The present invention relates to ginsenoside Rg1's content in the injection: 0.1824%

The present invention relates to ginsenoside Re's content in the injection: 0.1013%

The test of ginsenoside Rg1's average recovery

Numbering	Test sample weighing (g)	Pure product amount (mg) in the test sample	Pure product addition (mg)	Measured value (mg)	The response rate (%)
Numbering	Test sample weighing (g)	Pure product amount (mg) in the test sample	Pure product addition (mg)	Measured value (mg)	The response rate (%)	1 2 3 4 5	0.51384 0.50791 0.50326 0.51183 0.50745	0.9372 0.9264 0.9179 0.9336 0.9256	1.067 1.067 1.067 1.067 1.067	1.9851 1.9682 1.9746 1.9869 1.9545	98.21 97.64 99.03 98.72 96.43

Ginsenoside Rg1's average recovery rate=98.01%; RSD=1.05%

The test of ginsenoside Re's average recovery

Numbering	Test sample weighing (g)	Pure product amount (mg) in the test sample	Pure product addition (mg)	Measured value (mg)	Response rate %
Numbering	Test sample weighing (g)	Pure product amount (mg) in the test sample	Pure product addition (mg)	Measured value (mg)	Response rate %	1 2 3 4 5	0.51384 0.50791 0.50326 0.51183 0.50745	0.5205 0.5145 0.5098 0.5185 0.5140	0.602 0.602 0.602 0.602 0.602	1.1166 1.1042 1.1983 1.0972 1.0891	99.01 97.96 99.42 96.13 95.53

Ginsenoside Re's average recovery rate=97.61%; RSD=1.77%

Three batches of pilot scale sample sizes are measured

Get three batches in this product sample, press the described method of text and handle, sample introduction is measured.

Three batch sample assay results

Lot number	Ginsenoside Rg1's (mg/ bottle)	Ginsenoside Re's (mg/ bottle)
Lot number	Ginsenoside Rg1's (mg/ bottle)	Ginsenoside Re's (mg/ bottle)	1 2 3	1.36 1.41 1.32	0.68 0.64 0.71

Two, Radix Ginseng total saponins

Instrument, reagent

Instrument: the general logical TU-1810SPC ultraviolet/visible spectrophotometer of analysing

SARTORIUS BP211D electronic analytical balance

Reagent: vanillin analytical pure Tianjin recovery fine chemistry industry institute

Methanol chromatographically pure J.T.Baker

Glacial acetic acid analytical pure Shanghai reagent one factory

Chemical plant, prosperous source, perchloric acid analytical pure Tianjin

The pure water WAHAHA

Method and result

The selection precision that detects wavelength takes by weighing ginsenoside Rg1 6.11mg, put in the 100ml measuring bottle, add an amount of methanol supersound process (power 250W, frequency 33KHz) and make dissolving, add methanol to scale, shake up, precision is measured 1.2ml, puts in the 10ml tool plug test tube water bath method solvent, take out, the accurate 5% vanillin-glacial acetic acid solution 0.2ml that adds, perchloric acid 0.8ml shakes up, put in 60 ℃ of water-baths and heated 15 minutes, take out, in ice-water bath, cooled off 2 minutes immediately, the accurate glacial acetic acid 5ml that adds, shake up, scan in 700～400nm wave-length coverage, the result shows that the ginsenoside Rg1 has absorption maximum at the 547nm place, blank noiseless, therefore selecting 547nm is the detection wavelength that spectrophotometry the present invention relates to Radix Ginseng total saponins in the injection.

The investigation precision of linear relationship takes by weighing ginsenoside Rg1's reference substance 6.85mg, put in the 100ml measuring bottle, add an amount of supersound process of methanol (power 250W, frequency 33KHZ) and make dissolving, take out, put to room temperature, add methanol to scale, shake up, precision measures 0.4,0.8,1.2,1.6,2.0ml, split in the 10ml tool plug test tube, the water bath method solvent takes out, the accurate 5% vanillin-glacial acetic acid solution 0.2ml that adds, perchloric acid 0.8ml, shake up, put in 60 ℃ of water-baths and heated 15 minutes, take out, in ice-water bath, cooled off 2 minutes immediately, the accurate glacial acetic acid 5ml that adds shakes up, and is blank with the retinue solvent, according to spectrophotography (an appendix V of Chinese Pharmacopoeia version in 2000 A), measure trap at the wavelength place of 547nm.With the trap is vertical coordinate, and ginsenoside Rg1's amount (μ g) is an abscissa, the drawing standard curve.

Regression equation Y=0.0049X-0.0077; R=0.999

The ginsenoside Rg1 is linear good in 27.4～137.0 μ g scopes.

Ginsenoside Rg1's standard curve determination data

Numbering	Ginsenoside Rg1 (μ g)	Trap
Numbering	Ginsenoside Rg1 (μ g)	Trap	1 2 3 4 5	27.4 54.8 82.2 109.6 137.0	0.121 0.261 0.404 0.534 0.656

Precision test precision takes by weighing Radix Ginseng soap Re1 reference substance 6.85mg, puts in the 100ml measuring bottle, adds an amount of supersound process of methanol (power 250W, frequency 33KHZ) makes dissolving, take out, put to room temperature, add methanol to scale, shake up, precision is measured 1.2ml, put in the 10ml tool plug test tube, the water bath method solvent takes out, the accurate 5% vanillin-glacial acetic acid solution 0.2ml that adds, perchloric acid 0.8ml, shake up, put in 60 ℃ of water-baths and heated 15 minutes, take out, in ice-water bath, cooled off 2 minutes immediately, the accurate glacial acetic acid 5ml that adds shakes up, and is blank with the retinue solvent, press operation under the text algoscopy item, METHOD FOR CONTINUOUS DETERMINATION 5 times.

The experiment of Radix Ginseng total saponins precision

Numbering	1	2	3	4	5	X is average	RSD％
Numbering	1	2	3	4	5	X is average	RSD％	Absorbance	0.429	0.426	0.426	0.425	0.424	0.426	0.44

Stability test

The stability test precision of reference substance solution takes by weighing ginsenoside Rg1's reference substance 7.27g, puts in the 100ml measuring bottle, adds an amount of supersound process of methanol (power 250W, frequency 33KHZ) makes dissolving, take out, put to room temperature, add methanol to scale, shake up, precision is measured 1.2ml, put in the 10ml tool plug test tube, the water bath method solvent takes out, the accurate 5% vanillin-glacial acetic acid solution 0.2ml that adds, perchloric acid 0.8ml, shake up, put in 60 ℃ of water-baths and heated 15 minutes, take out, in ice-water bath, cooled off 2 minutes immediately, the accurate glacial acetic acid 5ml that adds shakes up, and is blank with the retinue solvent, promptly get and press operation under the text algoscopy item, respectively 0,10,20,40, measure in the time of 60 minutes.

Radix Ginseng total saponins reference substance solution stability experiment

Time (min)	0	10	20	40	60	X is average	RSD％
Time (min)	0	10	20	40	60	X is average	RSD％	Absorbance	0.453	0.453	0.450	0.451	0.449	0.451	0.40

The stability experiment of need testing solution is got under this product content uniformity item 5 bottles on powder pin, get content, porphyrize, precision takes by weighing 118.47mg, puts in the 10ml measuring bottle, adds water and makes dissolving and fixed to scale in right amount, shake up, precision is measured 0.2ml, by operating under the text algoscopy item, measures in the time of 0,10,20,40,60 minute respectively.

Radix Ginseng total saponins need testing solution stability experiment

Time (min)	0	10	20	40	60	X is average	RSD％
Time (min)	0	10	20	40	60	X is average	RSD％	Content (mg/ bottle)	30.13	29.67	29.87	30.05	30.02	29.95	0.61

Replica test is got under this product content uniformity item 5 bottles on powder pin, gets content, and porphyrize is therefrom got about 100mg, puts in the 10ml measuring bottle, adds water and makes dissolving in right amount and shake up to scale surely, and precision is measured 0.2ml, by operating under the text algoscopy item.

The Radix Ginseng total saponins repeated experiment

Numbering	1	2	3	4	5	X is average	RSD％
Numbering	1	2	3	4	5	X is average	RSD％	Content (mg/ bottle)	31.24	30.56	29.93	30.42	30.08	30.45	1.68

Recovery test adopts the application of sample absorption method, gets this product under the content uniformity item, gets content, and porphyrize is therefrom got about 50mg, and accurate the title decides; Other gets the about 2mg of ginsenoside Re's 1 reference substance, and accurate the title decides, and puts altogether in the 10ml measuring bottle, add water and make dissolving and fixed in right amount to scale, shake up, precision is measured 0.2ml, puts in the 10ml tool plug test tube, by operating under the text algoscopy item, with the retinue solvent is blank, measures trap in accordance with the law, reads the content of ginsenoside Rg1 the need testing solution from standard curve, calculate, promptly.

The present invention relates to the content of total saponins in the injection: 4.52%

The experiment of Radix Ginseng total saponins average recovery

Numbering	Test sample weighing (mg)	Total saponins amount (mg) in the test sample	Ginsenoside Rg1's addition (mg)	Measured value (mg)	The response rate (%)
Numbering	Test sample weighing (mg)	Total saponins amount (mg) in the test sample	Ginsenoside Rg1's addition (mg)	Measured value (mg)	The response rate (%)	1 2 3 4 5	52.46 50.38 49.42 51.13 52.08	2.3712 2.2772 2.2338 2.3111 2.3540	2.28 2.36 2.07 2.14 2.22	4.5978 4.6051 4.2276 4.4025 4.5327	97.66 98.64 96.32 97.73 98.14

Average recovery rate=97.70% RSD=0.88%

Three batches of pilot scale sample sizes are measured

Three batch sample assay results

Lot number	Radix Ginseng total saponins (mg/ bottle)
Lot number	Radix Ginseng total saponins (mg/ bottle)	1 2 3	30.27 31.04 28.59

Three, total flavones

1 instrument and reagent

(1) key instrument:

Ultraviolet spectrophotometer TU-1810SPC Beijing Puxi General Instrument Co., Ltd

Electronic analytical balance BP211D SARTORIUS

Ultrasonic washing unit KQ250DB Kunshan Ultrasonic Instruments Co., Ltd.

(2) reagent:

Rutin Nat'l Pharmaceutical ﹠ Biological Products Control Institute (lot number: 0080-9705)

Ethanol analytical pure atropic is Fine Chemical Co., Ltd now

Aluminum nitrate analytical pure Tianjin chemical reagent three factories

Sodium nitrite analytical pure Tianjin chemical reagent three factories

Northization glass purchase and sale center, sodium hydroxide analytical pure Tianjin

Control substance of Rutin is got in 2 selections that detect wavelength, operates by the preparation method of text reference substance solution, obtains reference substance solution.Get and the present invention relates to injection, operate, obtain need testing solution by the preparation method of need testing solution in the text algoscopy.Draw control substance of Rutin solution, need testing solution, press the text total flavones and measure item method suggested down, in 400～800nm wave-length coverage, scan, the result shows, reference substance solution and need testing solution all have absorption maximum at 510nm, and solvent is noiseless, therefore select 510nm to the present invention relates to the detection wavelength of general flavone content in the injection as spectrophotometry.

The preparation precision of 3 reference substance solution takes by weighing at the control substance of Rutin 20mg of 120 ℃ of drying under reduced pressure to constant weight, puts in the 100ml measuring bottle, adds an amount of supersound process of 60% alcoholic solution (power 250W, frequency 33KHz) makes dissolving, take out, put, add 60% alcoholic solution to scale to room temperature, shake up, precision is measured 25ml, puts in the 50ml measuring bottle, and thin up is to scale, shake up, promptly get (containing anhydrous rutin 0.1mg among every 1ml).

The about 50mg of this product powder is got in the preparation of 4 need testing solutions, and accurate the title decides, and puts in the 50ml measuring bottle, adds an amount of supersound process of 60% alcoholic solution (power 250W, frequency 33KHz) makes dissolving, take out, put to room temperature, add 60% alcoholic solution to scale, shake up, promptly.

5 algoscopy precisions are measured reference substance solution 3.0ml, need testing solution 1.0ml, split in the 10ml measuring bottle, respectively add 30% alcoholic solution and make into 5.0ml, precision adds sodium nitrite solution (1 → 20) 0.3ml and shakes up, and places 6 minutes, add aluminum nitrate solution (1 → 10) 0.3ml again, shake up, placed 6 minutes, hydro-oxidation sodium test solution 4.0ml, add water to scale again, shaking up, placed 15 minutes, is blank with the retinue solvent, according to spectrophotography (an appendix V of Chinese Pharmacopoeia version in 2000 B), wavelength place at 510nm measures trap respectively, calculates, promptly.

The preparation precision of 6 standard curves is measured control substance of Rutin solution (C=0.1064mg/ml) 0.0ml, 1.0ml, 2.0ml, 3.0ml, 4.0ml, 5.0ml, split in the 10ml measuring bottle, add 30% alcoholic solution respectively and make into 5.0ml, precision adds sodium nitrite solution (1 → 20) 0.3ml, shake up, placed 6 minutes, add aluminum nitrate solution (1 → 10) 0.3ml again, shake up, placed 6 minutes, hydro-oxidation sodium test solution 4.0ml adds water to scale again, shake up, placing 15 minutes, is blank with the retinue solvent, according to spectrophotography (an appendix V of Chinese Pharmacopoeia version in 2000 B), measure trap at 510nm wavelength place, with the trap is vertical coordinate, and concentration (mg/ml) is figure for abscissa, the drawing standard curve.

Total flavones standard curve determination result

Numbering	Rutin concentration (mg/ml)	Trap
Numbering	Rutin concentration (mg/ml)	Trap	1 2 3 4 5 6	0.000 0.0110 0.0210 0.0320 0.0430 0.0530	0.000 0.119 0.226 0.343 0.468 0.572

Regression equation: Y=10.818x-0.0005;

Correlation coefficient: γ=0.9999;

The result shows: rutin is good in 0.0110～0.0530mg/ml scope internal linear.

As calculated, the standard curve of rutin is crossed initial point, therefore selects for use one point external standard method to measure content of total flavone.

The accurate absorption of 7 precision test control substance of Rutin solution (concentration: 0.1064mg/ml) 3.0ml, put in the 10ml measuring bottle, press the method under the text algoscopy item, from " add 30% alcoholic solution and make into 5.0ml ", operation is in accordance with the law measured trap at the wavelength place of 510nm, measures 5 times.

The precision test

Numbering	1	2	3	4	5	RSD(％)
Numbering	1	2	3	4	5	RSD(％)	Trap	0.343	0.343	0.342	0.340	0.337	0.75

The result shows that reference substance solution precision is good.

8 stability tests

8.1 the accurate control substance of Rutin solution (concentration: 0.1064mg/ml) 3.0ml of drawing of reference substance solution stability test, put in the 10ml measuring bottle, press the method under the text algoscopy item, from " add 30% alcoholic solution and make into 5.0ml ", operation in accordance with the law, wavelength place at 510nm measures trap, respectively at 0,5,10,15,30 minute replication once.

The reference substance solution stability test

Time (min)	0	5	10	15	30	RSD(％)
Time (min)	0	5	10	15	30	RSD(％)	Trap	0.343	0.343	0.342	0.340	0.337	0.75

The result shows that reference substance solution has good stability.

8.2 the accurate need testing solution 1.0ml that draws of need testing solution stability test, put in the 10ml measuring bottle, press the method under the text algoscopy item, from " add 30% alcoholic solution and make into 5.0ml ", operation in accordance with the law, measure trap at the wavelength place of 510nm, respectively 0,5,10,15,30min measures trap.

The need testing solution stability test

Time (min)	0	5	10	15	30	RSD(％)
Time (min)	0	5	10	15	30	RSD(％)	Trap	0.352	0.349	0.351	0.346	0.354	0.87

The result shows that need testing solution is good at the 30min internal stability.

9 replica tests are got the about 50mg of this product powder (totally 5 parts), accurate claim fixed, by operating under the preparation of text need testing solution and the algoscopy item.

Replica test

Time (min)	0	5	10	15	30	Average	RSD(％)
Time (min)	0	5	10	15	30	Average	RSD(％)	Content (mg/ bottle)	19.64	20.03	19.21	19.73	19.55	19.63	1.51

The result shows that repeatability is good.

The application of sample absorption method is adopted in the test of 10 average recoveries; get the about 25mg of this product powder (totally 6 parts), and accurate the title decides, and splits in the 50ml measuring bottle; precision takes by weighing through the control substance of Rutin 10.18mg of 120 ℃ of drying under reduced pressure to constant weight; put in the 10ml measuring bottle, adds an amount of supersound process of 60% ethanol and makes dissolving, takes out; put to room temperature; shake up to scale with 60% ethanol is fixed, therefrom precision is measured 0.7ml respectively; 0.85ml; 1.0ml (each 2 parts); split in the above-mentioned 50ml measuring bottle; add an amount of supersound process of 60% alcoholic solution (power 250W, frequency 33KHz) and make dissolving, take out; put to room temperature; be diluted to scale with 60% alcoholic solution, shake up, and filtration; precision is measured subsequent filtrate 1.0ml; split in the 10ml measuring bottle is according to the method under the text algoscopy item, from " adding 30% alcoholic solution and make into 5.0ml " rise, operation in accordance with the law, wavelength place at 510nm measures trap, calculates, promptly.

The present invention relates to that content of total flavone is in the injection: 3.418%.

The average recovery test

Numbering	Test sample sample weighting amount (mg)	General flavone content in the test sample (mg)	Rutin addition (mg)	Measured value (mg)	The response rate (%)
Numbering	Test sample sample weighting amount (mg)	General flavone content in the test sample (mg)	Rutin addition (mg)	Measured value (mg)	The response rate (%)	1 2 3 4 5 6	24.71 26.25 25.86 24.67 25.09 26.34	0.8446 0.8972 0.8839 0.8432 0.8576 0.9003	0.7126 0.7126 0.8653 0.8653 1.018 1.018	1.5509 1.5907 1.7360 1.6874 1.8639 1.9084	99.12 97.32 98.47 97.56 98.85 99.03

Average recovery rate=98.39%, RSD=0.79%.

The assay of 11 samples is got this product three batch samples, according to the preparation of text need testing solution and the operation under the algoscopy item, working sample content.

Content of total flavone is measured

Lot number	Total flavones contains (mg/ bottle)
Lot number	Total flavones contains (mg/ bottle)	1 2 3	19.61 18.73 20.14

Four, scutellarin

1 instrument and reagent

1.1 key instrument:

High performance liquid chromatograph LC-2010AHT SHIMADZU

Ultraviolet/general all purpose instrument the company limited of analysing in visible spectrophotometer TU-1810SPC Beijing

Electronic analytical balance BP211D SARTORIUS

Ultrasonic washing unit KQ250DB Kunshan Ultrasonic Instruments Co., Ltd.

1.2 reagent:

Scutellarin Nat'l Pharmaceutical ﹠ Biological Products Control Institute

Methanol analytical pure Beijing Chemical Plant

Acetonitrile chromatographically pure Di Ma company

The pure Beijing of phosphate analysis chemical reagents corporation

2 scutellarins provide scutellarin by Nat'l Pharmaceutical ﹠ Biological Products Control Institute, and measuring purity through high performance liquid chromatography (normalization method) is 96.40%, meet assay with reference substance requirement (in mensuration, converting according to 96.40% purity).

It is an amount of that the scutellarin reference substance is got in 3 selections that detect wavelength, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 0.0463mg, in the interscan of 190～400nm wave-length coverage.The result shows that scutellarin has absorption maximum at 284nm and 335nm place, according to " the Sanitation Ministry medicine standard Chinese traditional patent formulation preparation " relevant kind and in conjunction with bibliographical information, selects the detection wavelength of 335nm as the scutellarin assay.

4 chromatographic conditions

Chromatograph: SHIMADZU LC-2010AHT;

Chromatographic column: Diamonsil ODS 250mm * 4.6mm 5 μ m;

Mobile phase: acetonitrile-0.1% phosphoric acid (20: 80);

Flow velocity: 1.0ml/min;

Column temperature: 30 ℃;

Sample size: 10 μ l;

Detect wavelength: 335nm.

It is an amount of that the preparation precision of reference substance solution takes by weighing the scutellarin reference substance, adds methanol and make the solution that every 1ml contains 0.05mg, promptly.

The about 40mg of this product powder is got in the preparation of need testing solution, the accurate title, decide, put in the 25ml measuring bottle, add an amount of supersound process of methanol (power 250W, frequency 33KHz) 5min, take out, put to room temperature, it is fixed to scale to add methanol, shakes up, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate as need testing solution.

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

Obtain scutellarin reference substance, test sample chromatogram according to above-mentioned chromatographic condition, its number of theoretical plate n is all greater than 5000, and the scutellarin chromatographic peak separates clear complete with close peak, and separating degree is all greater than 1.5, and solvent is noiseless.

The about 40mg of this product powder (totally 6 parts) is got in the selection of 5 extraction times, and accurate the title decides, and splits in the 25ml measuring bottle, it is an amount of to add methanol, respectively supersound process (power 250W, frequency 33KHz) 5,10,20min, take out, put, add methanol to scale to room temperature, shake up, filter with microporous filter membrane (0.45 μ m), the accurate subsequent filtrate 10 μ l that draw inject chromatograph of liquid, measure, promptly.

Extraction time is investigated

Extraction time (min)	Content (mg/ bottle)
Extraction time (min)	Content (mg/ bottle)	5 10 20	4.26 4.28 4.27

The result shows that supersound process 5min can extract fully.

6 linear relationships investigation precision is measured scutellarin reference substance solution (C=0.978mg/ml) 0.0,0.2,0.4,0.6,0.8,1.0ml, split in the 10ml measuring bottle, be diluted to scale with methanol, shake up, the therefrom accurate respectively 10 μ l that draw, inject chromatograph of liquid, measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).With the peak area is vertical coordinate, and the amount of scutellarin (μ g) is abscissa mapping, drawing standard curve.

Scutellarin standard curve determination result

Numbering	Scutellarin amount (μ g)	Conversion back (μ g)	Peak area
Numbering	Scutellarin amount (μ g)	Conversion back (μ g)	Peak area	1 2 3 4 5 6	0.0000 0.1956 0.3912 0.5868 0.7824 0.9780	0.0000 0.1886 0.3771 0.5657 0.7542 0.9428	0 603722 1224078 1835374 2505678 3038182

Regression equation: Y=3259091.50x-1830.06

Correlation coefficient: γ=0.9999

The result shows: scutellarin is good in 0.1886 μ g～0.9428 μ g scope internal linear.

As calculated, the standard curve of scutellarin is crossed initial point, therefore selects for use one point external standard method to measure the content of scutellarin.

Accurate scutellarin reference substance solution (0.0489mg/ml) the 10 μ l that draw of 7 precision experiment inject chromatograph of liquid, measure 5 times, investigate reference substance solution precision, and measurement result sees Table 33.

The precision test

Test number (TN)	1	2	3	4	5	Meansigma methods	RSD(％)
Test number (TN)	1	2	3	4	5	Meansigma methods	RSD(％)	Peak area	1572015	1570202	1566206	1559384	1557017	1564965	0.42

The result shows that reference substance solution precision is good.

8 stability experiments

8.1 accurate scutellarin reference substance solution (0.0489mg/ml) the 10 μ l that draw of the stability experiment of reference substance solution inject chromatograph of liquid, inject chromatograph of liquid, measure at 0,2,4,8,24 hour sample introduction respectively.

The reference substance solution stability test

Time (h)	0	2	4	8	24	Meansigma methods	RSD(％)
Time (h)	0	2	4	8	24	Meansigma methods	RSD(％)	Peak area	1572015	1570202	1566206	1559384	1557017	1564965	0.42

The result shows that 24 hours internal stabilities of reference substance solution are good.

8.2 accurate need testing solution (1.4013mg/ml) the 10 μ l that draw of the stability experiment of need testing solution inject chromatograph of liquid, measure at 0,2,4,8,24 hour sample introduction respectively.

The need testing solution stability experiment

Time (h)	0	2	4	8	24	Meansigma methods	RSD(％)
Time (h)	0	2	4	8	24	Meansigma methods	RSD(％)	Content (mg/ bottle)	4.21	4.22	4.18	4.24	4.23	4.22	0.55

The result shows that 24 hours internal stabilities of need testing solution are good.

9 repeated experiments are got this product, by operating under the preparation of text need testing solution and the algoscopy item.

Repeated experiment

Numbering	1	2	3	4	5	Meansigma methods	RSD(％)
Numbering	1	2	3	4	5	Meansigma methods	RSD(％)	Content (mg/ bottle)	4.26	4.31	4.24	4.38	4.28	4.29	1.27

The application of sample absorption method is adopted in the experiment of 10 average recoveries; get the about 100mg of this product powder (totally 6 parts); the accurate title decides; split in the 25ml measuring bottle; accurate respectively scutellarin reference substance solution (C=1.092mg/ml) 0.6ml, 0.8ml and the 1.0ml (each two parts) of adding; add an amount of supersound process of methanol (power 250W, frequency 33KHz) 5min, take out; put to room temperature; add methanol to scale, shake up, use microporous filter membrane (0.45 μ m) to filter; the accurate subsequent filtrate 10 μ l that draw; inject chromatograph of liquid, mensuration, promptly.

The content that the present invention relates to scutellarin in the injection is: 0.81%.

The average recovery experiment

Numbering	Test sample sample weighting amount (mg)	Pure product amount (mg) in the test sample	Scutellarin addition (mg)	Conversion back (mg)	Measured value (mg)	The response rate (%)
Numbering	Test sample sample weighting amount (mg)	Pure product amount (mg) in the test sample	Scutellarin addition (mg)	Conversion back (mg)	Measured value (mg)	The response rate (%)	1 2 3 4 5 6	109.60 114.11 102.88 108.57 119.11 106.59	0.8878 0.9243 0.8333 0.8794 0.9648 0.8634	0.6552 0.6552 0.8736 0.8736 1.092 1.092	0.6316 0.6316 0.8422 0.8422 1.0527 1.0527	1.4930 1.5346 1.6523 1.7059 1.9893 1.9077	95.82 96.63 97.25 98.14 97.32 99.20

Average recovery rate=97.39%, RSD=1.21%

11 sample sizes are measured and are got this product three batch samples, according to the preparation of text need testing solution and the operation under the algoscopy item.

The scutellarin assay

Lot number	Scutellarin content (mg/ bottle)
Lot number	Scutellarin content (mg/ bottle)	1 2 3	4.33 4.41 4.37

The specific embodiment

Embodiment 1 finger printing

Embodiment 2 finger printing

Embodiment 3 differentiates

It is an amount of to get each preparation, with n-butyl alcohol or ethanol or methanol extraction, filters, and filtrate is as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Get Radix Ginseng Rubra or Radix Ginseng control medicinal material, add the chloroform reflux, extract,, discard chloroform solution, residue adds water-saturated n-butanol and extracts, and extracting solution adds ammonia solution, and divide and get the upper strata, evaporate to dryness, residue is with methanol or dissolve with ethanol, medical material solution in contrast; Other gets ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rf's reference substance, adds methanol or ethanol respectively and makes the solution that every 1ml contains 2mg, in contrast product solution; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate or silica gel H lamellae or silica GF254 lamellae, lower floor's solution or n-butyl alcohol-ethyl acetate or the Ethyl formate-water 1～10: 0.2～2 placed below 5～40: 10～100: 5～50: 0.2～30 10 ℃ with chloroform one ethyl acetate or Ethyl formate-methanol-water: 1～15 upper solution is developing solvent, launch, take out, dry, spray is with 5～50% sulphuric acid ethanol reagent or 50% sulphate reagent, 90 ℃～130 ℃ dry by the fire to speckle colour developing clear, put respectively under daylight and the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, should show the speckle of same color, negative noiseless;

Embodiment 4 differentiates

Embodiment 5: assay

The assay of scutellarin in a, the injection:

Content of total flavone is measured in b, the injection:

C. ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's assay in the injection:

D. the assay of total saponins in the injection:

Ophiopogonin B, ophiopogonin D, ophiopogonin D ' assay in e, the injection:

Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin assay in f, the injection:

The assay of lobetyolin in g, the injection:

Embodiment 6: assay

The assay of scutellarin in a, the injection:

Content of total flavone is measured in b, the injection:

C. ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's assay in the injection:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's reference substance is contrast, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, acetonitrile-water is a mobile phase, gradient elution, solvent ratios is from 0 minute to 35 minutes, the ratio of acetonitrile is 19%, from 35 minutes to 55 minutes, the ratio of acetonitrile rises to 29% by 19%, and from 55 minutes to 70 minutes, the ratio of acetonitrile was 29%, from 70 minutes to 100 minutes, the ratio of acetonitrile rose to 4O% by 29%; Flow velocity is 1.0ml/min, detects wavelength 203nm, and column temperature is in 10～50 ℃ of scopes; Calculate with external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, the limit that the per unit amount contains the ginsenoside Rg1 must not be less than 1.0mg, the limit that contains the ginsenoside Re must not be less than 0.5mg, the limit that contains the ginsenoside Rb1 must not be less than 1.0mg, and the limit that contains ginsenoside Rg1, ginsenoside Re's summation must not be less than 2.5mg;

D. the assay of total saponins in the injection:

Ophiopogonin B, ophiopogonin D, ophiopogonin D ' assay in e, the injection:

Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin assay in f, the injection:

The assay of lobetyolin in g, the injection:

Claims

1, a kind of method of quality control with the Chinese medicine made Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis, it is characterized in that: this method comprises following all or part of content:

2, according to the described method of quality control of claim 1, it is characterized in that this method comprises one or more in the following finger printing with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis:

3, according to the described method of quality control of claim 2 with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis,, it is characterized in that this method comprises one or more in the following finger printing:

4, according to the described method of quality control with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis and the Chinese medicine made Radix Ophiopogonis of claim 1, it is characterized in that: the discrimination method of described injection is following all or part of content:

It is an amount of to get each preparation, puts in the measuring bottle, adds methanol or ethanol or mobile phase to scale, shakes up, and filters with microporous filter membrane, gets subsequent filtrate as need testing solution; Take by weighing other flavour of a drug and adjuvant in the prescription ratio, make negative control solution with method; Methanol solution with lobetyolin's reference substance is contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, 5%～80%: 95%～20% methanol or acetonitrile-water or 0.05%～5% glacial acetic acid aqueous solution or 0.02%～0.5% phosphate aqueous solution are mobile phase, detect wavelength and be in 200～410nm scope one or youngster, in 20～50 ℃ of scopes of column temperature; In the test sample chromatograph, answer tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless.

5, according to the described method of quality control with the Chinese medicine made Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis of claim 4, it is characterized in that: the discrimination method of described injection is following one or more:

6, according to the described method of quality control with the Chinese medicine made Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis of claim 1, it is characterized in that: the method for testing of described injection content should comprise one or more in following:

The assay of scutellarin in a, the injection:

Content of total flavone is measured in b, the injection:

C. ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's assay in the injection:

D. the assay of total saponins in the injection:

Ophiopogonin B, ophiopogonin D, ophiopogonin D ' assay in e, the injection:

Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin assay in f, the injection:

The assay of lobetyolin in g, the injection:

7, according to the described method of quality control with the Chinese medicine made Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis of claim 6, it is characterized in that: the method for testing of described injection content is following one or more methods:

The assay of scutellarin in a, the injection:

Content of total flavone is measured in b, the injection:

C. ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re's assay in the injection:

D. the assay of total saponins in the injection:

It is an amount of to get each preparation, put in the 10ml measuring bottle, adding distil water makes dissolving in right amount and decides and shakes up to scale, and precision is measured 0.6ml, to the 25ml measuring bottle, water bath method takes out immediately, and precision adds 5% vanillin glacial acetic acid solution 0.5ml, perchloric acid 2.0ml, shake up, heating is 15 minutes in 60 ℃ of water-baths, takes out, immediately with ice-water bath cooling 2 minutes, precision adds glacial acetic acid to 25ml, shakes up, and be blank with the retinue solvent, adopt the disclosed spectrophotography of Chinese Pharmacopoeia appendix, wavelength place at 547nm measures trap, calculates with external standard method or standard curve method, and each preparation is unit quantity to be equivalent to every day with output, the per unit amount contains total saponins in the ginsenoside Rg1, must not be less than 18.0mg;

Ophiopogonin B, ophiopogonin D, ophiopogonin D ' assay in e, the injection:

Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin assay in f, the injection:

The assay of lobetyolin in g, the injection:

8, according to claim 6 or 7 described with Herba Erigerontis, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, the method of quality control of the Chinese medicine of making Radix Ophiopogonis, it is characterized in that: calculate according to percentage by weight, all can be surveyed composition and comprise scutellarin in the described injection, the ginsenoside Rg1, the ginsenoside Rb1, the ginsenoside Re, total saponins, ophiopogonin B, ophiopogonin D, ophiopogonin D ', Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, lobetyolin, all or part of total content in the atractylenoide accounts for the total solid of deducting adjuvant amount and water quantities in the preparation and is not less than 25%.