CN1911395A

CN1911395A - Method for controlling quality of injection contg. traditional Chinese medicine

Info

Publication number: CN1911395A
Application number: CN 200510090432
Authority: CN
Inventors: 于文风
Original assignee: Union Xichuang Pharmaceutical Science & Tech Co Ltd Beijing
Current assignee: Union Xichuang Pharmaceutical Science & Tech Co Ltd Beijing
Priority date: 2005-08-12
Filing date: 2005-08-12
Publication date: 2007-02-14

Abstract

A quality control method for the Chinese-medicinal injection prepared from brevis capine, ginseng or red ginseng or pilose asiabell root, ophiopogon root and schisandra fruit includes all or part of the fingerprint test, differential test and content measurement of ginseng or red ginseng or piolse asiabell root, ophiopogon root, schisandra fruit, etc.

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 of the Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis, belong to the field of medical technology.

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 carries out breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis to have developed a kind of Chinese medicine after the assembly, comprises the injection that is directly used in drug administration by injection, needs to be used for after the dilution concentrated solution for injection of intravenous drip, directly for the glucose intravenous infusion of intravenous drip and sodium chloride intravenous infusion and the injectable sterile powder and the aseptic block that make with freeze-drying or spray drying method.Said preparation is achieved in that according to components by weight percent and calculates, it is by 10-500 part Radix Ophiopogonis, Radix Ginseng 1-300 part, Fructus Schisandrae Chinensis 1-300 part and breviscapine 0.1-30 part are made through extracting refining and adding suitable adjuvant, or adding suitable adjuvant by the breviscapine of corresponding weight portion medical material through extracting the extract that obtains after refining and corresponding weight portion is made, described have human body immunity improving power, the ejection preparation of treatment cardiovascular and cerebrovascular disease, calculate according to components by weight percent, it is by 50-200 part Radix Ophiopogonis, Radix Ginseng 10-100 part, Fructus Schisandrae Chinensis 10-100 part and breviscapine 0.3-15 part are made through extracting refining and adding suitable adjuvant, or adding suitable adjuvant by the breviscapine of corresponding weight portion medical material through extracting the extract that obtains after refining and corresponding weight portion is made and adds suitable adjuvant and be made, exactly, it is by 80～150 parts of Radix Ophiopogonis, 30～80 parts of Radix Ginsengs, 0.5～8 part of 30～80 parts of Fructus Schisandrae Chinensis and breviscapine are made through extracting refining and adding suitable adjuvant, or add suitable adjuvant and be made through extracting the extract that obtains after refining and corresponding weight portion breviscapine by corresponding weight portion medical material, the Radix Ginseng in the prescription can also be the Radix Ginseng Rubra or the Radix Codonopsis of same ratio.

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 Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis 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, active component and the breviscapine of not only having chosen Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis carry out assay, with the quality of how much judging of its content, have more set up finger printing, 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 of the Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis is provided, this method of utilizing the inventor to provide, can effectively control the quality of the Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis, 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:

The method of quality control of the Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis comprises following all or part of content:

(1) finger printing test, comprise based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics finger printing based on the finger printing of Radix Codonopsis composition characteristics with based on the finger printing of Fructus Schisandrae Chinensis composition characteristics;

(2) Radix Ginseng Rubra or ginseng crude drug, Radix Ophiopogonis medical material, schisandra chinensis medicinal material, codonopsis pilosula, scutellarin, ginsenoside Rg ₁, ginsenoside Rb ₁, 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, schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B, lobetyolin, atractylenoide;

(3) Radix Ginseng soap Rg ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf, ophiopogonin B, ophiopogonin D, ophiopogonin D ', the content test method of all or part of composition such as Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, schisandrin, schisantherin B, schisantherin A, deoxyschizandrin, schisandrin B, schisandrin C, total saponins, total lignans, lobetyolin, atractylenoide.

Described based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics finger printing based on the finger printing of Radix Codonopsis composition characteristics with based on the fingerprint test method of Fructus Schisandrae Chinensis composition characteristics be:

A, 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 that breviscapine, Radix Ginseng Rubra or Radix Ginseng, Radix Ophiopogonis, Radix Codonopsis and Fructus Schisandrae Chinensis make, and 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 in an amount of Radix Ginseng Rubra or Radix Ginseng and the Radix Ophiopogonis medical material, comprise the ginsenoside Rg ₁, ginsenoside Rb ₁, 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 0.2%～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 20～60 ℃;

(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;

(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, employing liquid chromatography are tested the finger printing based on the Fructus Schisandrae Chinensis composition characteristics:

(1) preparation of need testing solution: it is an amount of to get the Chinese medicine that breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis, Radix Codonopsis and Fructus Schisandrae Chinensis make, and adds water or methanol, 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 product in contrast in an amount of schisandra chinensis medicinal material, comprise in schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schizantherin the second grade one or more, water or methanol, ethanol dilution are 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: water or 0.01%～1% phosphoric acid solution or 0.2%～3% glacial acetic acid solution or 0.2%～3% formic acid solution, gradient elution, flow velocity are that 0.5～1.5ml/min, detection wavelength are 203-390nm, and column temperature is 20～50 ℃;

(4) based on the formulation of the standard finger-print of Fructus Schisandrae Chinensis 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 Fructus Schisandrae Chinensis ingredients fingerprint in the injection to be measured;

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

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

(1) preparation of need testing solution: it is an amount of to get the Chinese medicine that scutellarin, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis make, and 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 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 20～60 ℃;

(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;

(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.

The method of quality control of the described Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng, Radix Ophiopogonis and Fructus Schisandrae Chinensis comprises one or more in the following finger printing:

A, 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, with the microporous filter membrane filtration of 0.45 μ m, gets subsequent filtrate as need testing solution;

(2) preparation of object of reference solution: precision takes by weighing the ginsenoside Rg ₁In right amount, add 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;

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

(6) with pulse restoring 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 the finger printing of liquid chromatography for measuring based on the Fructus Schisandrae Chinensis composition characteristics:

(2) preparation of object of reference solution: it is an amount of that precision takes by weighing the schisandrin reference substance, adds water 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, gradient elution, solvent ratios are from 0 minute to 6 minutes, and the ratio of methanol is 68%, from 6 minutes to 8 minutes, the ratio of methanol rises to 76% by 68%, and from 8 minutes to 10 minutes, the ratio of methanol was 76%, from 10 minutes to 15 minutes, the ratio of methanol reduces to 68% by 76%, and from 15 minutes to 60 minutes, the ratio of methanol was 68%; Flow velocity is 1.0ml/min, and the detection wavelength is 250 ± 2nm, and column temperature is 30 ℃;

(4) based on the formulation of the standard finger-print of Fructus Schisandrae Chinensis composition characteristics: accurate respectively each the 10 μ l of reference substance solution and need testing solution that draw, inject chromatograph of liquid respectively, according to measured collection of illustrative plates, the formulation standard finger-print;

(5) with the means of testing of said method as Fructus Schisandrae Chinensis ingredients fingerprint in the pulse restoring injection to be measured;

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

(2) preparation of object of reference solution: get the main active atractylenoide in an amount of codonopsis pilosula, be diluted to suitable concn, as object of reference solution with dissolve with methanol;

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler: mobile phase is 67%: 33% methanol-water, and flow velocity is that .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.

The discrimination method of the described Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis 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 equal solvent and extracts, and filters, and filtrate volatilizes, and residue is with the dissolving of methanol equal solvent, 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 the methanol equal solvent 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 four kinds of solution, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica gel G F ₂₅₄Lamellae, 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, 105 ℃ 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 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 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 200～410nm, 20～50 ℃ of column temperatures; 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;

C. the thin layer chromatography discrimination method of schisandra chinensis medicinal material, schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B in the injection:

It is an amount of to get each preparation, adds chloroform or ether or ethyl acetate or normal hexane or 10～95% ethanol equal solvents and extracts, and filters, and filtrate volatilizes, and residue adds chloroform or the dissolving of ethyl acetate equal solvent, 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 Fructus Schisandrae Chinensis control medicinal material, shines medical material solution in pairs with legal system; Get schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B reference substance again, add chloroform respectively or the ethyl acetate equal solvent is made 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 ten kinds of solution, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica gel G F ₂₅₄Lamellae, with petroleum ether (30～60 ℃) or petroleum ether (60～90 ℃)-Ethyl formate or ethyl acetate-formic acid or acetic acid 2～40: 0.2～15: 0.1～5 upper solution or benzene or toluene-ethyl acetate or Ethyl formate 1～30: 0.2～10 or normal hexane-benzene or toluene-Ethyl formate or ethyl acetate-formic acid or acetic acid 0.2～5: 0.5～10: 1～15: 0.1～5 are developing solvent, launch, take out, dry, putting uviol lamp 365nm or 254nm inspects or sprays with 2～20% chromotropic acids-concentrated sulfuric acid solution or 2～20% phosphomolybdic acid ethanol solutions or anisaldehyde sulfuric acid solution, 80 ℃～160 ℃ are dried by the fire the clear or smoked colour developing of iodine to the speckle colour developing, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, the speckle that should show same color, negative noiseless;

The liquid chromatograph discrimination method of schisandrin, deoxyschizandrin, schisandrin B in d, 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 schisandrin, deoxyschizandrin, schisandrin B 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 200～410nm, 20～50 ℃ of column temperatures; 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 e, the injection Radix Ophiopogonis medical material the thin layer chromatography discrimination method:

It is an amount of to get each preparation, adds the n-butyl alcohol equal solvent and extracts, 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, adds chloroform-methanol or ethyl acetate or n-butyl alcohol equal solvent and extracts, and filters, and evaporate to dryness, residue add chloroform dissolving, medical material solution in contrast; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned three kinds of 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 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 ℃～120 ℃ 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 f, the injection, ophiopogonin D, ophiopogonin D ' the thin layer chromatography discrimination method:

It is an amount of to get each preparation, and the water equal solvent extracts back n-butyl alcohol and ether or the extraction of ethyl acetate equal solvent, and the extract evaporate to dryness is with the dissolving of methanol equal solvent, as need testing solution; Other get ophiopogonin B, ophiopogonin D, ophiopogonin D ', add the methanol equal solvent 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 four liquors, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica gel G F ₂₅₄On the 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 ℃～120 ℃ 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 g, the injection, ophiopogonin D, ophiopogonin D ' the liquid chromatograph discrimination method:

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 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, detecting wavelength is 200～410nm, 20～50 ℃ of column temperatures; 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 h, 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 the dissolving of ethyl acetate equal solvent, as need testing solution.Other gets butchers broom soap former times unit, 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 1～30 μ l of above-mentioned four liquid, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica gel G F ₂₅₄On the 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 ℃～120 ℃ 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 discrimination method of Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin in i, the injection:

It is an amount of to get each preparation, puts in the round-bottomed flask, is dissolved in water, and adds 0.5～38% sulphuric acid or hydrochloric acid hydrolysis, take out, put, transfer pH, extract with the chloroform equal solvent to neutral to room temperature, the extracting solution evaporate to dryness, residue filters with microporous filter membrane with the dissolving of methanol equal solvent, 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, detecting wavelength is 200～410nm, 20～50 ℃ of column temperatures; 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 Rg in j, the injection ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf the thin layer chromatography discrimination method:

It is an amount of to get each preparation, extracts with n-butyl alcohol or ethanol equal solvent, filters, and filtrate is as need testing solution; Get Radix Ginseng Rubra or Radix Ginseng control medicinal material, add the chloroform reflux, extract,, discard chloroform solution, residue adds the water-saturated n-butanol equal solvent and extracts, and extracting solution adds ammonia solution, and divide and get the upper strata, evaporate to dryness, residue dissolves with the methanol equal solvent, in contrast medical material solution; Other gets the ginsenoside Rg ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf's reference substance, add dissolvings such as methanol respectively and make 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 7 kinds of solution, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica gel G F ₂₅₄On the 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 strata is developing solvent, launch, take out, dry, spray is with 5～50% sulphuric acid ethanol reagent or 50% sulphate reagent, 90 ℃～120 ℃ 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;

K. ginsenoside Rg in the injection ₁, ginsenoside Rb ₁, the ginsenoside Re the liquid chromatograph discrimination method:

It is an amount of to get each preparation, puts in the measuring bottle, adds water or 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; With the ginsenoside Rg ₁, ginsenoside Re's reference substance methanol solution for the contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels, 5%～40%: 95%～60% 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, the detection wavelength is in 200～350nm scope or evaporate the photodetector detection, 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 L, the injection:

It is an amount of to get each preparation, is dissolved in water, and with the extraction of n-butyl alcohol equal solvent, extract is concentrated into dried, use dissolve with methanol,, or put in C18 or the C8 solid phase extraction column, use 5%～50% methanol, methanol-eluted fractions successively as need testing solution, 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; Other gets lobetyolin's reference substance, adds methanol 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 four kinds of 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 m, 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 200～410nm, 20～50 ℃ of column temperatures; 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 discrimination method of the described Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng, Radix Ophiopogonis and Fructus Schisandrae Chinensis 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 the methanol equal solvent 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 four kinds of solution, put respectively in 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 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 or alcoholic solution with the scutellarin reference substance are contrast, adopt liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-0.1% phosphate aqueous solution (20%: 80%) is a mobile phase, 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;

C. the thin layer chromatography discrimination method of Fructus Schisandrae Chinensis, schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B in the injection:

It is an amount of to get each preparation, adds chloroform and extracts, and filters, and filtrate volatilizes, and residue adds the chloroform dissolution with solvents, 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 Fructus Schisandrae Chinensis control medicinal material, shine medical material solution in pairs with legal system, get schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B reference substance again, add chloroform respectively or the ethyl acetate equal solvent is made the solution that every 1ml contains 1mg, adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 2 μ l of above-mentioned ten kinds of solution, put in same silica gel G F respectively ₂₅₄Lamellae, with petroleum ether (30～60 ℃)-upper solution of 15: 5: 1 of Ethyl formate-formic acid is developing solvent, launch, take out, dry, put uviol lamp 254nm 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 schisandrin, deoxyschizandrin, schisandrin B reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-water is a mobile phase at 65: 35, and the detection wavelength is 250nm, 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 e, the injection:

It is an amount of to get each preparation, adds chloroform-methanol (7: 3) mixed solution and extracts, and filters, and evaporate to dryness, residue add 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, adds chloroform-methanol and extracts, and filters, and evaporate to dryness, residue add chloroform dissolving, medical material solution in contrast; Adopt the disclosed thin layer chromatography test of Chinese Pharmacopoeia appendix, draw each 10 μ l of above-mentioned three kinds of solution, put respectively in same silicon silica gel G F ₂₅₄On the lamellae, be at 80: 5: 0.1 developing solvent, launch, inspect under the 254nm 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; Other get ophiopogonin B, ophiopogonin D, ophiopogonin D ', add the methanol equal solvent 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 four kinds of 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, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-water is a mobile phase at 90: 10, detecting 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.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 four liquid, 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, detecting 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;

Get this product, use n-butanol extraction, filter, filtrate is as need testing solution; 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 the ginsenoside Rg ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf's reference substance, add dissolvings such as methanol respectively and make 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 7 kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform one ethyl acetate-methanol-water 15: 40: 22: lower floor's solution of placing below 1010 ℃ was developing solvent, launch, take 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; With the ginsenoside Rg ₁, ginsenoside Re's reference substance methanol solution be contrast, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, 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 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, answer 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 that other gets lobetyolin's reference substance, adds methanol and make 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 (7: 1: 0.5) is developing solvent, launch to take out, dry, spray is with 10% ethanol solution of sulfuric acid, 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, answer tool and the reference substance chromatograph consistent chromatographic peak of time of withing a hook at the end, negative noiseless.

The method of testing of the described Chinese medicine injection agent content made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis 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 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 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 200～410nm, 20～50 ℃ 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;

B. ginsenoside Rg in the injection ₁, ginsenoside Rb ₁, ginsenoside Re's assay:

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; With the ginsenoside Rg ₁, ginsenoside Re's reference substance methanol solution for the contrast, adopt liquid chromatography, chromatographic column is a filler with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels, 5%～40%: 95%～60% 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, the detection wavelength is in 200～350nm scope or evaporate the photodetector detection, 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, and the per unit amount contains the ginsenoside Rg ₁Limit must not be less than 1.0mg, the limit that contains the ginsenoside Re must not be less than 0.5mg, contains ginsenoside Rb ₁Limit must not be less than 1.0mg, contain the ginsenoside Rg ₁, the ginsenoside Re the limit of summation must not be less than 2.5mg;

C. the assay of total saponins in the injection:

It is an amount of to get each preparation, puts in the 10ml measuring bottle, and adding distil water makes dissolving and fixed to scale in right amount, shake up, 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 0.2～4ml, perchloric acid 0.5～5ml shakes up, and heating is 15 minutes in 60 ℃ of water-baths, take out, with ice-water bath cooling 2 minutes, precision added glacial acetic acid to 25ml immediately, shake up, as need testing solution; With the ginsenoside Rg ₁Or ginsenoside Rb ₁Or the product of 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, according to spectrophotography, measures trap at the wavelength place of 547 ± 10nm, calculates with external standard method or standard curve method, and each preparation is unit quantity to be equivalent to every day with output, and the per unit amount contains total saponins with the ginsenoside Rg ₁Or ginsenoside Rb ₁Or ginsenoside Re or ginsenoside Rf or ophiopogonin B or ophiopogonin D or ophiopogonin D ' meter, must not be less than 18.0mg;

Total lignans assay in d, the injection:

It is an amount of to get each preparation, puts in the measuring bottle, and it is fixed to scale to add water, behind salt Acid precipitation saponin component, use ethyl acetate extraction and extracting solution also, water bath method, and the residue dissolve with methanol is as need testing solution; With schisandrin product in contrast, get reference substance solution with legal system.With the retinue solvent is blank, according to spectrophotography, measures trap at the wavelength place of 254 ± 10nm, calculate with external standard method or standard curve method, each preparation is unit quantity to be equivalent to every day with output, and the per unit amount contains total lignans in schisandrin, must not be less than 4.0mg;

Schisandrin, deoxyschizandrin, schisandrin B 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 schisandrin, deoxyschizandrin, schisandrin B 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 200～410nm, 20～50 ℃ 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 schisandrin must not be less than 0.3mg, and the limit that contains deoxyschizandrin must not be less than 0.3mg; The limit that contains schisandrin B must not be less than 0.3mg; The limit that contains the summation of schisandrin, deoxyschizandrin, schisandrin B must not be less than 0.9mg;

Ophiopogonin B, ophiopogonin D, ophiopogonin D ' assay in f, 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, detecting wavelength is that 200～410nm or evaporation photodetector detect 20～50 ℃ 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 g, the injection:

It is an amount of to get each preparation, puts in the round-bottomed flask, is dissolved in water, and adds 0.5～38% sulphuric acid or hydrochloric acid hydrolysis, take out, put, transfer pH, use chloroform extraction to neutral to room temperature, the extracting solution evaporate to dryness, 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, detecting wavelength is that 200～410nm or evaporation photodetector detect 20～50 ℃ 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 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 h, 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 200～410nm, 20～50 ℃ 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.

The method of testing of the described Chinese medicine injection agent content made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis 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 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, acetonitrile-0.1% phosphate aqueous solution is a mobile phase at 20: 80, the detection wavelength is 335nm, 30 ℃ of column temperatures; Calculate with one point external standard 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;

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; With the ginsenoside Rg ₁, ginsenoside Re's reference substance methanol solution be contrast, adopt liquid chromatography, the chromatographic column octadecylsilane chemically bonded silica is a filler, 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 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 one point external standard method, each preparation is unit quantity to be equivalent to every day with output, and the per unit amount contains the ginsenoside Rg ₁Limit must not be less than 1.0mg, the limit that contains the ginsenoside Re must not be less than 0.5mg, contains ginsenoside Rb ₁Limit must not be less than 1.0mg, contain the ginsenoside Rg ₁, the ginsenoside Re the limit of summation must not be less than 2.5mg;

C. the assay of total saponins in the injection:

It is an amount of to get each preparation, puts in the 10ml measuring bottle, and adding distil water makes dissolving and fixed to scale in right amount, shake up, precision is measured 0.6ml, to the 25ml measuring bottle, water bath method, take out immediately, 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, shaking up, is blank with the retinue solvent, according to spectrophotography, wavelength place at 547nm measures trap, calculate with one point external standard method, each preparation is unit quantity to be equivalent to every day with output, and the per unit amount contains total saponins with the ginsenoside Rg ₁Meter must not be less than 18.0mg;

Total lignans assay in d, the injection

It is an amount of to get each preparation, puts in the measuring bottle, and it is fixed to scale to add water, behind salt Acid precipitation saponin component, use ethyl acetate extraction and and extracting solution, water bath method, the residue dissolve with methanol is a blank with the retinue solvent, according to spectrophotography, wavelength place at 254nm measures trap, calculates with one point external standard method, and each preparation is unit quantity to be equivalent to every day with output, the per unit amount contains total lignans in schisandrin, must not be less than 4.0mg;

Schisandrin, deoxyschizandrin, schisandrin B 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 schisandrin, deoxyschizandrin, schisandrin B reference substance is contrast, adopts liquid chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filler, and methanol-water is a mobile phase at 65: 35, and the detection wavelength is 250nm, 30 ℃ of column temperatures; Calculate with one point external standard method, each preparation is unit quantity to be equivalent to every day with output, and the limit that the per unit amount contains schisandrin must not be less than 0.3mg, and the limit that contains deoxyschizandrin must not be less than 0.3mg; The limit that contains schisandrin B must not be less than 0.3mg; The limit that contains the summation of schisandrin, deoxyschizandrin, schisandrin B must not be less than 0.9mg;

Ophiopogonin B, ophiopogonin D, ophiopogonin D ' assay in f, 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, the chromatographic column octadecylsilane chemically bonded silica is a filler, and acetonitrile-water is a mobile phase at 90: 10, detecting wavelength is 203nm, 30 ℃ of column temperatures; Calculate with one point external standard 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 g, the injection:

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, put to room temperature, transfer pH, use chloroform extraction and extracting solution also to neutral, evaporate to dryness, 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 (90: 10) is a mobile phase, detecting 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, 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 h, 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, 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;

Compared with prior art, the present invention's quality of the Chinese medicine injection products made with breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis of perfect control more.The Chinese medicine ingredients complexity, if only with wherein one, two kind of composition illustrate its inherent quality, has certain one-sidedness, more can't judge the index components of no drug effect.Therefore the applicant formulated based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics finger printing based on the finger printing of Radix Codonopsis composition characteristics with control the quality of injection based on the finger printing of Fructus Schisandrae Chinensis composition characteristics comprehensively.But because contained complex chemical composition between each medical material in the breviscapus pulse-engendering 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 of breviscapus pulse-engendering preparation is not that the finger printing simple superposition of breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and schisandra chinensis medicinal material or preparation is just getable, because several medical material ingredients interference effect each other in the prescription, cause the finger printing characteristic peak of Radix Ginseng Rubra in the breviscapus pulse-engendering preparation or Radix Ginseng and Radix Ophiopogonis part, Radix Codonopsis part, Fructus Schisandrae Chinensis part to 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 more effective to the quality control of the Chinese medicine injection products made with breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis, and method precision, precision, stability are all higher.

Experimental example 1 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.

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

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

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 is based on the preparation of the finger printing of Fructus Schisandrae Chinensis composition characteristics

A, experimental apparatus, reagent and sample

Reference substance: schisandrin: 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 5400 (calculating with the object of reference schisandrin).So finally selecting Diamonsil ODS chromatographic column (250mm * 4.6mm, 5 μ m) for use is the experimentation post.

2. the selection of mobile phase

(1) (20: 80) have been investigated in the research process respectively, (2) acetonitrile-water (10: 80), (3) (the gradient elution volume proportion is from 0 minute to 6 minutes to acetonitrile-water (gradient elution) (4) methanol-water, the ratio of methanol is 68%, from 6 minutes to 8 minutes, the ratio of methanol rises to 76% by 68%, from 8 minutes to 10 minutes, the ratio of methanol is 76%, from 10 minutes to 15 minutes, the ratio of methanol reduces to 68% by 76%, and from 15 minutes to 60 minutes, the ratio of methanol was 68%) 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

In the research under methanol-water (gradient elution) mobile phase condition, investigated the chromatographic peak situation under different-waveband typical wavelengths 203,210,230,250,300nm respectively, the result shows that chromatographic peak is more under 250nm, peak shape is better, so finally select for use 250nm 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); 30 ℃ of column temperatures, flow velocity 1.0ml/min.

5. the preparation of need testing solution

6. the preparation of object of reference solution

Schisandrin is one of main active in the Fructus Schisandrae Chinensis, 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 schisandrin is as object of reference.

7. finger printing and technical parameter

Experimental example 3 is differentiated and assay

The thin layer chromatography discrimination method of Herba Erigerontis in the breviscapus pulse-engendering 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) reference substance is expanded to the forward position

Silica gel g thin-layer plate

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

The silica gel H lamellae

Ethyl acetate-acetic acid-water (10-5-1) reference substance is expanded to the forward position

Silica gel g thin-layer plate

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

Silica gel g thin-layer plate

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

Silica gel g thin-layer plate

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

Silica gel g thin-layer plate

Determine condition: it is clear that toluene-ethyl acetate-acetic acid-water (10-5-1-0.5) separates, and the moderate negative nothing of Rf value is done

Silica gel g thin-layer plate is disturbed

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 liquid chromatograph discrimination method of scutellarin in the breviscapus pulse-engendering 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) feminine gender has interference

Eight alkyl silane bonded silica gels

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,

The octadecylsilane chemically bonded silica symmetry, negative 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 thin layer chromatography discrimination method of schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B in the breviscapus pulse-engendering injection

Feature for outstanding Fructus Schisandrae Chinensis, selected schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B is as its feature speckle, but owing to exist more and schisandrin in the medical material, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, the schisantherin B structure is 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 schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B launches:

Condition question

Normal hexane-benzene-Ethyl formate-formic acid (5-5-10-4) silica gel H lamellae reference substance is expanded to the forward position

Normal hexane-benzene-Ethyl formate-formic acid (5-5-5-3) reference substance does not separate, and feminine gender has interference

The silica gel H lamellae

Be expanded to the forward position with petroleum ether (30～60 ℃)-Ethyl formate-formic acid (10: 10: 3) reference substance

The upper solution silica gel g thin-layer plate

Do not separate with petroleum ether (60～90 ℃)-Ethyl formate-formic acid (15: 5: 1) reference substance, feminine gender has interference

Upper solution silica gel G F ₂₅₄Lamellae

Toluene-ethyl acetate (10-5) silica gel H lamellae reference substance does not separate, and feminine gender has interference

Toluene-Ethyl formate (15-5) silica gel g thin-layer plate reference substance does not separate, and feminine gender has interference

It is clear to separate with petroleum ether (30～60 ℃)-Ethyl formate-formic acid (15: 5: 1), and the moderate feminine gender of Rf value is noiseless

Upper solution silica gel G F ₂₅₄Lamellae

Through screening, determined with the silica gel g thin-layer plate to be immobile phase, upper solution with petroleum ether (30～60 ℃)-Ethyl formate-formic acid (15: 5: 1) is developing solvent, with this understanding, the Rf value of schisandrin, schisantherin B, deoxyschizandrin, schisandrin B, schisandrin C, schisantherin A, schisantherin B is moderate, it is clear to separate with other speckle, negative noiseless.

The liquid chromatograph discrimination method of schisandrin, deoxyschizandrin, schisandrin B in the breviscapus pulse-engendering injection

Feature for outstanding Fructus Schisandrae Chinensis, selected schisandrin, deoxyschizandrin, schisandrin B as its characteristic component, but because have in the medical material that more and schisandrin, deoxyschizandrin, schisandrin B 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 schisandrin, deoxyschizandrin, schisandrin B are separated:

Condition question

Methanol-water (80: 20) octadecylsilane chemically bonded silica appearance time is too fast

Acetonitrile-water (40: 60) octadecylsilane chemically bonded silica feminine gender has interference

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

Methanol-0.2% phosphate aqueous solution (70: 30) octadecylsilane chemically bonded silica peak shape is slightly asymmetric

Acetonitrile-0.2% phosphate aqueous solution (70: 30) octadecylsilane chemically bonded silica peak shape is slightly asymmetric

Methanol-water (50: 50) eight alkyl silane bonded silica gel feminine genders have interference

Methanol-water (65: 35) octadecylsilane chemically bonded silica retention time is moderate, the capable point in peak

Sharp, symmetry, negative noiseless

Through screening, determined with the octadecylsilane chemically bonded silica to be immobile phase, methanol-water (65: 35) be a mobile phase, and with this understanding, schisandrin, deoxyschizandrin, schisandrin B retention time are moderate, and the peak is capable sharp-pointed, symmetry, feminine gender is noiseless.

The thin layer chromatography discrimination method of Radix Ophiopogonis in the breviscapus pulse-engendering 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.

Ophiopogonin B in the breviscapus pulse-engendering 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 plate reference substance is expanded to the forward position

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) silicon, and the moderate feminine gender of Rf value is noiseless

Glue G lamellae

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.

Ophiopogonin B in the breviscapus pulse-engendering 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 thin layer chromatography discrimination method of Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin in the breviscapus pulse-engendering injection

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, 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 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 liquid chromatograph discrimination method of Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin in the breviscapus pulse-engendering injection

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

Methanol-2% glacial acetic acid aqueous solution (95: 5) eight alkyl silane bonded silica gel feminine genders have interference

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

There is bifurcated at acetonitrile-1% glacial acetic acid aqueous solution (95: 5) eight alkyl silane bonded silica gel peaks,

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

Acetonitrile-water (90: 10) octadecylsilane chemically bonded silica retention time is moderate, the peak

Row is sharp-pointed, symmetry, the moon

Property 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.

Ginsenoside Rg in the breviscapus pulse-engendering 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.

Ginsenoside Rg in the breviscapus pulse-engendering 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

1 0.510 150.04

2 1.275 324.27

3 2.550 856.87

4 3.060 1024.83

5 6.120 2098.79

6 10.200 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

1 0.536 129.65

2 1.340 283.75

3 2.680 759.26

4 3.216 902.30

5 6.432 1840.48

6 10.720 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.

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 1, 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) 12345 meansigma methods RSD (%)

Ginsenoside Rg1's peak area 859.78 847.26 846.53 856.40 845.30 851.054 0.772%

Ginsenoside Re's peak area 746.19 774.10 740.15 736.24 730.54 740.044 0.135%

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

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 times (h) 06 12 24 48 average RSD (%)

Ginsenoside Rg1's peak area 856.87 861.83 853.35 863.07 860.64 859.152 0.464%

Ginsenoside Re's peak area 759.26 755.05 755.69 759.75 756.70 757.29 0.279%

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

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 (mg/ bottle) ginsenoside Re (mg/ bottle)

1 1.14 0.63

2 1.26 0.59

3 1.18 0.61

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 of Radix Ginseng total saponins in the spectrophotometry breviscapus pulse-engendering 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 (appendix VA of Chinese Pharmacopoeia version in 2000), 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

1 27.4 0.121

2 54.8 0.261

3 82.2 0.404

4 109.6 0.534

5 137.0 0.656

Precision test precision takes by weighing ginsenoside Rg1's 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

Number the average RSD% of 12345 X

Absorbance 0.429 0.426 0.426 0.425 0.424 0.426 0.44

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 Radix Ginseng total saponins (mg/ bottle)

1 27.62

2 29.52

3 28.33

Three, schisandrin

Instrument and reagent

Instrument: the Agilent1100 high performance liquid chromatograph, the chemstationsys work station,

The TU-1800SPC ultraviolet spectrophotometer, the AE240 electronic balance.

Reagent: schisandrin: Nat'l Pharmaceutical ﹠ Biological Products Control Institute (110857-200203);

Detect the selection of wavelength

Precision takes by weighing schisandrin reference substance 15.00, puts in the 50ml measuring bottle, adds methanol and is diluted to scale, shakes up, and gets the solution that every 1ml contains schisandrin 0.30mg, in 200-400nm wave-length coverage scanning (scanning spectra is as follows).Schisandrin has absorption maximum at the 250nm place, therefore selects the detection wavelength of 250nm as assay.

Chromatographic condition: Dikma OHS (4.6 * 250mm, 5um); Mobile phase: methanol-water (65: 35) is a mobile phase; The detection wavelength is 250nm; Flow velocity: 1.0ml/min.

The system suitability test

The preparation of reference substance solution

Precision takes by weighing schisandrin reference substance 3.04mg, puts in the 10ml measuring bottle, adds methanol and is diluted to scale, shakes up, and gets the solution that every 1ml contains schisandrin 0.304mg.

The preparation of need testing solution

Get the about 0.5g of this product, the accurate title, decide, and puts in the 10ml measuring bottle, adds methanol to scale, shakes up, and filters, promptly.

The preparation of negative need testing solution

Take by weighing the negative sample that does not contain Fructus Schisandrae Chinensis, according to the preparation method preparation of need testing solution, as negative need testing solution.

Accurate respectively reference substance solution, need testing solution, each 10 μ l of negative need testing solution of drawing inject chromatograph of liquid, measure the record chromatogram.As seen from the figure, test sample schisandrin peak separates good, and the schisandrin peak is noiseless in the negative test sample.

The linear relationship of schisandrin

Precision takes by weighing schisandrin reference substance 15.06mg, put and add methanol in the 50ml measuring bottle and make dissolving and fixed in right amount to scale, shake up, accurate 1 μ l, 2 μ l, 4 μ l, 6 μ l, 8 μ l, the 10 μ l of drawing, inject chromatograph of liquid, with the peak area is vertical coordinate, and the amount of schisandrin (μ g) is figure for abscissa, the drawing standard curve.

Regression equation: Y=1540.47X+38.26

The coefficient of determination: r=0.998

Schisandrin is good in 0.3012～3.0120 μ g scope internal linear relation.

The schisandrin linear relationship is investigated

Numbering schisandrin (μ g) peak area

1 0.3012 437.66

2 0.6024 980.45

3 1.2048 1879.71

4 1.8072 2957.46

5 2.4096 3757.37

6 3.0120 4600.71

The precision test

Get schisandrin reference substance solution (0.3012mg/ml) under the standard curve item, the accurate 10 μ l that draw inject chromatograph of liquid, measure 5 times, investigate the precision of reference substance solution.

The test of schisandrin reference substance precision

Test number (TN) 12345 average RSD (%)

Peak area 4692.34 4691.17 4698.78 4691.29 4705.46 4695.81 0.13

The result shows that schisandrin reference substance solution precision is good.

Stability test

Get schisandrin reference substance solution (0.3012mg/ml) under the standard curve item, the accurate 10 μ l that draw inject chromatograph of liquid, measure at 0,6,12,24,48 hour sample introduction respectively.

The reference substance solution stability test

Testing times (h) 06 12 24 48 average RSD (%)

Peak area 4692.34 4691.17 4698.78 4691.29 4705.46 4695.81 0.13

The result shows that the schisandrin reference substance solution has good stability.

The test agent schisandrin is measured in three batches

Three batch sample schisandrin assay results

Lot number 123

Schisandrin content (mg/ bottle) 0.227 0.236 0.255

Four, scutellarin

Instrument and reagent

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.

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

Scutellarin provides scutellarin by Nat'l Pharmaceutical ﹠ Biological Products Control Institute, and measuring purity through high performance liquid chromatography (normalization method) is 96.40%, meets 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 the selection of detection 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.。

Chromatographic condition

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.

Linear relationship 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

1 0.0000 0.0000 0

2 0.1956 0.1886 603722

3 0.3912 0.3771 1224078

4 0.5868 0.5657 1835374

5 0.7824 0.7542 2505678

6 0.9780 0.9428 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 precision experiment inject chromatograph of liquid, measure 5 times, investigate reference substance solution precision.

The precision test

Test number (TN) 12345 meansigma methods RSD (%)

Peak area 1,572,015 1,570,202 1,566,206 1,559,384 1,557,017 1,564,965 0.42

The result shows that reference substance solution precision is good.

Sample size is measured and to be 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)

1 4.07

2 3.91

3 4.22

Total lignans

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:

Schisandrin Nat'l Pharmaceutical ﹠ Biological Products Control Institute

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

Detect the selection of wavelength and get the schisandrin reference substance, operate, obtain reference substance solution by the preparation method of text reference substance solution.Get the breviscapus pulse-engendering injection, operate, obtain need testing solution by the preparation method of need testing solution in the text algoscopy.Draw the schisandrin reference substance solution, need testing solution, press the text total lignans and measure item method suggested down, in 200～400nm wave-length coverage, scan, the result shows, reference substance solution and need testing solution all have absorption maximum at 250nm, and solvent is noiseless, therefore select the detection wavelength of 250nm as total lignans content in the spectrophotometry breviscapus pulse-engendering injection.

The preparation precision of reference substance solution takes by weighing at the schisandrin reference substance 15mg of 60 ℃ of drying under reduced pressure to constant weight, put in the 50ml measuring bottle, add an amount of supersound process of methanol solution (power 250W, frequency 33KHz) makes dissolving, take out, put, add methanol solution to scale to room temperature, shake up, promptly get (containing schisandrin 0.3mg among every 1ml).

The preparation precision of standard curve is measured reference substance solution (C=0.2908mg/ml) 0.1ml, 0.2ml, 0.4ml, 0.6ml 0.8ml, 1.0ml split in the 10ml measuring bottle, add methanol to scale, shaking up, is blank with the retinue solvent, according to spectrophotography (an appendix V of Chinese Pharmacopoeia version in 2000 B), measure trap at 250nm wavelength place, with the trap is vertical coordinate, and concentration (μ g/ml) is figure for abscissa, the drawing standard curve.

Total lignans standard curve determination result

Numbering schisandrin concentration (μ g/ml) trap

1 2.908 0.118

2 5.816 0.245

3 11.632 0.491

4 17.448 0.732

5 23.264 0.971

6 29.080 1.217

Regression equation: Y=0.0418x+0.0003;

Correlation coefficient: γ=0.9999;

The result shows: schisandrin is good in 2.908～29.080 μ g/ml scope internal linear.

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

The accurate absorption of precision test schisandrin reference substance solution (concentration: 0.2908mg/ml) 0.6ml, put in the 10ml measuring bottle, press the method under the text algoscopy item, measure 5 times.

The precision test

Number 12345 RSD (%)

Trap 0.732 0.733 0.731 0.734 0.732 0.56

The result shows that reference substance solution precision is good.

The assay of sample 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.

The assay of total lignans

Lot number total lignans content (mg/ bottle)

1 3.26

2 3.05

3 3.22

The specific embodiment

Embodiment 1: finger printing

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

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

Embodiment 2: finger printing

(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler: mobile phase is the 67%:33% methanol-water, and flow velocity is that .0ml/min, evaporation photodetector detect, 30 ℃ of drift tube temperatures, and carrier gas flux 2.2L/min, column temperature are 30 ℃;

Embodiment 3: differentiate

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 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 200～410nm, 20～50 ℃ of column temperatures; 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;

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 the dissolving of ethyl acetate equal solvent, as need testing solution.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 1～30 μ l of above-mentioned four liquid, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica gel G F ₂₅₄On the 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 ℃～120 ℃ 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;

It is an amount of to get each preparation, extracts with n-butyl alcohol or ethanol equal solvent, filters, and filtrate is as need testing solution; Get Radix Ginseng Rubra or Radix Ginseng control medicinal material, add the chloroform reflux, extract,, discard chloroform solution, residue adds the water-saturated n-butanol equal solvent and extracts, and extracting solution adds ammonia solution, and divide and get the upper strata, evaporate to dryness, residue dissolves with the methanol equal solvent, in contrast medical material solution; Other gets the ginsenoside Rg ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf's reference substance, add dissolvings such as methanol respectively and make 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 7 kinds of solution, put respectively in same silica gel g thin-layer plate or silica gel H lamellae or silica gel G F ₂₅₄On the 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: 0.2～3010 ℃ with chloroform-ethyl acetate or Ethyl formate-methanol-water: 1～15 upper strata is developing solvent, launch, take out, dry, spray is with 5～50% sulphuric acid ethanol reagent or 50% sulphate reagent, 90 ℃～120 ℃ 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: differentiate

Get this product, use n-butanol extraction, filter, filtrate is as need testing solution; 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 the ginsenoside Rg ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf's reference substance, add dissolvings such as methanol respectively and make 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 7 kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-methanol-water 15: 40: 22: lower floor's solution of placing below 1010 ℃ was developing solvent, launch, take 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;

Embodiment 5: assay

The assay of scutellarin in a, the injection:

C. the assay of total saponins in the injection:

Total lignans assay in d, the injection:

Schisandrin, deoxyschizandrin, schisandrin B assay in e, the injection:

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

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

The assay of lobetyolin in h, the injection:

Embodiment 6: assay

The assay of scutellarin in a, the injection:

C. the assay of total saponins in the injection:

Total lignans assay in d, the injection

Schisandrin, deoxyschizandrin, schisandrin B assay in e, the injection:

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

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

The assay of lobetyolin in h, 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, 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.

Claims

1, a kind of method of quality control of the traditional medicine Injectio made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis, it is characterized in that: this method comprises following all or part of content:

(1) finger printing test, comprise based on Radix Ginseng Rubra or Radix Ginseng and Radix Ophiopogonis composition characteristics finger printing, based on all or part of test in the finger printing of Radix Codonopsis composition characteristics and the finger printing based on the Fructus Schisandrae Chinensis composition characteristics;

(3) ginsenoside Rg ₁, ginsenoside Rb ₁, ginsenoside Re, ginsenoside Rf, ophiopogonin B, ophiopogonin D, ophiopogonin D ', the content test method of all or part of composition such as Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, schisandrin, schisantherin B, schisantherin A, deoxyschizandrin, schisandrin B, schisandrin C, total saponins, total lignans, lobetyolin, atractylenoide.

2, according to the method for quality control of the described Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis of claim 1, it is characterized in that this method comprises one or more in the following finger printing:

3, according to the method for quality control of the described Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis of claim 2, it is characterized in that this method comprises one or more in the following finger printing:

4, according to the method for quality control of the described Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis of claim 1, it is characterized in that: the discrimination method of described injection comprises following all or part of content:

5, according to the method for quality control of the described Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis of claim 4, it is characterized in that: the discrimination method of described injection comprises following one or more:

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

The assay of scutellarin in a, the injection:

C. the assay of total saponins in the injection:

Total lignans assay in d, the injection:

Schisandrin, deoxyschizandrin, schisandrin B assay in e, the injection:

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

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

The assay of lobetyolin in h, the injection:

7, according to the method for quality control of the described Chinese medicine made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis of claim 6, it is characterized in that: the method for testing of described injection content comprise following one or more:

The assay of scutellarin in a, the injection:

C. the assay of total saponins in the injection:

Total lignans assay in d, the injection

Schisandrin, deoxyschizandrin, schisandrin B assay in e, the injection:

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

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

The assay of lobetyolin in h, the injection:

8, according to the method for quality control of claim 6 or the 7 described Chinese medicines made from breviscapine, Radix Ginseng Rubra or Radix Ginseng or Radix Codonopsis, Radix Ophiopogonis and Fructus Schisandrae Chinensis, it is characterized in that: the assay result of described injection, calculate scutellarin, ginsenoside Rg according to percentage by weight ₁, ginsenoside Rb ₁, ginsenoside Re, total saponins, total lignans, schisandrin, deoxyschizandrin, schisandrin B, ophiopogonin B, ophiopogonin D, ophiopogonin D ', the total content of all or part of material in the Ruscus aculeatus L. sapogenin, diosgenin, ruscogenin, lobetyolin, atractylenoide accounts for deduction adjuvant and outer more than 25% of total solid of moisture.