CN1739655B - Quality control method of Qingkailing injection preparation - Google Patents

Abstract

The quality control method for Qingkailing injection preparation includes character identification, inspection and content measurement. The character identification adopts silicone gel G thin-layer plate and ethyl acetate, acetone, formic acid and water as developing solvent. The jasminoidin content measurement has the contrast dissolved in methanol; the cholic acid and hyodesoxycholic acid content measurement is in efficient liquid phase chromatographic process with evaporating dispersion detector to obtain accurate result; and the nitrogen content, insoluble particle, etc. are also detected to avoid negative reaction. The quality control method for Qingkailing injection preparation can ensure the curative effect of the medicine preparation and reach effective control of the medicine preparation quality.

Description

The method of quality control of Qingkailing injection preparation

Technical field: the present invention relates to a kind of method of quality control of Qingkailing injection preparation, belong to the technical field of medicine being carried out quality control.

Technical background: Qingkailing preparation is got by the changing agent of China's famous and precious Chinese patent drug of tradition " Angong Niuhuang Wan ", is made up of cholic acid, mother-of-pearl, hyodesoxycholic acid, cape jasmine, cornu bubali, Radix Isatidis, scutelloside and honeysuckle; Have clearing heat and detoxicating, as to keep away dirty sensible, tranquilizing and allaying excitement function, cure mainly that heat evil is caved in, coma and delirium, irritated, the diseases such as frightened Jue and acute infantile convulsion wind of twitching of body heat; Modern medicine is mainly used in the treatment infection of the upper respiratory tract, multiple pyreticosis and acute disease etc., and it is evident in efficacy; Yu Wenyong has applied for that on April 10th, 2003 patent name is: each method of the system of qingkailing injections or injection powder injection and method of quality control thereof, application number is: 03109422.8 patent, this patent disclosure scutelloside, cholic acid, hyodesoxycholic acid, Gardenoside, chlorogenic acid contents assay method in qingkailing injections or the powder ampoule agent for injection, and the discrimination method of qingkailing injections or powder ampoule agent for injection.Though the method for quality control of this patent disclosure is played a role to the quality control of Qingkailing preparation, find that in our research process still there is certain problem in the quality in order to last method control Qingkailing preparation.Be embodied in: this patented claim is adopted the GF254 prefabricated board in the method for differentiating cape jasmine, the while, presss from both sides blue aldehyde-concentrated sulfuric acid solution with perfume (or spice) and makes developer as developping agent with ethyl acetate-absolute ethyl alcohol-glacial acetic acid; We discover that the specificity as a result of coming out with the discriminating of this method is strong inadequately, and the spot colour developing is good inadequately; And use the GF254 fluorescent plate generally speaking, be meant at ultraviolet lamp, observations under the 254nm condition, and this patented claim does not clearly illustrate this, so the discrimination method of Gardenoside also has unclear place in this patented claim; In the content assaying method of Gardenoside, the Gardenoside reference substance adopts the methanol-water dissolving, makes the Gardenoside solute effect good inadequately in addition, and gained Gardenoside reference substance solution is difficult for preserving; In this patented claim, adopt high performance liquid chromatography in the content assaying method of cholic acid and hyodesoxycholic acid in addition, use UV-detector, ultraviolet is terminal to be absorbed because cholic acid and hyodesoxycholic acid only have, and makes measurement result not accurate enough.

Summary of the invention: the objective of the invention is to: the method for quality control that a kind of Qingkailing injection preparation is provided.

This ejection preparation comprises freeze-dried powder, parenteral solution and great transfusion preparation.

Among the present invention, in the discrimination method of Gardenoside, we adopt silica gel g thin-layer plate, and as developping agent, the specificity of differentiating with this method as a result is strong with ethyl acetate-acetone-formic acid-water, and spot develops the color, and naked eyes are observed easily; In the content assaying method of Gardenoside, the Gardenoside reference substance adopts dissolve with methanol, and its solute effect is good, and gained Gardenoside reference substance solution is preserved easily; Among the present invention, adopt high performance liquid chromatography in the content assaying method of cholic acid and hyodesoxycholic acid, use evaporation light dispersion dispersion, its testing result is accurate, ultraviolet is terminal to be absorbed because cholic acid and hyodesoxycholic acid only have, use UV-detector, its measurement result is not accurate enough, and evaporation light dispersion dispersion is for no uv absorption or only there is the macromolecular organic compound of the terminal absorption of ultraviolet to have great superiority; In the product research process, we find: cholic acid and hyodesoxycholic acid adopt high performance liquid chromatography, and using evaporation light dispersion dispersion is to measure cholic acid and the advanced detection method of hyodesoxycholic acid at present; Owing to contain effective constituent amino acid in the Qingkailing preparation, so the present invention also carries out assay to contained nitrogen in the Qingkailing preparation, index as amino acid content in the control Qingkailing preparation, this is never to have in the prior art, we find that the used content assaying method of the present invention also has the advantage of favorable reproducibility in the product research process in addition.

Qingkailing injection preparation is the commercially available prod described in the present invention, comprising: freeze-dried powder, parenteral solution and great transfusion preparation.

Described method of quality control mainly comprises the part or all of of projects such as proterties, discriminating, inspection, assay; Wherein differentiate comprise furfurol reaction, with the Gardenoside be the thin layer of reference substance differentiate partly or entirely; Inspection comprise particulate matter, pH value, protein, tannin, resin, oxalates, heavy metal, arsenic salt, potassium ion, undue toxicity, haemolysis and cohesion, barium ion, residue on ignition inspection, moisture and other should meet in every regulation under the injection item partly or entirely; Assay comprises with Gardenoside reference substance, scutelloside reference substance, chlorogenic acid reference substance, cholic acid reference substance, the hyodesoxycholic acid reference substance detection index as the said preparation content assaying method, and nitrogen content measure partly or entirely.

Described method of quality control is:

Proterties:

For freeze-dried powder: product is light yellow to pale brown look loose block.

For parenteral solution: product is pale brown look or henna clear liquid.

For infusion solutions: product is yellow clear liquid to pale brown look.

Differentiate:

(1) get Qingkailing, parenteral solution or infusion solutions respectively, the dissolving of freeze-dried powder water adds furfural solution and sulfuric acid solution mixing, the water-bath heating, and solution shows gray purple.

(2) get Qingkailing, parenteral solution or infusion solutions, wherein the preparation method of freeze-dried powder need testing solution is: get the content of freeze-dried powder, add dissolve with ethanol, get supernatant promptly; The preparation method of parenteral solution or infusion solutions need testing solution is: get parenteral solution 5-20ml or infusion solutions 50-200ml, put evaporate to dryness in the water-bath, put coldly, residue adds ethanol makes dissolving, gets supernatant promptly.Other gets the Gardenoside reference substance, adds dissolve with ethanol, in contrast product solution.Reference substance solution and need testing solution are put respectively on same silica gel g thin-layer plate, with ethyl acetate or phosphoric acid: acetone: formic acid: water=2-10: 2-10: 0.52: 0.5-2 is a developping agent, with 10% ethanol solution of sulfuric acid is developer, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Check:

Particulate matter is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds the 50-100ml that purifies waste water makes dissolving, makes blank of purifying waste water, and measures according to Chinese Pharmacopoeia appendix of version in 2005 " particulate matter inspection technique " first method, should be up to specification.

PH value is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 1-10ml makes dissolving, surveys its pH value respectively, and its pH value all should be 5-9.

Protein is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 1-10ml makes dissolving, gets 1ml respectively and puts in the test tube, drips tannic acid test solution 1-5 and drips, and must not produce muddiness.

Tannin is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 1-10ml makes dissolving, gets 1ml, adds the physiological saline 1-10ml that contains the 1-5% Ovum Gallus domesticus album of new preparation, places 5-20 minute, muddiness or precipitation must not occur.

Resin is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 1-10ml makes dissolving, gets above-mentioned solution 5ml respectively, put in the separating funnel, add chloroform 5-20ml jolting and extract, divide and get chloroform solution, evaporate to dryness, residue adds glacial acetic acid 1-5ml makes dissolving, put in the tool plug test tube, add water 1-5ml, mixing, placed 20-50 minute, and should not have floccus and separate out.

Oxalates is got Qingkailing, parenteral solution or infusion solutions, freeze-dried powder 200mg adds water 1-10ml makes dissolving, get above-mentioned solution 5ml respectively, regulate pH value to 1～3 with watery hydrochloric acid, 30-70 ℃ is incubated 5-20 minute, insulation elimination precipitation, regulate pH value to 4～7, get 1-5ml, add 1～5 of 1-5% calcium chloride solution, placed 5-20 minute, and muddiness or precipitation must not occur.

Heavy metal is got Qingkailing, parenteral solution or infusion solutions, freeze-dried powder 200mg adds water 1-10ml makes dissolving, draw 1-2ml respectively, evaporate to dryness, slowly baking is to complete approximately ashing, puts coldly, and control tube is got standard lead solution 1ml, measure according to Chinese Pharmacopoeia appendix of version in 2005 " heavy metal inspection technique " second method, contain heavy metal in the 1ml aqueous solution and must not cross 10/1000000ths.

Arsenic salt is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 1-10ml makes dissolving, draws 1ml respectively, evaporate to dryness, add 1-5% magnesium nitrate ethanolic solution 1-5ml, light the afterburnt, burn with little heated earlier and make charing, again 500～600 ℃ blazing to fully ashing, put coldly, add hydrochloric acid 1-10ml and water 10-30ml makes dissolving, measure according to appendix of Chinese Pharmacopoeia version in 2005 " arsenic salt inspection technique " first method, arsenic content must not cross 2/1000000ths in the 1ml aqueous solution.

Potassium ion is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 1-10ml makes dissolving, draws 1-5ml respectively, and evaporate to dryness is measured according to appendix of Chinese Pharmacopoeia version in 2005 " injection related substance inspection technique ", should be up to specification.

Undue toxicity is got Qingkailing, parenteral solution or infusion solutions, freeze-dried powder adds the chlorination sodium injection and makes the solution that contains 30-50mg among every 1ml, check according to Chinese Pharmacopoeia two appendix of version in 2005 " undue toxicity inspection technique ", press the intravenous injection administration, should be up to specification.

Haemolysis is respectively got rabbit blood or sheep blood with the system of cohesion 2% red blood cell suspension, put into the conical flask that fills beaded glass, jolting 5-20 minute, remove fibrinogen, make to become to take off fine blood, add physiological sodium chloride solution washing 1-5 time that 5-20 doubly measures, till the apparent redness of supernatant, the gained red blood cell is made into 2% suspension with physiological sodium chloride solution, promptly.

Test method is got 6 in test tube, add 2% red blood cell suspension and physiological sodium chloride solution successively by proportional quantity in the last table, behind the mixing, in 37 ℃ of constant temperature ovens, placed 10-60 minute, the soup that adds different amounts respectively, wherein freeze-dried powder is got 200mg, with physiological sodium chloride solution dissolving and be diluted to 5-40ml, is control tube to be numbered 6 the 6th pipe; After shaking up, put in 37 ℃ of constant temperature ovens, beginning was observed 1 time every 15 minutes, after 1 hour, observed 1 time every 1 hour, observed altogether 2 hours.

Test method is got 6 in test tube, add 2% red blood cell suspension and physiological sodium chloride solution successively by proportional quantity in the last table, behind the mixing, in 37 ℃ of constant temperature ovens, placed 10-60 minute, the soup that adds different amounts respectively, wherein freeze-dried powder is got 200mg, with physiological sodium chloride solution dissolving and be diluted to 5-40ml, is control tube to be numbered 6 the 6th pipe; After shaking up, put in 37 ℃ of constant temperature ovens, beginning was observed 1 time every 15 minutes, after 1 hour, observed 1 time every 1 hour, observed altogether 2 hours.

By last method inspection, be as the criterion to be numbered 3 the 3rd pipe, this product haemolysis and red blood cell condensation must not occur in 2 hours.

Because barium ion is harmful, also barium ion is checked in the inspection item of the present invention that avoid it to enter human body, the inspection method of barium ion is:

Barium ion method 1: get platinum filament, with hydrochloric acid moistening after, dip in and get Qingkailing, parenteral solution or infusion solutions, in colourless flame, burn, flame shows yellow green, by the green glass perspective, it is blue that flame shows.

Method 2: get Qingkailing, parenteral solution or infusion solutions, freeze-dried powder 200mg dissolves with water for injection 1-5ml, and taking liquid drips dilute sulfuric acid respectively, promptly generates white precipitate, separates, and is deposited in all dissolvings in hydrochloric acid or the nitric acid.

The present invention causes bad reaction to produce for avoiding in Qingkailing injection preparation and the blood plasma calcium ion to react, and also can check following project:

Get Qingkailing, parenteral solution or infusion solutions, freeze-dried powder 200mg dissolves with water for injection 1-10ml water for injection, and taking liquid drips the CaCl that 1-50 drips 0.01-0.2% respectively ₂Solution or Ca (OH) ₂Solution must not become turbid or precipitate.

The present invention checks to residue on ignition also that for inorganics impurity content in the Qingkailing preparation is controlled its inspection method is:

Qingkailing, parenteral solution or infusion solutions are got in the residue on ignition inspection, and freeze-dried powder 200mg dissolves with 1-10ml water for injection, and taking liquid 1-5ml checks according to the bright residue inspection technique of Chinese Pharmacopoeia respectively, should be less than 2%.

The present invention is directed to freeze-dried powder in addition and also carry out the moisture inspection, its detection method is:

Moisture is got freeze-dried powder, measures according to a Chinese Pharmacopoeia appendix of version " aquametry " in 2005 three therapeutic methods of traditional Chinese medicine, must not cross 5.0%.

Other should meet relevant every regulation under an appendix injection of Chinese Pharmacopoeia version in 2005 item

Assay:

Gardenoside adopts high performance liquid chromatography, and chromatographic column is C18 or C4 or C8 post, and with methyl alcohol or acetonitrile: water=20～40: 80～60 is moving phase, and flow velocity is 0.6-1.2ml/min, and column temperature: 20-50 ℃, the detection wavelength is 205～350nm; Precision takes by weighing the Gardenoside reference substance, adds dissolve with methanol, shakes up, and promptly gets reference substance solution; Get Qingkailing and dissolve with methanol-water solution, filter membrane filters, and promptly gets need testing solution, or get qingkailing injections or QINGKAILING infusion solutions, and put evaporate to dryness in the water-bath earlier, put cold, residue dissolves with methanol-water solution, filter membrane filters, or with chloroform extraction, puts cold, the extract evaporate to dryness, residue adds the methanol-water dissolving, and filter membrane filters, and promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination content, in this Qingkailing preparation, Gardenoside content should be less than 0.8% in the freeze-dried powder, Gardenoside content should be less than that Gardenoside content should be less than 0.04mg/ml in 0.4mg/ml, the infusion solutions in the parenteral solution.

Scutelloside is according to high effective liquid chromatography for measuring, and chromatographic column is C18 or C4 or C8 post, and with acetonitrile or methyl alcohol: 0.3～0.5% phosphoric acid=10～40: 90～60 is moving phase, and flow velocity is 0.6-1.2ml/min, and column temperature: 20-50 ℃, the detection wavelength is 205～350nm; Precision takes by weighing the scutelloside reference substance, adds dissolve with methanol, shakes up, and promptly gets reference substance solution; Get Qingkailing and dissolve with methanol-water solution, filter membrane filters, and promptly gets need testing solution, or get qingkailing injections or QINGKAILING infusion solutions, and put evaporate to dryness in the water-bath earlier, put cold, residue dissolves with methanol-water solution, filter membrane filters, or extracts with the mixed solution of ethyl acetate and methyl alcohol, puts cold, the extract evaporate to dryness, residue adds the methanol-water dissolving, and filter membrane filters, and promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination content, in this Qingkailing preparation, content of baicalin should be 5%～17.5% in the freeze-dried powder, content of baicalin should be that content of baicalin should be 0.24～2.1mg/ml in 2～7mg/ml, the infusion solutions in the parenteral solution.

Chlorogenic acid is according to high effective liquid chromatography for measuring, and chromatographic column is C18 or C4 or C8 post, and with acetonitrile or methyl alcohol: 0.2～0.7% phosphoric acid=5～25: 95～75 is moving phase, and flow velocity is 0.6-1.2ml/min, and column temperature: 20-50 ℃, the detection wavelength is 245～600nm; Precision takes by weighing the chlorogenic acid reference substance, adds the methanol-water dissolving, shakes up, and promptly gets reference substance solution; Get Qingkailing, with the dissolving of methanol-water solution, filter membrane filters, and promptly get need testing solution, or gets qingkailing injections or infusion solutions, puts evaporate to dryness in the water-bath earlier, puts coldly, and residue is with the dissolving of methanol-water solution, and the filter membrane filtration promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination content, in this Qingkailing preparation, chlorogenic acid content should be less than 0.4% in the freeze-dried powder, chlorogenic acid content should be less than that chlorogenic acid content should be less than 0.018mg/ml in 0.15mg/ml, the infusion solutions in the parenteral solution.

Cholic acid is according to high effective liquid chromatography for measuring, chromatographic column is C18 or C4 or C8 post, moving phase is: methyl alcohol or acetonitrile: water=40-85: 60-15, or be methyl alcohol or acetonitrile: water: glacial acetic acid=40-85: 60-15: 0.01-0.99, or be methyl alcohol or acetonitrile: water: formic acid=40-85: 60-15: 0.01-0.5, flow rate of mobile phase is 0.6-1.2ml/min, column temperature: 20-50 ℃, the evaporative light-scattering detector drift tube temperature is 25-115 ℃, atomization gas is air or nitrogen, and gas flow rate is 1.0-4.0L/min; Get the cholic acid reference substance, add dissolve with methanol, shake up, promptly get reference substance solution; Get Qingkailing, add methanol-water solution sonicated 5-30 minute, put cold, add the dissolving of methanol-water solution, shake up, filter membrane filters, promptly get need testing solution, or get qingkailing injections or infusion solutions, with methanol-water solution sonicated 5-30 minute, put cold, add the dissolving of methanol-water solution, shake up, filter membrane filters, and promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination, press external standard two-point method logarithmic equation and calculate content, in this Qingkailing preparation, cholic acid content should be in 2.5%-10%, the parenteral solution cholic acid content and should be that cholic acid content should be 0.12-1.2mg/ml in 1-4mg/ml, the infusion solutions in the freeze-dried powder.

Hyodesoxycholic acid is according to high effective liquid chromatography for measuring, chromatographic column is C18 or C4 or C8 post, moving phase is: methyl alcohol or acetonitrile: water=40-85: 60-15, or be methyl alcohol or acetonitrile: water: glacial acetic acid=40-85: 60-15: 0.01-0.99, or be methyl alcohol or acetonitrile: water: formic acid=40-85: 60-15: 0.01-0.5, flow rate of mobile phase is 0.6-1.2ml/min, column temperature: 20-50 ℃, the evaporative light-scattering detector drift tube temperature is 25-115 ℃, atomization gas is air or nitrogen, and gas flow rate is 1.0-4.0L/min; Get the hyodesoxycholic acid reference substance, add dissolve with methanol, shake up, promptly get reference substance solution; Get Qingkailing, add methanol-water solution sonicated 5-30 minute, put cold, add the dissolving of methanol-water solution, shake up, filter membrane filters, promptly get need testing solution, or get qingkailing injections or infusion solutions, with methanol-water solution sonicated 5-30 minute, put cold, add the dissolving of methanol-water solution, shake up, filter membrane filters, and promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination, press external standard two-point method logarithmic equation and calculate content, in this Qingkailing preparation, hyodesoxycholic acid content should be in 2.5%-10%, the parenteral solution hyodesoxycholic acid content and should be that hyodesoxycholic acid content should be 0.12-1.2mg/ml in 1-4mg/ml, the infusion solutions in the freeze-dried powder.

The nitrogenous Qingkailing preparation of measuring is measured according to the Chinese Pharmacopoeia n2 method, and wherein the freeze-dried powder nitrogen content is that 4%-10%, parenteral solution nitrogen content are that 1.6-4mg/ml, infusion solutions nitrogen content are 0.18-1.2mg/ml.

Compared with the prior art, among the present invention, in the discrimination method of Gardenoside, we adopt silica gel g thin-layer plate, and as developping agent, its identification result specificity is strong with ethyl acetate-acetone-formic acid-water, and spot develops the color, and naked eyes are observed easily; In the content assaying method of Gardenoside, the Gardenoside reference substance adopts dissolve with methanol, and solute effect is good, and gained Gardenoside reference substance solution is preserved easily; In the product research process, find in addition, evaporation light dispersion dispersion has great superiority for the detection of cholic acid and hyodesoxycholic acid, be to detect cholic acid and the advanced detection method of hyodesoxycholic acid at present, so adopt high performance liquid chromatography in the content assaying method of cholic acid and hyodesoxycholic acid among the present invention, use evaporation light dispersion dispersion, its testing result is accurate; The present invention also carries out assay to contained nitrogen in the Qingkailing preparation, as the index of amino acid content in the control Qingkailing preparation; Simultaneously, the present invention also checks projects such as particulate matter, pH value, protein, tannin, resin, oxalates, heavy metal, arsenic salt, potassium ion, undue toxicity, haemolysis and the cohesion of Qingkailing injection preparation, barium ion, residue on ignition inspection, moisture, the content of impurity in the preparation in effective control QINGKAILING, avoid bad reaction to produce, so quality with the inventive method control Qingkailing preparation, not only the discrimination method specificity is strong, the content assaying method favorable reproducibility; Simultaneously, inspection method of the present invention is comprehensive, can effectively avoid adverse drug reaction to produce, and the assay project can be guaranteed the curative effect of medicine, has reached the goal of the invention of the quality of effective control medicine.

Specific embodiment:

The embodiment of the invention 1: the method for quality control of freeze-dried powder of the present invention is:

Proterties: product is light yellow to pale brown look loose block.

Differentiate: get commercially available Qingkailing 200mg and add water 10ml dissolving, get furfural solution (1 → 100) 2ml and sulfuric acid solution (get sulfuric acid 50ml, add water 50ml, mix) 10ml that 1ml adds new system, mixing heats in 60 ℃ of water-baths, and solution shows gray purple.

Check:

Particulate matter is got Qingkailing 200mg and is added the 50ml that purifies waste water and make dissolving, makes blank of purifying waste water, and measures according to Chinese Pharmacopoeia appendix of version in 2005 " particulate matter inspection technique " first method, should be up to specification.

PH value is got Qingkailing 200mg and is added water 10ml and make dissolving, surveys its pH value, and its pH value should be 5.0～8.0.

Protein is got Qingkailing 200mg and is added water 10ml and make dissolving, gets 1ml and puts in the test tube, drips tannic acid test solution 2-5 and drips, and must not produce muddiness.

Tannin is got Qingkailing 200mg and is added water 1ml and make dissolving, gets 1ml, adds the physiological saline 1ml that contains 5% Ovum Gallus domesticus album of new preparation, places 5 minutes, muddiness or precipitation must not occur.

Resin is got Qingkailing 200mg and is added water 1ml and make dissolving, gets above-mentioned solution 5ml, puts in the separating funnel, adds chloroform 5ml jolting and extracts, divide and get chloroform solution, evaporate to dryness, residue add glacial acetic acid 1ml makes dissolving, puts in the tool plug test tube, add water 1ml, mixing was placed 20 minutes, should not have floccus and separated out.

Oxalates is got Qingkailing 200mg and is added water 10ml and make dissolving, gets above-mentioned solution 5ml, regulates pH value to 1～3,30 ℃ insulation 20 minutes with watery hydrochloric acid, insulation elimination precipitation is regulated pH value to 4～6, gets 5ml, add 1～3 of 5% calcium chloride solution, placed 20 minutes, muddiness or precipitation must not occur.

Heavy metal is got Qingkailing 200mg and is added water 1ml and make dissolving, draw 1ml, evaporate to dryness, slowly baking is to complete approximately ashing, put cold, control tube is got standard lead solution 1ml, measures according to Chinese Pharmacopoeia appendix of version in 2005 " heavy metal inspection technique " second method, contains heavy metal in the 1ml aqueous solution and must not cross 10/1000000ths.

Arsenic salt is got Qingkailing 200mg and is added water 10ml and make dissolving, draw 1ml respectively, evaporate to dryness adds 1% magnesium nitrate ethanolic solution 5ml, light, in the afterburnt, burning with little heated earlier makes charing, blazing to ashing fully at 500～600 ℃ again, put cold, add hydrochloric acid 10ml and water 30ml makes dissolving, measure according to appendix of Chinese Pharmacopoeia version in 2005 " arsenic salt inspection technique " first method, arsenic content must not cross 2/1000000ths in the 1ml aqueous solution.

Undue toxicity is got Qingkailing and is added the chlorination sodium injection and make the solution that contains 50mg among every 1ml, checks according to Chinese Pharmacopoeia two appendix of version in 2005 " undue toxicity inspection technique ", presses the intravenous injection administration, should be up to specification.

Get Qingkailing 200mg and dissolve with water for injection 10ml water for injection, dropping 30-50 drips 0.01% CaCl ₂Solution must not become turbid or precipitate.

Moisture is got freeze-dried powder, measures according to a Chinese Pharmacopoeia appendix of version " aquametry " in 2005 three therapeutic methods of traditional Chinese medicine, must not cross 5.0%.

Other should meet relevant every regulation under an appendix injection of Chinese Pharmacopoeia version in 2005 item.

Assay:

Gardenoside is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C4 post, and with acetonitrile: water=20: 80 is moving phase, and flow velocity is 0.6ml/min, column temperature: 20 ℃, the detection wavelength is 350nm; Theoretical cam curve is calculated by the Gardenoside peak should be not less than 1500.

The preparation precision of reference substance solution takes by weighing the Gardenoside reference substance, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 30 μ g, shakes up, promptly.

Qingkailing 30mg is got in the preparation of need testing solution, adds 80% dissolve with methanol and is diluted to 5ml, shakes up, and filters with 0.45 μ m filter membrane, promptly.

Accurate reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and Gardenoside content should be less than 0.8% in the Qingkailing.

Chlorogenic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C8 post, and with methyl alcohol: 0.2% phosphoric acid=5: 95 is moving phase, and flow velocity is 0.6ml/min, column temperature: 50 ℃, the detection wavelength is 245nm; Theoretical cam curve is calculated by the chlorogenic acid peak should be not less than 1000.

The preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, adds 20% dissolve with methanol and makes the solution that every 1ml contains chlorogenic acid 10 μ g, shakes up, promptly.

Qingkailing 200mg is got in the preparation of need testing solution, adds 20% dissolve with methanol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, promptly.

Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and chlorogenic acid content should be less than 0.4% in the Qingkailing.

Scutelloside is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C4 post, and with methyl alcohol: 0.4% phosphoric acid=30: 70 is moving phase, and flow velocity is 0.8ml/min, column temperature: 35 ℃, the detection wavelength is 285nm; Theoretical cam curve is calculated by the scutelloside peak should be not less than 2500.

The preparation precision of reference substance solution takes by weighing the scutelloside reference substance, adds dissolve with methanol and makes the solution that every 1ml contains scutelloside 100 μ g, shakes up, promptly.

Qingkailing 15mg is got in the preparation of need testing solution, with 60% dissolve with methanol solution and be diluted to 20ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of determination method inject hplc determination, and content of baicalin should be 5%～17.5% in the Qingkailing.

Cholic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C4 post, acetonitrile: water: glacial acetic acid=85: 15: 0.99 is a moving phase; Flow velocity is 0.8ml/mln, column temperature: 40 ℃, the evaporative light-scattering detector drift tube temperature is 75 ℃, and atomization gas is a nitrogen, and gas flow rate is 3.0L/min, and theoretical cam curve is calculated by the cholic acid peak should be not less than 2000.

The cholic acid reference substance is got in the preparation of reference substance solution, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 0.2mg, shakes up, promptly.

Qingkailing 20mg is got in the preparation of need testing solution, adds 10% methanol solution 5ml sonicated 20 minutes, puts coldly, adds 70% methyl alcohol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution 10 μ l, 20 μ l and the need testing solution 10 μ l of drawing of determination method inject liquid chromatograph, measure, and press external standard two-point method logarithmic equation and calculate content, and cholic acid content should be 2.5%-10% in the Qingkailing.

The nitrogenous Qingkailing of measuring is measured according to a Chinese Pharmacopoeia appendix of version " nitrogen content " in 2005 second method, and the freeze-dried powder nitrogen content is 4%-10%.

Described Qingkailing is that Guizhou Yibai Pharmaceutical Co., Ltd produces.

The embodiment of the invention 2: the method for quality control of parenteral solution of the present invention is:

Proterties: product is pale brown look or henna clear liquid;

Differentiate: get qingkailing injections 1ml, add furfural solution (1 → 100) 1ml and sulfuric acid solution (get sulfuric acid 10ml, add water 15ml, the mix) 10ml of new system, mixing heats in 50 ℃ of water-baths, and solution shows gray purple.

Check:

PH value is got qingkailing injections, surveys its pH value, and its pH value should be 6.0～9.0.

Protein is got qingkailing injections 1ml and is put in the test tube, drips tannic acid test solution 1-3 and drips, and must not produce muddiness.

Tannin is got qingkailing injections 1ml, adds the physiological saline 7ml that contains 3% Ovum Gallus domesticus album of new preparation, places 15 minutes, muddiness or precipitation must not occur.

Oxalates is got qingkailing injections 5ml, regulates pH value to 2～3,70 ℃ insulation 5 minutes with watery hydrochloric acid, and insulation elimination precipitation is regulated pH value to 5～7, gets 1ml, adds 3～5 of 1% calcium chloride solutions, places 5 minutes, muddiness or precipitation must not occur.

Potassium ion is got qingkailing injections 1ml, and evaporate to dryness is measured according to appendix of Chinese Pharmacopoeia version in 2005 " injection related substance inspection technique ", should be up to specification.

Undue toxicity is got qingkailing injections, checks according to Chinese Pharmacopoeia two appendix of version in 2005 " undue toxicity inspection technique ", presses the intravenous injection administration, should be up to specification.

Get qingkailing injections 1ml, dropping 1-10 drips 0.2% Ca (OH) ₂Solution must not become turbid or precipitate.

Qingkailing injections 1ml is got in the residue on ignition inspection, measures according to Chinese Pharmacopoeia appendix of version in 2005 " residue on ignition inspection technique ", should be less than 2%.

Other should meet relevant every regulation under an appendix injection of Chinese Pharmacopoeia version in 2005 item.

Assay:

Gardenoside is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C18 post, and with acetonitrile: water=40: 60 is moving phase, and flow velocity is 1.2ml/min, column temperature: 50 ℃, the detection wavelength is 350nm; Theoretical cam curve is calculated by the Gardenoside peak should be not less than 1500.

The preparation precision of reference substance solution takes by weighing the Gardenoside reference substance, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 100 μ g, shakes up, promptly.

Qingkailing injections 20ml is got in the preparation of need testing solution, puts in the separating funnel, uses chloroform extraction three times, each 20ml, the combined chloroform extract is put evaporate to dryness in the water-bath, puts coldly, and residue adds 20% dissolve with methanol and is diluted to 10ml, shake up, filter with 0.45 μ m filter membrane, promptly.

Accurate reference substance solution and each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and Gardenoside content should be less than 0.4mg/ml in the qingkailing injections.

Scutelloside is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C4 post, and with methyl alcohol: 0.3% phosphoric acid=10: 90 is moving phase, and flow velocity is 0.6ml/min, column temperature: 20 ℃, the detection wavelength is 205nm; Theoretical cam curve is calculated by the scutelloside peak should be not less than 2500.

The preparation precision of reference substance solution takes by weighing the scutelloside reference substance, adds dissolve with methanol and makes the solution that every 1ml contains scutelloside 40 μ g, shakes up, promptly.

Qingkailing injections 20ml is got in the preparation of need testing solution, and with the mixed solution sonicated of ethyl acetate 20ml and methyl alcohol 5ml 10 minutes, extract evaporate to dryness, residue added 20% dissolve with methanol solution, and 0.45 μ m filter membrane filters, and promptly gets need testing solution;

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject hplc determination, and baicalin in Qingkailing Injection content should be 2～7mg/ml.

Chlorogenic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C4 post, and with acetonitrile: 0.5% phosphoric acid=25: 75 is moving phase, and flow velocity is 1.2ml/min, column temperature: 20 ℃, the detection wavelength is 600nm; Theoretical cam curve is calculated by the chlorogenic acid peak should be not less than 1500.

The preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, adds 70% dissolve with methanol and makes the solution that every 1ml contains chlorogenic acid 30 μ g, shakes up, promptly.

Qingkailing injections 5ml is got in the preparation of need testing solution, puts evaporate to dryness in the water-bath earlier, puts coldly, and residue adds 70% dissolve with methanol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, promptly.

Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and chlorogenic acid content should be less than 0.15mg/ml in the qingkailing injections.

Cholic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C4 post, methyl alcohol: water=75: 25 is moving phase; Flow velocity is 0.6ml/min, column temperature: 20 ℃, the evaporative light-scattering detector drift tube temperature is 25 ℃, and atomization gas is a nitrogen, and gas flow rate is 1.0L/min, and theoretical cam curve is calculated by the cholic acid peak should be not less than 2000.

The cholic acid reference substance is got in the preparation of reference substance solution, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 1mg, shakes up, promptly.

Qingkailing injections 5ml is got in the preparation of need testing solution, adds 20% methanol solution 5ml sonicated 15 minutes, puts coldly, adds 80% methyl alcohol and is diluted to 10ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution 5 μ l, l0 μ l and the need testing solution 5 μ l of drawing of determination method inject liquid chromatograph, measure, and press external standard two-point method logarithmic equation and calculate content, and cholic acid content should be 1-4mg/ml in the qingkailing injections.

Hyodesoxycholic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C4 post, acetonitrile: water=85: 15 is moving phase; Flow velocity is 0.6ml/min, column temperature: 20 ℃, the evaporative light-scattering detector drift tube temperature is 25 ℃, and atomization gas is an air, and gas flow rate is 4.0L/min, and theoretical cam curve is calculated by the cholic acid peak should be not less than 2000.

The hyodesoxycholic acid reference substance is got in the preparation of reference substance solution, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 2mg, shakes up, promptly.

Qingkailing injections 2ml is got in the preparation of need testing solution, adds 10% methanol solution 5ml sonicated 15 minutes, puts coldly, adds 70% methyl alcohol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution 5 μ l, 10 μ l and the need testing solution l of drawing of determination method injects liquid chromatograph, measures, and presses external standard two-point method logarithmic equation and calculates content, and hyodesoxycholic acid content should be 1-4mg/ml in the qingkailing injections.

Described qingkailing injections is that Shijiazhuang martial prowess medicine company is produced.

The embodiment of the invention 3: the method for quality control of infusion solutions of the present invention is:

Proterties: product is yellow clear liquid to pale brown look.

Differentiate: get infusion solutions 200ml, put evaporate to dryness in the water-bath, put coldly, residue adds ethanol 2.5ml makes dissolving, gets supernatant and promptly gets need testing solution; Other gets the Gardenoside reference substance, adds ethanol and makes the solution that every lml contains 10mg, in contrast product solution; Draw each 10 μ l of reference substance solution and need testing solution, put respectively on same silica gel g thin-layer plate, with phosphoric acid-acetone-formic acid-water=10: 2: 0.5: 2 be developping agent, launches, and takes out, and dries, and spray was dried by the fire about 15 minutes at 105 ℃ with 10% ethanol solution of sulfuric acid.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Check:

Particulate matter is got infusion solutions, calculates a method according to Chinese Pharmacopoeia appendix of version in 2005 " particulate matter inspection technique " and measures, should be up to specification.

Resin is got QINGKAILING infusion solutions 5ml, puts in the separating funnel, adds chloroform 10ml jolting and extracts, and divides and gets chloroform solution, and evaporate to dryness, residue add glacial acetic acid 3ml makes dissolving, puts in the tool plug test tube, adds water 4ml, and mixing was placed 40 minutes, should not have floccus and separated out.

Oxalates is got QINGKAILING infusion solutions 5ml, regulates pH value to 1～2,50 ℃ insulation 10 minutes with watery hydrochloric acid, and insulation elimination precipitation is regulated pH value to 5～6, gets 2ml, adds 2～3 of 3% calcium chloride solutions, places 10 minutes, muddiness or precipitation must not occur.

Heavy metal is got QINGKAILING infusion solutions 2ml, evaporate to dryness, and slowly baking is to complete approximately ashing, put cold, control tube is got standard lead solution 1ml, measures according to Chinese Pharmacopoeia appendix of version in 2005 " heavy metal inspection technique " second method, contains heavy metal in the 1ml aqueous solution and must not cross 10/1000000ths.

Arsenic salt is got QINGKAILING infusion solutions 1ml, evaporate to dryness, add 5% magnesium nitrate ethanolic solution 1ml, light the afterburnt, burn with little heated earlier and make charing, again 500～600 ℃ blazing to fully ashing, put coldly, add hydrochloric acid 1ml and water 10ml makes dissolving, measure according to appendix of Chinese Pharmacopoeia version in 2005 " arsenic salt inspection technique " first method, arsenic content must not cross 2/1000000ths in the 1ml aqueous solution.

Haemolysis and the system of cohesion 2% red blood cell suspension are respectively got rabbit blood or sheep blood number milliliter, put into the conical flask that fills beaded glass, jolting 5 minutes, remove fibrinogen, make to become to take off fine blood, the physiological sodium chloride solution that adds 5 times of amounts washs 1～3 time, till the apparent redness of supernatant, the gained red blood cell is made into 2% suspension with physiological sodium chloride solution, promptly.

Test method is got 6 in test tube, adds 2% red blood cell suspension and physiological sodium chloride solution successively by proportional quantity in the last table, behind the mixing, places 10 minutes in 37 ℃ of constant temperature ovens, adds the soup of different amounts respectively, and the 6th pipe is control tube; After shaking up, put in 37 ℃ of constant temperature ovens, beginning was observed 1 time every 15 minutes, after 1 hour, observed 1 time every 1 hour, observed altogether 2 hours.

By last method inspection, be as the criterion with the 3rd pipe, this product haemolysis and red blood cell condensation must not occur in 2 hours.

Barium ion is got platinum filament, with hydrochloric acid moistening after, dip in and get the QINGKAILING infusion solutions, in colourless flame, burn, flame shows yellow green, by the green glass perspective, it is blue that flame shows.

Get QINGKAILING infusion solutions 10ml, dropping 10-30 drips 0.15% CaCl ₂Solution must not become turbid or precipitate.

Other should meet relevant every regulation under an appendix injection of Chinese Pharmacopoeia version in 2005 item.

Assay:

Chlorogenic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C18 post, and with methyl alcohol: 0.7% phosphoric acid=20: 80 is moving phase, and flow velocity is 1.0ml/min, column temperature: 40 ℃, the detection wavelength is 500nm; Theoretical cam curve is calculated by the chlorogenic acid peak should be not less than 1000.

The preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, adds 85% dissolve with methanol and makes the solution that every 1ml contains chlorogenic acid 50 μ g, shakes up, promptly.

The system of need testing solution is respectively got QINGKAILING infusion solutions 20ml, puts evaporate to dryness in the water-bath earlier, puts coldly, and residue adds 50% dissolve with methanol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, promptly.

Accurate respectively reference substance solution and each the 20 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and chlorogenic acid content should be less than 0.018mg/ml in the QINGKAILING infusion solutions.

Hyodesoxycholic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C8 post, methyl alcohol: water: formic acid=40: 60: 0.01 is moving phase; Flow velocity is 1.2ml/min, column temperature: 50 ℃, the evaporative light-scattering detector drift tube temperature is 115 ℃, and atomization gas is a nitrogen, and gas flow rate is 1.0L/min, and theoretical cam curve is calculated by the cholic acid peak should be not less than 2000.

The hyodesoxycholic acid reference substance is got in the preparation of reference substance solution, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 1mg, shakes up, promptly.

QINGKAILING infusion solutions 20ml is got in the preparation of need testing solution, adds 3% methanol solution 10ml sonicated 5 minutes, puts coldly, adds 2096 methyl alcohol and is diluted to 10ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution 5 μ l, 10 μ l and the need testing solution 5 μ l of drawing of determination method inject liquid chromatograph, measure, and press external standard two-point method logarithmic equation and calculate content, and hyodesoxycholic acid content should be 0.12-1.2mg/ml in the QINGKAILING infusion solutions.

The nitrogenous QINGKAILING infusion solutions of measuring is measured according to a Chinese Pharmacopoeia appendix of version " nitrogen content " in 2005 second method, and the infusion solutions nitrogen content is 0.18-1.2mg/ml.

Described QINGKAILING infusion solutions is according to the patent of having applied for, name is called: the preparation method of Qingkailing freeze-drying powder preparation, application number: 200510003126.5 described method preparations.

The embodiment of the invention 4: the method for quality control of freeze-dried powder of the present invention is:

Proterties: product is light yellow to pale brown look loose block.

Differentiate: get the content 200mg of freeze-dried powder, add ethanol 5ml and make dissolving, get supernatant and promptly get need testing solution; Other gets the Gardenoside reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Draw each 10 μ l of reference substance solution and need testing solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water=2: 10: 1: 0.5 was developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, about 10 minutes of 105 ℃ of bakings.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Check:

Particulate matter is got Qingkailing 200mg and is added the 100ml that purifies waste water and make dissolving, makes blank of purifying waste water, and measures according to Chinese Pharmacopoeia appendix of version in 2005 " particulate matter inspection technique " first method, should be up to specification.

Protein is got Qingkailing 200mg and is added water 1ml and make dissolving, gets 1ml and puts in the test tube, drips tannic acid test solution 1-5 and drips, and must not produce muddiness.

Tannin is got Qingkailing 200mg and is added water 10ml and make dissolving, gets 1ml, adds the physiological saline 10ml that contains 2% Ovum Gallus domesticus album of new preparation, places 20 minutes, muddiness or precipitation must not occur.

Resin is got Qingkailing 200mg and is added water 10ml and make dissolving, gets above-mentioned solution 5ml respectively, puts in the separating funnel, adds chloroform 20ml jolting and extracts, divide and get chloroform solution, evaporate to dryness, residue add glacial acetic acid 5ml makes dissolving, puts in the tool plug test tube, add water 5ml, mixing was placed 50 minutes, should not have floccus and separated out.

Heavy metal is got Qingkailing 200mg and is added water 10ml and make dissolving, draw 1ml respectively, evaporate to dryness, slowly baking is to complete approximately ashing, put cold, control tube is got standard lead solution 1ml, measures according to Chinese Pharmacopoeia appendix of version in 2005 " heavy metal inspection technique " second method, contains heavy metal in the 1ml aqueous solution and must not cross 10/1000000ths.

Potassium ion is got Qingkailing 200mg and is added water 10ml and make dissolving, draws 5ml respectively, and evaporate to dryness is measured according to appendix of Chinese Pharmacopoeia version in 2005 " injection related substance inspection technique ", should be up to specification.

Haemolysis and the system of cohesion 2% red blood cell suspension are respectively got rabbit blood or sheep blood number milliliter, put into the conical flask that fills beaded glass, jolting 20 minutes, remove fibrinogen, make to become to take off fine blood, the physiological sodium chloride solution that adds 20 times of amounts washs 3～5 times, till the apparent redness of supernatant, the gained red blood cell is made into 2% suspension with physiological sodium chloride solution, promptly.

Test method is got 6 in test tube, add 2% red blood cell suspension and physiological sodium chloride solution successively by proportional quantity in the last table, behind the mixing, in 37 ℃ of constant temperature ovens, placed 60 minutes, the soup that adds different amounts respectively, wherein freeze-dried powder is got 200mg, and with physiological sodium chloride solution dissolving and be diluted to 40ml, the 6th pipe is control tube; After shaking up, put in 37 ℃ of constant temperature ovens, beginning was observed 1 time every 15 minutes, after 1 hour, observed 1 time every 1 hour, observed altogether 2 hours.

By last method inspection, be as the criterion with the 3rd pipe, this product haemolysis and red blood cell condensation must not occur in 2 hours.

Barium ion is got Qingkailing 200mg and is dissolved with water for injection 1ml, and taking liquid drips dilute sulfuric acid, promptly generates white precipitate, separates, and is deposited in all dissolvings in hydrochloric acid or the nitric acid.

The residue on ignition inspection is got Qingkailing 200mg and is dissolved with 10ml water for injection, and taking liquid 5ml measures according to Chinese Pharmacopoeia appendix of version in 2005 " residue on ignition inspection technique " respectively, should be less than 2%.

Moisture is got freeze-dried powder, measures according to a Chinese Pharmacopoeia appendix of version " aquametry " in 2005 three therapeutic methods of traditional Chinese medicine, must not cross 5.0%.

Other should meet relevant every regulation under an appendix injection of Chinese Pharmacopoeia version in 2005 item.

Assay:

Gardenoside is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C8 post, and with methyl alcohol: water=25: 75 is moving phase, and flow velocity is 0.8ml/min, column temperature: 30 ℃, the detection wavelength is 300nm; Theoretical cam curve is calculated by the Gardenoside peak should be not less than 1500.

Each precision of the system of reference substance solution takes by weighing the Gardenoside reference substance, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 80 μ g, shakes up, promptly.

Qingkailing 50mg is got in the preparation of need testing solution, adds 40% dissolve with methanol and is diluted to 5ml, shakes up, and filters with 0.45 μ m filter membrane, promptly.

Accurate reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and Gardenoside content should be less than 0.8% in the Qingkailing.

Scutelloside is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C8 post, and with methyl alcohol: 0.5% phosphoric acid=40: 60 is moving phase, and flow velocity is 1.2ml/min, column temperature: 50 ℃, the detection wavelength is 350nm; Theoretical cam curve is calculated by the scutelloside peak should be not less than 2500.

The preparation precision of reference substance solution takes by weighing the scutelloside reference substance, adds dissolve with methanol and makes the solution that every 1ml contains scutelloside 100 μ g, shakes up, promptly.

Qingkailing 50mg is got in the preparation of need testing solution, with 80% dissolve with methanol solution and be diluted to 50ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject hplc determination, and content of baicalin should be 5%～17.5% in the Qingkailing.

Cholic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C8 post, methyl alcohol: water: formic acid=75: 25: 0.01 is moving phase; Flow velocity is 1.2ml/min, column temperature: 50 ℃, the evaporative light-scattering detector drift tube temperature is 115 ℃, and atomization gas is an air, and gas flow rate is 4.0L/min, and theoretical cam curve is calculated by the cholic acid peak should be not less than 2000.

The cholic acid reference substance is got in the preparation of reference substance solution, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 0.3mg, shakes up, promptly.

Qingkailing 100mg is got in the preparation of need testing solution, adds 50% methanol solution 10ml sonicated 5min, puts coldly, adds 50% methyl alcohol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution 5 μ l, 10 μ l and the need testing solution 5 μ l of drawing of determination method inject liquid chromatograph, measure, and press external standard two-point method logarithmic equation and calculate content, and cholic acid content should be 2.5%-10% in the Qingkailing.

Described Qingkailing is according to the patent of having applied for, name is called: the preparation method of Qingkailing freeze-drying powder preparation, application number: 200510003126.5 described method productions.

The embodiment of the invention 5: the method for quality control of described Qingkailing injection preparation is:

Proterties:

For freeze-dried powder: product is light yellow to pale brown look loose block.

For parenteral solution: product is pale brown look or henna clear liquid.

For infusion solutions: product is yellow clear liquid to pale brown look.

Differentiate:

(1) get Qingkailing, parenteral solution or infusion solutions respectively, freeze-dried powder 200mg adds water 5ml dissolving, gets furfural solution (1 → 100) 1ml and the sulfuric acid solution that 0.5ml adds new system and (gets sulfuric acid 50ml, add water 65ml, mix) 10ml, mixing, heat in 70 ℃ of water-baths, solution shows gray purple.

(2) get Qingkailing, parenteral solution or infusion solutions, wherein the preparation method of freeze-dried powder need testing solution is: get the content 400mg of freeze-dried powder, add ethanol 2.5ml and make dissolving, get supernatant promptly; The preparation method of parenteral solution or infusion solutions need testing solution is: get parenteral solution 5ml or infusion solutions 50ml, put evaporate to dryness in the water-bath, put coldly, residue adds ethanol 1.25ml makes dissolving, gets supernatant promptly.Other gets the Gardenoside reference substance, adds ethanol and makes the solution that every 1ml contains 4mg, in contrast product solution.Draw each 5 μ l of reference substance solution and need testing solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water=5: 5: 1: 1 be developping agent, launches, and takes out, and dries, and spray was dried by the fire about 10 minutes at 105 ℃ with 10% ethanol solution of sulfuric acid.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Check:

Particulate matter is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds the 80ml that purifies waste water makes dissolving, makes blank of purifying waste water, and measures according to Chinese Pharmacopoeia appendix of version in 2005 " particulate matter inspection technique " first method, should be up to specification.

PH value is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 5ml makes dissolving, surveys its pH value respectively, and its pH value all should be 6.5～8.0.

Protein is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 5ml makes dissolving, gets 1ml respectively and puts in the test tube, drips tannic acid test solution 1-3 and drips, and must not produce muddiness.

Tannin is got QINGKAILING and is frozen in powder, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 5ml makes dissolving, gets 1ml, adds the physiological saline 5ml that contains 1% Ovum Gallus domesticus album of new preparation, places 10 minutes, muddiness or precipitation must not occur.

Resin is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 5ml makes dissolving, gets above-mentioned solution 5ml respectively, put in the separating funnel, add chloroform 10ml jolting and extract, divide and get chloroform solution, evaporate to dryness, residue adds glacial acetic acid 2ml makes dissolving, put in the tool plug test tube, add water 3ml, mixing, placed 30 minutes, and should not have floccus and separate out.

Oxalates is got Qingkailing, parenteral solution or infusion solutions, freeze-dried powder 200mg adds water 5ml makes dissolving, get above-mentioned solution 5ml respectively, regulate pH value to 1～2,50 ℃ insulation 10 minutes with watery hydrochloric acid, insulation elimination precipitation, regulate pH value to 5～6, get 2ml, add 2～3 of 3% calcium chloride solutions, placed 10 minutes, and muddiness or precipitation must not occur.

Heavy metal is got Qingkailing, parenteral solution or infusion solutions, freeze-dried powder 200mg adds water 5ml makes dissolving, draw 1ml respectively, evaporate to dryness, slowly baking is to complete approximately ashing, puts coldly, and control tube is got standard lead solution 1ml, measure according to Chinese Pharmacopoeia appendix of version in 2005 " heavy metal inspection technique " second method, contain heavy metal in the 1ml aqueous solution and must not cross 10/1000000ths.

Arsenic salt is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 5ml makes dissolving, draws 1ml respectively, evaporate to dryness, add 2% magnesium nitrate ethanolic solution 3ml, light the afterburnt, burn with little heated earlier and make charing, again 500～600 ℃ blazing to fully ashing, put coldly, add hydrochloric acid 5ml and water 21ml makes dissolving, measure according to appendix of Chinese Pharmacopoeia version in 2005 " arsenic salt inspection technique " first method, arsenic content must not cross 2/1000000ths in the 1ml aqueous solution.

Potassium ion is got Qingkailing, parenteral solution or infusion solutions, and freeze-dried powder 200mg adds water 5ml makes dissolving, draws 2ml respectively, and evaporate to dryness is measured according to appendix of Chinese Pharmacopoeia version in 2005 " injection related substance inspection technique ", should be up to specification.

Undue toxicity is got Qingkailing, parenteral solution or infusion solutions, freeze-dried powder adds the chlorination sodium injection and makes the solution that contains 40mg among every 1ml, check according to Chinese Pharmacopoeia two appendix of version in 2005 " undue toxicity inspection technique ", press the intravenous injection administration, should be up to specification.

Rabbit blood or sheep blood number milliliter are got in haemolysis and the preparation of cohesion 2% red blood cell suspension, put into the conical flask 212 that fills beaded glass, jolting 10 minutes, remove fibrinogen, make to become to take off fine blood, the physiological sodium chloride solution that adds about 10 times of amounts washs 2～3 times, till the apparent redness of supernatant, the gained red blood cell is made into 2% suspension with physiological sodium chloride solution, promptly.

Test method is got 6 in test tube, add 2% red blood cell suspension and physiological sodium chloride solution successively by proportional quantity in the last table, behind the mixing, in 37 ℃ of constant temperature ovens, placed 30 minutes, the soup that adds different amounts respectively, wherein freeze-dried powder is got 200mg, and with physiological sodium chloride solution dissolving and be diluted to 20ml, the 6th pipe is control tube; After shaking up, put in 37 ℃ of constant temperature ovens, beginning was observed 1 time every 15 minutes, after 1 hour, observed 1 time every 1 hour, observed altogether 2 hours.

By last method inspection, be as the criterion with the 3rd pipe, this product haemolysis and red blood cell condensation must not occur in 2 hours.

Barium ion is got platinum filament, with hydrochloric acid moistening after, dip in and get Qingkailing, parenteral solution or infusion solutions, in colourless flame, burn, flame shows yellow green, by the green glass perspective, it is blue that flame shows.

Get Qingkailing, parenteral solution or infusion solutions, freeze-dried powder 200mg is with the dissolving of water for injection 5ml water for injection, taking liquid respectively, the CaCl of Dropwise 5-10 droplet 0.1% ₂Solution or Ca (OH) ₂Solution must not become turbid or precipitate.

Qingkailing, parenteral solution or infusion solutions are got in the residue on ignition inspection, and freeze-dried powder 200mg dissolves with 5ml water for injection, and taking liquid 2ml measures according to Chinese Pharmacopoeia appendix of version in 2005 " residue on ignition inspection technique " respectively, should be less than 2%.

Moisture is got freeze-dried powder, measures according to a Chinese Pharmacopoeia appendix of version " aquametry " in 2005 three therapeutic methods of traditional Chinese medicine, must not cross 5.0%.

Other should meet relevant every regulation under an appendix injection of Chinese Pharmacopoeia version in 2005 item.

Assay:

Gardenoside is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C18 post, and with methyl alcohol: water=30: 70 is moving phase, and flow velocity is 1ml/min, column temperature: 40 ℃, the detection wavelength is 237nm; Theoretical cam curve is calculated by the Gardenoside peak should be not less than 1500.

The preparation precision of reference substance solution takes by weighing the Gardenoside reference substance, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 60 μ g, shakes up, promptly.

Qingkailing 30mg is got in the preparation of need testing solution, adds 50% dissolve with methanol and is diluted to 5ml, shakes up, and filters with 0.45 μ m filter membrane, promptly; Or get qingkailing injections 0.75ml or QINGKAILING infusion solutions 2.5ml, and put evaporate to dryness in the water-bath earlier, to put coldly, residue adds 50% dissolve with methanol and is diluted to 5ml, shakes up, and filters with 0.45 μ m filter membrane, promptly.

Accurate reference substance solution and each the 10 μ l of need testing solution of drawing of determination method, inject liquid chromatograph, measure, in this Qingkailing preparation, Gardenoside content should be less than 0.8% in the freeze-dried powder, Gardenoside content should be less than that Gardenoside content should be less than 0.04mg/ml in 0.4mg/ml, the infusion solutions in the parenteral solution.

Scutelloside is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C18 post, and with acetonitrile: 0.4% phosphoric acid=26: 74 is moving phase, and flow velocity is 1ml/min, column temperature: 40 ℃, the detection wavelength is 277nm; Theoretical cam curve is calculated by the scutelloside peak should be not less than 2500.

The preparation precision of reference substance solution takes by weighing the scutelloside reference substance, adds dissolve with methanol and makes the solution that every 1ml contains scutelloside 60 μ g, shakes up, promptly.

The system of need testing solution is respectively got Qingkailing 12.5mg, with 50% dissolve with methanol solution and be diluted to 25ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution; Or get qingkailing injections 1.25ml or QINGKAILING infusion solutions 4ml, and put evaporate to dryness in the water-bath earlier, to put coldly, residue is with 50% dissolve with methanol solution and be diluted to 100ml, shakes up, and 0.45 μ m filter membrane filters, promptly.

Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of determination method, inject hplc determination, in this Qingkailing preparation, content of baicalin should be 5%～17.5% in the freeze-dried powder, content of baicalin should be that content of baicalin should be 0.24～2.1mg/ml in 2～7mg/ml, the infusion solutions in the parenteral solution.

Chlorogenic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C18 post, and with acetonitrile: 0.4% phosphoric acid=10: 90 is moving phase, and flow velocity is 1.0ml/min, column temperature: 40 ℃, the detection wavelength is 327nm; Theoretical cam curve is calculated by the chlorogenic acid peak should be not less than 1000.

The preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, adds 50% dissolve with methanol and makes the solution that every 1ml contains chlorogenic acid 20 μ g, shakes up, promptly.

The system of need testing solution is respectively got Qingkailing 150mg, adds 50% dissolve with methanol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, promptly; Or get qingkailing injections 3.75ml or infusion solutions 12.5ml, and put evaporate to dryness in the water-bath earlier, to put coldly, residue adds 50% dissolve with methanol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, promptly.

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method, inject liquid chromatograph, measure, in this Qingkailing preparation, chlorogenic acid content should be less than 0.4% in the freeze-dried powder, chlorogenic acid content should be less than that chlorogenic acid content should be less than 0.018mg/ml in 0.15mg/ml, the infusion solutions in the parenteral solution.

Cholic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C18 post, methyl alcohol: water: glacial acetic acid=80: 20: 0.01 is a moving phase; Flow velocity is 1.0ml/min, column temperature: 40 ℃, the evaporative light-scattering detector drift tube temperature is 105 ℃, and atomization gas is an air, and gas flow rate is 2.2L/min, and theoretical cam curve is calculated by the cholic acid peak should be not less than 2000.

The cholic acid reference substance is got in the preparation of reference substance solution, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 0.5mg, shakes up, promptly.

Qingkailing 50mg is got in the preparation of need testing solution, adds 5% methanol solution 3ml sonicated 10 minutes, puts coldly, adds 50% methyl alcohol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution; Or get qingkailing injections 1.25ml or infusion solutions 10ml, and add 5% methanol solution 3ml sonicated 10 minutes, put coldly, add 50% methyl alcohol and be diluted to 5ml, shake up, 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution 5 μ l, 10 μ l and the need testing solution 5 μ l of drawing of determination method, inject liquid chromatograph, measure, press external standard two-point method logarithmic equation and calculate content, in this Qingkailing preparation, cholic acid content should be in 2.5%-10%, the parenteral solution cholic acid content and should be that cholic acid content should be 0.12-1.2mg/ml in 1-4mg/ml, the infusion solutions in the freeze-dried powder.

Hyodesoxycholic acid is according to high effective liquid chromatography for measuring.

Chromatographic condition and system suitability test chromatographic column is the C18 post, methyl alcohol: water: glacial acetic acid=80: 20: 0.01 is a moving phase; Flow velocity is 1.0ml/min, column temperature: 40 ℃, the evaporative light-scattering detector drift tube temperature is 105 ℃, and atomization gas is an air, and gas flow rate is 2.2L/min, and theoretical cam curve is calculated by the cholic acid peak should be not less than 2000.

The hyodesoxycholic acid reference substance is got in the preparation of reference substance solution, adds dissolve with methanol and makes the reference substance solution that every 1ml contains 0.5mg, shakes up, promptly.

Qingkailing 50mg is got in the preparation of need testing solution, adds 5% methanol solution 3ml sonicated 10 minutes, puts coldly, adds 50% methyl alcohol and is diluted to 5ml, shakes up, and 0.45 μ m filter membrane filters, and promptly gets need testing solution; Or get qingkailing injections 1.25ml or infusion solutions 10ml, and add 5% methanol solution 3ml sonicated 10 minutes, put coldly, add 50% methyl alcohol and be diluted to 5ml, shake up, 0.45 μ m filter membrane filters, and promptly gets need testing solution.

Accurate respectively reference substance solution 5 μ l, 10 μ l and the need testing solution 5 μ l of drawing of determination method, inject liquid chromatograph, measure, press external standard two-point method logarithmic equation and calculate content, in this Qingkailing preparation, hyodesoxycholic acid content should be in 2.5%-10%, the parenteral solution hyodesoxycholic acid content and should be that hyodesoxycholic acid content should be 0.12-1.2mg/ml in 1-4mg/ml, the infusion solutions in the freeze-dried powder.

Nitrogenous Qingkailing, parenteral solution or the infusion solutions of measuring, measure according to a Chinese Pharmacopoeia appendix of version " nitrogen content " in 2005 second method, wherein the freeze-dried powder nitrogen content is that 4%-10%, parenteral solution nitrogen content are that 1.6-4mg/ml, infusion solutions nitrogen content are 0.18-1.2mg/ml.

Described Qingkailing preparation is according to the patent of having applied for, name is called: the preparation method of Qingkailing freeze-drying powder preparation, application number: 200510003126.5 described method preparations.

Claims (5)

1. the detection method of a Qingkailing injection preparation, described ejection preparation is freeze-dried powder, parenteral solution or infusion solutions, described detection method is proterties, discriminating, inspection and assay; Wherein discriminating project is: furfurol reaction, be that the thin layer of reference substance is differentiated with the Gardenoside; Inspection item is: particulate matter, pH value, protein, tannin, resin, oxalates, heavy metal, arsenic salt, potassium ion, undue toxicity, haemolysis and cohesion, barium ion, residue on ignition inspection, moisture and other should meet the every regulation under the injection item: assay is: with Gardenoside reference substance, scutelloside reference substance, chlorogenic acid reference substance, cholic acid reference substance, the hyodesoxycholic acid reference substance detection index as the said preparation content assaying method, and nitrogen content is measured; It is characterized in that: content assaying method is:

Gardenoside adopts high performance liquid chromatography, and chromatographic column is C18 or C4 or C8 post, and with methyl alcohol or acetonitrile: water=20～40: 80～60 is moving phase, and flow velocity is 0.6-1.2ml/min, and column temperature: 20-50 ℃, the detection wavelength is 205～350nm; Precision takes by weighing the Gardenoside reference substance, adds dissolve with methanol, shakes up, promptly get reference substance solution: get Qingkailing and dissolve with methanol-water solution, filter membrane filters, and promptly gets need testing solution, or get qingkailing injections or QINGKAILING infusion solutions, and put evaporate to dryness in the water-bath earlier, put cold, residue dissolves with methanol-water solution, and filter membrane filters, or with chloroform extraction, put coldly, extract evaporate to dryness, residue add methanol-water dissolving, filter membrane filters, and promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination content, in this Qingkailing preparation, Gardenoside content should be less than 0.8% in the freeze-dried powder, Gardenoside content should be less than that Gardenoside content should be less than 0.04mg/ml in 0.4mg/ml, the infusion solutions in the parenteral solution;

Scutelloside is according to high effective liquid chromatography for measuring, and chromatographic column is C18 or C4 or C8 post, and with acetonitrile or methyl alcohol: 0.3～0.5% phosphoric acid=10～40: 90～60 is moving phase, and flow velocity is 0.6-1.2ml/min, and column temperature: 20-50 ℃, the detection wavelength is 205～350nm; Precision takes by weighing the scutelloside reference substance, adds dissolve with methanol, shakes up, and promptly gets reference substance solution; Get Qingkailing and dissolve with methanol-water solution, filter membrane filters, and promptly gets need testing solution, or get qingkailing injections or QINGKAILING infusion solutions, and put evaporate to dryness in the water-bath earlier, put cold, residue dissolves with methanol-water solution, filter membrane filters, or extracts with the mixed solution of ethyl acetate and methyl alcohol, puts cold, the extract evaporate to dryness, residue adds the methanol-water dissolving, and filter membrane filters, and promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination content, in this Qingkailing preparation, content of baicalin should be 5%～17.5% in the freeze-dried powder, content of baicalin should be that content of baicalin should be 0.24～2.1mg/ml in 2～7mg/ml, the infusion solutions in the parenteral solution;

Chlorogenic acid is according to high effective liquid chromatography for measuring, and chromatographic column is C18 or C4 or C8 post, and with acetonitrile or methyl alcohol: 0.2～0.7% phosphoric acid=5～25: 95～75 is moving phase, and flow velocity is 0.6-1.2ml/min, and column temperature: 20-50 ℃, the detection wavelength is 245～600nm; Precision takes by weighing the chlorogenic acid reference substance, adds the methanol-water dissolving, shakes up, and promptly gets reference substance solution; Get Qingkailing, with the dissolving of methanol-water solution, filter membrane filters, and promptly get need testing solution, or gets qingkailing injections or infusion solutions, puts evaporate to dryness in the water-bath earlier, puts coldly, and residue is with the dissolving of methanol-water solution, and the filter membrane filtration promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination content, in this Qingkailing preparation, chlorogenic acid content should be less than 0.4% in the freeze-dried powder, chlorogenic acid content should be less than that chlorogenic acid content should be less than 0.018mg/ml in 0.15mg/ml, the infusion solutions in the parenteral solution;

Cholic acid is according to high effective liquid chromatography for measuring, chromatographic column is C18 or C4 or C8 post, moving phase is: methyl alcohol or acetonitrile: water=40-85: 60-15, or be methyl alcohol or acetonitrile: water: glacial acetic acid=40-85: 60-15: 0.01-0.9g, or be methyl alcohol or acetonitrile: water: formic acid=40-85: 60-15: 0.01-0.5, flow rate of mobile phase is 0.6-1.2ml/min, column temperature: 20-50 ℃, the evaporative light-scattering detector drift tube temperature is 25-115 ℃, atomization gas is air or nitrogen, and gas flow rate is 1.0-4.0L/min; Get the cholic acid reference substance, add dissolve with methanol, shake up, promptly get reference substance solution; Get Qingkailing, add methanol-water solution sonicated 5-30 minute, put cold, add the dissolving of methanol-water solution, shake up, filter membrane filters, promptly get need testing solution, or get qingkailing injections or infusion solutions, with methanol-water solution sonicated 5-30 minute, put cold, add the dissolving of methanol-water solution, shake up, filter membrane filters, and promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination, press external standard two-point method logarithmic equation and calculate content, in this Qingkailing preparation, cholic acid content should be in 2.5%-10%, the parenteral solution cholic acid content and should be that cholic acid content should be 0.12-1.2mg/ml in 1-4mg/ml, the infusion solutions in the freeze-dried powder;

Hyodesoxycholic acid is according to high effective liquid chromatography for measuring, chromatographic column is C18 or C4 or C8 post, moving phase is: methyl alcohol or acetonitrile: water=40-85: 60-15, or be methyl alcohol or acetonitrile: water: glacial acetic acid=40-85: 60-15: 0.01-0.99, or be methyl alcohol or acetonitrile: water: formic acid=40 85: 60-15: 0.01-0.5, flow rate of mobile phase is 0.6-1.2ml/min, column temperature: 20-50 ℃, the evaporative light-scattering detector drift tube temperature is 25-115 ℃, atomization gas is air or nitrogen, and gas flow rate is 1.0-4.0L/min; Get the hyodesoxycholic acid reference substance, add dissolve with methanol, shake up, promptly get reference substance solution; Get Qingkailing, add methanol-water solution sonicated 5-30 minute, put cold, add the dissolving of methanol-water solution, shake up, filter membrane filters, promptly get need testing solution, or get qingkailing injections or infusion solutions, with methanol-water solution sonicated 5-30 minute, put cold, add the dissolving of methanol-water solution, shake up, filter membrane filters, and promptly gets need testing solution; Accurate reference substance solution and the need testing solution drawn, inject hplc determination, press external standard two-point method logarithmic equation and calculate content, in this Qingkailing preparation, hyodesoxycholic acid content should be in 2.5%-10%, the parenteral solution hyodesoxycholic acid content and should be that hyodesoxycholic acid content should be 0.12-1.2mg/ml in 1-4mg/ml, the infusion solutions in the freeze-dried powder;

The nitrogenous Qingkailing preparation of measuring is measured according to the Chinese Pharmacopoeia n2 method, and wherein the freeze-dried powder nitrogen content is that 4%-10%, parenteral solution nitrogen content are that 1.6-4mg/ml, infusion solutions nitrogen content are 0.18-1.2mg/ml.

2. according to the detection method of the described Qingkailing injection preparation of claim 1, it is characterized in that: discrimination method is:

(1) get Qingkailing, parenteral solution or infusion solutions respectively, the dissolving of freeze-dried powder water adds furfural solution and sulfuric acid solution mixing, the water-bath heating, and solution shows gray purple;

(2) get Qingkailing, parenteral solution or infusion solutions, wherein the preparation method of freeze-dried powder need testing solution is: get the content of freeze-dried powder, add dissolve with ethanol, get supernatant promptly; The preparation method of parenteral solution or infusion solutions need testing solution is: get parenteral solution 5-20ml or infusion solutions 50-200ml, put evaporate to dryness in the water-bath, put coldly, residue adds ethanol makes dissolving, gets supernatant promptly; Other gets the Gardenoside reference substance, adds dissolve with ethanol, in contrast product solution; Reference substance solution and need testing solution are put respectively on same silica gel g thin-layer plate, with ethyl acetate or phosphoric acid: acetone: formic acid: water=2-10: 2-10: 0.5-2: 0.5-2 is a developping agent, with 10% ethanol solution of sulfuric acid is developer, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

3. according to the detection method of the described Qingkailing injection preparation of claim 1, it is characterized in that: described proterties is: freeze-dried powder: product is light yellow to pale brown look loose block; Parenteral solution: product is pale brown look or henna clear liquid; Great transfusion preparation: product is yellow clear liquid to pale brown look.

4. according to the detection method of the described Qingkailing injection preparation of claim 1, it is characterized in that: inspection method is:

Particulate matter is got Qingkailing, parenteral solution or infusion solutions, wherein freeze-dried powder 200mg adds the 50-100ml that purifies waste water and makes dissolving, make blank of purifying waste water, measure according to Chinese Pharmacopoeia appendix of version in 2005 " particulate matter inspection technique " first method, should be up to specification;

PH value is got Qingkailing, parenteral solution or infusion solutions, and wherein freeze-dried powder 200mg adds water 1-10ml and makes dissolving, surveys its pH value respectively, and its pH value all should be 5-9;

Protein is got Qingkailing, parenteral solution or infusion solutions, and wherein freeze-dried powder 200mg adds water 1-10ml and makes dissolving, gets 1ml respectively and puts in the test tube, drips tannic acid test solution 1-5 and drips, and must not produce muddiness;

Tannin is got Qingkailing, parenteral solution or infusion solutions, and wherein freeze-dried powder 200mg adds water 1-10ml and makes dissolving, gets 1ml, adds the physiological saline 1-10ml that contains the 1-5% Ovum Gallus domesticus album of new preparation, places 5-20 minute, muddiness or precipitation must not occur;

Resin is got Qingkailing, parenteral solution or infusion solutions, and wherein freeze-dried powder 200mg adds water 1-10ml and makes dissolving, gets above-mentioned solution 5ml respectively, put in the separating funnel, add chloroform 5-20ml jolting and extract, divide and get chloroform solution, evaporate to dryness, residue adds glacial acetic acid 1-5ml makes dissolving, put in the tool plug test tube, add water 1-5ml, mixing, placed 20-50 minute, and should not have floccus and separate out;

Oxalates is got Qingkailing, parenteral solution or infusion solutions, wherein freeze-dried powder 200mg adds water 1-10ml and makes dissolving, get above-mentioned solution 5ml respectively, regulate pH value to 1～3 with watery hydrochloric acid, 30-70 ℃ is incubated 5-20 minute, insulation elimination precipitation, regulate pH value to 4～7, get 1-5ml, add 1～5 of 1-5% calcium chloride solution, placed 5-20 minute, and muddiness or precipitation must not occur;

Heavy metal is got Qingkailing, parenteral solution or infusion solutions, wherein freeze-dried powder 200mg adds water 1-10ml and makes dissolving, draw 1-2ml respectively, evaporate to dryness, slowly baking is to fully ashing, puts coldly, and control tube is got standard lead solution 1ml, measure according to Chinese Pharmacopoeia appendix of version in 2005 " heavy metal inspection technique " second method, contain heavy metal in the 1ml aqueous solution and must not cross 10/1000000ths;

Arsenic salt is got Qingkailing, parenteral solution or infusion solutions, wherein freezes to add water 1-10ml in powder 200mg and make dissolving, draws 1ml respectively, evaporate to dryness, add 1-5% magnesium nitrate ethanolic solution 1-5ml, light the afterburnt, burn with little heated earlier and make charing, again 500～600 ℃ blazing to fully ashing, put coldly, add hydrochloric acid 1-10ml and water 10-30ml makes dissolving, measure according to appendix of Chinese Pharmacopoeia version in 2005 " arsenic salt inspection technique " first method, arsenic content must not cross 2/1000000ths in the 1ml aqueous solution;

Potassium ion is got Qingkailing, parenteral solution or infusion solutions, and wherein freeze-dried powder 200mg adds water 1-10ml and makes dissolving, draws 1-5ml respectively, and evaporate to dryness is measured according to appendix of Chinese Pharmacopoeia version in 2005 " injection related substance inspection technique ", should be up to specification;

Undue toxicity is got Qingkailing, parenteral solution or infusion solutions, wherein freeze-dried powder adds the chlorination sodium injection and makes the solution that contains 30-50mg among every 1ml, check according to Chinese Pharmacopoeia two appendix of version in 2005 " undue toxicity inspection technique ", press the intravenous injection administration, should be up to specification;

Haemolysis is got rabbit blood or sheep blood with the preparation of cohesion 2% red blood cell suspension, put into the conical flask that fills beaded glass, jolting 5-20 minute, remove fibrinogen, make to become to take off fine blood, add physiological sodium chloride solution washing 1-5 time that 5-20 doubly measures, till the apparent redness of supernatant, the gained red blood cell is made into 2% suspension with physiological sodium chloride solution, promptly;

Test tube numbering 1,2% red blood cell suspension 2.5ml, physiological sodium chloride solution 2.0ml, soup 0.5ml;

Numbering 2,2% red blood cell suspension 2.5ml, physiological sodium chloride solution 2.1ml, soup 0.4ml;

Numbering 3,2% red blood cell suspension 2.5ml, physiological sodium chloride solution 2.2ml, soup 0.3ml;

Numbering 4,2% red blood cell suspension 2.5ml, physiological sodium chloride solution 2.3ml, soup 0.2ml;

Numbering 5,2% red blood cell suspension 2.5ml, physiological sodium chloride solution 2.4ml, soup 0.1ml;

Numbering 6,2% red blood cell suspension 2.5ml, physiological sodium chloride solution 2.5ml, soup 0ml;

Test method is got 6 in test tube, add 2% red blood cell suspension and physiological sodium chloride solution successively by the said ratio amount, behind the mixing, in 37 ℃ of constant temperature ovens, placed 10-60 minute, the soup that adds different amounts respectively, wherein freeze-dried powder is got 200mg, and with physiological sodium chloride solution dissolving and be diluted to 5-40ml, the 6th pipe of numbering 6 is control tube; After shaking up, put in 37 ℃ of constant temperature ovens, beginning was observed 1 time every 15 minutes, after 1 hour, observed 1 time every 1 hour, observed altogether 2 hours;

By last method inspection, be as the criterion to number 3 the 3rd pipe, this product haemolysis and red blood cell condensation must not occur in 2 hours;

Barium ion is got platinum filament, with hydrochloric acid moistening after, dip in and get Qingkailing, parenteral solution or infusion solutions, in colourless flame, burn, flame shows yellow green, by the green glass perspective, it is blue that flame shows;

Get Qingkailing, parenteral solution or infusion solutions, wherein freeze-dried powder 200mg dissolves with water for injection 1-10ml water for injection, and taking liquid drips the CaCl that 1-50 drips 0.01-0.2% respectively ₂Solution or Ca (OH) ₂Solution must not become turbid or precipitate;

Qingkailing, parenteral solution or infusion solutions are got in the residue on ignition inspection, and wherein freeze-dried powder 200mg dissolves with 1-10ml water for injection, and taking liquid 1-5ml checks according to the bright residue inspection technique of Chinese Pharmacopoeia respectively, should be less than 2%;

Moisture is got freeze-dried powder, measures according to a Chinese Pharmacopoeia appendix of version " aquametry " in 2005 three therapeutic methods of traditional Chinese medicine, must not cross 5.0%;

Other should meet relevant every regulation under an appendix injection of Chinese Pharmacopoeia version in 2005 item.

5. according to the detection method of the described Qingkailing injection preparation of claim 4, it is characterized in that: the inspection method of barium ion is: get Qingkailing, parenteral solution or infusion solutions, wherein freeze-dried powder 200mg dissolves with water for injection 1-5ml, the difference taking liquid, drip dilute sulfuric acid, promptly generate white precipitate, separate, be deposited in all dissolvings in hydrochloric acid or the nitric acid.