CN102879495B

CN102879495B - Antidotal capsule of Tibetan medicine compound and quality detection method of preparation of antidotal capsule

Info

Publication number: CN102879495B
Application number: CN201210365507.8A
Authority: CN
Inventors: 马振元; 陈芹利; 任松鹏; 王海苹; 董玉波
Original assignee: Shandong Arura Pharmaceutical Research & Development Co Ltd
Current assignee: JINHE TIBETAN MEDICINE CO., LTD.
Priority date: 2012-09-27
Filing date: 2012-09-27
Publication date: 2014-07-16
Anticipated expiration: 2032-09-27
Also published as: CN102879495A

Abstract

The invention discloses an antidotal capsule of a Tibetan medicine compound and a quality detection method of a preparation of the antidotal capsule. Content of strychnine of semen strychni is controlled by a high performance liquid chromatography (HPLC) process; iron content in iron powder is controlled by means of titration; cholic acid in calculus bovis artifactus and hydroxy saffionin A in safflower are detected quantitatively by the high performance liquid liquid-phase chromatography; so that efficacy is guaranteed while hidden dangers to health of patients when the patients take excessive antidotal capsules are avoided, and product quality is improved, product safety, stability and quality controllability are guaranteed.

Description

The quality determining method of a kind of Tibetan medicinal composition detoxicating capsule and preparation thereof

Technical field

The present invention relates to a kind of quality determining method of Tibetan medicinal composition preparation, particularly the quality determining method of a kind of Tibetan medicinal composition detoxicating capsule and preparation thereof.

Background technology

Detoxicating capsule comes from Tibetan medicine's ancient books and records < < sweet dew storehouse > >, is the good recipe that Tibetan medicine treats various nosotoxicosis.Record in Chinese patent drug provincial standard rising national standard part, standard number: WS-10537(ZD-0537)-2002.Detoxicating capsule is comprised of 35 taste medicines such as vomiting nut (system), safflower, iron powder (system), calculus bovis factitius, Phytolacca acinosa, dry and soft, tsaoko, hooker winghead roots, has heat reliving and toxin-eliminating, the effect of putrefaction-removing granulation-promoting.For various toxications, outmoded pyreticosis, some contact dermatitis etc.

Detoxicating capsule contains Strychnos nux-vomica subclass toxicity medicinal material, and its toxic component is mainly alkaloids composition, and have and remove obstruction in channels to relieve pain, the effect of mass dissipating and swelling eliminating, poisoning but overdose may cause, the various bad reactions such as even dead, should not too much take or clothes for a long time.In prescription, contain iron powder, < < Kaibao Bencao > > records iron powder and has tranquilizing mind simultaneously, hard marrow, the effects such as profit skin.Iron is the trace element of human body, has the vital role such as the red blood cell of promotion generation.In Tibetan medicinal composition detoxicating capsule and preparation prescription thereof, iron powder used is to be used as medicine after concocting, and is more conducive to absorb.But iron is taken in, too much may cause acute iron poisoning, be detrimental to health.

Detoxicating capsule proper mass standard only has the discriminating of cholic acid thin layer, gallic acid content to measure item, and, to vomiting nut Poisoning composition alkaloid, iron content does not carry out quality control, makes said preparation have potential safety hazard.

Summary of the invention

For the deficiencies in the prior art, the object of this invention is to provide the quality determining method of a kind of Tibetan medicinal composition detoxicating capsule and preparation thereof.

Summary of the invention

The invention provides the control method of iron content in the peace strychnia, iron powder of vomiting nut sergeant in a kind of Tibetan medicinal composition detoxicating capsule and preparation thereof; the content assaying method of hydroxyl radical carthamin yellow carthamus A in cholic acid, safflower in calculus bovis factitius; guarantee that strychnine, strychnia and iron content are all inexcessive; make said preparation in treatment disease, guaranteed the safe and effective of medication.

Term explanation:

Detoxicating capsule is the nomenclature of drug that national standard for traditional Chinese medicines compilation (Chinese patent drug provincial standard rising national standard part) is recorded.

Detoxicating capsule and preparation thereof comprise other preparations of preparing with detoxicating capsule bulk drug formula, comprise detoxicating capsule.The proportioning of each bulk drug can be identical with detoxicating capsule proportioning, also can within the scope of drug effect, suitably adjust.

Vomiting nut (system), borax (system), pearl (system), lazurite (system), turquoise (system), iron powder (system), sound copper (system), parmelia saxatilis (system), it is the routine techniques term of Tibetan medicine, " system " in bracket all concocted according to the inner method of the Tibetan medicine concocted specification > > of < < Qinghai Province (Qinghai Province food and medicine Surveillance Authority compiles, the versions in 2010 that the Qinghai People's Press publishes).

For achieving the above object, the present invention adopts following technical proposals:

The quality determining method of a kind of Tibetan medicinal composition detoxicating capsule and preparation thereof, the bulk drug of said preparation consists of animal treasured, Radix Brassicae rapae, earthworm fruit mustard, vomiting nut (system), German tamarisk, borax (system), pearl (system), coral, lazurite (system), turquoise (system), iron powder (system), ring copper (system), parmelia saxatilis (system), ichor cream, nutmeg, tsaoko, Tabasheer, safflower, cloves, myrobalan, Phytolacca acinosa, santal, santal, Moschus, calculus bovis factitius, whorlleaf oxytropis herb, grass-leaved sweetflag, cardamom, hooker winghead root, west szechwan pyrethrum, the rhizome of davallia, rhizoma nardostachyos, Corallodiscus flabellatus (Franch.) Burtt, alpine gentian herb, muscicolous woodbetony root, it is characterized in that, quality determining method comprises one or more in following content assaying method:

A. the assay of vomiting nut: get respectively strychnine reference substance, strychnia reference substance is contrast, octadecylsilane chemically bonded silica is filling agent; The volume parts of take is mobile phase than acetonitrile-0.01mol/l sodium heptanesulfonate of 15-25:85-75 and 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution (by volume parts than 10% phosphorus acid for adjusting pH value 2.8); The high performance liquid chromatography that detection wavelength is 250-270nm;

B. the assay of iron powder: by after Chinese medicine preparation ashing to be determined, dissolve with concentrated hydrochloric acid, add methyl orange indicator solution, drip stannous chloride solution by Fe ³⁺be reduced to Fe ²⁺, solution becomes pale red, then take diphenylamine sulfonic acid sodium salt as indicator, is titrated to solution is purple and is terminal with potassium dichromate standard solution, calculates the content of iron powder with the amount of known potassium dichromate standard solution;

C. the assay of safflower: get hydroxyl safflower anthocyanidin A reference substance and add volume parts and be mixed with reference substance solution than 25% methanol aqueous solution, get preparation to be detected and add volume parts after ultrasonic dissolution, removal of impurities, to be mixed with need testing solution than 25% methanol aqueous solution, take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than methyl alcohol-volume parts of 20-30:80-70 than 0.5% aqueous formic acid, the high performance liquid chromatography that detection wavelength is 400-410nm;

D. the assay of calculus bovis factitius: get cholic acid reference substance and add methyl alcohol and be mixed with reference substance solution, get preparation to be detected and add methyl alcohol to be mixed with need testing solution after ultrasonic dissolution, removal of impurities, take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than methyl alcohol-volume parts of 80-70:20-30 than 1% glacial acetic acid aqueous solution, the high performance liquid chromatography of evaporative light-scattering detector.

As a further improvement on the present invention, the assay concrete steps of described vomiting nut are:

According to high performance liquid chromatography (appendix VI D of < < Chinese Pharmacopoeia > > version in 2010), measure;

Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than acetonitrile-0.01mol/l sodium heptanesulfonate of 15-25:85-75 and 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution (by volume parts than 10% phosphorus acid for adjusting pH value 2.8); Detection wavelength is 250-270nm; Number of theoretical plate calculates and should be not less than 5000 by strychnine peak;

The preparation of reference substance solution: get strychnine reference substance 6mg, strychnia reference substance 3mg, accurately weighed, put respectively in 100ml measuring bottle, add methenyl choloride and make in right amount to dissolve and be diluted to scale, shake up.Precision measures 2ml respectively, puts in same 10ml measuring bottle, with methyl alcohol, is diluted to scale, shakes up, and obtains (every 1ml is containing strychnine 12 μ g, strychnia 6 μ g);

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content or preparation powder 10-14g, accurately weighed, put in tool plug conical flask, hydro-oxidation sodium test solution 10ml, mix, place 30 minutes, precision adds methenyl choloride 25ml, close plug, weighed weight, put in water-bath refluxing extraction 2 hours, let cool, weighed weight again, with methenyl choloride, supply the weight of less loss, shake up, divide and get methenyl choloride liquid, with the filter paper that is covered with a small amount of anhydrous sodium sulfate, filter, discard just filtrate, precision measures subsequent filtrate 5ml, put in 10ml measuring bottle, add methyl alcohol to scale, shake up, obtain,

Determination method: draw respectively each 5-10 μ l of reference substance solution and need testing solution, injection liquid chromatography, measures, and obtains.

As a further improvement on the present invention, the content assaying method concrete steps of described iron powder are:

Get Tibetan medicinal composition detoxicating capsule content or preparation powder 6 ~ 10g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning, blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 0.5-2ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, in 500-700 ℃ of blazing 3-5 hour, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, at 60-70 ℃ of left and right heating for dissolving 3-5 hour, after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 0-15ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, can add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 2-10ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, use immediately dichromate titration liquid (0.002mol/l) titration.Every 1ml dichromate titration liquid (0.002mol/l) is equivalent to the iron of 0.6702mg.

As a further improvement on the present invention, the concrete steps of described safflower are:

Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than methyl alcohol-volume parts of 20-30:80-70 than 0.5% aqueous formic acid; Detection wavelength is 400-410nm; Number of theoretical plate calculates and should be not less than 3000 by hydroxyl radical carthamin yellow carthamus A peak;

The preparation of reference substance solution: get hydroxyl safflower anthocyanidin A reference substance appropriate, accurately weighed, put in brown bottle, add volume parts and make every 1ml containing the solution of 6-10 μ g than 25% methanol aqueous solution, obtain;

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content or preparation powder 4-6g, accurately weighed, put in tool plug conical flask, precision adds volume parts than 25% methanol aqueous solution 20-30ml, close plug, weighed weight, ultrasonic 40 minutes, let cool, more weighed weight, the weight of less loss by volume parts, than 25% methanol aqueous solution, supplied, shake up, filter, get subsequent filtrate, obtain;

As a further improvement on the present invention, the concrete steps of described calculus bovis factitius assay are:

Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than methyl alcohol-volume parts of 80-70:20-30 than 1% glacial acetic acid aqueous solution; Evaporative light-scattering detector; Number of theoretical plate calculates and should be not less than 5000 by cholic acid peak;

The preparation of reference substance solution: it is appropriate that precision takes cholic acid reference substance, adds methyl alcohol and makes every 1ml containing the solution of 0.4-0.5mg, obtains;

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content or preparation powder 6-8g, accurately weighed, to put in the conical flask of tool plug, precision adds methyl alcohol 40-60ml, close plug, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, with methyl alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate 20-30ml, 40 ℃ of following recovered under reduced pressure, are transferred in 5ml measuring bottle, add methanol constant volume to scale, filter, get subsequent filtrate, obtain;

Determination method: precision is drawn reference substance solution 10 μ l, 20 μ l respectively, need testing solution 10 μ l, injection liquid chromatography, measures respectively, with external standard two-point method logarithmic equation, calculates, and obtains.

The present invention detoxicating capsule and preparation thereof that preferably bulk drug forms below: precious 4.0 weight portions of animal, Radix Brassicae rapae 4.0 weight portions, earthworm fruit mustard 30.4 weight portions, vomiting nut (system) 1.6 weight portions, German tamarisk 30.4 weight portions, borax (system) 0.8 weight portion, pearl (system) 3.2 weight portions, coral 1.6 weight portions, lazurite (system) 1.6 weight portions, turquoise (system) 1.6 weight portions, iron powder (system) 2.4 weight portions, ring copper (system) 0.8 weight portion, parmelia saxatilis (system) 0.8 weight portion, ichor cream 4.0 weight portions, nutmeg 1.6 weight portions, tsaoko 1.6 weight portions, Tabasheer 1.6 weight portions, safflower 1.6 weight portions, cloves 1.6 weight portions, myrobalan's 2.4 weight portions, Phytolacca acinosa 1.6 weight portions, santal 30.4 weight portions, santal 27.9 weight portions, Moschus 1.6 weight portions, calculus bovis factitius 2.4 weight portions, whorlleaf oxytropis herb 30.4 weight portions, grass-leaved sweetflag 1.6 weight portions, cardamom 1.6 weight portions, hooker winghead root 30.4 weight portions, west szechwan pyrethrum 30.4 weight portions, the rhizome of davallia 3.2 weight portions, rhizoma nardostachyos 1.6 weight portions, Corallodiscus flabellatus (Franch.) Burtt 30.4 weight portions, alpine gentian herb 30.4 weight portions, muscicolous woodbetony root 30.4 weight portions.

For above-mentioned preferred formulation, described quality determining method is:

A. the assay of vomiting nut:

Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than acetonitrile-0.01mol/l sodium heptanesulfonate of 20:80 and 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution (by volume parts than 10% phosphorus acid for adjusting pH value 2.8); Detection wavelength is 260nm; Number of theoretical plate calculates and should be not less than 5000 by strychnine peak;

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content or preparation powder 12g, accurately weighed, put in tool plug conical flask, hydro-oxidation sodium test solution 10ml, mix, place 30 minutes, precision adds methenyl choloride 25ml, close plug, weighed weight, put in water-bath refluxing extraction 2 hours, let cool, weighed weight again, with methenyl choloride, supply the weight of less loss, shake up, divide and get methenyl choloride liquid, with the filter paper that is covered with a small amount of anhydrous sodium sulfate, filter, discard just filtrate, precision measures subsequent filtrate 5ml, put in 10ml measuring bottle, add methyl alcohol to scale, shake up, obtain,

Determination method: draw respectively each 10 μ l of reference substance solution and need testing solution, injection liquid chromatography, measures, and obtains.

B. the assay of iron powder:

Get Tibetan medicinal composition detoxicating capsule content or preparation powder 8g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning, blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 1ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, blazing 4 hours in 600 ℃, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, 65 ℃ of left and right heating for dissolving 4 hours, after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 5ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, can add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 4ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, use immediately dichromate titration liquid (0.002mol/l) titration.Every 1ml dichromate titration liquid (0.002mol/l) is equivalent to the iron of 0.6702mg.

C. the assay of safflower:

Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than methyl alcohol-volume parts of 25:75 than 0.5% aqueous formic acid; Detection wavelength is 403nm; Number of theoretical plate calculates and should be not less than 3000 by hydroxyl radical carthamin yellow carthamus A peak;

The preparation of reference substance solution: get hydroxyl safflower anthocyanidin A reference substance appropriate, accurately weighed, put in brown bottle, add volume parts and make every 1ml containing the solution of 8 μ g than 25% methanol aqueous solution, obtain;

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content or preparation powder 5g, accurately weighed, put in tool plug conical flask, precision adds volume parts than 25% methanol aqueous solution 25ml, close plug, weighed weight, ultrasonic 40 minutes, let cool, more weighed weight, the weight of less loss by volume parts, than 25% methanol aqueous solution, supplied, shake up, filter, get subsequent filtrate, obtain;

D. the assay of calculus bovis factitius:

Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than methyl alcohol-volume parts of 76:24 than 1% glacial acetic acid aqueous solution; Evaporative light-scattering detector; Number of theoretical plate calculates and should be not less than 5000 by cholic acid peak;

The preparation of reference substance solution: it is appropriate that precision takes cholic acid reference substance, adds methyl alcohol and makes every 1ml containing the solution of 0.45mg, obtains;

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content or preparation powder 7g, accurately weighed, to put in the conical flask of tool plug, precision adds methyl alcohol 50ml, close plug, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, with methyl alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate 25ml, 40 ℃ of following recovered under reduced pressure, are transferred in 5ml measuring bottle, add methanol constant volume to scale, filter, get subsequent filtrate, obtain;

In to the quality determining method of preferred formulation, in described vomiting nut content, vomiting nut content is with strychnine (C ₃₁h ₂₂n ₂o ₂) meter, should be 0.043mg/g-0.080mg/g; With strychnia (C ₂₃h ₂₆n ₂o ₁) meter, must not be less than 0.028mg/g.In the assay of described iron powder, iron content is 0.409mg/g-0.682mg/g.In described safflower content mensuration, safflower content is with hydroxyl radical carthamin yellow carthamus A (C ₂₇h ₃₀o ₁₅) meter, must not be less than 0.027mg/g.In described calculus bovis factitius assay, calculus bovis factitius content is with cholic acid

(C ₂₄h ₄₀o ₅) meter, must not be less than 0.252mg/g.

The unit corresponding relation of the weight portion described in this instructions and parts by volume is g/ml or kg/l.

Safe and effective for medication in order to guarantee patient, primary standard urgently needs to improve, the toxic component alkaloid that vomiting nut in detoxicating capsule is contained carries out content control, and the content of iron in iron powder is controlled comprehensively, increases the assay of effective constituent cholic acid, hydroxyl radical carthamin yellow carthamus A simultaneously.

The invention discloses the quality determining method of a kind of Tibetan medicinal composition detoxicating capsule and preparation thereof; iron content in vomiting nut sergeant's peace strychnia content, iron powder has been carried out to quality control; hydroxyl radical carthamin yellow carthamus A in cholic acid, safflower in calculus bovis factitius has been carried out to quantitative detection; not only guaranteed drug effect but also guaranteed to be unlikely to take too much to cause healthy hidden danger; improve product quality, guaranteed product safety, stable, quality controllable.This law also can be used for other preparations of Tibetan medicinal composition detoxicating capsule simultaneously, as detoxification particles, detoxifying pills, Jiedu Tablets etc.

Following experimental example and embodiment are used for further illustrating but are not limited to the present invention.

Experimental example 1: the assay experiment of vomiting nut

1. instrument, reagent and test sample

Instrument: L-2100 type Hitachi high performance liquid chromatograph, AuW220D type Shimadzu electronic analytical balance.

Reference substance: strychnine reference substance (Nat'l Pharmaceutical & Biological Products Control Institute), lot number: 110705-200306; Strychnia reference substance (Nat'l Pharmaceutical & Biological Products Control Institute), lot number: 110706-200505.

Sample: detoxicating capsule (Qinghai gold is scolded Tibetan medicine medicine company incorporated company and provided), lot number: 20100701,20100702,20100703.

2. detect the selection of wavelength

Get strychnine, strychnia mixing reference substance solution, in 190-400nm wavelength coverage, scan, according to ultraviolet absorpting spectrum, selected 260nm is for detecting wavelength.

3. mobile phase is selected

Selecting acetonitrile-0.01mol/l sodium heptanesulfonate and 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution (by volume parts than 10% phosphorus acid for adjusting pH value 2.8) is mobile phase.Separating effect when acetonitrile-0.01mol/l sodium heptanesulfonate and 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution (by volume parts than 10% phosphorus acid for adjusting pH value 2.8) the different volumes ratio of having investigated is respectively mobile phase, volume parts all can reach better separating effect during for mobile phase than acetonitrile-0.01mol/l sodium heptanesulfonate of 15-25:85-75 and 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution (by volume parts than 10% phosphorus acid for adjusting pH value 2.8), and acetonitrile-0.01mol/l sodium heptanesulfonate and 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution (by volume parts than 10% phosphorus acid for adjusting pH value 2.8) volume parts is when for 20:80, strychnine peak retention time is 21min, strychnia peak retention time is 17min, separating effect is best.

4. system suitability and negative interference test

Under above-mentioned chromatographic condition, accurate absorption mixed contrast solution, need testing solution, each 10 μ l of negative control solution respectively, and injection liquid chromatography, records chromatogram.Result shows, mix the degree of separation that strychnine, strychnia in reference substance, test sample chromatogram be adjacent chromatographic peak and be all greater than 1.5, negative control solution at strychnine, that strychnia goes out peak position is noiseless.The results are shown in Figure 1, Fig. 2 and Fig. 3.

5. mix reference substance preparation

Get strychnine reference substance 6mg, strychnia reference substance 3mg, accurately weighed, put respectively in 100ml measuring bottle, add methenyl choloride and make in right amount to dissolve and be diluted to scale, shake up.Precision measures 2ml respectively, puts in same 10ml measuring bottle, with methyl alcohol, is diluted to scale, shakes up, and obtains (every 1ml is containing strychnine 12 μ g, strychnia 6 μ g).

6. test sample preparation

6.1 extract the investigation of quantity of solvent

By method under assay item, need testing solution is detected.By the method under the preparation of need testing solution, prepare 3 parts, precision adds methenyl choloride 20ml, 25ml, 50ml respectively.The content of strychnine in every gram of medicine, strychnia of take is determined the methenyl choloride amount that adds as index.The results are shown in Table 1.

Table 1 extracts quantity of solvent and investigates test findings

Result shows, the content that adds strychnine, strychnia in methenyl choloride 20ml, 25ml, every gram of medicine of 50ml gained is basically identical, for guaranteeing that strychnine in medicine, strychnia extract completely, therefore select methenyl choloride to extract quantity of solvent, is 25ml.

The investigation of 6.2 extraction times

By method under assay item, need testing solution is detected.By the method under the preparation of need testing solution, prepare 3 parts, put respectively in water-bath refluxing extraction 1 hour, 2 hours, 3 hours.The content of strychnine in every gram of medicine, strychnia of take is determined extraction time as index.The results are shown in Table 2.

Table 2 extraction time investigation test findings

Result shows, extract 2 hours and every gram of medicine of 3 hours gained in the content of strychnine, strychnia basic identical, therefore select reflux extracting time, be 2 hours.

7. quantitative limit

Get strychnine reference substance appropriate, accurately weighed, add methyl alcohol and make every 1ml containing the solution of strychnine 0.630 μ g.Get strychnia reference substance appropriate, accurately weighed, add methyl alcohol and make every 1ml containing the solution of strychnia 0.315 μ g.Precision is drawn 10 μ l, injection liquid chromatography respectively.Result shows, the signal to noise ratio (S/N ratio) of strychnine, strychnia chromatographic peak is all about 10:1, strychnine be quantitatively limited to 6.30ng, strychnia be quantitatively limited to 3.15ng.

8. the preparation of typical curve and the investigation of linear relationship

Precision measures the mixing reference substance stock solution solution of strychnine, strychnia, and (strychnine content is 25.2 μ g/ml, strychnia content is 12.6 μ g/ml) 1ml, 3ml, 5ml, 8ml, 10ml, put respectively in 10ml volumetric flask, methyl alcohol is diluted to scale, shake up, each accurate sample introduction 10 μ l, carry out linear regression with peak area (A) to reference substance concentration (C), obtain strychnine regression equation: A=9174.6C+174.47, related coefficient: R=0.9998; Strychnia regression equation: A=9170.1C+633.28, related coefficient: R=0.9999.Result shows, strychnine is within the scope of 2.52 μ g/ml-25.2 μ g/ml, and the peak area of strychnine (A) is good with reference substance concentration (C) linear relationship; Strychnia is within the scope of 1.26 μ g/ml-12.6 μ g/ml, and the peak area of strychnia (A) is good with reference substance concentration (C) linear relationship.The results are shown in Table 3, table 4.

Table 3 strychnine linear relationship is investigated result

Table 4 strychnia linear relationship is investigated result

9. precision test

The accurate mixing reference substance solution 10 μ l that draw strychnine, strychnia, injection liquid chromatography, each METHOD FOR CONTINUOUS DETERMINATION 6 times, records peak area and calculates relative standard deviation.Result shows, instrument precision is good.The results are shown in Table 5, table 6.

Table 5 strychnine Precision test result

Table 6 strychnia Precision test result

10. stability test

After prepared by need testing solution, the accurate 10 μ l that draw, injection liquid chromatography, records peak area, measures once every 2 hours later, investigates 8 hours, records peak area and calculates relative standard deviation.Result shows, in need testing solution, strychnine, strychnia measurement result in 8 hours is stable.The results are shown in Table 7, table 8.

Table 7 strychnine stability test result

Table 8 strychnia stability test result

11. replica tests

Get with a collection of detoxicating capsule sample (product batch number: 20100701) 12g, accurately weighed, totally 6 parts, by the method under the preparation of need testing solution, prepare need testing solution, precision is drawn 10 μ l, injection liquid chromatography, the content of strychnine, strychnia in calculation sample respectively.Result shows, this analytical approach repeatability is good.The results are shown in Table 9, table 10.

Table 9 strychnine replica test result

Table 10 strychnia replica test result

12. recovery tests

Precision takes with a collection of detoxicating capsule sample (product batch number: 20100701) 6 parts, each precision adds strychnine, strychnia reference substance, measures its content, calculate recovery rate.Result shows, this assay method measurement result is accurate.The results are shown in Table 11, table 12.

Table 11 strychnine recovery test result

Table 12 strychnia recovery test result

13. sample determinations

Get three batches of Tibetan medicinal composition detoxicating capsules, measure and calculate strychnine, strychnia content.The results are shown in Table 13.

Table 13 sample size measurement result

Experimental example 2: the assay experiment of iron powder

1. instrument, reagent and test sample

Instrument: BT125D type Sai Duolisi electronic analytical balance (German Sai Duolisi sartorius company); 2.5-10 type chamber type electric resistance furnace (Beijing is bright medical apparatus and instruments factory forever); HH-4 type digital display thermostat water bath (Changzhou Rong Guan experimental analysis instrument plant).

Main agents: concentration expressed in percentage by weight 5% stannous chloride solution (takes stannous chloride and is dissolved in 40ml concentrated hydrochloric acid, be diluted with water to 100ml.Dilute with water is one times before use, obtains); Methyl orange indicator solution (get methyl orange 0.1g and be dissolved in 100ml water, obtain); Diphenylamine sulfonic acid sodium salt indicator solution (get diphenylamine sulfonic acid sodium salt 0.2g and be dissolved in 100ml water, obtain); Mixture of sulfuric phosphoric acid solution (the 150ml mass percentage concentration 95%-98% concentrated sulphuric acid is under agitation slowly injected to 500ml water, add again 150ml phosphoric acid after cooling, be diluted with water to 1000ml, mix, obtain).Potassium dichromate standard solution (standard reagent in 150 ~ 160 ℃ dry 2 hours, be stored in exsiccator, accurately take 0.0980g potassium dichromate in 250ml beaker, add after suitable quantity of water is dissolved and quantitatively proceed in 1000ml volumetric flask, be diluted with water to scale, shake up, its concentration is 0.002mol/l).

Sample: precious imperial restoring consciouness capsule (Qinghai gold is scolded Tibetan medicine medicine company incorporated company and provided), lot number is respectively: 20090516,20090908,20110426.

2. the ashing of sample

The crucible that fills sample is put on electric furnace to slowly vehement burning (avoiding burning, expanding, overflow), blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature.Drip the mass percentage concentration 95%-98% concentrated sulphuric acid appropriate, make carbide all moistening, take 0.5ml to 2ml as excellent, wherein 1ml is optimum.Continue to be heated to steam and eliminate, white cigarette emits to the greatest extent, in 500-700 ℃ of blazing 3-5 hour.Wherein take 600 ℃ within blazing 4 hours, be optimum.Use calcination charing, then add sulfuric acid, then the method object of ashing is: remove the interference of other flavour of a drug in Tibetan medicinal preparation, improved the content of iron simultaneously, make iron be converted into Fe3+, be convenient to measure.

3. the selection of concentration of hydrochloric acid

Reaction is carried out in hydrochloric acid (HCl) medium, reduction Fe ³⁺time hydrochloric acid (HCl) concentration with 4mol/l for well, Sn while being greater than 6mol/l ²⁺the methyl orange of first reducing is colourless, makes it cannot indicate Fe ³⁺reduction, Cl simultaneously ^-excessive concentration also may consume K ₂cr ₂o ₇, Methyl Orange Discoloration is slow lower than 2mol/l for HCl concentration.

4. the selection of molten sample temperature

Molten sample temperature has considerable influence to the measurement result of iron content.Molten sample temperature is during lower than 55 ℃, and molten sample is slow; Surpass 75 ℃ of HCl volatilizations too fast, concentration reduces, and molten sample is incomplete.Experiment shows that molten sample temperature is controlled at 65 ℃ of left and right best results.

5. the control of redox temperature

When reducing with stannous chloride, temperature should be controlled at 70-80 ℃, and constantly shakes.And in last oxidimetric titration, solution temperature should be controlled at 20～30 ℃ for well, so that reaction can comparatively fast be carried out.

6. the selection of mixture of sulfuric phosphoric acid solution

The Fe generating in titration process ³⁺be yellow, affect the observation of terminal, if add phosphoric acid (H in solution ₃pO ₄), H ₃pO ₄with Fe ³⁺generate colourless Fe (HPO4) ^2-, can shelter Fe ³⁺.Simultaneously due to Fe (HPO4) ^2-generation, make Fe ³⁺/ Fe ²⁺the right conditional potential of electricity reduces, and titration jump increases, and indicator can variable color in mutation range, thereby reduces titration error.

7. principle

This product sample is after sample ashing, dissolving with hydrochloric acid, and iron is wherein converted into Fe ³⁺.Under strong acidic condition, Fe ³⁺can pass through SnCl ₂be reduced to Fe ²⁺.Sn ²⁺by Fe ³⁺after reduction, methyl orange also can be by Sn ²⁺be reduced into hydrogenation methyl orange and fade, thereby methyl orange can be indicated Fe ³⁺reduction terminal.Sn ²⁺can also continue to make hydrogenation methyl orange to be reduced into N, N-dimethyl-p-phenylenediamine and sodium sulfanilate.Its reaction equation is:

(CH ₃) ₂NC ₆H ₄N=NC ₆H ₄SO ₃Na+2e ^-+2H ⁺→(CH ₃) ₂NC ₆H ₄NH-NHC ₆H ₄SO ₃Na

(CH ₃) ₂NC ₆H ₄NH-NHC ₆H ₄SO ₃Na＋2e _-＋2H ⁺→(CH ₃) ₂NC ₆H ₄NH ₂+NH ₂C ₆H ₄SO ₃Na

So, slightly excessive Sn ²⁺also be eliminated.Because above-mentioned reaction is irreversible, so the reduzate of methyl orange does not consume K ₂cr ₂o ₇.After having reacted, take diphenylamine sulfonic acid sodium salt as indicator, use K ₂cr ₂o ₇standard solution is titrated to solution and is purple and is terminal, and key reaction formula is as follows:

2FeCl ₄ ^-+SnCl ^2-+2Cl ^-=2FeCl ₄ ^2-+SnCl ₆ ^2-

6Fe ²⁺+Cr ₂O ₇ ^2-+14H ⁺=6Fe ³⁺+2Cr ³⁺+7H ₂O

8. the assay of iron in sample

Get Tibetan medicinal composition detoxicating capsule content powder 14g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning (avoiding burning, expanding, overflow), blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 1ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, blazing 4 hours in 600 ℃, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, 65 ℃ of left and right heating for dissolving, within 4 hours, (frequently shake, avoid boiling), after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 5ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, can add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 4ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, use immediately dichromate titration liquid (0.002mol/l) titration.Every 1ml dichromate titration liquid (0.002mol/l) is equivalent to the iron of 0.6702mg.The results are shown in Table 14.

Iron content measurement result in table 14 detoxicating capsule

9. reappearance test

Get with a collection of detoxicating capsule sample (product batch number: 20100701), accurately weighed, totally 6 parts, according to iron content determination method, measure respectively.Result shows, the method repeatability is good.The results are shown in Table 15.

In table 15 detoxicating capsule, iron content is measured replica test

10. recovery test

In the ratio autogamy of Tibetan medicinal composition detoxicating capsule prescription consumption, not containing the sample of iron powder, get the about 14g of this blank sample, accurately weighed, the about 0.1g of standard iron powder that precision adds known content, carries out ashing processing, measures as stated above content calculate recovery rate.Result shows, this assay method measurement result is accurate.The results are shown in Table 16.

Table 16 recovery test measurement result

experimental example 3: the assay experiment of safflower

1. instrument, reagent and test sample

Instrument: 1220 type Agilent high performance liquid chromatographs; AuW220D type Shimadzu electronic analytical balance.

Reference substance: hydroxyl radical carthamin yellow carthamus A reference substance (Nat'l Pharmaceutical & Biological Products Control Institute), lot number: 111637-200905.

2. detect the selection of wavelength

Get hydroxyl radical carthamin yellow carthamus A reference substance solution, in 190-500nm wavelength coverage, scan, according to ultraviolet absorpting spectrum, selected 403nm is for detecting wavelength.

3. mobile phase is selected

Research finds, the volume parts of take during as mobile phase, all can reach good chromatographic resolution effect than 0.5% aqueous formic acid than methyl alcohol-volume parts of 20-30:80-70.Methyl alcohol-the volume parts of wherein take is more optimum as 25:75 as mobile phase than the volume ratio of 0.5% formic acid solution, relative retention time 10min left and right.

4. system suitability and negative interference test

Under above-mentioned chromatographic condition, precision is drawn reference substance solution, need testing solution, each 10 μ l of negative control solution respectively, and injection liquid chromatography, records chromatogram.Result shows, the degree of separation that in reference substance, test sample chromatogram, hydroxyl radical carthamin yellow carthamus A is adjacent chromatographic peak is all greater than 1.5, and it is noiseless that negative control solution goes out peak position at hydroxyl radical carthamin yellow carthamus A.The results are shown in Figure 4, Fig. 5 and Fig. 6.

5. reference substance preparation

Get hydroxyl safflower anthocyanidin A reference substance appropriate, accurately weighed, put in brown bottle, add volume parts and make every 1ml containing the solution of 8 μ g than 25% methanol aqueous solution, obtain.

6. test sample preparation

The investigation of 6.1 extracting method

By method under assay item, need testing solution is detected.By the method under the preparation of need testing solution, prepare 3 parts, ultrasonic, backflow, Zhen Oscillating processes 40 minutes respectively.The content of hydroxyl radical carthamin yellow carthamus A in every gram of medicine of take is determined extracting method as index.The results are shown in Table 17.

Table 17 extracting method is investigated test findings

Result shows, in every gram of medicine of ultrasonic extraction gained, the content of hydroxyl radical carthamin yellow carthamus A is the highest, therefore select extracting method, is ultrasonic extraction.

6.2 extract the investigation of solvent

By method under assay item, need testing solution is detected.By the method under the preparation of need testing solution, prepare 3 parts, precision adds water, volume parts than 25% methanol aqueous solution, each 25ml of methyl alcohol respectively.The content of hydroxyl radical carthamin yellow carthamus A of take in every gram of medicine is that index is determined and extracted solvent.The results are shown in Table 18.

Table 18 extracts solvent and investigates test findings

Result shows, the hydroxyl radical carthamin yellow carthamus A content that water, volume parts are surveyed than 25% methanol aqueous solution and methyl alcohol is basically identical, owing to using water as, extracts solvent filter difficulty, and pure methyl alcohol toxicity is large, therefore select, to extract solvent be that volume parts is than 25% methanol aqueous solution.

The investigation of 6.3 extraction times

By method under assay item, need testing solution is detected.By the method under the preparation of need testing solution, prepare 3 parts, ultrasonic processing is 30 minutes, 40 minutes, 50 minutes respectively.The content of hydroxyl radical carthamin yellow carthamus A in every gram of medicine of take is determined extraction time as index.The results are shown in Table 19.

Table 19 extraction time investigation test findings

Result shows, in ultrasonic extraction 40 minutes and every gram of medicine of 50 minutes gained, the content of hydroxyl radical carthamin yellow carthamus A is basically identical, therefore select extraction time, is 40 minutes.

7. the preparation of typical curve and the investigation of linear relationship

Precision measures hydroxyl radical carthamin yellow carthamus A reference substance stock solution solution (hydroxyl radical carthamin yellow carthamus A content is 16.2 μ g/ml) 1ml, 3ml, 5ml, 8ml, 10ml, put respectively in 10ml volumetric flask, volume parts is diluted to scale than 25% methanol aqueous solution, shake up, each accurate sample introduction 10 μ l, with peak area (A), reference substance concentration (C) is carried out to linear regression, obtain hydroxyl radical carthamin yellow carthamus A regression equation: A=21.006C-4.0763, related coefficient: R=0.9998.Result shows, hydroxyl radical carthamin yellow carthamus A is within the scope of 1.62 μ g/ml-16.2 μ g/ml, and the peak area of hydroxyl radical carthamin yellow carthamus A (A) is good with reference substance concentration (C) linear relationship.The results are shown in Table 20.

Table 20 hydroxyl radical carthamin yellow carthamus A linear relationship is investigated result

8. precision test

The accurate hydroxyl radical carthamin yellow carthamus A reference substance solution 10 μ l that draw, injection liquid chromatography, each METHOD FOR CONTINUOUS DETERMINATION 6 times, records peak area and calculates relative standard deviation.Result shows, instrument precision is good.The results are shown in Table 21.

Table 21 hydroxyl radical carthamin yellow carthamus A Precision test result

9. stability test

After prepared by need testing solution, the accurate 10 μ l that draw, injection liquid chromatography, records peak area, measures once every 2 hours later, investigates 8 hours, records peak area and calculates relative standard deviation.Result shows, in need testing solution, hydroxyl radical carthamin yellow carthamus A measurement result in 8 hours is stable.The results are shown in Table 22.

Table 22 hydroxyl radical carthamin yellow carthamus A stability test result

10. replica test

Get with a collection of detoxicating capsule sample (product batch number: 20100701) 5g, accurately weighed, totally 6 parts, by the method under the preparation of need testing solution, prepare need testing solution, precision is drawn 10 μ l, injection liquid chromatography, the content of hydroxyl radical carthamin yellow carthamus A in calculation sample respectively.Result shows, this analytical approach repeatability is good.The results are shown in Table 23.

Table 23 hydroxyl radical carthamin yellow carthamus A replica test result

11. recovery tests

Precision takes with a collection of detoxicating capsule sample (product batch number: 20100701) 6 parts, each precision adds hydroxyl radical carthamin yellow carthamus A reference substance, measures its content, calculate recovery rate.Result shows, this assay method measurement result is accurate.The results are shown in Table 24.

Table 24 hydroxyl radical carthamin yellow carthamus A recovery test result

12. sample determinations

Get three batches of Tibetan medicinal composition detoxicating capsules, measure and calculate hydroxyl radical carthamin yellow carthamus A content.The results are shown in Table 25.

Table 25 sample size measurement result

Experimental example 4: the assay experiment of calculus bovis factitius

1. instrument, reagent and test sample

Reference substance: cholic acid reference substance (Nat'l Pharmaceutical & Biological Products Control Institute) lot number: 100078-200414.

2. mobile phase is selected

The assay of cholic acid adopts methyl alcohol-glacial acetic acid system more, research finds take that volume parts all can reach content inspection requirements than 1% glacial acetic acid aqueous solution as mobile phase than methyl alcohol-volume parts of 80-70:20-30, wherein methyl alcohol-volume parts is more best than 1% glacial acetic acid aqueous solution (76:24) separating effect, relative retention time 13min left and right.

4. system suitability

Under above-mentioned chromatographic condition, precision is drawn reference substance solution, need testing solution, each 10 μ l of negative control solution respectively, and injection liquid chromatography, records chromatogram.Result shows, the degree of separation that in reference substance, test sample chromatogram, cholic acid is adjacent chromatographic peak is all greater than 1.5, and it is noiseless that negative control solution goes out peak position at cholic acid.The results are shown in Figure 7, Fig. 8 and Fig. 9.

5. reference substance preparation

It is appropriate that precision takes cholic acid reference substance, adds methyl alcohol and make every 1ml containing the solution of 0.45mg, obtains.

6. test sample preparation

The investigation of 6.1 extracting method

By method under assay item, need testing solution is detected.By the method under the preparation of need testing solution, prepare 3 parts, ultrasonic, backflow, Zhen Oscillating processes 30 minutes respectively.The content of cholic acid in every gram of medicine of take is determined extracting method as index.The results are shown in Table 26.

Table 26 extracting method is investigated test findings

Result shows, in every gram of medicine of ultrasonic extraction gained, the content of cholic acid is the highest, therefore select extracting method, is ultrasonic extraction.

6.2 extract the investigation of solvent

By method under assay item, need testing solution is detected.By the method under the preparation of need testing solution, prepare 3 parts, precision adds methyl alcohol, ethanol, volume parts than each 50ml of 60% ethanol water respectively.The content of cholic acid of take in every gram of medicine is that index is determined and extracted solvent.The results are shown in Table 27.

Table 27 extracts solvent and investigates test findings

Result shows, in the measured every gram of medicine of methyl alcohol, the content of cholic acid is the highest, therefore select, to extract solvent be methyl alcohol.

The investigation of 6.3 extraction times

By method under assay item, need testing solution is detected.By the method under the preparation of need testing solution, prepare 3 parts, ultrasonic processing is 20 minutes, 30 minutes, 40 minutes respectively.The content of cholic acid in every gram of medicine of take is determined extraction time as index.The results are shown in Table 28.

Table 28 extraction time investigation test findings

Result shows, in ultrasonic extraction 30 minutes and every gram of medicine of 40 minutes gained, the content of cholic acid is basically identical, therefore select extraction time, is 30 minutes.

Precision measures cholic acid reference substance stock solution solution (cholic acid content is 892.8 μ g/ml) 1ml, 3ml, 5ml, 8ml, 10ml, put respectively in 10ml volumetric flask, methyl alcohol is diluted to scale, shake up, each accurate sample introduction 10 μ l, logarithm (Log A) with peak area carries out linear regression to the logarithm of reference substance concentration (Log C), obtains cholic acid regression equation: Log A=1.0191Log C+3.4350, related coefficient: R=0.9995.Result shows, cholic acid is within the scope of 89.28 μ g/ml-892.8 μ g/ml, and the logarithm of cholic acid peak area (LogA) is good with logarithm (Log C) linear relationship of reference substance concentration.The results are shown in Table 29.

Table 29 cholic acid linear relationship is investigated result

8. precision test

The accurate cholic acid reference substance solution 10 μ l that draw, injection liquid chromatography, each METHOD FOR CONTINUOUS DETERMINATION 6 times, records peak area and calculates relative standard deviation.Result shows, instrument precision is good.The results are shown in Table 30.

Table 30 cholic acid Precision test result

9. stability test

After prepared by need testing solution, the accurate 10 μ l that draw, injection liquid chromatography, records peak area, measures once every 2 hours later, investigates 8 hours, records peak area and calculates relative standard deviation.Result shows, in need testing solution, cholic acid measurement result in 8 hours is stable.The results are shown in Table 31.

Table 31 cholic acid stability test result

10. replica test

Get with a collection of detoxicating capsule sample (product batch number: 20100701) 7g, accurately weighed, totally 6 parts, by the method under the preparation of need testing solution, prepare need testing solution, respectively accurate 10 μ l, injection liquid chromatography, the content of cholic acid in calculation sample drawn.Result shows, this analytical approach repeatability is good.The results are shown in Table 32.

Table 32 cholic acid replica test result

11. recovery tests

Precision takes with a collection of detoxicating capsule sample (product batch number: 20100701) 6 parts, each precision adds cholic acid reference substance, measures its content, calculate recovery rate.Result shows, this assay method measurement result is accurate.The results are shown in Table 33.

Table 33 cholic acid recovery test result

12. sample determinations

Get three batches of Tibetan medicinal composition detoxicating capsules, measure and calculate cholic acid content.The results are shown in Table 34.

Table 34 sample size measurement result

Following embodiment all can realize the effect described in above-mentioned experimental example.

Accompanying drawing explanation

Fig. 1 is that strychnine, strychnia mix contrast high-efficient liquid phase chromatogram;

Fig. 2 is Tibetan medicinal composition detoxicating capsule vomiting nut test sample high-efficient liquid phase chromatogram;

Fig. 3 is that Tibetan medicinal composition detoxicating capsule lacks vomiting nut negative control high-efficient liquid phase chromatogram;

Fig. 4 is hydroxyl radical carthamin yellow carthamus A contrast high-efficient liquid phase chromatogram;

Fig. 5 is Tibetan medicinal composition detoxicating capsule safflower test sample high-efficient liquid phase chromatogram;

Fig. 6 is that Tibetan medicinal composition detoxicating capsule lacks safflower negative control high-efficient liquid phase chromatogram;

Fig. 7 is cholic acid contrast high-efficient liquid phase chromatogram;

Fig. 8 is Tibetan medicinal composition detoxicating capsule calculus bovis factitius test sample high-efficient liquid phase chromatogram;

Fig. 9 is that Tibetan medicinal composition detoxicating capsule lacks calculus bovis factitius negative control high-efficient liquid phase chromatogram;

Wherein, the horizontal ordinate of Fig. 1, Fig. 2, Fig. 3, Fig. 7, Fig. 8, Fig. 9 is the time, unit: minute (Minutes); Ordinate is voltage, unit: volt (Volts).The horizontal ordinate of Fig. 4, Fig. 5, Fig. 6 is the time, unit: minute (min); Ordinate is absorbance (mAU).

Embodiment

Below in conjunction with embodiment, the present invention is done to detailed elaboration, but be not limited to these concrete embodiment recording.The Tibetan medicinal composition detoxicating capsule detecting is that Qinghai gold is scolded Tibetan medicine medicine company incorporated company and produced and to sell.

Embodiment 1: detoxicating capsule quality determining method

Detoxicating capsule comprises that cholic acid thin layer is differentiated, gallic acid content is measured (also can not comprise these mensuration), also comprises following mensuration:

A. the assay of vomiting nut:

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content powder 12g, accurately weighed, put in tool plug conical flask, hydro-oxidation sodium test solution 10ml, mixes, and places 30 minutes, precision adds methenyl choloride 25ml, close plug, weighed weight, puts in water-bath refluxing extraction 2 hours, let cool, weighed weight again, supplies the weight of less loss with methenyl choloride, shake up, divide and get methenyl choloride liquid, with the filter paper that is covered with a small amount of anhydrous sodium sulfate, filter, discard just filtrate, precision measures subsequent filtrate 5ml, put in 10ml measuring bottle, add methyl alcohol to scale, shake up, obtain;

This product vomiting nut content is with strychnine (C ₃₁h ₂₂n ₂o ₂) meter, should be 0.043mg/g-0.080mg/g; With strychnia (C ₂₃h ₂₆n ₂o ₁) meter, must not be less than 0.028mg/g.

B. the assay of iron powder:

Get Tibetan medicinal composition detoxicating capsule content powder 8g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning, blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 1ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, blazing 4 hours in 600 ℃, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, 65 ℃ of left and right heating for dissolving 4 hours, after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 5ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, can add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 4ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, use immediately dichromate titration liquid (0.002mol/l) titration.Every 1ml dichromate titration liquid (0.002mol/l) is equivalent to the iron of 0.6702mg.

This product iron content is 0.409mg/g-0.682mg/g.

C. the assay of safflower:

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content powder 5g, accurately weighed, put in tool plug conical flask, precision adds volume parts than 25% methanol aqueous solution 25ml, close plug, weighed weight, ultrasonic 40 minutes, let cool, more weighed weight, the weight of less loss by volume parts, than 25% methanol aqueous solution, supplied, shake up, filter, get subsequent filtrate, obtain;

This product safflower content is with hydroxyl radical carthamin yellow carthamus A (C ₂₇h ₃₀o ₁₅) meter, must not be less than 0.027mg/g.

D. the assay of calculus bovis factitius:

The preparation of need testing solution: get Tibetan medicinal composition detoxicating capsule content powder 7g, accurately weighed, to put in the conical flask of tool plug, precision adds methyl alcohol 50ml, close plug, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, with methyl alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate 25ml, 40 ℃ of following recovered under reduced pressure, are transferred in 5ml measuring bottle, add methanol constant volume to scale, filter, get subsequent filtrate, obtain;

This product calculus bovis factitius content is with cholic acid (C ₂₄h ₄₀o ₅) meter, must not be less than 0.252mg/g.

Embodiment 2: detoxification particles quality determining method

Detoxification particles comprises that cholic acid thin layer is differentiated, gallic acid content is measured (also can not comprise these mensuration), also comprises following mensuration:

A. the assay of vomiting nut:

The preparation of need testing solution: get Tibetan medicinal composition detoxification particles powder 12g, accurately weighed, put in tool plug conical flask, hydro-oxidation sodium test solution 10ml, mixes, and places 30 minutes, precision adds methenyl choloride 25ml, close plug, weighed weight, puts in water-bath refluxing extraction 2 hours, let cool, weighed weight again, supplies the weight of less loss with methenyl choloride, shake up, divide and get methenyl choloride liquid, with the filter paper that is covered with a small amount of anhydrous sodium sulfate, filter, discard just filtrate, precision measures subsequent filtrate 5ml, put in 10ml measuring bottle, add methyl alcohol to scale, shake up, obtain;

This product vomiting nut content is with strychnine (C ₃₁h22N ₂o ₂) meter, should be 0.043mg/g-0.080mg/g; With strychnia (C ₂₃h ₂₆n ₂o ₁) meter, must not be less than 0.028mg/g.

B. the assay of iron powder:

Get Tibetan medicinal composition detoxification particles powder 8g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning, blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 1ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, blazing 4 hours in 600 ℃, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, 65 ℃ of left and right heating for dissolving 4 hours, after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 5ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, can add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 4ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, use immediately dichromate titration liquid (0.002mol/l) titration.Every 1ml dichromate titration liquid (0.002mol/l) is equivalent to the iron of 0.6702mg.

This product iron content is 0.409mg/g-0.682mg/g.

C. the assay of safflower:

The preparation of need testing solution: get Tibetan medicinal composition detoxification particles powder 5g, accurately weighed, put in tool plug conical flask, precision adds volume parts than 25% methanol aqueous solution 25ml, close plug, weighed weight, ultrasonic 40 minutes, let cool, more weighed weight, the weight of less loss by volume parts, than 25% methanol aqueous solution, supplied, shake up, filter, get subsequent filtrate, obtain;

D. the assay of calculus bovis factitius:

The preparation of need testing solution: get Tibetan medicinal composition detoxification particles powder 7g, accurately weighed, to put in the conical flask of tool plug, precision adds methyl alcohol 50ml, close plug, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, with methyl alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate 25ml, 40 ℃ of following recovered under reduced pressure, are transferred in 5ml measuring bottle, add methanol constant volume to scale, filter, get subsequent filtrate, obtain;

Described Tibetan medicinal composition detoxification particles refers to the precious 4.0g of detoxicating capsule bulk drug formula animal, Radix Brassicae rapae 4.0g, earthworm fruit mustard 30.4g, vomiting nut (system) 1.6g, German tamarisk 30.4g, borax (system) 0.8g, pearl (system) 3.2g, coral 1.6g, lazurite (system) 1.6g, turquoise (system) 1.6g, iron powder (system) 2.4g, ring copper (system) 0.8g, parmelia saxatilis (system) 0.8g, ichor cream 4.0g, nutmeg 1.6g, tsaoko 1.6g, Tabasheer 1.6g, safflower 1.6g, cloves 1.6g, myrobalan 2.4g, Phytolacca acinosa 1.6g, santal 30.4g, santal 27.9g, Moschus 1.6g, calculus bovis factitius 2.4g, whorlleaf oxytropis herb 30.4g, grass-leaved sweetflag 1.6g, cardamom 1.6g, hooker winghead root 30.4g, west szechwan pyrethrum 30.4g, rhizome of davallia 3.2g, rhizoma nardostachyos 1.6g, Corallodiscus flabellatus (Franch.) Burtt 30.4g, alpine gentian herb 30.4g, muscicolous woodbetony root 30.4g, add conventional auxiliary material, the granule of preparing according to common process.

Embodiment 3: detoxifying pills quality determining method

Detoxifying pills comprises that cholic acid thin layer is differentiated, gallic acid content is measured (also can not comprise these mensuration), also comprises following mensuration:

A. the assay of vomiting nut:

The preparation of need testing solution: get Tibetan medicinal composition detoxifying pills powder 12g, accurately weighed, put in tool plug conical flask, hydro-oxidation sodium test solution 10ml, mixes, and places 30 minutes, precision adds methenyl choloride 25ml, close plug, weighed weight, puts in water-bath refluxing extraction 2 hours, let cool, weighed weight again, supplies the weight of less loss with methenyl choloride, shake up, divide and get methenyl choloride liquid, with the filter paper that is covered with a small amount of anhydrous sodium sulfate, filter, discard just filtrate, precision measures subsequent filtrate 5ml, put in 10ml measuring bottle, add methyl alcohol to scale, shake up, obtain;

B. the assay of iron powder:

Get Tibetan medicinal composition detoxifying pills powder 8g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning, blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 1ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, blazing 4 hours in 600 ℃, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, 65 ℃ of left and right heating for dissolving 4 hours, after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 5ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, can add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 4ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, use immediately dichromate titration liquid (0.002mol/l) titration.Every 1ml dichromate titration liquid (0.002mol/l) is equivalent to the iron of 0.6702mg.

This product iron content is 0.409mg/g-0.682mg/g.

C. the assay of safflower:

The preparation of need testing solution: get Tibetan medicinal composition detoxifying pills powder 5g, accurately weighed, put in tool plug conical flask, precision adds volume parts than 25% methanol aqueous solution 25ml, close plug, weighed weight, ultrasonic 40 minutes, let cool, more weighed weight, the weight of less loss by volume parts, than 25% methanol aqueous solution, supplied, shake up, filter, get subsequent filtrate, obtain;

D. the assay of calculus bovis factitius:

The preparation of need testing solution: get Tibetan medicinal composition detoxifying pills powder 7g, accurately weighed, to put in the conical flask of tool plug, precision adds methyl alcohol 50ml, close plug, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, with methyl alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate 25ml, 40 ℃ of following recovered under reduced pressure, are transferred in 5ml measuring bottle, add methanol constant volume to scale, filter, get subsequent filtrate, obtain;

Described Tibetan medicinal composition detoxifying pills refers to the precious 4.0g of detoxicating capsule bulk drug formula animal, Radix Brassicae rapae 4.0g, earthworm fruit mustard 30.4g, vomiting nut (system) 1.6g, German tamarisk 30.4g, borax (system) 0.8g, pearl (system) 3.2g, coral 1.6g, lazurite (system) 1.6g, turquoise (system) 1.6g, iron powder (system) 2.4g, ring copper (system) 0.8g, parmelia saxatilis (system) 0.8g, ichor cream 4.0g, nutmeg 1.6g, tsaoko 1.6g, Tabasheer 1.6g, safflower 1.6g, cloves 1.6g, myrobalan 2.4g, Phytolacca acinosa 1.6g, santal 30.4g, santal 27.9g, Moschus 1.6g, calculus bovis factitius 2.4g, whorlleaf oxytropis herb 30.4g, grass-leaved sweetflag 1.6g, cardamom 1.6g, hooker winghead root 30.4g, west szechwan pyrethrum 30.4g, rhizome of davallia 3.2g, rhizoma nardostachyos 1.6g, Corallodiscus flabellatus (Franch.) Burtt 30.4g, alpine gentian herb 30.4g, muscicolous woodbetony root 30.4g, add conventional auxiliary material, the pill of preparing according to common process.

Embodiment 4: Jiedu Tablets quality determining method

Jiedu Tablets comprises that cholic acid thin layer is differentiated, gallic acid content is measured (also can not comprise these mensuration), also comprises following mensuration:

A. the assay of vomiting nut:

The preparation of need testing solution: get Tibetan medicinal composition Jiedu Tablets powder 12g, accurately weighed, put in tool plug conical flask, hydro-oxidation sodium test solution 10ml, mixes, and places 30 minutes, precision adds methenyl choloride 25ml, close plug, weighed weight, puts in water-bath refluxing extraction 2 hours, let cool, weighed weight again, supplies the weight of less loss with methenyl choloride, shake up, divide and get methenyl choloride liquid, with the filter paper that is covered with a small amount of anhydrous sodium sulfate, filter, discard just filtrate, precision measures subsequent filtrate 5ml, put in 10ml measuring bottle, add methyl alcohol to scale, shake up, obtain;

B. the assay of iron powder:

Get Tibetan medicinal composition Jiedu Tablets powder 8g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning, blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 1ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, blazing 4 hours in 600 ℃, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, 65 ℃ of left and right heating for dissolving 4 hours, after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 5ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, can add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 4ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, use immediately dichromate titration liquid (0.002mol/l) titration.Every 1ml dichromate titration liquid (0.002mol/l) is equivalent to the iron of 0.6702mg.

This product iron content is 0.409mg/g-0.682mg/g.

C. the assay of safflower:

The preparation of need testing solution: get Tibetan medicinal composition Jiedu Tablets powder 5g, accurately weighed, put in tool plug conical flask, precision adds volume parts than 25% methanol aqueous solution 25ml, close plug, weighed weight, ultrasonic 40 minutes, let cool, more weighed weight, the weight of less loss by volume parts, than 25% methanol aqueous solution, supplied, shake up, filter, get subsequent filtrate, obtain;

D. the assay of calculus bovis factitius:

The preparation of need testing solution: get Tibetan medicinal composition Jiedu Tablets powder 7g, accurately weighed, to put in the conical flask of tool plug, precision adds methyl alcohol 50ml, close plug, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, with methyl alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate 25ml, 40 ℃ of following recovered under reduced pressure, are transferred in 5ml measuring bottle, add methanol constant volume to scale, filter, get subsequent filtrate, obtain;

Described Tibetan medicinal composition Jiedu Tablets refers to the precious 4.0g of detoxicating capsule bulk drug formula animal, Radix Brassicae rapae 4.0g, earthworm fruit mustard 30.4g, vomiting nut (system) 1.6g, German tamarisk 30.4g, borax (system) 0.8g, pearl (system) 3.2g, coral 1.6g, lazurite (system) 1.6g, turquoise (system) 1.6g, iron powder (system) 2.4g, ring copper (system) 0.8g, parmelia saxatilis (system) 0.8g, ichor cream 4.0g, nutmeg 1.6g, tsaoko 1.6g, Tabasheer 1.6g, safflower 1.6g, cloves 1.6g, myrobalan 2.4g, Phytolacca acinosa 1.6g, santal 30.4g, santal 27.9g, Moschus 1.6g, calculus bovis factitius 2.4g, whorlleaf oxytropis herb 30.4g, grass-leaved sweetflag 1.6g, cardamom 1.6g, hooker winghead root 30.4g, west szechwan pyrethrum 30.4g, rhizome of davallia 3.2g, rhizoma nardostachyos 1.6g, Corallodiscus flabellatus (Franch.) Burtt 30.4g, alpine gentian herb 30.4g, muscicolous woodbetony root 30.4g, add conventional auxiliary material, the tablet of preparing according to common process.

Claims

1. the quality determining method of a Tibetan medicinal composition detoxicating capsule, the bulk drug of said preparation consists of animal treasured, Radix Brassicae rapae, earthworm fruit mustard, Semen Strychni (processed), German tamarisk, borax processed, pearl processed, coral, lazurite processed, turquoise processed, Magnetite (processed), system is rung copper, parmelia saxatilis processed, ichor cream, nutmeg, tsaoko, Tabasheer, safflower, cloves, myrobalan, Phytolacca acinosa, santal, santal, Moschus, calculus bovis factitius, whorlleaf oxytropis herb, grass-leaved sweetflag, cardamom, hooker winghead root, west szechwan pyrethrum, the rhizome of davallia, rhizoma nardostachyos, Corallodiscus flabellatus (Franch.) Burtt, alpine gentian herb, muscicolous woodbetony root, it is characterized in that, quality determining method comprises following content assaying method:

A. the assay of vomiting nut: get respectively strychnine reference substance, strychnia reference substance is contrast, octadecylsilane chemically bonded silica is filling agent; Take volume parts than acetonitrile-0.01mol/l sodium heptanesulfonate of 15-25:85-75 with 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution and be mobile phase by volume parts than 10% phosphorus acid for adjusting pH value 2.8; The high performance liquid chromatography that detection wavelength is 250-270nm;

B. the assay of iron powder: by after Chinese medicine preparation ashing to be determined, with concentrated hydrochloric acid, dissolve, add methyl orange indicator solution, drip stannous chloride solution Fe3+ is reduced to Fe2+, solution becomes pale red, then take diphenylamine sulfonic acid sodium salt as indicator, with potassium dichromate standard solution, be titrated to solution and be purple and be terminal, with the amount of known potassium dichromate standard solution, calculate the content of iron powder;

C. the assay of safflower: get hydroxyl safflower anthocyanidin A reference substance and add volume parts and be mixed with reference substance solution than 25% methanol aqueous solution, get preparation powder to be detected and add volume parts after ultrasonic dissolution, removal of impurities, to be mixed with need testing solution than 25% methanol aqueous solution, take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than methyl alcohol-volume parts of 20-30:80-70 than 0.5% aqueous formic acid, the high performance liquid chromatography that detection wavelength is 400-410nm;

D. the assay of calculus bovis factitius: get cholic acid reference substance and add methyl alcohol and be mixed with reference substance solution, get preparation powder to be detected and add methyl alcohol to be mixed with need testing solution after ultrasonic dissolution, removal of impurities, take octadecylsilane chemically bonded silica as filling agent; The volume parts of take is mobile phase than methyl alcohol-volume parts of 80-70:20-30 than 1% glacial acetic acid aqueous solution, the high performance liquid chromatography of evaporative light-scattering detector;

The content assaying method concrete steps of described iron powder are:

Get preparation powder to be detected 6～10g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning, blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 0.5-2ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, in 500-700 ℃ of blazing 3-5 hour, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, at 60-70 ℃ of left and right heating for dissolving 3-5 hour, after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 0-15ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, can add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 2-10ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, use immediately 0.002mol/l potassium dichromate solution titration titration.

2. quality determining method as claimed in claim 1, is characterized in that, the assay concrete steps of described vomiting nut are:

According to high effective liquid chromatography for measuring;

Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take volume parts than acetonitrile-0.01mol/l sodium heptanesulfonate of 15-25:85-75 with 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution and be mobile phase by volume parts than 10% phosphorus acid for adjusting pH value 2.8; Detection wavelength is 250-270nm; Number of theoretical plate calculates and should be not less than 5000 by strychnine peak;

The preparation of reference substance solution: get strychnine reference substance 6mg, strychnia reference substance 3mg, accurately weighed, put respectively in 100ml measuring bottle, add methenyl choloride and make in right amount to dissolve and be diluted to scale, shake up; Precision measures 2ml respectively, puts in same 10ml measuring bottle, with methyl alcohol, is diluted to scale, shakes up, and obtains;

The preparation of need testing solution: get preparation powder 10-14g to be detected, accurately weighed, put in tool plug conical flask, hydro-oxidation sodium test solution 10ml, mixes, and places 30 minutes, precision adds methenyl choloride 25ml, close plug, weighed weight, puts in water-bath refluxing extraction 2 hours, let cool, weighed weight again, supplies the weight of less loss with methenyl choloride, shake up, divide and get methenyl choloride liquid, with the filter paper that is covered with a small amount of anhydrous sodium sulfate, filter, discard just filtrate, precision measures subsequent filtrate 5ml, put in 10ml measuring bottle, add methyl alcohol to scale, shake up, obtain;

3. quality determining method as claimed in claim 1, is characterized in that, the assay concrete steps of described safflower are:

According to high effective liquid chromatography for measuring;

The preparation of need testing solution: get preparation powder 4-6g to be detected, accurately weighed, put in tool plug conical flask, precision adds volume parts than 25% methanol aqueous solution 20-30ml, close plug, weighed weight, ultrasonic 40 minutes, let cool, more weighed weight, the weight of less loss by volume parts, than 25% methanol aqueous solution, supplied, shake up, filter, get subsequent filtrate, obtain;

4. quality determining method as claimed in claim 1, is characterized in that, the concrete steps of described calculus bovis factitius assay are:

According to high effective liquid chromatography for measuring;

The preparation of need testing solution: get preparation powder 6-8g to be detected, accurately weighed, to put in the conical flask of tool plug, precision adds methyl alcohol 40-60ml, close plug, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, with methyl alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate 20-30ml, 40 ℃ of following recovered under reduced pressure, are transferred in 5ml measuring bottle, add methanol constant volume to scale, filter, get subsequent filtrate, obtain;

5. quality determining method as claimed in claim 1, is characterized in that, the bulk drug of described preparation consists of: precious 4.0 weight portions of animal, Radix Brassicae rapae 4.0 weight portions, earthworm fruit mustard 30.4 weight portions, Semen Strychni (processed) 1.6 weight portions, German tamarisk 30.4 weight portions, borax 0.8 weight portion processed, pearl 3.2 weight portions processed, coral 1.6 weight portions, lazurite 1.6 weight portions processed, turquoise 1.6 weight portions processed, Magnetite (processed) 2.4 weight portions, system is rung copper 0.8 weight portion, parmelia saxatilis 0.8 weight portion processed, ichor cream 4.0 weight portions, nutmeg 1.6 weight portions, tsaoko 1.6 weight portions, Tabasheer 1.6 weight portions, safflower 1.6 weight portions, cloves 1.6 weight portions, myrobalan's 2.4 weight portions, Phytolacca acinosa 1.6 weight portions, santal 30.4 weight portions, santal 27.9 weight portions, Moschus 1.6 weight portions, calculus bovis factitius 2.4 weight portions, whorlleaf oxytropis herb 30.4 weight portions, grass-leaved sweetflag 1.6 weight portions, cardamom 1.6 weight portions, hooker winghead root 30.4 weight portions, west szechwan pyrethrum 30.4 weight portions, the rhizome of davallia 3.2 weight portions, rhizoma nardostachyos 1.6 weight portions, Corallodiscus flabellatus (Franch.) Burtt 30.4 weight portions, alpine gentian herb 30.4 weight portions, muscicolous woodbetony root 30.4 weight portions,

Described quality determining method is:

A. the assay of vomiting nut:

According to high effective liquid chromatography for measuring;

Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take volume parts than acetonitrile-0.01mol/l sodium heptanesulfonate of 20:80 with 0.02mol/l potassium dihydrogen phosphate mixed in equal amounts solution and be mobile phase by volume parts than 10% phosphorus acid for adjusting pH value 2.8; Detection wavelength is 260nm; Number of theoretical plate calculates and should be not less than 5000 by strychnine peak;

The preparation of need testing solution: get preparation powder 12g to be detected, accurately weighed, put in tool plug conical flask, hydro-oxidation sodium test solution 10ml, mixes, and places 30 minutes, precision adds methenyl choloride 25ml, close plug, weighed weight, puts in water-bath refluxing extraction 2 hours, let cool, weighed weight again, supplies the weight of less loss with methenyl choloride, shake up, divide and get methenyl choloride liquid, with the filter paper that is covered with a small amount of anhydrous sodium sulfate, filter, discard just filtrate, precision measures subsequent filtrate 5ml, put in 10ml measuring bottle, add methyl alcohol to scale, shake up, obtain;

Determination method: draw respectively each 10 μ l of reference substance solution and need testing solution, injection liquid chromatography, measures, and obtains;

B. the assay of iron powder:

Get Tibetan medicinal composition detoxicating capsule content or preparation powder 8g, accurately weighed, the crucible that fills sample is put on electric furnace to slowly vehement burning, blazingly to the whole charings of test sample, be black, and no longer smolder, let cool to room temperature, drip mass percentage concentration 95%-98% concentrated sulphuric acid 1ml, make carbide all moistening, continuing to be heated to steam eliminates, white cigarette emits to the greatest extent, blazing 4 hours in 600 ℃, let cool, with 50ml mass percentage concentration 36%-38% concentrated hydrochloric acid gradation washing, move in 250ml tool plug conical flask, 65 ℃ of left and right heating for dissolving 4 hours, after cooling, solution is moved in 250ml volumetric flask, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, be diluted with water to scale, shake up, filter, precision measures subsequent filtrate 25ml, put in 250ml conical flask, add mass percentage concentration 36%-38% concentrated hydrochloric acid 5ml, be heated to 70-80 ℃, add after 6 methyl orange indicator solutions, while shaking, drip while hot concentration expressed in percentage by weight 5% stannous chloride solution to solution and become pale pink, if shaking rear pink colour takes off, add 1 methyl orange to solution and present stable pale pink, with cold water, be cooled to after room temperature rapidly, add distilled water 50ml, mixture of sulfuric phosphoric acid solution 4ml, 15 of diphenylamine sulfonic acid sodium salt indicator solutions, fixed with 0.002mol/l dichromate titration drop immediately,

C. the assay of safflower:

According to high effective liquid chromatography for measuring;

D. the assay of calculus bovis factitius:

According to high effective liquid chromatography for measuring;

The preparation of need testing solution: get preparation powder 7g to be detected, accurately weighed, to put in the conical flask of tool plug, precision adds methyl alcohol 50ml, close plug, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, with methyl alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate 25ml, 40 ℃ of following recovered under reduced pressure, are transferred in 5ml measuring bottle, add methanol constant volume to scale, filter, get subsequent filtrate, obtain;