CN102323345A

CN102323345A - Detection method of Houttuynia herb injection

Info

Publication number: CN102323345A
Application number: CN201110208803A
Authority: CN
Inventors: 毕开顺; 吴飞驰; 潘红炬; 李清; 仇萍; 朱雅宁; 李苏翠; 吴建国
Original assignee: ZHENGQING PHARMACEUTICAL GROUP CORP Ltd HUNAN PROV; Yaan Sanjiu Pharmaceutical Co Ltd
Current assignee: ZHENGQING PHARMACEUTICAL GROUP CORP Ltd HUNAN PROV; Yaan Sanjiu Pharmaceutical Co Ltd
Priority date: 2011-07-25
Filing date: 2011-07-25
Publication date: 2012-01-18

Abstract

The invention discloses a quality detection method of a Houttuynia herb injection. The method comprises content measurement, identification and/or fingerprint detection processes. The method is a gas chromatography, comprising the following steps of: taking a 4-terpineol reference substance, an alpha-terpinol reference substance, a bronyl acetate reference substance and methylnnonylketone, respectively adding n-hexane to prepare a reference substance solution; adding the n-hexane in the Houttuynia herb injection, adding anhydrous sulfate to prepare a test sample solution; respectively absorbing the reference substance solution and the test sample solution; injecting in the gas chromatography to test; and detecting according to the method of content measurement items, and recording a chromatogram, wherein the similarity of the test sample fingerprint and the reference fingerprint is not less than 0.85. The invention further provides a detection method of polysorbate 80 limit test in the Houttuynia herb injection, an identification method of thin-layer chromatography and a detection method of essential oil content in Houttuynia herb medical materials or injections. The quality detection method provided by the invention has good specificity, good linear relation, good instrument precision and good repeatability.

Description

A kind of detection method of houttuynia cordata injection

Technical field

The present invention relates to a kind of quality determining method of parenteral solution, particularly a kind of quality determining method of houttuynia cordata injection.

Background technology

Houttuynia is Saururaceae perennial herb Houttuynia Houttuynia? Cordata? Thunb.'s Whole plant with roots, also known as ear root, purple Ji, pig nose, nine lotus, with its fresh, pungent after crushing plant stench, widely distributed in Central, Southeast and Southwest provinces, especially in Hunan, Hubei, Sichuan, Jiangsu and other provinces majority.That cordate houttuynia has is clearing heat and detoxicating, the effect of the carbuncle that disappears apocenosis, inducing diuresis for treating strangurtia, the clinical illnesss such as lung carbuncle pyemesis, phlegm heat panting are coughed, hot dysentery, heat pouring, carbuncle sore tumefacting virus that are used for.Research shows that the volatile ingredient that contains in the cordate houttuynia has compositions such as flavonoids, organic acid, sterols, alkaloids.Houttuynia cordata injection is to get through adding auxiliary material sterilization preparation behind the steam distillation, and principal ingredient is its volatile ingredient.

Houttuynia cordata injection is the sterile water solution that bright cordate houttuynia is processed through steam distillation.Official quality standands comes from the 17 WS3-B-3264-98 of ministerial standard " Chinese traditional patent formulation preparation "; And national drug standards revision official written reply 2001ZB0086; The assay item is only measured a kind of composition of methylnonanone in the official quality standands; And only stipulated lower lower limit; Standard do not record the control item of traditional Chinese medicine fingerprint, the oil mass of volatilizing control, differentiate that item has only recorded that two specificity not strong chemical reactions are differentiated and the thin layer discrimination test of a single component, existing legal quality fails effectively the physicochemical property and the validity of product are control effectively.

Summary of the invention

The object of the invention is to disclose a kind of quality determining method of houttuynia cordata injection.

The present invention seeks to realize through following scheme:

The quality determining method of houttuynia cordata injection of the present invention comprises following assay, discriminating and/or fingerprint atlas detection method:

A. the content assaying method that 4 kinds of compositions detect simultaneously in the houttuynia cordata injection is: chromatographic condition and system suitability test, and 30m * 0.25mm, 0.25 μ mDB-1 capillary column is a chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes; Per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID; Detector temperature is 280 ℃, and carrier gas is N2, and flow rate of carrier gas is 1.0ml/min; Split ratio is 5～15: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 1～4mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation respectively, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 8～12mg, 4～6mg, 6～10mg, 1～4mg, precision is measured each reference substance storing solution respectively, adds normal hexane and processes the mixed solution that every 1ml contains 1mg, 0.5mg, 0.8mg, 1mg approximately; As mixing the reference substance storing solution, get and mix reference substance storing solution 0.5ml, put in the 2ml measuring bottle; The accurate 0.2ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get; Houttuynia cordata injection 50ml is got in the need testing solution preparation, puts in the round-bottomed flask, connects volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1ml contains alpha-terpineol (C ₈H ₁₈O) should be 2.0～10.0 μ g, methylnonanone (C ₁₁H ₂₂O) should be 6.0～20.0 μ g, 4-terpilenol (C ₈H ₁₈O) should be not less than 5.0 μ g, Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 0.8 μ g;

B. the content assaying method that 4 kinds of compositions detect simultaneously in the cordate houttuynia medicinal material is: chromatographic condition and system suitability test, and 30m * 0.25mm, 0.25 μ mDB-1 capillary column is as chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes; Per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID; Detector temperature is 280 ℃, and carrier gas is N2, and flow rate of carrier gas is 1.0ml/min; Split ratio is 5～15: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 1～4mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 0.5～1.5mg, 0.2～1mg, 6～10mg, 8～12mg; Precision pipettes each reference substance storing solution 2ml, 1ml, 1ml, 4ml in the 10ml measuring bottle respectively, and the accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale; Shake up, promptly get; The cordate houttuynia medicinal material is got in the need testing solution preparation, shreds, and gets 45～55g, and accurate the title decides; Put in the 250ml round-bottomed flask, add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, be heated to and boil, keep little and boiled 4 hours, be cooled to room temperature, obtain the normal hexane layer; Add the about 0.3～0.5g of anhydrous sodium sulfate, jolting, normal hexane liquid moves in the 10ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1g contains alpha-terpineol (C ₈H ₁₈O) should be not less than 1.5 μ g, contain methylnonanone (C ₁₁H ₂₂O) should be not less than 120 μ g, contain 4-terpilenol (C ₈H ₁₈O) should be not less than 3.0 μ g, contain Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 12.0 μ g;

C. houttuynia cordata injection fingerprint atlas detection method: detect the record chromatogram by method under the assay item; Integral parameter is set to excise preceding 4 minutes solvent peaks; Mark peak in removing, slope (Slope Sensitivity) is 5, peak width (Peak Width) is 0.1; Smallest peaks area (Area reject) is 4; Minimum peak height (Height reject) is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.85; Finger-print has 11 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.465 ± 0.080, No. 2 peaks 0.486 ± 0.049, No. 1 peak; 0.673 ± 0.067, No. 5 peaks 0.712 ± 0.071,0.557 ± 0.056, No. 4 peaks, No. 3 peaks; 0.810 ± 0.081, No. 8 peaks 0.828 ± 0.082,0.741 ± 0.074, No. 7 peaks, No. 6 peaks; No. 9 peaks 0.991 ± 0.099,1.000 ± 0.000, No. 10 peaks 1.554 ± 0.156, S peak; Relative peak area is 0.041 ± 0.004, No. 3 peaks 0.072 ± 0.007,0.062 ± 0.006, No. 2 peaks, No. 1 peak; 0.031 ± 0.003, No. 6 peaks 0.024 ± 0.002,0.066 ± 0.007, No. 5 peaks, No. 4 peaks; 0.463 ± 0.046, No. 9 peaks 0.185 ± 0.019,1.541 ± 0.154, No. 8 peaks, No. 7 peaks; 1.000 ± 0.000, No. 10 peaks 0.021 ± 0.002, S peak;

D. cordate houttuynia medicinal materials fingerprint detection method: detect the record chromatogram by method under the assay item; Integral parameter is set to excise preceding 4 minutes solvent peaks; Mark peak in removing, Slope Sensitivity is 5, Peak Width is 0.1; Area reject is 4; Height reject is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.90; Finger-print has 22 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.383 ± 0.038, No. 2 peaks 0.397 ± 0.040, No. 1 peak; 0.466 ± 0.047, No. 6 peaks 0.486 ± 0.049,0.457 ± 0.046, No. 5 peaks, 0.418 ± 0.042, No. 4 peaks, No. 3 peaks; 0.566 ± 0.057, No. 10 peaks 0.587 ± 0.059,0.556 ± 0.056, No. 9 peaks, 0.533 ± 0.053, No. 8 peaks, No. 7 peaks; 0.668 ± 0.067, No. 14 peaks 0.807 ± 0.081,0.661 ± 0.066, No. 13 peaks, 0.607 ± 0.061, No. 12 peaks, No. 11 peaks; 0.826 ± 0.083, No. 16 peaks 0.989 ± 0.099, No. 15 peaks, 1.000 ± 0.000, No. 17 peaks 1.088 ± 0.101, S peak; 1.346 ± 0.135, No. 21 peaks 1.567 ± 157,1.230 ± 0.123, No. 20 peaks, 1.134 ± 0.113, No. 19 peaks, No. 18 peaks; Relative peak area is 0.090 ± 0.009, No. 4 peaks 0.444 ± 0.044,0.606 ± 0.060, No. 3 peaks, 0.009 ± 0.001, No. 2 peaks, No. 1 peak; 0.023 ± 0.002, No. 8 peaks 0.354 ± 0.035,0.493 ± 0.049, No. 7 peaks, 1.113 ± 0.111, No. 6 peaks, No. 5 peaks; 0.036 ± 0.004, No. 12 peaks 0.021 ± 0.002,0.005 ± 0.001, No. 11 peaks, 0.022 ± 0.002, No. 10 peaks, No. 9 peaks; 0.011 ± 0.001, No. 16 peaks 0.136 ± 0.014,0.078 ± 0.008, No. 15 peaks, 0.012 ± 0.001, No. 14 peaks, No. 13 peaks; 0.017 ± 0.002, No. 18 peaks 0.061 ± 0.006,1.000 ± 0.000, No. 17 peaks, S peak; 0.134 ± 0.013, No. 21 peaks 0.029 ± 0.003,0.035 ± 0.004, No. 20 peaks, No. 19 peaks;

E. the detection method of polyoxyethylene sorbitan monoleate limit examine is in the houttuynia cordata injection: precision pipettes houttuynia cordata injection 1ml and puts in the conical flask; Get cobalt nitrate 5～8g, ammonium thiocyanate 35～45g is dissolved in water and is diluted to 200ml, shakes up; Process ammonium cobalt thiocyanate salt solution, 10～20ml puts in the conical flask with ammonium cobalt thiocyanate salt solution, and the accurate methylene chloride 8～12ml that adds claims to decide weight; With oscillator vibration 15 minutes, take out, supply the weight that subtracts mistake with methylene chloride, move in the separating funnel; Left standstill 15 minutes, and obtained dichloromethane layer solution, as need testing solution; Other gets the polyoxyethylene sorbitan monoleate reference substance to the 10ml measuring bottle, and accurate the title decides, and adds redistilled water and processes the solution that every 1ml contains 2.5mg, as reference substance solution; Get need testing solution and reference substance solution respectively, put into ultraviolet spectrophotometer, under the 623nm wavelength, measure absorbance log, calculate, promptly get; Contain polyoxyethylene sorbitan monoleate and must not cross 0.27%;

F. the discrimination method of cordate houttuynia medicinal material thin-layer chromatography is: get cordate houttuynia medicinal material 125g, shred, put in the 250ml round-bottomed flask, add water 150ml; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml, connect reflux condensing tube; Slowly be heated to and boil, and keep little and boiled 4 hours, place half an hour, get the normal hexane layer; To the 2ml measuring bottle, add normal hexane and be diluted to scale, as need testing solution; Other gets 4-terpilenol reference substance, alpha-terpineol reference substance and methylnonanone reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 0.8～1.5mg, 0.3～0.8mg, 1.0～3.0mg, as reference substance solution; Drawing above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, is to be developping agent at 15～21: 3 with cyclohexane-ethyl acetate ratio; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

G. houttuynia cordata injection or wherein between the discrimination method of product thin-layer chromatography be: get houttuynia cordata injection 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate, jolting; Normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, as need testing solution; Other gets methylnonanone, alpha-terpineol reference substance and 4-terpilenol reference substance reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 0.8～1.5mg, 0.3～0.8mg, 1.0～3.0mg, as reference substance solution; Drawing above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, is to be developping agent at 15～21: 3 with the ratio of cyclohexane-ethyl acetate; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

H. the detection method of volatile oil content is in the cordate houttuynia medicinal material: get cordate houttuynia medicinal material 500g, the accurate title, decide, and puts in the 2000ml round-bottomed flask; Add water 1000ml and beaded glass number, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Connect reflux condensing tube, slowly be heated to and boil, keep little and boiled 5 hours; Be cooled to room temperature, calculate the total amount that these article contain volatile oil; Contain volatile oil and must not be less than 0.03% (ml/g);

I. houttuynia cordata injection or wherein between in the product detection method of volatile oil content be: get houttuynia cordata injection or wherein between product 500ml, put in the 2000ml round-bottomed flask, add beaded glass number; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube; Slowly be heated to and boil; Keep little and boiled 35～45 minutes, be cooled to room temperature, calculate the total amount that these article contain volatile oil.

The quality determining method of houttuynia cordata injection of the present invention, preferred following assay, discriminating and/or fingerprint atlas detection method:

A. the content assaying method that 4 kinds of compositions detect simultaneously in the houttuynia cordata injection is: chromatographic condition and system suitability test, and 30m * 0.25mm, 0.25 μ mDB-1 capillary column is a chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes; Per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID; Detector temperature is 280 ℃, and carrier gas is N2, and flow rate of carrier gas is 1.0ml/min; Split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation respectively, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 10mg, 5mg, 8mg, 2mg, precision is measured each reference substance storing solution respectively, adds normal hexane and processes the mixed solution that every 1ml contains 1mg, 0.5mg, 0.8mg, 1mg approximately; As mixing the reference substance storing solution, get and mix reference substance storing solution 0.5ml, put in the 2ml measuring bottle; The accurate 0.2ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get; Houttuynia cordata injection 50ml is got in the need testing solution preparation, puts in the round-bottomed flask, connects volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1ml contains alpha-terpineol (C ₈H ₁₈O) should be 2.0～10.0 μ g, methylnonanone (C ₁₁H ₂₂O) should be 6.0～20.0 μ g, 4-terpilenol (C ₈H ₁₈O) should be not less than 5.0 μ g, Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 0.8 μ g;

B. the content assaying method that 4 kinds of compositions detect simultaneously in the cordate houttuynia medicinal material is: chromatographic condition and system suitability test, and 30m * 0.25mm, 0.25 μ mDB-1 capillary column is as chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes; Per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID; Detector temperature is 280 ℃, and carrier gas is N2, and flow rate of carrier gas is 1.0ml/min; Split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 1mg, 0.5mg, 8mg, 10mg; Precision pipettes each reference substance storing solution 2ml, 1ml, 1ml, 4ml in the 10ml measuring bottle respectively, and the accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale; Shake up, promptly get; The cordate houttuynia medicinal material is got in the need testing solution preparation, shreds, and gets 50g, and accurate the title decides; Put in the 250ml round-bottomed flask, add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, be heated to and boil, keep little and boiled 4 hours, be cooled to room temperature, obtain the normal hexane layer; Add the about 0.4g of anhydrous sodium sulfate, jolting, normal hexane liquid moves in the 10ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1g contains alpha-terpineol (C ₈H ₁₈O) should be not less than 1.5 μ g, contain methylnonanone (C ₁₁H ₂₂O) should be not less than 120 μ g, contain 4-terpilenol (C ₈H ₁₈O) should be not less than 3.0 μ g, contain Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 12.0 μ g;

C. houttuynia cordata injection fingerprint atlas detection method: detect the record chromatogram by method under the assay item; Integral parameter is set to excise preceding 6 minutes solvent peaks; Mark peak in removing, slope (Slope Sensitivity) is 5, peak width (Peak Width) is 0.1; Smallest peaks area (Area reject) is 4; Minimum peak height (Height reject) is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.85; Finger-print has 11 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.465, No. 2 peak 0.486, No. 1 peak; 0.673, No. 5 peak 0.712,0.557, No. 4 peak, No. 3 peaks; 0.810, No. 8 peak 0.828,0.741, No. 7 peak, No. 6 peaks; No. 9 peaks 0.991,1.000, No. 10 peaks 1.554, S peak; Relative peak area is 0.463, No. 9 peak 0.185,1.541, No. 8 peaks, 0.024, No. 7 peak, 0.031, No. 6 peak, 0.066, No. 5 peak, 0.072, No. 4 peak, 0.041, No. 3 peak, 0.062, No. 2 peak, No. 1 peak, 1.000, No. 10 peaks 0.021, S peak (concrete numerical value is seen table 1, accompanying drawing 1);

Table 1 houttuynia cordata injection finger-print each peak relative retention time and relative peak area

D. cordate houttuynia medicinal materials fingerprint detection method: detect the record chromatogram by method under the assay item; Integral parameter is set to excise preceding 6 minutes solvent peaks; Mark peak in removing, Slope Sensitivity is 5, Peak Width is 0.1; Area reject is 4; Height reject is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.90; Finger-print has 22 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.397, No. 3 peak 0.418,0.383, No. 2 peak, No. 1 peak; 0.533, No. 8 peak 0.556,0.486, No. 7 peak, 0.466, No. 6 peak, 0.457, No. 5 peak, No. 4 peaks; 0.661, No. 13 peak 0.668,0.607, No. 12 peak, 0.587, No. 11 peak, 0.566, No. 10 peak, No. 9 peaks; 0.826, No. 16 peak 0.989,0.807, No. 15 peak, No. 14 peaks, 1.000, No. 17 peaks 1.088, S peak; 1.346, No. 21 peaks 1.567,1.230, No. 20 peaks, 1.134, No. 19 peaks, No. 18 peaks; Relative peak area is 1.113, No. 6 peaks 0.493,0.444, No. 5 peak, 0.090, No. 4 peak, 0.606, No. 3 peak, 0.009, No. 2 peak, No. 1 peak; 0.036, No. 12 peak 0.021,0.005, No. 11 peak, 0.022, No. 10 peak, 0.354, No. 9 peak, 0.023, No. 8 peak, No. 7 peaks; 0.011, No. 16 peak 0.136,0.078, No. 15 peak, 0.012, No. 14 peak, No. 13 peaks, S peak 1.000; 0.134, No. 21 peak 0.029,0.035, No. 20 peak, 0.061, No. 19 peak, 0.017, No. 18 peak, No. 17 peaks (concrete numerical value is seen table 2, Fig. 2);

Table 2 houttuynia cordata injection finger-print each peak relative retention time and relative peak area

E. the detection method of polyoxyethylene sorbitan monoleate limit examine is in the houttuynia cordata injection: precision pipettes houttuynia cordata injection 1ml and puts in the conical flask; Get cobalt nitrate 6g, ammonium thiocyanate 40g is dissolved in water and is diluted to 200ml, shakes up; Process ammonium cobalt thiocyanate salt solution, 15ml puts in the conical flask with ammonium cobalt thiocyanate salt solution, and the accurate methylene chloride 10ml that adds claims to decide weight; With the oscillator 15min that vibrates, take out, supply the weight that subtracts mistake with methylene chloride, move in the separating funnel; Leave standstill 15min, obtain dichloromethane layer solution, as need testing solution; Other gets the polyoxyethylene sorbitan monoleate reference substance to the 10ml measuring bottle, and accurate the title decides, and adds redistilled water and processes the solution that every 1ml contains 2.5mg, as reference substance solution; Get need testing solution and reference substance solution respectively, put into ultraviolet spectrophotometer, under the 623nm wavelength, measure absorbance log, calculate, promptly get;

F. the discrimination method of cordate houttuynia medicinal material thin-layer chromatography is: get cordate houttuynia medicinal material 125g, shred, put in the 250ml round-bottomed flask, add water 150ml; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml, connect reflux condensing tube; Slowly be heated to and boil, and keep little and boiled 4 hours, place half an hour, get the normal hexane layer; To the 2ml measuring bottle, add normal hexane and be diluted to scale, as need testing solution; Other gets 4-terpilenol reference substance, alpha-terpineol reference substance and methylnonanone reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 10.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate ratio be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

H. the detection method of volatile oil content is in the cordate houttuynia medicinal material: get cordate houttuynia medicinal material 500g; The accurate title, decide; Put in the 2000ml round-bottomed flask, add water 1000ml and bead number, connect volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part; Connect reflux condensing tube, slowly be heated to and boil, keep little and boiled 5 hours; Be cooled to room temperature, calculate the total amount that this product contains volatile oil;

I. houttuynia cordata injection or wherein between in the product detection method of volatile oil content be: get houttuynia cordata injection or wherein between product 500ml, put in the 2000ml round-bottomed flask, add beaded glass number; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube; Slowly be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, calculate the total amount that these article contain volatile oil.

Houttuynia cordata injection content assaying method of the present invention is easy and simple to handle with the volatile oil extractor method for distilling, cost is low, extracts the composition to be determined in the parenteral solution with volatile oil extractor, and reference substance and inclusion-free peak, interior mark retention time place disturb; Other compositions are noiseless to measuring in the need testing solution; The method specificity is good, and each component is good in its respective range internal linear relation, and instrument precision is good; Repeatability is good, and need testing solution is stable in 24h.The finger-print of cordate houttuynia medicinal material of the present invention and parenteral solution detects; Instrument precision is good; Have good reappearance, RSD＜5%, need testing solution are stable in 48h; With methylnonanone is interior reference peak, and the relative retention time and the relative peak area of each fingerprint peaks of each batch medicinal material all have reappearance preferably.The limit examine of polyoxyethylene sorbitan monoleate in the houttuynia cordata injection, reference substance solution and need testing solution scan respectively in 400～700nm wavelength coverage after chromogenic reaction; The result finds that the polyoxyethylene sorbitan monoleate reference substance solution is consistent with houttuynia cordata injection need testing solution peak shape; Maximum absorption wavelength is consistent, is 623nm, and houttuynia cordata injection and reference substance all have better absorption; Show that other component is noiseless to this test determination result in the houttuynia cordata injection; Negative control solution and blank solution do not have absorption basically in the 623nm wavelength, and polyoxyethylene sorbitan monoleate is good linear relationship with absorbance in 1.02～5.10mg scope, and instrument precision is good; Repeatability is good, and is stable in the 3h after colour developing according to article solution and need testing solution; This method is quick, easy, can effectively control the quality of polyoxyethylene sorbitan monoleate in the houttuynia cordata injection, in being suitable for houttuynia cordata injection, the assay of polyoxyethylene sorbitan monoleate, also is suitable for the traditional Chinese medicine that other contain polyoxyethylene sorbitan monoleate simultaneously.Cordate houttuynia medicinal material of the present invention and parenteral solution thin layer identification color spectrometry have fast, characteristics simply and easily.In the detection method of volatile oil content, methylnonanone content is tangible positive correlation in volatile oil content and the intermediate product in cordate houttuynia medicinal material of the present invention or the parenteral solution, and the volatile oil content of re-distilled liquid should cross 0.08%.

Description of drawings:

Fig. 1: houttuynia cordata injection reference fingerprint;

Fig. 2: cordate houttuynia medicinal material reference fingerprint;

Fig. 3: the GC/MS total ion current collection of illustrative plates of the bright careless volatile ingredient of cordate houttuynia;

Fig. 4: houttuynia injection sample original two dimensional schematic diagram data

Fig. 5: houttuynia cordata injection assay specificity chromatogram

A: blank solvent chromatogram B: mix reference substance+interior mark chromatogram C: sample+interior mark chromatogram IS: undecylene 1:4-terpenol 2: alpha-terpineol 3: Bronyl acetate 4: methylnonanone

Fig. 6: houttuynia cordata injection assay linear relationship curve

Fig. 7: specificity chromatogram

Fig. 7-1: blank solvent chromatogram Fig. 7-2: mix reference substance+interior mark chromatogram Fig. 7-3: sample+interior mark chromatogram

IS: undecylene 1:4-terpilenol 2: alpha-terpineol 3: Bronyl acetate 4: methylnonanone

The bright cordate houttuynia assay of Fig. 8 component lines sexual intercourse curve

Fig. 9: typical color spectrogram

A:HP-5 capillary column B:DB-1 capillary column C:DB-17 capillary column

Figure 10: 2 times of time chromatograms

Figure 11: the cordate houttuynia medicinal materials fingerprint relatively

Figure 12: the houttuynia cordata injection finger-print relatively

Figure 13: polyoxyethylene sorbitan monoleate limit examine blank, negative control, reference substance and test sample spectrogram

Figure 14: the linear line curve map that closes of polyoxyethylene sorbitan monoleate limit examine

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

The content assaying method of experimental example 1 cordate houttuynia medicinal material and parenteral solution

1.1 chemical constitution study

(bright grass is produced in the Hunan Huaihua to the bright grass of cordate houttuynia; Providing by Zhengqing Pharmaceutical Group Corp., Ltd., Hunan Prov.) the pure article volatile ingredient that under the regulation test condition, extracts carried out the GC-MS analysis; The total ion current collection of illustrative plates is seen Fig. 3, as can be seen from the figure, and the Herba Houttuyniae volatile oil complicated component; Contain nearly up to a hundred chromatographic peaks, and the bigger chromatographic peak of content just reaches tens.

Fig. 4 is the finger-print GC-MS collection of illustrative plates of houttuynia cordata injection finished product; Outflow chromatogram information that component can be provided based on the coupling chromatogram and mass spectrum information (its mass spectrum of structural property decision of material distributes) in different retention times; Utilize Chemical Measurement resolution techniques such as derivation formula evolution characteristic projection directly perceived and subwindow factorial analysis to obtain the mass spectrum of the bigger chromatographic component of content respectively; And then confirm its chemistry ownership, and provided the bigger component information of content proportion.

Table 3 houttuynia cordata injection GC-MS analysis bank submeter

1.2 solid phase extraction detects the houttuynia injection liquid hold-up

1.2.1 content assaying method

Chromatographic condition and system suitability test (14%) cyanogen propyl group phenyl-dimethyl polysiloxane capillary column (30m * 0.53m * 1.0um); Column temperature is temperature programme: initial temperature is 90 ℃, keeps 10 minutes, is warming up to 115 ℃ with the speed of 1 ℃ of per minute, and the speed with 20 ℃ of per minutes is warming up to 220 ℃ again, keeps 10 minutes; Carrier gas: nitrogen, flow velocity are per minute 6ml; Number of theoretical plate calculates by the methylnonanone peak and is not less than 20000.

It is an amount of that 4-methyl isophthalic acid-isopropyl-3-cyclohexene-1-alcohol, 2-(4-methyl-3-cyclohexenyl group) isopropyl alcohol, methylnonanone reference substance are got in the preparation of reference substance solution; The accurate title, decide; Add acetonitrile and process every 1ml and contain the solution that 4-methyl isophthalic acid-isopropyl-3-cyclohexene-1-alcohol, 2-(4-methyl-3-cyclohexenyl group) isopropyl alcohol, methylnonanone are respectively 250 μ g, 50 μ g, 30 μ g, promptly get.

The preparation precision of need testing solution is measured these article 25ml, places the C that handles well ₈Solid phase extraction column (100mg/ml is earlier with the activation of 10ml acetonitrile, water 10ml balance again) is gone up the speed wash-out with per minute 2～3ml, collects eluent 4.8ml, puts in the 5ml measuring bottle, adds acetonitrile to scale, shakes up, and filters, and gets subsequent filtrate, promptly gets.

Accurate respectively test sample and each the 1 μ l of reference substance solution of drawing of determination method, inject gas chromatograph is measured, and promptly gets.

1.2.2C ₁₈, C ₈The column extracting ability is investigated

1.2.2.1C ₁₈The column extracting ability is investigated

Test method: get the houttuynia cordata injection sample,, use C for the first time according to sample preparation in the assay ₁₈After solid phase extraction column (200mg/3ml) is crossed post, use C to filtrating again ₁₈Solid phase extraction column is crossed post for the second time, for the third time, and preparation supplies test agent, each accurate 1 μ l inject gas chromatograph respectively of drawing, and the record chromatogram, other gets reference substance solution 1 μ l, injects liquid chromatograph, calculates content with external standard method.

Table 4 is crossed post absorption situation for three times

Mark: component A-4 methyl isophthalic acid-isopropyl-3-cyclohexene-1-alcohol

B component-2 (4-methyl-3-cyclohexenyl group) isopropyl alcohol

Component C-methylnonanone

Result: C ₁₈Solid phase extraction column is once crossed post absorption not exclusively, and crossing post for the second time can adsorb fully, crosses post for the third time and can not survey content.C ₁₈Solid phase extraction column is strong to component C absorption affinity, almost can adsorb fully for the first time crossing post, to B component, C absorption affinity a little less than, can adsorb about 90% for the first time.

C ₁₈The solid phase extraction column durability is investigated

Test method: get the houttuynia cordata injection sample, according to sample preparation in the assay, with same C ₁₈Solid phase extraction column carries out repeatedly crossing the post preparation to sample and supplies test agent, and each accurate 1 μ l inject gas chromatograph respectively of drawing writes down each peak area.

Table 5C ₁₈The SPE peak area

Result: C ₁₈Solid phase extraction column uses 6 times result to show that to the 5th time, peak area obviously reduces, so C ₁₈Solid phase extraction column uses 4 times better.

1.2.2.2C ₈The column extracting ability is investigated

Test method: get houttuynia cordata injection sample (lot number: d907603),, use C for the first time according to sample preparation in the assay ₈After solid phase extraction column (100mg/ml) is crossed post, use C to filtrating again ₈Solid phase extraction column is crossed post for the second time, for the third time, and preparation supplies test agent, each accurate 1 μ l inject gas chromatograph respectively of drawing, and the record chromatogram, other gets reference substance solution 1 μ l, injects liquid chromatograph, calculates content with external standard method.

Table 6C ₈Column extracting absorption affinity situation

Result: C ₈Solid phase extraction column is the strongest to component C absorption affinity, almost can adsorb fully for the first time crossing post, and is stronger to B component, C absorption affinity, can adsorb 98% for the first time.

Get with batch pilot product (lot number: d907603), use C ₁₈Solid-phase extraction column pillar (200mg/3ml) substitutes C ₈Solid-phase extraction column pillar (100mg/ml) extracts, and the samely carries out for the first time, crosses post for the second time, for the third time, and preparation supplies test agent; Each accurate 1 μ l inject gas chromatograph respectively of drawing, the record chromatogram, other gets reference substance solution 1 μ l; Inject liquid chromatograph, calculate content with external standard method.

Table 7C ₁₈The solid-phase extraction column pillar is crossed the post situation three times

Brief summary: from above table result, find out C ₁₈Solid phase extraction column (200mg/3ml) is lower to the percentage extraction of two kinds of terpilenols in three kinds of major components in the houttuynia cordata injection, and residual quantity is higher in the waste liquid behind the single extraction, even still has small amount of residual in the waste liquid of reextraction.But C ₈Solid phase extraction column (100mg/ml) is all higher to three kinds of major component extracting powers in the houttuynia cordata injection, and all oneself surpasses 98% to the percentage extraction of single extraction, and C is described ₈Solid phase extraction column (100mg/ml) is than C ₁₈Solid phase extraction column (200mg/3ml) more is applicable to the assay of a plurality of compositions of houttuynia cordata injection,

C ₈The solid phase extraction column durability is investigated

Test method: get the houttuynia cordata injection sample, according to sample preparation in the assay, with same C ₈Solid phase extraction column carries out repeatedly crossing the post preparation to sample and supplies test agent, and each accurate 1 μ l inject gas chromatograph respectively of drawing writes down each peak area.

Table 8C ₈The solid phase extraction column result

Different manufacturers C ₈Post is investigated

Get pilot product (lot number: d907603), in accordance with the law use the C of three different labels respectively ₈Solid phase extraction column (100mg/1ml) extraction makes 3 parts of need testing solutions.Accurate each 1 μ l inject gas chromatograph respectively of drawing, the record chromatogram calculates content and RSD with external standard method.

The different label C of table 9 ₈The solid phase extraction column extracting power is measured the result

Brief summary: on show to show and find out, use the C of the different labels of same model ₈The solid phase extraction column extracting power differs greatly, and especially the difference of component A, B component is very big.

1.3. steam distillation method detects the content of houttuynia cordata injection

1.3.1. chromatographic condition

Chromatographic column: DB-1 capillary column (30m * 0.25mm, 0.25 μ m); Temperature programme: initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes, and per minute rises to 250 ℃ for 20 ℃; Injector temperature: 230 ℃; Detecting device: FID; Detector temperature: 280 ℃; Carrier gas: N ₂Flow rate of carrier gas: 1.0ml/min; Split ratio: 10: 1; Sample introduction: 1 μ l, number of theoretical plate calculate by the methylnonanone peak should be not less than 10000.

1.3.2. the preparation of solution

1.3.2.1. inner mark solution preparation

It is an amount of to get positive undecylene, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg.

1.3.2.2. reference substance solution preparation

It is an amount of to get 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance respectively, accurately claims surely, adds normal hexane respectively and processes the reference substance storing solution that every 1ml contains 10mg, 5mg, 8mg, 2mg; It is an amount of that precision is measured each reference substance storing solution respectively, adds normal hexane and process the mixed solution that every 1ml contains 1mg, 0.5mg, 0.8mg, 1mg approximately, as mixing the reference substance storing solution.Get and mix reference substance storing solution 0.5ml, put in the 2ml measuring bottle, the accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets.

1.3.2.3. need testing solution preparation

Get houttuynia cordata injection 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate, jolting; Normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate in right amount with normal hexane, and washing lotion is incorporated in the same measuring bottle, the accurate 0.2ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get.

1.3.3. parenteral solution assay condition is investigated:

1.3.3.1. extraction time: according to " 1.3.2.3 " need testing solution preparation, setting extraction time is 20min, 40min, 60min, 90min.

The different extraction times extraction of table 10 result

The result shows that extraction time, extraction effect was best when being 40min.

1.3.3.2. extracting mode: this experiment is investigated respectively and is adopted volatile oil extractor to extract C ₈Two kinds of methods of solid phase extraction column enrichment are extracted the volatile oil in the houttuynia cordata injection.

Table 11 Different Extraction Method is extracted the result

Explain: when investigating extracting mode.Volatile oil extractor extracts parenteral solution 50ml, solid phase extraction column applied sample amount parenteral solution 25ml.

The result shows that the volatile oil extractor extraction effect is better, because the volatile oil extractor method for distilling is easy and simple to handle, cost is low, so select for use volatile oil extractor to extract the composition to be determined in the parenteral solution.

1.3.4. methodology checking

1.3.4.1. specificity is investigated

Get blank solution (normal hexane), reference substance solution and need testing solution record the gas chromatogram (see figure 5), and the result is illustrated in reference substance and inclusion-free peak, interior mark retention time place disturbs, and other compositions are noiseless to measuring in the need testing solution, and the method specificity is good.

1.3.4.2. linearity and scope

Precision is measured and is mixed reference substance solution 0.05ml respectively, 0.1ml, and 0.25ml, 0.5ml, 1.0ml, 1.5ml put in the 2ml measuring bottle, and the accurate 0.2ml inner mark solution that adds adds the normal hexane constant volume, promptly gets serial mixing contrast solution, and sample introduction 1 μ l measures.Measuring the result is ordinate with reference substance and interior mark peak area ratio (A), is horizontal ordinate with each reference substance concentration (C), carries out linear regression, and each regression equation such as table 12, shown in Figure 6 show that each component is good in its respective range internal linear relation.

Table 12 regression equation data

1.3.4.3. precision test

The accurate reference substance solution of drawing, inject gas chromatograph, continuous sample introduction 5 times records the 4-terpenol; Alpha-terpineol, Bronyl acetate, the RSD of the peak area of methylnonanone and interior mark peak area ratio is respectively 0.3%, 0.7%; 1.4%, 1.6%, the result shows that instrument precision is good.

The test of table 13 instrument precision

1.3.4.4. replica test

Get 6 parts of same lot sample article respectively, every part of 50.0ml is equipped with solution according to " 2.3 " below legal system, sample introduction 1 μ l, and the RSD that records each component content is respectively 1.8%, 1.4%, 1.4%, 2.1%, shows that method repeatability is good.

Table 14 reperformance test

1.3.4.5 stability test

Get same test sample, measure respectively at 0h, 1h, 1.5h, 12h, 15h, 24h, the RSD of each composition peak area to be determined is respectively 1.9%, 2.3%, 0.8%, 0.5%.Show that need testing solution is stable in 24h.

Table 15 stability test

1.3.4.6. recovery test

Get 9 parts of same batch of parenteral solutions, every part of precision is measured 25ml, and it is an amount of to add the reference substance storing solution respectively, is equipped with solution according to " 2.3 " below legal system, and sample introduction 1 μ l calculates the recovery of each component.

Table 16 recovery data

1.3.5. sample determination

Get two producer's houttuynia cordata injections, totally 21 batches, be equipped with solution by " 1.2.3 " below legal system, sample introduction is measured, and calculates 4 kinds of component contents of each batch.

4 kinds of component contents of table 17 houttuynia cordata injection

1.4. cordate houttuynia medicinal material assay

1.4.1. chromatographic condition and system suitability test

30m * 0.25mm, 0.25 μ mDB-1 capillary column are as chromatographic column, and initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃; Kept 5 minutes, per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃; Detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000.

1.4.2. the preparation of solution

1.4.2.1. inner mark solution preparation

Get positive undecylene, the accurate title, decide, and adds normal hexane and process the solution that every 1ml contains 2mg.

1.4.2.2. reference substance solution preparation

Get 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance; The accurate title, decide, and adds normal hexane respectively and process the reference substance storing solution that every 1ml contains 1mg, 0.5mg, 8mg, 10mg, and precision pipettes each reference substance storing solution 2ml, 1ml, 1ml, 4ml in the 10ml measuring bottle respectively; The accurate 1ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get;

14.2.3. need testing solution preparation

Get the cordate houttuynia medicinal material, shred, get 50g; The accurate title, decide, and puts in the 250ml round-bottomed flask, adds water 150ml; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add n-hexane 1ml; Connect reflux condensing tube; Be heated to and boil, keep little 4h that boils, be cooled to room temperature; Obtain the n-hexane layer; Add the about 0.4g of anhydrous sodium sulfate, jolting, n-hexane liquid moves in the 10ml measuring bottle; And wash anhydrous sodium sulfate with n-hexane; Washing lotion is incorporated in the same measuring bottle, and the accurate 1ml inner mark solution that adds adds n-hexane and is diluted to scale; Shake up, promptly get; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets.

1.4.3. measure

Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets.

1.4.4. medicinal material assay condition is investigated

1.4.4.1. extraction time

Prepare need testing solution down according to 4.2.2.3 preparation item, extraction time is set at 3h respectively, 4h, and 5h, three kinds of composition peak areas to be measured see Table 18.

Table 18 three kinds of composition peak areas to be measured of different extraction times

Table 18 shows that cordate houttuynia is best at 4 hours extraction effects, so confirm that extraction time is 4 hours.

1.4.4.2. amount of water

Prepare need testing solution down according to 4.2.2.3 preparation item, amount of water is set at 3 times of water gagings (150ml), 4 times of water gagings (200ml), and three kinds of composition peak areas to be measured of 5 times of water gagings (250ml) are seen table 19.

Three kinds of composition peak areas to be measured of the different amount of water of table 19

Table 19 shows that cordate houttuynia medicinal material amount of water is that 3 times of amount extraction effects are best.

1.4.4.3. specificity is investigated

Get blank solution (normal hexane), reference substance solution and need testing solution record the gas chromatogram (see figure 7), and the result is illustrated in reference substance and inclusion-free peak, interior mark retention time place disturbs, and other composition is noiseless to measuring in the need testing solution, and the method specificity is good.

1.4.4.4. linearity and scope

Measure respectively and mix reference substance solution 0.1ml, 0.2ml, 0.4ml, 0.5ml, 1ml, 1.5ml put in the 2ml volumetric flask, add the 0.2ml inner mark solution, and the normal hexane constant volume promptly gets serial mixing reference substance solution.With reference substance and interior mark peak area ratio (A) is ordinate, and each reference substance concentration (C) is horizontal ordinate, and (table 20-22, Fig. 8), the result shows that each composition to be determined is good in its respective range internal linear relation to carry out linear regression.

1.4.4.5 precision test

The accurate reference substance solution of drawing, continuous sample introduction 5 times records 4-methyl isophthalic acid-isopropyl-3-cyclohexene-1-alcohol; 2-(4-methyl-3-cyclohexenyl group) isopropyl alcohol, Bronyl acetate, the RSD of the peak area of methylnonanone and interior mark peak area ratio is respectively 0.8%; 1.7%; 0.8%, 1.1%, the result shows that instrument precision is good.

Each composition peak area to be measured of table 20 and interior mark peak area are as shown in the table

Each component lines sexual intercourse result to be determined of table 21

Table 22 typical curve drawing data and linear equation are following

1.4.4.6. replica test

Get 6 parts respectively with a collection of medicinal material, every part of 50g, the accurate title, decide, and is equipped with solution according to " 4.2.2.3 " below legal system, and the RSD that records each component concentration is respectively 2.4%, 1.5%, 1.9%, 2.3%, and the result shows the method good reproducibility, sees table 23.

Each composition peak area to be measured of table 23 and interior mark peak area are as shown in the table

1.4.4.7. stability test

Get same need testing solution, respectively at 0h, 1h, 4h, 8h, 12h, 24h measures, and the RSD that records each component content is respectively 2.6%, 1.7%, 1.2%, 1.4%, shows that need testing solution is stable in 24h, sees table 24.

Each composition peak area to be measured of table 24 and interior mark peak area are as shown in the table

1.4.4.8. recovery test

Get with 6 parts of a collection of medicinal materials, every part of precision takes by weighing 25g, and it is an amount of to add the reference substance storing solution respectively; Be equipped with solution according to " 2.2.3 " below legal system; Each composition recovery is respectively 94.6%～97.4%, and 93.0%～98.4%, 95.1%～100.9%; Between 94.0%～100.8%, RSD is all less than 3% (seeing table 25).

4 kinds of composition determination of recovery rates of table 25 cordate houttuynia medicinal material result

1.4.4.9. sample determination

Get Yaan three nine-day periods after the winter solstice, the just clear cordate houttuynia medicinal material in Hunan is respectively 8 batches and 10 batches, prepares solution by " 4.2.3 " method, and sample introduction is measured, and calculates 4 kinds of component contents, and the result sees the following form 26 and table 27.

Three nine-day periods after the winter solstice cordate houttuynia medicinal material 4 kinds of component contents in table 26 Yaan are measured result (unit: μ gg ^-1)

Table 27 Hunan 4 kinds of component contents of just clear cordate houttuynia medicinal material are measured result (unit: μ gg ^-1)

The fingerprint atlas detection method research of experimental example 2 cordate houttuynia medicinal materials and parenteral solution

2.1. cordate houttuynia medicinal material detection method

2.1.1. confirming of chromatographic column

Investigated HP-5 respectively, DB-1, DB-17 capillary column, its typical color spectrogram (see figure 9).

It is best that the result is illustrated on the DB-1 chromatographic column separating effect, therefore adopts the chromatographic column of DB-1 capillary column as finger-print research.

2.1.2. confirming of temperature programme

Selected multiple temperature-programmed mode in the experimentation:

1.70℃(12min)-10℃/min-250℃

2.70℃(10min)-20℃/min-130℃-5℃/min-140℃-20℃/min-230℃

3.70℃(10min)-5℃/min-130℃-2℃/min-140℃-20℃/min-230℃

4.70℃(10min)-5℃/min-140℃-10℃/min-230℃

5.70℃(5min)-5℃/min-140℃-10℃/min-250℃

6.70℃(10min)-5℃/min-140℃(5min)-10℃/min-250℃

7.70℃(10min)-5℃/min-140℃(5min)-5℃/min-250℃(10min)

The result shows, the 6th, 7 kind of temperature-programmed mode be best, and each peak degree of separation is good, and the binding analysis time confirms to adopt the temperature-programmed mode of the 7th kind of mode as finger-print.

Figure 10 is 2 times of time chromatograms of cordate houttuynia medicinal material, and from figure, can find out 1 hour does not have chromatographic peak later on, and it is complete to explain that medicine goes out the peak.

2.2. cordate houttuynia medicinal materials fingerprint test method checking

2.2.1. precision test

Medicinal material with lot number 090906 prepares need testing solution, and continuous sample introduction 5 times writes down the retention time and the peak area at each total peak, is reference with the retention time and the peak area of methylnonanone, converses the relative retention time and the relative peak area at each total peak.The result shows that the RSD of relative retention time and relative peak area shows that all less than 3% (seeing table 28 and table 29) instrument precision is good.

2.2.2. replica test

Get 5 parts of the medicinal materials of lot number 090906,, investigate the relative retention time of each chromatographic peak, the consistance of relative peak area by preparing need testing solution with quadrat method.The result shows that the relative retention time at each total peak and relative peak area have good reappearance, RSD＜5% (seeing table 30 and table 31).

2.2.3. stability test

With lot number 090906 is that medicinal material prepares need testing solution; Respectively at 0h, 1h, 4h, 8h, 24h and 48h sample introduction; Writing down the retention time and the peak area at each total peak, is reference with the methylnonanone, converses the relative retention time and the relative peak area at each total peak.The result shows RSD＜5% (seeing table 32 and table 33) of need testing solution relative retention time and relative peak area, shows that need testing solution is stable in 48h.

Table 28 precision relative retention time

Table 29 precision relative peak area

The repeated relative retention time of table 30

The repeated relative peak area of table 31

The stable relative retention time of table 32

The stable relative peak area of table 33

2.3. cordate houttuynia medicinal material GC finger-print and technical parameter

2.3.1 finger-print is selected and the demarcation of characteristic peak

According to 17 batches of medicinal material test sample GC finger-prints, set up common pattern.Methylnonanone is the principal ingredient in the medicinal material, can find out that from spectrogram the area percentage of methylnonanone is bigger, therefore selects methylnonanone as interior reference peak.Interior reference peak methylnonanone label is S, other total peaks successively respectively label be 1-21.Pointed out wherein 8 total peaks, wherein No. 2 peaks are the α firpene, and No. 5 peaks are beta pinene, and No. 6 peaks are laurene, and No. 8 peaks are citrene, and No. 14 peaks are the 4-terpenol, and No. 15 peaks are alpha-terpineol, and No. 16 peaks are Bronyl acetate, and S is an interior reference peak methylnonanone.

2.3.2 characteristic fingerprint peak relative retention time and relative peak area

With methylnonanone is interior reference peak, and the relative retention time and the relative peak area of each fingerprint peaks of each batch medicinal material (22 peaks) all have reappearance preferably.

2.3.3 finger-print similarity

Calculate through " traditional Chinese medicine fingerprint similarity software for calculation ", the collection of illustrative plates that obtains each batch is seen table 34, Figure 11 with the similarity of contrast collection of illustrative plates:

Collection of illustrative plates of each batch of table 34 and the similarity that contrasts collection of illustrative plates

2.4. houttuynia cordata injection finger-print test method checking

2.4.1. precision test

Parenteral solution with lot number 201011607 prepares need testing solution, and continuous sample introduction 5 times writes down the retention time and the peak area at each total peak, is reference with the retention time and the peak area of methylnonanone, calculates each total peak relative retention time and relative peak area.The result shows that the relative retention time at each total peak and relative peak area meet the requirements, and shows that instrument precision is good.

Table 35 precision relative retention time:

Table 36 precision relative peak area:

2.4.2. replica test

Get 5 parts of the parenteral solutions of lot number 201011607,, investigate relative retention time, the relative peak area of each chromatographic peak by preparing need testing solution with quadrat method.The result shows that the relative retention time at each total peak and relative peak area meet the requirements, and have good reappearance.

The repeated relative retention time of table 37

The repeated relative peak area of table 38

2.4.3. stability test

Parenteral solution with lot number 201011607 prepares need testing solution, respectively at 0h, and 1h; 2h, 8h, 24h sample introduction; Writing down the retention time and the integral area at each total peak, is reference with the retention time and the peak area of methylnonanone, converses the relative retention time and the relative peak area at each total peak.The result shows that the RSD of need testing solution relative retention time and relative peak area meets the requirements.Show that parenteral solution is stable in 24h.

The stable relative retention time of table 39

The stable relative peak area of table 40

2.5. houttuynia cordata injection GC finger-print and technical parameter

2.5.1. finger-print is selected and the demarcation of characteristic peak

Generate common pattern with 19 batches of parenteral solution test sample GC finger-prints, methylnonanone is the principal ingredient in the parenteral solution, can find out that from spectrogram the area percentage of methylnonanone is bigger, therefore selects methylnonanone as interior reference peak.

Interior reference peak methylnonanone label is S, other total peaks successively respectively label be 1-10.Having pointed out 7 total peaks, is respectively No. 1 peak nopinene, and No. 2 peaks are laurene, and No. 3 peaks are citrene, No. 7 peak 4-terpenols, and No. 8 peaks are alpha-terpineol, and No. 9 peaks are Bronyl acetate, and S is an interior reference peak methylnonanone.

2.5.2. characteristic fingerprint peak relative retention time and relative peak area

With methylnonanone is interior reference peak, and the relative retention time and the relative peak area of 19 batches of each fingerprint peakses of parenteral solution (11 peaks) all have reappearance preferably.

2.5.3. confirming of finger-print similarity

19 batches of parenteral solution test sample data are carried out cluster analysis through SPSS16.0 software, set up common pattern with 19 batches of parenteral solutions.The finger-print and the common pattern finger-print of the present invention of 19 lot sample article are compared; Calculate through " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) (2.0 editions) "; Obtain the similarity of collection of illustrative plates with the contrast collection of illustrative plates of each batch, the result sees table 41, Figure 12.

Collection of illustrative plates of each batch of table 41 and the similarity that contrasts collection of illustrative plates

The limit examine of polyoxyethylene sorbitan monoleate in experimental example 3 houttuynia cordata injections

Li Lianda academician operating room is through safety testing research proof; Polyoxyethylene sorbitan monoleate is the main matter that causes the houttuynia cordata injection anaphylactoid reaction; And has a dose-effect relationship; Low concentration polyoxyethylene sorbitan monoleate (0.3%) is safe, therefore needs the content of polyoxyethylene sorbitan monoleate in the houttuynia cordata injection is carried out the strict control of limiting the quantity of.

3.1. instrument and reagent:

UV-265FW ultraviolet spectrophotometer (day island proper Tianjin); The polyoxyethylene sorbitan monoleate reference substance; Houttuynia cordata injection (lot number: 090916-090935,0907401-0907406,0907410,0904401,0904404,0904407), cordate houttuynia distillate (lot number 090910).Cobalt nitrate (the special chemicals in Rui Jin, Tianjin company limited); Ammonium thiocyanate (Tianjin Botong chemical industry company limited), methylene chloride (Tianjin Da Mao chemical reagent factory) is analyzed pure; Water is the self-control redistilled water, and oscillator (Jintan City the earth robot factory) is used in the HY-4 speed governing more.

3.2. method and result

The polyoxyethylene sorbitan monoleate chemical constitution is a polyoxyethylene sorbitol acid anhydride monoleate; Polyethoxy in its structure and ammonium cobalt rhodanide reaction form blue compound; This compound dissolves in methylene chloride, separates with water-solubility impurity, available colorimetric method for determining polyoxyethylene sorbitan monoleate content.

3.2.1. the selection of developing time

Precision is measured houttuynia cordata injection 1ml, and totally 12 parts, put in the conical flask, per 3 parts one group, add sulphur cyanogen cobalt ammonium salt solution 15ml respectively; The accurate methylene chloride 10ml that adds claims to decide weight, how puts speed governing with vibrating 15,30,45 on the oscillator respectively; 60min (120r/min), take out, and supplies the weight that subtracts mistake with methylene chloride by room temperature; Move in the separating funnel, leave standstill 15min, obtain dichloromethane layer solution, measure absorbance in the 623nm wavelength.The result shows that developing time develops the color when being 15min fully, so the selection developing time is 15min.

The different developing time absorbances of table 42

3.2.2. the selection of developer sulphur cyanogen cobalt ammonium salt solution consumption

Precision is measured houttuynia cordata injection 1ml, totally 12 parts, adds sulphur cyanogen cobalt

ammonium salt solution

5,10 respectively; 15,20ml, the accurate methylene chloride 10ml that adds claims to decide weight; It is many with the 15min that vibrates on the oscillator (room temperature) to put speed governing, takes out, and supplies the weight that subtracts mistake with methylene chloride, moves in the separating funnel; Leave standstill 15min, obtain dichloromethane layer solution, measure absorbance in the 623nm wavelength; With the methylene chloride is blank, and the result shows that absorbance reached maximum when sulphur cyanogen cobalt ammonium salt solution addition was 15ml, and selecting the addition of sulphur cyanogen cobalt ammonium salt solution is 15ml.

The different developer consumption of table 43 absorbance

3.2.3. the preparation of reference substance solution

Get the polyoxyethylene sorbitan monoleate reference substance in right amount to the 10ml measuring bottle, accurately claim surely, increase the weight of to steam redistilled water and process the solution that every 1ml contains 2.5mg.

3.2.4. the preparation of need testing solution

Precision pipettes houttuynia cordata injection 1ml and puts in the conical flask, and (get cobalt nitrate 6g, ammonium thiocyanate 40g is dissolved in water and is diluted to 200ml to add ammonium cobalt thiocyanate salt solution 15ml; Shake up, promptly get), the accurate methylene chloride 10ml that adds claims to decide weight; Put that speed governing is many takes out with the oscillator 15min that vibrates, supply the weight that subtracts mistake, move in the separating funnel with methylene chloride; Leave standstill 15min, obtain dichloromethane layer solution, as need testing solution.

3.2.5. measure the selection of wavelength

Reference substance solution and need testing solution scan respectively in 400～700nm wavelength coverage after chromogenic reaction, and the result finds that the polyoxyethylene sorbitan monoleate reference substance solution is consistent with houttuynia cordata injection need testing solution peak shape, and maximum absorption wavelength is consistent, is 623nm.

3.2.6. methodology checking

3.2.6.1. specificity test

Get blank solution (water), the cordate houttuynia re-distilled liquid (does not add polyoxyethylene sorbitan monoleate; As negative control), reference substance polyoxyethylene sorbitan monoleate, houttuynia cordata injection; By comparing absorbance behind " preparation of need testing solution " below Faxian look, the result is in the 623nm wavelength, and the absorbance of blank solution is 0.002; The absorbance of negative control solution is 0.001; Houttuynia cordata injection and reference substance all have better absorption, show that other component is noiseless to this test determination result in the houttuynia cordata injection, and negative control solution and blank solution do not have absorption basically in the 623nm wavelength.See Figure 13.

3.2.6.2. the typical curve and the range of linearity

Precision is measured polyoxyethylene sorbitan monoleate reference substance solution 0.2,0.4,0.6,0.8; 1.0ml put in the conical flask, add sulphur cyanogen cobalt ammonium salt solution 15ml respectively, the accurate methylene chloride 10ml that adds claims to decide weight; It is many with vibration (room temperature) 15min on the oscillator to put speed governing, and taking-up is supplied the weight that subtracts mistake with methylene chloride, moves in the separating funnel; Leave standstill 15min, obtain lower floor's solution, measure absorbance in the 623nm wavelength; With the methylene chloride is blank, with absorbance (A) polyoxyethylene sorbitan monoleate reference substance amount (C) is carried out linear regression, drawing standard curve (seeing table 44, Figure 14).The result shows that polyoxyethylene sorbitan monoleate is good linear relationship with absorbance in 1.02～5.10mg scope, and regression equation is: A=0.1749C-0.0018, r=0.9994.

Table 44 linear regression numerical value

3.2.6.3. precision test

Get the houttuynia cordata injection need testing solution under same " supplying the preparation of examination solution " item, under the 623nm wavelength, measure 5 times, mensuration RSD as a result is 0.2%, shows that instrument precision is good.

Table 45 precision data

3.2.6.4. replica test

Get with 6 parts of a collection of houttuynia cordata injections, by preparation need testing solution under " supplying the preparation of examination solution " item, measure in the 623nm wavelength, the RSD of the polyoxyethylene sorbitan monoleate content of surveying is 2.2%, shows this method repeatability well.

Table 46 polyoxyethylene sorbitan monoleate content

3.2.6.5. stability test

Get the houttuynia cordata injection need testing solution under " preparation of need testing solution " item; Every interval 0.5h measures an absorbance under the 623nm wavelength; METHOD FOR CONTINUOUS DETERMINATION 7 times, mensuration RSD as a result is 2.7%, shows that reference substance solution and need testing solution are stable in the 3h after colour developing.

Table 47 different time absorbance

3.2.6.6. recovery test

The houttuynia cordata injection 0.5ml that precision is measured own knowledge content puts in the conical flask, and totally 9 parts, per 3 parts 1 group; Accurate respectively polyoxyethylene sorbitan monoleate reference substance solution 0.11,0.22, the 0.33ml of adding; Method by under " preparation of need testing solution " item prepares need testing solution; Under the 623nm wavelength, measure, the average recovery rate of measuring polyoxyethylene sorbitan monoleate is 100.3%, and RSD is 2.1% (n=9).

The average recovery rate of table 48 polyoxyethylene sorbitan monoleate

3.2.7. sample determination

Precision pipettes each batch houttuynia cordata injection 1ml, puts in the conical flask, under 2.2, carries out chromogenic reaction, under the 623nm wavelength, measures absorbance.Polyoxyethylene sorbitan monoleate content to houttuynia cordata injection (totally 20 batches) is measured.

Table 49 polyoxyethylene sorbitan monoleate content

This method is quick, easy, can effectively control the quality of polyoxyethylene sorbitan monoleate in the houttuynia cordata injection, in being suitable for houttuynia cordata injection, the assay of polyoxyethylene sorbitan monoleate, also is suitable for the traditional Chinese medicine that other contain polyoxyethylene sorbitan monoleate simultaneously.

Experimental example 4 houttuynia cordata injection thin layer discrimination tests

Get houttuynia cordata injection 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate, jolting; Normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, as need testing solution.Other gets methylnonanone reference substance, alpha-terpineol reference substance and 4-terpilenol reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution.Drawing above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with cyclohexane-ethyl acetate (17: 3), launches, and takes out, and dries, and sprays 5% vanillic aldehyde ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Each detects injection products and to be no less than ten lot sample article, all with the corresponding position of reference substance chromatogram on, the spot of apparent same color.

The inspection of experimental example 5 cordate houttuynia medicinal material volatile oil

5.1 the mensuration of cordate houttuynia medicinal material volatile oil

5.1.1 crude drug source: just clear cordate houttuynia base, Pharmacy stock Co., Ltd Huaihua Yangchuan village, Hunan, totally 10 batches.

5.1.2 essential oil extraction method research: get the bright grass of cordate houttuynia and clean, shred, take by weighing 500g; Put in the 2000ml round-bottomed flask, add water 1000ml and beaded glass number, connect volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube, slowly be heated to and boil; Keep little and boiled 5 hours, be cooled to room temperature, calculate the total amount that these article contain volatile oil.

5.1.2.1 confirming of sampling amount

Get bright cordate houttuynia 250g respectively, 500g, 1000g carries out trial test, extracts according to the 5.1.2 essential oil extraction method, measures volatile oil.

Table 50 Herba Houttuyniae volatile oil amount

Result by last table can confirm that the sampling amount of cordate houttuynia is 500g.

5.1.2.2 confirming of amount of water

Extract according to the 5.1.2 essential oil extraction method, press the sampling amount that 5.1.2.1. confirms, amount of water is set at 500ml, 1000ml, 1500ml respectively, i.e. the water of one times of amount of medicinal material, qdx, triplication.

The volatilization oil mass of the different amount of water of table 51

The result shows that amount of water is the best results that 1000ml (medicinal material qdx) extracts.

5.1.2.3 confirming of distillation time

Get cordate houttuynia 500g, amount of water is 1000ml, extracts according to the 5.1.2 essential oil extraction method.The control return velocity is about 2 droplets/second, and volatile oil volume number was read in every distillation in 30 minutes, collected fully to volatile oil to finish.Confirm volatile oil extraction time with this.

The different distillation time volatile oil of table 52 volume number

Confirm that according to last table result extraction time is 5 hours.

According to above result of study, confirm the detection method of following cordate houttuynia medicinal material volatile oil: get the bright grass of cordate houttuynia and clean, shred, get about 500g; Accurate claim surely, put in the 2000ml round-bottomed flask, add water 1000ml and beaded glass number, connection volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube, slowly be heated to and boil; Keep little and boiled 5 hours, be cooled to room temperature, calculate the total amount that these article contain volatile oil.

5.1.3 the volatile oil testing result of cordate houttuynia medicinal material: the volatile oil to 10 cordate houttuynia medicinal materials detects.

The volatile oil content of table 53 cordate houttuynia medicinal material

5.2 the limit examine of cordate houttuynia re-distilled liquid volatile oil

When methylnonanone content is approximately higher than 20 μ g/ml in the cordate houttuynia re-distilled liquid; Opalescence or muddiness, deposited phenomenon can appear after the formulation products sterilization; And proper the being proportionate property of methylnonanone content in re-distilled liquid volatile oil content and the houttuynia cordata injection intermedium; Therefore to cordate houttuynia re-distilled liquid volatile oil limit examine, thereby effectively control the clarity of finished product.

5.2.1 confirming of distillation time

Get cordate houttuynia re-distilled liquid 2000ml, put in the 2000ml round-bottomed flask, add beaded glass number; Connect volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube, slowly be heated to and boil; Keep little and boiled 2 hours, respectively the 10th minute, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 60 minutes, 1.5 hours, 2 hours reading volatilization oil masses.

The different distillation time volatilization of table 54 oil mass

Find out that from last table distillation is complete basically for volatile oil when being distilled to 40 minutes, confirms that therefore the volatile oil inspection method is in the cordate houttuynia re-distilled liquid: get cordate houttuynia re-distilled liquid 500ml; Put in the 2000ml round-bottomed flask, add beaded glass number, connect volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube, slowly be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, calculate the total amount that these article contain volatile oil.

5.2.2 use the result that method is measured 9 batches of cordate houttuynia re-distilled liquid samples.

Methylnonanone content in the different lot number cordate houttuynia of table 55 re-distilled liquid volatile oil content and the houttuynia cordata injection intermediate product

The result shows that methylnonanone content is tangible positive correlation in volatile oil content and the intermediate product, and calculating the intermediate product methylnonanone content of above-mentioned nine batches of products and the mean ratio of re-distilled liquid volatile oil content is 290 (RSD=13.4%).Test through technical study; When methylnonanone content is approximately higher than 30 μ g/ml in the re-distilled liquid; To cause preparation sterilization back product opalescence or muddiness, deposited phenomenon to occur; The content that should control methylnonanone in the re-distilled liquid therefore, according to the correlativity of component D content in volatile oil content and the finished product in the above-mentioned table 23, the volatile oil content of re-distilled liquid should be crossed 0.08% (being to contain volatile oil 0.40ml in the 500ml test sample).

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

4 kinds of content assaying methods that composition detects simultaneously in embodiment 1 houttuynia cordata injection

Chromatographic condition and system suitability test, 30m * 0.25mm, 0.25 μ mDB-1 capillary column is a chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes; Per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID; Detector temperature is 280 ℃, and carrier gas is N2, and flow rate of carrier gas is 1.0ml/min; Split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation respectively, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 10mg, 5mg, 8mg, 2mg, precision is measured each reference substance storing solution respectively, adds normal hexane and processes the mixed solution that every 1ml contains 1mg, 0.5mg, 0.8mg, 1mg approximately; As mixing the reference substance storing solution, get and mix reference substance storing solution 0.5ml, put in the 2ml measuring bottle; The accurate 0.2ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get; Houttuynia cordata injection 50ml is got in the need testing solution preparation, puts in the round-bottomed flask, connects volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1ml contains alpha-terpineol (C ₈H ₁₈O) should be 2.0～10.0 μ g, methylnonanone (C ₁₁H ₂₂O) should be 6.0～20.0 μ g, 4-terpilenol (C ₈H ₁₈O) should be not less than 5.0 μ g, Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 0.8 μ g.

4 kinds of content assaying methods that composition detects simultaneously in the embodiment 2 cordate houttuynia medicinal materials

Chromatographic condition and system suitability test, 30m * 0.25mm, 0.25 μ mDB-1 capillary column is as chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes; Per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID; Detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 1mg, 0.5mg, 8mg, 10mg; Precision pipettes each reference substance storing solution 2ml, 1ml, 1ml, 4ml in the 10ml measuring bottle respectively, and the accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale; Shake up, promptly get; The cordate houttuynia medicinal material is got in the need testing solution preparation, shreds, and gets 50g, and accurate the title decides; Put in the 250ml round-bottomed flask, add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, be heated to and boil, keep little 4h that boils, be cooled to room temperature, obtain the normal hexane layer; Add the about 0.4g of anhydrous sodium sulfate, jolting, normal hexane liquid moves in the 10ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1g contains alpha-terpineol (C ₈H ₁₈O) should be not less than 1.5 μ g, contain methylnonanone (C ₁₁H ₂₂O) should be not less than 120 μ g, contain 4-terpilenol (C ₈H ₁₈O) should be not less than 3.0 μ g, contain Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 12.0 μ g.

Embodiment 3 houttuynia cordata injection fingerprint atlas detection methods

Detect the record chromatogram by method under the assay item of the present invention; Integral parameter is set to excise preceding 6 minutes solvent peaks; Mark peak in removing, Slope Sensitivity is 5, Peak Width is 0.1; Area reject is 4; Height reject is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.85; Finger-print has 11 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.465, No. 2 peak 0.486, No. 1 peak; 0.673, No. 5 peak 0.712,0.557, No. 4 peak, No. 3 peaks; 0.810, No. 8 peak 0.828,0.741, No. 7 peak, No. 6 peaks; No. 9 peaks 0.991,1.000, No. 10 peaks 1.554, S peak; Relative peak area is 0.463, No. 9 peak 0.185,1.541, No. 8 peaks, 0.024, No. 7 peak, 0.031, No. 6 peak, 0.066, No. 5 peak, 0.072, No. 4 peak, 0.041, No. 3 peak, 0.062, No. 2 peak, No. 1 peak, 1.000, No. 10 peaks 0.021, S peak.

Embodiment 4 cordate houttuynia medicinal materials fingerprint detection methods

Detect the record chromatogram by method under the assay item of the present invention; Integral parameter is set to excise preceding 6 minutes solvent peaks; Mark peak in removing, Slope Sensitivity is 5, Peak Width is 0.1; Area reject is 4; Height reject is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.90; Finger-print has 22 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.397, No. 3 peak 0.418,0.383, No. 2 peak, No. 1 peak; 0.533, No. 8 peak 0.556,0.486, No. 7 peak, 0.466, No. 6 peak, 0.457, No. 5 peak, No. 4 peaks; 0.661, No. 13 peak 0.668,0.607, No. 12 peak, 0.587, No. 11 peak, 0.566, No. 10 peak, No. 9 peaks; 0.826, No. 16 peak 0.989,0.807, No. 15 peak, No. 14 peaks, 1.000, No. 17 peaks 1.088, S peak; 1.346, No. 21 peaks 1.567,1.230, No. 20 peaks, 1.134, No. 19 peaks, No. 18 peaks; Relative peak area is 1.113, No. 6 peaks 0.493,0.444, No. 5 peak, 0.090, No. 4 peak, 0.606, No. 3 peak, 0.009, No. 2 peak, No. 1 peak; 0.036, No. 12 peak 0.021,0.005, No. 11 peak, 0.022, No. 10 peak, 0.354, No. 9 peak, 0.023, No. 8 peak, No. 7 peaks; 0.011, No. 16 peak 0.136,0.078, No. 15 peak, 0.012, No. 14 peak, No. 13 peaks, S peak 1.000; 0.134, No. 21 peak 0.029,0.035, No. 20 peak, 0.061, No. 19 peak, 0.017, No. 18 peak, No. 17 peaks.

The detection method of polyoxyethylene sorbitan monoleate limit examine in embodiment 5 houttuynia cordata injections

Precision pipettes houttuynia cordata injection 1ml and puts in the conical flask; Get cobalt nitrate 6g, ammonium thiocyanate 40g is dissolved in water and is diluted to 200ml, shakes up; Process ammonium cobalt thiocyanate salt solution, 15ml puts in the conical flask with ammonium cobalt thiocyanate salt solution, and the accurate methylene chloride 10ml that adds claims to decide weight; With the oscillator 15min that vibrates, take out, supply the weight that subtracts mistake with methylene chloride, move in the separating funnel; Leave standstill 15min, obtain dichloromethane layer solution, as need testing solution; Other gets the polyoxyethylene sorbitan monoleate reference substance to the 10ml measuring bottle, and accurate the title decides, and adds redistilled water and processes the solution that every 1ml contains 2.5mg, as reference substance solution; Get need testing solution and reference substance solution respectively, put into ultraviolet spectrophotometer, under the 623nm wavelength, measure absorbance log, calculate, promptly get; Contain polyoxyethylene sorbitan monoleate and must not cross 0.27%.

The discrimination test detection method of thin-layer chromatography in embodiment 6 houttuynia cordata injections

Get houttuynia cordata injection 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate, jolting; Normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, as need testing solution; Other gets methylnonanone, alpha-terpineol reference substance and 4-terpilenol reference substance reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with the ratio of cyclohexane-ethyl acetate be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

The discrimination test detection method of thin-layer chromatography in the embodiment 7 houttuynia cordata injection intermediate product

Get houttuynia cordata injection intermediate product 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate, jolting; Normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, as need testing solution; Other gets methylnonanone, alpha-terpineol reference substance and 4-terpilenol reference substance reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with the ratio of cyclohexane-ethyl acetate be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

The discrimination test detection method of thin-layer chromatography in the embodiment 8 cordate houttuynia medicinal materials

Get cordate houttuynia medicinal material 125g, shred, put in the 250ml round-bottomed flask, add water 150ml; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml, connect reflux condensing tube; Slowly be heated to and boil, and keep little and boiled 4 hours, place half an hour, get the normal hexane layer; To the 2ml measuring bottle, add normal hexane and be diluted to scale, as need testing solution; Other gets 4-terpilenol reference substance, alpha-terpineol reference substance and methylnonanone reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate ratio be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

The detection method of volatile oil content in the embodiment 9 cordate houttuynia medicinal materials

Get cordate houttuynia medicinal material 500g, the accurate title, decide, and puts in the 2000ml round-bottomed flask; Add water 1000ml and beaded glass number, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Connect reflux condensing tube, slowly be heated to and boil, keep little and boiled 5 hours; Be cooled to room temperature, calculate the total amount that these article contain volatile oil; Contain volatile oil and must not be less than 0.03% (ml/g).

The detection method of volatile oil content in embodiment 10 houttuynia cordata injections

Get houttuynia cordata injection or wherein between product 500ml, put in the 2000ml round-bottomed flask, add beaded glass number; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube; Slowly be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, calculate the total amount that these article contain volatile oil.

Embodiment 11 houttuynia cordata injection quality determining methods

A. the content assaying method that 4 kinds of compositions detect simultaneously in the houttuynia cordata injection is a vapor-phase chromatography, chromatographic condition and system suitability test, and 30m * 0.25mm, 0.25 μ mDB-1 capillary column is a chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes, and per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation respectively, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 10mg, 5mg, 8mg, 2mg, precision is measured each reference substance storing solution respectively, adds normal hexane and processes the mixed solution that every 1ml contains 1mg, 0.5mg, 0.8mg, 1mg approximately; As mixing the reference substance storing solution, get and mix reference substance storing solution 0.5ml, put in the 2ml measuring bottle; The accurate 0.2ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get; Houttuynia cordata injection 50ml is got in the need testing solution preparation, puts in the round-bottomed flask, connects volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets;

B. the content assaying method that 4 kinds of compositions detect simultaneously in the cordate houttuynia medicinal material is a vapor-phase chromatography, chromatographic condition and system suitability test, 30m * 0.25mm; 0.25 μ mDB-1 capillary column is as chromatographic column, initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃; Kept 5 minutes, per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃; Detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 1mg, 0.5mg, 8mg, 10mg; Precision pipettes each reference substance storing solution 2ml, 1ml, 1ml, 4ml in the 10ml measuring bottle respectively, and the accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale; Shake up, promptly get; The cordate houttuynia medicinal material is got in the need testing solution preparation, shreds, and gets 50g, and accurate the title decides; Put in the 250ml round-bottomed flask, add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, be heated to and boil, keep little 4h that boils, be cooled to room temperature, obtain the normal hexane layer; Add the about 0.4g of anhydrous sodium sulfate, jolting, normal hexane liquid moves in the 10ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets;

C. the houttuynia cordata injection fingerprint atlas detection method detects by method under the assay item, the record chromatogram; Integral parameter is set to excise preceding 4 minutes solvent peaks; Mark peak in removing, slope (Slope Sensitivity) is 5, peak width (Peak Width) is 0.1; Smallest peaks area (Area reject) is 4; Minimum peak height (Height reject) is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.85; Finger-print has 11 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.465, No. 2 peak 0.486, No. 1 peak; 0.673, No. 5 peak 0.712,0.557, No. 4 peak, No. 3 peaks; 0.810, No. 8 peak 0.828,0.741, No. 7 peak, No. 6 peaks; No. 9 peaks 0.991,1.000, No. 10 peaks 1.554, S peak; Relative peak area is 0.463, No. 9 peak 0.185,1.541, No. 8 peaks, 0.024, No. 7 peak, 0.031, No. 6 peak, 0.066, No. 5 peak, 0.072, No. 4 peak, 0.041, No. 3 peak, 0.062, No. 2 peak, No. 1 peak, 1.000, No. 10 peaks 0.021, S peak; Relative peak area is 0.041 ± 0.004, No. 3 peaks 0.072 ± 0.007,0.062 ± 0.006, No. 2 peaks, No. 1 peak; 0.031 ± 0.003, No. 6 peaks 0.024 ± 0.002,0.066 ± 0.007, No. 5 peaks, No. 4 peaks; 0.463 ± 0.046, No. 9 peaks 0.185 ± 0.019,1.541 ± 0.154, No. 8 peaks, No. 7 peaks; 1.000 ± 0.000, No. 10 peaks 0.021 ± 0.002, S peak;

D. cordate houttuynia medicinal materials fingerprint detection method detects by method under the assay item, the record chromatogram; Integral parameter is set to excise preceding 4 minutes solvent peaks; Mark peak in removing, Slope Sensitivity is 5, Peak Width is 0.1; Area reject is 4; Height reject is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.90; Finger-print has 22 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.397, No. 3 peak 0.418,0.383, No. 2 peak, No. 1 peak; 0.533, No. 8 peak 0.556,0.486, No. 7 peak, 0.466, No. 6 peak, 0.457, No. 5 peak, No. 4 peaks; 0.661, No. 13 peak 0.668,0.607, No. 12 peak, 0.587, No. 11 peak, 0.566, No. 10 peak, No. 9 peaks; 0.826, No. 16 peak 0.989,0.807, No. 15 peak, No. 14 peaks, 1.000, No. 17 peaks 1.088, S peak; 1.346, No. 21 peaks 1.567,1.230, No. 20 peaks, 1.134, No. 19 peaks, No. 18 peaks; Relative peak area is 1.113, No. 6 peaks 0.493,0.444, No. 5 peak, 0.090, No. 4 peak, 0.606, No. 3 peak, 0.009, No. 2 peak, No. 1 peak; 0.036, No. 12 peak 0.021,0.005, No. 11 peak, 0.022, No. 10 peak, 0.354, No. 9 peak, 0.023, No. 8 peak, No. 7 peaks; 0.011, No. 16 peak 0.136,0.078, No. 15 peak, 0.012, No. 14 peak, No. 13 peaks, S peak 1.000; 0.134, No. 21 peak 0.029,0.035, No. 20 peak, 0.061, No. 19 peak, 0.017, No. 18 peak, No. 17 peaks (concrete numerical value is seen table 2, Fig. 2);

E. the detection method of polyoxyethylene sorbitan monoleate limit examine does in the houttuynia cordata injection, and precision pipettes houttuynia cordata injection 1ml and puts in the conical flask; Get cobalt nitrate 6g, ammonium thiocyanate 40g is dissolved in water and is diluted to 200ml, shakes up; Process ammonium cobalt thiocyanate salt solution, 15ml puts in the conical flask with ammonium cobalt thiocyanate salt solution, and the accurate methylene chloride 10ml that adds claims to decide weight; With the oscillator 15min that vibrates, take out, supply the weight that subtracts mistake with methylene chloride, move in the separating funnel; Leave standstill 15min, obtain dichloromethane layer solution, as need testing solution; Other gets the polyoxyethylene sorbitan monoleate reference substance to the 10ml measuring bottle, and accurate the title decides, and adds redistilled water and processes the solution that every 1ml contains 2.5mg, as reference substance solution; Get need testing solution and reference substance solution respectively, put into ultraviolet spectrophotometer, under the 623nm wavelength, measure absorbance log, calculate, promptly get;

F. the discrimination method of cordate houttuynia medicinal material thin-layer chromatography does, gets cordate houttuynia medicinal material 125g, shreds, and puts in the 250ml round-bottomed flask; Add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, slowly be heated to and boil, and keep little and boiled 4 hours, place half an hour; Get the normal hexane layer, to the 2ml measuring bottle, add normal hexane and be diluted to scale, as need testing solution; Other gets 4-terpilenol reference substance, alpha-terpineol reference substance and methylnonanone reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate ratio be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

G. houttuynia cordata injection or wherein between the discrimination method of product thin-layer chromatography do, get houttuynia cordata injection or wherein between product 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, as need testing solution; Other gets methylnonanone, alpha-terpineol reference substance and 4-terpilenol reference substance reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with the ratio of cyclohexane-ethyl acetate be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

H. the detection method of volatile oil content does in the cordate houttuynia medicinal material; Get cordate houttuynia medicinal material 500g, the accurate title, decide, and puts in the 2000ml round-bottomed flask; Add water 1000ml and bead number; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube; Slowly be heated to and boil; Keep little and boiled 5 hours, be cooled to room temperature, calculate the total amount that this product contains volatile oil;

I. houttuynia cordata injection or wherein between in the product detection method of volatile oil content do, get houttuynia cordata injection or wherein between product 500ml, put in the 2000ml round-bottomed flask; Add beaded glass number, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Connect reflux condensing tube, slowly be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, calculate the total amount that these article contain volatile oil.

Embodiment 12 houttuynia cordata injection quality determining methods

A. the content assaying method that 4 kinds of compositions detect simultaneously in the houttuynia cordata injection is a vapor-phase chromatography, chromatographic condition and system suitability test, and 30m * 0.25mm, 0.25 μ mDB-1 capillary column is a chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes, and per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation respectively, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 10mg, 5mg, 8mg, 2mg, precision is measured each reference substance storing solution respectively, adds normal hexane and processes the mixed solution that every 1ml contains 1mg, 0.5mg, 0.8mg, 1mg approximately; As mixing the reference substance storing solution, get and mix reference substance storing solution 0.5ml, put in the 2ml measuring bottle; The accurate 0.2ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get; Houttuynia cordata injection 50ml is got in the need testing solution preparation, puts in the round-bottomed flask, connects volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1ml contains alpha-terpineol (C ₈H ₁₈O) should be 2.0～10.0 μ g, methylnonanone (C ₁₁H ₂₂O) should be 6.0～20.0 μ g, 4-terpilenol (C ₈H ₁₈O) should be not less than 5.0 μ g, Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 0.8 μ g;

B. the houttuynia cordata injection fingerprint atlas detection method detects by method under the assay item, the record chromatogram; Integral parameter is set to excise preceding 4 minutes solvent peaks; Mark peak in removing, slope (Slope Sensit1vity) is 5, peak width (Peak Width) is 0.1; Smallest peaks area (Area reject) is 4; Minimum peak height (Height reject) is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.85; Finger-print has 11 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.465, No. 2 peak 0.486, No. 1 peak; 0.673, No. 5 peak 0.712,0.557, No. 4 peak, No. 3 peaks; 0.810, No. 8 peak 0.828,0.741, No. 7 peak, No. 6 peaks; No. 9 peaks 0.991,1.000, No. 10 peaks 1.554, S peak; Relative peak area is 0.066, No. 5 peak 0.031,0.072, No. 4 peak, 0.041, No. 3 peak, 0.062, No. 2 peak, No. 1 peak; 0.463, No. 9 peak 0.185,1.541, No. 8 peaks, 0.024, No. 7 peak, No. 6 peaks; 1.000, No. 10 peak 0.021 relative peak areas in S peak are 0.041 ± 0.004, No. 3 peaks 0.072 ± 0.007,0.062 ± 0.006, No. 2 peaks, No. 1 peak; 0.024 ± 0.002, No. 7 peaks 1.541 ± 0.154,0.031 ± 0.003, No. 6 peaks, 0.066 ± 0.007, No. 5 peaks, No. 4 peaks; 0.463 ± 0.046, No. 9 peaks 0.185 ± 0.019, No. 8 peaks, 1.000 ± 0.000, No. 10 peaks 0.021 ± 0.002, S peak;

C. the detection method of polyoxyethylene sorbitan monoleate limit examine does in the houttuynia cordata injection, and precision pipettes houttuynia cordata injection 1ml and puts in the conical flask; Get cobalt nitrate 6g, ammonium thiocyanate 40g is dissolved in water and is diluted to 200ml, shakes up; Process ammonium cobalt thiocyanate salt solution, 15ml puts in the conical flask with ammonium cobalt thiocyanate salt solution, and the accurate methylene chloride 10ml that adds claims to decide weight; With the oscillator 15min that vibrates, take out, supply the weight that subtracts mistake with methylene chloride, move in the separating funnel; Leave standstill 15min, obtain dichloromethane layer solution, as need testing solution; Other gets the polyoxyethylene sorbitan monoleate reference substance to the 10ml measuring bottle, and accurate the title decides, and adds redistilled water and processes the solution that every 1ml contains 2.5mg, as reference substance solution; Get need testing solution and reference substance solution respectively, put into ultraviolet spectrophotometer, under the 623nm wavelength, measure absorbance log, calculate, promptly get; Contain polyoxyethylene sorbitan monoleate and must not cross 0.25%;

D. houttuynia cordata injection or wherein between the discrimination method of product thin-layer chromatography do, get houttuynia cordata injection or wherein between product 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, as need testing solution; Other gets methylnonanone, alpha-terpineol reference substance and 4-terpilenol reference substance reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with the ratio of cyclohexane-ethyl acetate be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

E. houttuynia cordata injection or wherein between in the product detection method of volatile oil content do, get houttuynia cordata injection or wherein between product 500ml, put in the 2000ml round-bottomed flask; Add beaded glass number, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Connect reflux condensing tube, slowly be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, calculate the total amount that these article contain volatile oil.

The quality determining method of embodiment 13 cordate houttuynia medicinal materials

A. the content assaying method that 4 kinds of compositions detect simultaneously in the cordate houttuynia medicinal material is a vapor-phase chromatography, chromatographic condition and system suitability test, 30m * 0.25mm; 0.25 μ mDB-1 capillary column is as chromatographic column, initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃; Kept 5 minutes, per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃; Detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 1mg, 0.5mg, 8mg, 10mg; Precision pipettes each reference substance storing solution 2ml, 1ml, 1ml, 4ml in the 10ml measuring bottle respectively, and the accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale; Shake up, promptly get; The cordate houttuynia medicinal material is got in the need testing solution preparation, shreds, and gets 50g, and accurate the title decides; Put in the 250ml round-bottomed flask, add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, be heated to and boil, keep little 4h that boils, be cooled to room temperature, obtain the normal hexane layer; Add the about 0.4g of anhydrous sodium sulfate, jolting, normal hexane liquid moves in the 10ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1g contains alpha-terpineol (C ₈H ₁₈O) should be not less than 1.5 μ g, contain methylnonanone (C ₁₁H ₂₂O) should be not less than 120 μ g, contain 4-terpilenol (C ₈H ₁₈O) should be not less than 3.0 μ g, contain Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 12.0 μ g;

B. cordate houttuynia medicinal materials fingerprint detection method detects by method under the assay item, the record chromatogram; Integral parameter is set to excise preceding 4 minutes solvent peaks; Mark peak in removing, Slope Sensitivity is 5, Peak Width is 0.1; Area reject is 4; Height reject is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.90; Finger-print has 22 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.397, No. 3 peak 0.418,0.383, No. 2 peak, No. 1 peak; 0.533, No. 8 peak 0.556,0.486, No. 7 peak, 0.466, No. 6 peak, 0.457, No. 5 peak, No. 4 peaks; 0.661, No. 13 peak 0.668,0.607, No. 12 peak, 0.587, No. 11 peak, 0.566, No. 10 peak, No. 9 peaks; 0.826, No. 16 peak 0.989,0.807, No. 15 peak, No. 14 peaks, 1.000, No. 17 peaks 1.088, S peak; 1.346, No. 21 peaks 1.567,1.230, No. 20 peaks, 1.134, No. 19 peaks, No. 18 peaks; Relative peak area is 1.113, No. 6 peaks 0.493,0.444, No. 5 peak, 0.090, No. 4 peak, 0.606, No. 3 peak, 0.009, No. 2 peak, No. 1 peak; 0.036, No. 12 peak 0.021,0.005, No. 11 peak, 0.022, No. 10 peak, 0.354, No. 9 peak, 0.023, No. 8 peak, No. 7 peaks; 0.011, No. 16 peak 0.136,0.078, No. 15 peak, 0.012, No. 14 peak, No. 13 peaks, S peak 1.000; 0.134, No. 21 peak 0.029,0.035, No. 20 peak, 0.061, No. 19 peak, 0.017, No. 18 peak, No. 17 peaks;

C. the discrimination method of cordate houttuynia medicinal material thin-layer chromatography does, gets cordate houttuynia medicinal material 125g, shreds, and puts in the 250ml round-bottomed flask; Add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, slowly be heated to and boil, and keep little and boiled 4 hours, place half an hour; Get the normal hexane layer, to the 2ml measuring bottle, add normal hexane and be diluted to scale, as need testing solution; Other gets 4-terpilenol reference substance, alpha-terpineol reference substance and methylnonanone reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate ratio be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

D. the detection method of volatile oil content does in the cordate houttuynia medicinal material, gets cordate houttuynia medicinal material 500g, and accurate the title decides; Put in the 2000ml round-bottomed flask, add water 1000ml and beaded glass number, connect volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, connect reflux condensing tube, slowly be heated to and boil; Keep little and boiled 5 hours, be cooled to room temperature, calculate the total amount that these article contain volatile oil; Contain volatile oil and must not be less than 0.03% (ml/g).

Claims

1. the quality determining method of a houttuynia cordata injection is characterized in that this method comprises following assay, discriminating and/or fingerprint atlas detection method:

A. the content assaying method that 4 kinds of compositions detect simultaneously in the houttuynia cordata injection is: vapor-phase chromatography, and chromatographic condition and system suitability test, 30m * 0.25mm, 0.25 μ mDB-1 capillary column is a chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes, and per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 5～15: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 1～4mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation respectively, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 8～12mg, 4～6mg, 6～10mg, 1～4mg, precision is measured each reference substance storing solution respectively, adds normal hexane and processes the mixed solution that every 1ml contains 1mg, 0.5mg, 0.8mg, 1mg approximately; As mixing the reference substance storing solution, get and mix reference substance storing solution 0.5ml, put in the 2ml measuring bottle; The accurate 0.2ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get; Houttuynia cordata injection 50ml is got in the need testing solution preparation, puts in the round-bottomed flask, connects volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Contain alpha-terpineol (C in every 1ml houttuynia cordata injection ₈H ₁₈0) should be 2.0～10.0 μ g, methylnonanone (C ₁₁H ₂₂O) should be 6.0～20.0 μ g, 4-terpilenol (C ₈H ₁₈O) should be not less than 5.0 μ g, Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 0.8 μ g;

B. the content assaying method that 4 kinds of compositions detect simultaneously in the cordate houttuynia medicinal material is: vapor-phase chromatography, chromatographic condition and system suitability test, 30m * 0.25mm; 0.25 μ mDB-1 capillary column is as chromatographic column, initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃; Kept 5 minutes, per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃; Detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 5～15: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 1～4mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 0.5～1.5mg, 0.2～1mg, 6～10mg, 8～12mg; Precision pipettes each reference substance storing solution 2ml, 1ml, 1ml, 4ml in the 10ml measuring bottle respectively, and the accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale; Shake up, promptly get; The cordate houttuynia medicinal material is got in the need testing solution preparation, shreds, and gets 45～55g, and accurate the title decides; Put in the 250ml round-bottomed flask, add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, be heated to and boil, keep little and boiled 4 hours, be cooled to room temperature, obtain the normal hexane layer; Add the about 0.3～0.5g of anhydrous sodium sulfate, jolting, normal hexane liquid moves in the 10ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1g cordate houttuynia medicinal material contains alpha-terpineol (C ₈H ₁₈O) should be not less than 1.5 μ g, contain methylnonanone (C ₁₁H ₂₂O) should be not less than 120 μ g, contain 4-terpilenol (C ₈H ₁₈O) should be not less than 3.0 μ g, contain Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 12.0 μ g;

G. houttuynia cordata injection or wherein between the discrimination method of product thin-layer chromatography be: get houttuynia cordata injection or wherein between product 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate, jolting; Normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, as need testing solution; Other gets methylnonanone, alpha-terpineol reference substance and 4-terpilenol reference substance reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 0.8～1.5mg, 0.3～0.8mg, 1.0～3.0mg, as reference substance solution; Drawing above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, is to be developping agent at 15～21: 3 with the ratio of cyclohexane-ethyl acetate; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

H. the detection method of volatile oil content is in the cordate houttuynia medicinal material: get cordate houttuynia medicinal material 500g, the accurate title, decide, and puts in the 2000ml round-bottomed flask; Add water 1000ml and beaded glass number, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Connect reflux condensing tube, slowly be heated to and boil, keep little and boiled 5 hours; Be cooled to room temperature, calculate the total amount that these article contain volatile oil; Contain volatile oil and must not be less than 0.03%ml/g;

2. the quality determining method of houttuynia cordata injection as claimed in claim 1 is characterized in that this method comprises following assay, discriminating and/or fingerprint atlas detection method:

A. the content assaying method that 4 kinds of compositions detect simultaneously in the houttuynia cordata injection is: vapor-phase chromatography, and chromatographic condition and system suitability test, 30m * 0.25mm, 0.25 μ mDB-1 capillary column is a chromatographic column; Initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃, keeps 5 minutes, and per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃, and detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation respectively, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 10mg, 5mg, 8mg, 2mg, precision is measured each reference substance storing solution respectively, adds normal hexane and processes the mixed solution that every 1ml contains 1mg, 0.5mg, 0.8mg, 1mg approximately; As mixing the reference substance storing solution, get and mix reference substance storing solution 0.5ml, put in the 2ml measuring bottle; The accurate 0.2ml inner mark solution that adds; Add normal hexane and be diluted to scale, shake up, promptly get; Houttuynia cordata injection 50ml is got in the need testing solution preparation, puts in the round-bottomed flask, connects volatile oil determination apparatus; Add water from the analyzer upper end and be full of the scale part, add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil; Keep little and boiled 40 minutes, be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate; Jolting, normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1ml houttuynia cordata injection contains alpha-terpineol (C ₈H ₁₈O) should be 2.0～10.0 μ g, methylnonanone (C ₁₁H ₂₂O) should be 6.0～20.0 μ g, 4-terpilenol (C ₈H ₁₈O) should be not less than 5.0 μ g, Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 0.8 μ g;

B. the content assaying method that 4 kinds of compositions detect simultaneously in the cordate houttuynia medicinal material is: vapor-phase chromatography, chromatographic condition and system suitability test, 30m * 0.25mm; 0.25 μ mDB-1 capillary column is as chromatographic column, initial 70 ℃ kept 5 minutes, and per minute rises to 140 ℃ for 5 ℃; Kept 5 minutes, per minute rises to 250 ℃ for 20 ℃, and injector temperature is 230 ℃; Detecting device is FID, and detector temperature is 280 ℃, and carrier gas is N ₂, flow rate of carrier gas is 1.0ml/min, and split ratio is 10: 1, and sample introduction is 1 μ l, and number of theoretical plate calculates by the methylnonanone peak should be not less than 10000; Positive undecylene is got in the inner mark solution preparation, and accurate the title decides, and adds normal hexane and processes the solution that every 1ml contains 2mg; 4-terpilenol reference substance, alpha-terpineol reference substance, Bronyl acetate reference substance, methylnonanone reference substance are got in the reference substance solution preparation, and accurate the title decides; Add normal hexane respectively and process the reference substance storing solution that every 1ml contains 1mg, 0.5mg, 8mg, 10mg; Precision pipettes each reference substance storing solution 2ml, 1ml, 1ml, 4ml in the 10ml measuring bottle respectively, and the accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale; Shake up, promptly get; The cordate houttuynia medicinal material is got in the need testing solution preparation, shreds, and gets 50g, and accurate the title decides; Put in the 250ml round-bottomed flask, add water 150ml, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml; Connect reflux condensing tube, be heated to and boil, keep little and boiled 4 hours, be cooled to room temperature, obtain the normal hexane layer; Add the about 0.4g of anhydrous sodium sulfate, jolting, normal hexane liquid moves in the 10ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 1ml inner mark solution that adds adds normal hexane and is diluted to scale, shakes up, and promptly gets; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing, inject gas chromatograph is measured, and promptly gets; Every 1g cordate houttuynia medicinal material contains alpha-terpineol (C ₈H ₁₈O) should be not less than 1.5 μ g, contain methylnonanone (C ₁₁H ₂₂O) should be not less than 120 μ g, contain 4-terpilenol (C ₈H ₁₈O) should be not less than 3.0 μ g, contain Bronyl acetate (C ₁₂H ₂₀O ₂) should be not less than 12.0 μ g;

C. houttuynia cordata injection fingerprint atlas detection method: detect the record chromatogram by method under the assay item; Integral parameter is set to excise preceding 4 minutes solvent peaks; Mark peak in removing, slope (SlopeSensitivity) is 5, peak width (Peak Width) is 0.1; Smallest peaks area (Area reject) is 4; Minimum peak height (Height reject) is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.85; Finger-print has 11 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.465, No. 2 peak 0.486, No. 1 peak; 0.673, No. 5 peak 0.712,0.557, No. 4 peak, No. 3 peaks; 0.810, No. 8 peak 0.828,0.741, No. 7 peak, No. 6 peaks; No. 9 peaks 0.991,1.000, No. 10 peaks 1.554, S peak; Relative peak area is 0.463, No. 9 peak 0.185,1.541, No. 8 peaks, 0.024, No. 7 peak, 0.031, No. 6 peak, 0.066, No. 5 peak, 0.072, No. 4 peak, 0.041, No. 3 peak, 0.062, No. 2 peak, No. 1 peak, 1.000, No. 10 peaks 0.021, S peak;

D. cordate houttuynia medicinal materials fingerprint detection method: detect the record chromatogram by method under the assay item; Integral parameter is set to excise preceding 4 minutes solvent peaks; Mark peak in removing, Slope Sensitivity is 5, Peak Width is 0.1; Area reject is 4; Height reject is 0.1, calculates similarity with " chromatographic fingerprints of Chinese materia medica similarity evaluation system (2009 editions) ", and test sample finger-print and reference fingerprint similarity should be lower than 0.90; Finger-print has 22 characteristic peaks, is with reference to the peak with methylnonanone RT=17.20min, and each peak relative retention time is 0.397, No. 3 peak 0.418,0.383, No. 2 peak, No. 1 peak; 0.533, No. 8 peak 0.556,0.486, No. 7 peak, 0.466, No. 6 peak, 0.457, No. 5 peak, No. 4 peaks; 0.661, No. 13 peak 0.668,0.607, No. 12 peak, 0.587, No. 11 peak, 0.566, No. 10 peak, No. 9 peaks; 0.826, No. 16 peak 0.989,0.807, No. 15 peak, No. 14 peaks, 1.000, No. 17 peaks 1.088, S peak; 1.346, No. 21 peaks 1.567,1.230, No. 20 peaks, 1.134, No. 19 peaks, No. 18 peaks; Relative peak area is 1.113, No. 6 peaks 0.493,0.444, No. 5 peak, 0.090, No. 4 peak, 0.606, No. 3 peak, 0.009, No. 2 peak, No. 1 peak; 0.036, No. 12 peak 0.021,0.005, No. 11 peak, 0.022, No. 10 peak, 0.354, No. 9 peak, 0.023, No. 8 peak, No. 7 peaks; 0.011, No. 16 peak 0.136,0.078, No. 15 peak, 0.012, No. 14 peak, No. 13 peaks, S peak 1.000; 0.134, No. 21 peak 0.029,0.035, No. 20 peak, 0.061, No. 19 peak, 0.017, No. 18 peak, No. 17 peaks;

E. the detection method of polyoxyethylene sorbitan monoleate limit examine is in the houttuynia cordata injection: precision pipettes houttuynia cordata injection 1ml and puts in the conical flask; Get cobalt nitrate 6g, ammonium thiocyanate 40g is dissolved in water and is diluted to 200ml, shakes up; Process ammonium cobalt thiocyanate salt solution, 15ml puts in the conical flask with ammonium cobalt thiocyanate salt solution, and the accurate methylene chloride 10ml that adds claims to decide weight; With oscillator vibration 15 minutes, take out, supply the weight that subtracts mistake with methylene chloride, move in the separating funnel; Left standstill 15 minutes, and obtained dichloromethane layer solution, as need testing solution; Other gets the polyoxyethylene sorbitan monoleate reference substance to the 10ml measuring bottle, and accurate the title decides, and adds redistilled water and processes the solution that every 1ml contains 2.5mg, as reference substance solution; Get need testing solution and reference substance solution respectively, put into ultraviolet spectrophotometer, under the 623nm wavelength, measure absorbance log, calculate, promptly get; Contain polyoxyethylene sorbitan monoleate and must not cross 0.27%;

F. the discrimination method of cordate houttuynia medicinal material thin-layer chromatography is: get cordate houttuynia medicinal material 125g, shred, put in the 250ml round-bottomed flask, add water 150ml; Connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part, add normal hexane 1ml, connect reflux condensing tube; Slowly be heated to and boil, and keep little and boiled 4 hours, place half an hour, get the normal hexane layer; To the 2ml measuring bottle, add normal hexane and be diluted to scale, as need testing solution; Other gets 4-terpilenol reference substance, alpha-terpineol reference substance and methylnonanone reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate ratio be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

G. houttuynia cordata injection or wherein between the discrimination method of product thin-layer chromatography be: get houttuynia cordata injection or wherein between product 50ml, put in the round-bottomed flask, connect volatile oil determination apparatus, add water from the analyzer upper end and be full of the scale part; Add normal hexane 0.5ml, connect reflux condensing tube, be heated to and boil, keep little and boiled 40 minutes; Be cooled to room temperature, obtain the normal hexane layer, add the about 0.4g of anhydrous sodium sulfate, jolting; Normal hexane liquid moves in the 2ml measuring bottle, and washs anhydrous sodium sulfate with normal hexane, and washing lotion is incorporated in the same measuring bottle; The accurate 0.2ml inner mark solution that adds adds normal hexane and is diluted to scale, as need testing solution; Other gets methylnonanone, alpha-terpineol reference substance and 4-terpilenol reference substance reference substance, adds normal hexane respectively and processes the solution that every 1ml contains 1.0mg, 0.5mg, 2.0mg, as reference substance solution; Draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with the ratio of cyclohexane-ethyl acetate be 17: 3 be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;