CN101401880B - Quality control method for Ditong rhinitis drop and mist - Google Patents
Quality control method for Ditong rhinitis drop and mist Download PDFInfo
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Abstract
The invention discloses a method for controlling the quality of rhinitis-treating drop medicine and spray. The method carries out TLC identification to biond magnolia flower and/or grassleaf sweelflag rhizome, and/or carries out content determination to ephedrine hydrochloride which is a main active constituent of ephedra, and establishes a microbial limit examination method at the same time. The method for controlling the quality of rhinitis-treating drop medicine and spray can be favorable for effectively ensuring the internal quality of products in industrial production and further guarantee the efficacy of medicine.
Description
Technical field
The invention belongs to technical field of Chinese medicines, be specifically related to the quality determining method of DITONGBIYAN SHUIDIJI and spray.
Background technology
DITONG BIYAN SHUI prescription and method for preparing are recorded in the 5th 199 pages in the Sanitation Ministry medicine standard Chinese traditional patent formulation preparation; Standard code is WS3-B-1061-91; Prescription is made up of Herba Taraxaci 120g, Radix Scutellariae 60g, Herba Ephedrae 50g, Fructus Xanthii 50g, Flos Magnoliae 25g, Radix Angelicae Dahuricae 25g, Herba Asari 5g, Rhizoma Acori Graminei 60g, and above-mentioned raw materials is processed 1000ml altogether.Spray is identical with prescription, method for making and the effect of drop, is same medical substance, and just packaging material are different.DITONG BIYAN SHUI has expelling wind and clearing away heat, the sensible effect of lung qi dispersing, and it is sick to be used for treatment cold nasal obstruction, stuffy nose (chronic rhinitis), nose allergic rhinitis, nasal sinusitis (sinusitis) etc., uses for many years clinically, obtains more gratifying curative effect.
Former drop quality standard has only carried out the test tube qualitative identification and the Radix Angelicae Dahuricae has been carried out the thin layer chromatography discriminating volatile oil, alkaloid, is difficult to guarantee pharmaceutical effectiveness.
Summary of the invention
The technical problem that the present invention will solve provides a kind of further the guarantee DITONGBIYAN SHUIDIJI of pharmaceutical effectiveness and the quality determining method of spray.
The present invention solves the problems of the technologies described above with following technical scheme:
The quality determining method of DITONGBIYAN SHUIDIJI and spray, one or more in may further comprise the steps:
1. with thin layer chromatography the Flos Magnoliae in the medicine is carried out qualitative identification: get these article 50ml, put in the separatory funnel, with dichloromethane, ethyl acetate or ether extraction 1～3 time; Each 10～30ml, merge extractive liquid,, evaporate to dryness; Residue solubilizer methanol, ethanol or ethyl acetate 1ml make dissolving, and the neutral alumina post that is added on internal diameter 5～15mm is (on 2～4g), with methanol or ethanol 10～30ml eluting; Collect eluent, be concentrated into 1ml, as need testing solution.Other gets Flos Magnoliae control medicinal material 1g, adds methylene chloride, ethyl acetate or ether 10～30ml supersound process or reflux 20～60 minutes; Filter, filtrating is concentrated into about 1ml, and the neutral alumina post that is added on internal diameter 5～15mm is (on 2～4g); With methanol or ethanol 10～40ml eluting; Collect eluent, be concentrated into 1ml, as need testing solution.Get the magnelin reference substance again, add methanol, ethanol or ethyl acetate and process the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; Chloroform-ether with 10-4: 3-1 is developing solvent; Launch, take out, dry; Spray is with 2% vanillin sulphuric acid or 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing in 105 ℃.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.
2. with thin layer chromatography the Rhizoma Acori Graminei in the medicine is carried out qualitative identification: get these article 20ml, put in the separatory funnel, with chloroform, ether or ethyl acetate extraction 1～3 time; Each 10～40ml, merge extractive liquid, adds anhydrous sodium sulfate 2g; Jolting filters, the filtrating evaporate to dryness; Residue adds methanol, ethanol, dehydrated alcohol or ethyl acetate 1ml makes dissolving, as need testing solution.Other gets Rhizoma Acori Graminei control medicinal material 0.5g, adds methanol, ethanol or dehydrated alcohol 10～50ml, and reflux or supersound process 10～60 minutes filter, and filtrating is concentrated into about 2ml, as control medicinal material solution.According to thin layer chromatography test, draw need testing solution 5 μ l, control medicinal material solution 2 μ l, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With normal hexane or cyclohexane extraction-dichloromethane or chloroform-ethyl acetate (16～8: 4～1: 4～0.5) be developing solvent; Launch, take out, dry; Spray is with 2% vanillin sulphuric acid or 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing in 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
3. with Determination of Ephedrine Hydrochloride in this medicine of high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filler; Methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH2-3) (0.8～1.2: 10) be mobile phase; The detection wavelength is 210 ± 2nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 4000.
The preparation of reference substance solution: it is an amount of to get the ephedrine hydrochloride reference substance, and accurate the title decides, and adds mobile phase or methanol and processes the solution that contains 30 μ g among every 1ml, promptly gets.
The preparation of need testing solution: get these article, the accurate 4.0～12.0ml that draws puts in the 250ml round-bottomed flask; Add sodium chloride 20g, 8% sodium hydroxide solution, 120～180ml shakes up; (heat up in a steamer speed 0.2～1.0ml/min), receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 5～10ml, thin up is to scale with distillation; Shake up, ultrasonic 10 minutes, promptly get.
Algoscopy: accurate respectively reference substance solution and each 10～20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure.The every 1ml of these article contains Herba Ephedrae with ephedrine hydrochloride (C
10H
15NOHCl) meter must not be less than 0.30mg.
4. check the microorganism of these article by following method:
These article of getting 10ml adds the aseptic sodium chloride-peptone buffer agent of pH7.0 to 100ml, and mixing was as 1: 10 test liquid.
Bacterial population is got 1: 10 test liquid 1ml of these article and is annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Mycete and yeast count are got 1: 10 test liquid 1ml of these article and are annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Escherichia coli is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
Staphylococcus aureus is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml nutrient broth medium, in accordance with the law inspection.
Pseudomonas aeruginosa is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
The every 1ml of these article bacterial population must not cross 100, and the every 1ml of mycete and yeast count must not cross 10, and the every 1ml of escherichia coli must not detect, and staphylococcus aureus, the every 1ml of Pseudomonas aeruginosa must not detect.
During optimized technical scheme comprises the following steps one or more:
1 usefulness thin layer chromatography is carried out qualitative identification to the Flos Magnoliae in the medicine: get these article 50ml, put in the separatory funnel, with ethyl acetate extraction 2 times; Each 15ml, merge extractive liquid,, evaporate to dryness; Residue adds ethyl acetate 1ml makes dissolving, and the neutral alumina post that is added on internal diameter 9mm is (on 2～4g), with methanol 15ml eluting; Collect eluent, be concentrated into about 1ml, as need testing solution.Other gets Flos Magnoliae control medicinal material 1g, adds ethyl acetate 15ml, and supersound process 30 minutes filters, and filtrating is concentrated into about 1ml, and the neutral alumina post that is added on internal diameter 9mm (on 2～4g), with methanol 15ml eluting, is collected eluent, is concentrated into about 1ml, as need testing solution.Get the magnelin reference substance again, add methanol and process the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With 5: 1 chloroform-ether was developing solvent, launched, and took out; Dry, spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing in 105 ℃.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.
2 usefulness thin layer chromatography are carried out qualitative identification to the Rhizoma Acori Graminei in the medicine: get these article 20ml, put in the separatory funnel, and with ether extraction 2 times, each 20ml; Merge extractive liquid, adds anhydrous sodium sulfate 2g, and jolting filters; The filtrating evaporate to dryness, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution.Other gets Rhizoma Acori Graminei control medicinal material 0.5g, adds dehydrated alcohol 25ml, and reflux 20 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution.According to thin layer chromatography test, draw need testing solution 5 μ 1, control medicinal material solution 2 μ l, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With cyclohexane extraction-chloroform-ethyl acetate (8: 2: 1) is developing solvent; Launch, take out, dry; Spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing in 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
3. with the Determination of Ephedrine Hydrochloride in this medicine epheday intermedia medical material of high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filler; Methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH2.5) (1: 10) is mobile phase; The detection wavelength is 210nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 4000.
It is an amount of that the ephedrine hydrochloride reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds mobile phase and processes the solution that contains 30 μ g among every 1ml, promptly gets.
These article are got in the preparation of need testing solution, and the accurate 8.0ml that draws puts in the 250ml round-bottomed flask; Add sodium chloride 20g, 8% sodium hydroxide solution 150ml shakes up; With distillation (heating up in a steamer fast 0.5ml/min), receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 10ml, thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure.The every 1ml of these article contains Herba Ephedrae with ephedrine hydrochloride (C
10H
16NOHCl) meter must not be less than 0.30mg.
4. check the microorganism of these article by following method:
These article of getting 10ml adds the aseptic sodium chloride-peptone buffer agent of pH7.0 to 100ml, and mixing was as 1: 10 test liquid.
Bacterial population is got 1: 10 test liquid 1ml of these article and is annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Mycete and yeast count are got 1: 10 test liquid 1ml of these article and are annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Escherichia coli is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
Staphylococcus aureus is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml nutrient broth medium, in accordance with the law inspection.
Pseudomonas aeruginosa is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
Per 1 ml of these article bacterial population must not cross 100, and the every 1ml of mycete and yeast count must not cross 10, and the every 1ml of escherichia coli must not detect, and staphylococcus aureus, the every 1ml of Pseudomonas aeruginosa must not detect.
Method of the present invention has versatility to the quality control of DITONGBIYAN SHUIDIJI and spray, is one or more in the following method of increase on former drop method of quality control basis: with thin layer chromatography Flos Magnoliae is carried out qualitative identification; With thin layer chromatography Rhizoma Acori Graminei is carried out qualitative identification; Microbial limit is checked; With the HPLC method to the Herba Ephedrae characteristic component---ephedrine hydrochloride carries out assay, helps in suitability for industrialized production effectively guaranteeing the clinical efficacy of DITONGBIYAN SHUIDIJI and spray.
The specific embodiment
Method of quality control of the present invention is the preferred plan that obtains through a large amount of screenings, and following experimental study is a preferred process of the present invention.
One, the thin layer chromatography Study on Identification of Flos Magnoliae
The method for preparing of Flos Magnoliae need testing solution compares:
Method one is got these article 50ml, puts in the separatory funnel, and with ethyl acetate extraction 2 times, each 15ml, combined ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution.Lack the Flos Magnoliae negative controls with the method preparation.
Method two is got these article 50ml, puts in the separatory funnel, and with chloroform extraction 2 times, each 15ml, combined chloroform liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Lack the Flos Magnoliae negative controls with the method preparation.
Method three is got these article 20ml, puts in the separatory funnel, and with ether extraction 2 times, each 15ml merges ether solution, and evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Lack the Flos Magnoliae negative controls with the method preparation.
Method four is got these article 50ml, puts in the separatory funnel, and with dichloromethane extraction 2 times, each 15ml, merge extractive liquid,, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Lack the Flos Magnoliae negative controls with the method preparation.
Method five is got these article 50ml, puts in the separatory funnel, with ethyl acetate extraction 2 times; Each 15ml, merge extractive liquid,, evaporate to dryness; Residue adds ethyl acetate 1ml makes dissolving, and the neutral alumina post that is added on internal diameter 9mm is (on 2～4g), with methanol 15ml eluting; Collect eluent, be concentrated into about 1ml, as need testing solution.Lack the Flos Magnoliae negative controls with the method preparation.
Above-mentioned five kinds of methods are compared, and in the method one～five, test sample and control medicinal material, reference substance chromatograph all have corresponding speckle as a result; Negative control does not have corresponding speckle; But have impurity to disturb in method one, two, four chromatographs, method three speckles a little less than, best with method five.
The comparison of Flos Magnoliae development system: (1) chloroform-ether (5: 1); (2) benzene-ethyl acetate (9: 1); (3) toluene-ethyl acetate-glacial acetic acid (8: 2: 1); (4) chloroform-ether (4: 1); (5) chloroform-ether (6: 1).Result (2), (3) Rf value are all higher, (1) (4) (5) launch effect all can, Rf value is moderate, Thin-layer separation is good, clear spot.
The comparison of Flos Magnoliae inspection knowledge condition: 2% vanillin sulphuric acid liquid and 10% sulphuric acid ethanol liquid all can identify this speckle, and 2% vanillin sulphuric acid liquid colour developing color is more bright-coloured.
Two, the thin layer chromatography Study on Identification of Rhizoma Acori Graminei
The method for preparing of Rhizoma Acori Graminei need testing solution compares:
Method one is got these article 20ml, puts in the separatory funnel, and with ether extraction 2 times, each 20ml, merge extractive liquid, adds anhydrous sodium sulfate 2g, and jolting filters, the filtrating evaporate to dryness, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution.Lack the Rhizoma Acori Graminei negative controls with the method preparation.
Method two is got these article 20ml, puts in the separatory funnel, and with ethyl acetate extraction 2 times, each 20ml, merge extractive liquid,, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution.Lack the Rhizoma Acori Graminei negative controls with the method preparation.
Method three is got these article 20ml, puts in the separatory funnel, and with ether extraction 2 times, each 20ml, merge extractive liquid,, evaporate to dryness, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution.Lack the Rhizoma Acori Graminei negative controls with the method preparation.
Method four is got these article 20ml, puts in the volatile oil extractor, adds water 300ml, extracts 2 hours, and volatile oil absorbs with ethyl acetate 2ml, as need testing solution.Lack the Rhizoma Acori Graminei negative controls with the method preparation.
Method five is got these article 20ml, puts in the round-bottomed flask, adds petroleum ether 40ml, and reflux 1 hour is put coldly, obtains petroleum ether liquid, volatilizes, and residue adds petroleum ether 1ml makes dissolving, as need testing solution.Lack the Rhizoma Acori Graminei negative controls with the method preparation.
Above-mentioned five kinds of methods are compared, and in the method one～five, test sample and control medicinal material all have corresponding speckle as a result; Negative control does not have corresponding speckle; But all have small amount of impurities to disturb in the method two～five colors spectrum, and method four, five complicated operations, time are long, best with method one.
The comparison of Rhizoma Acori Graminei development system: (1) cyclohexane extraction-chloroform-ethyl acetate (8: 2: 1); (2) normal hexane-dichloromethane-ethyl acetate (10: 3: 1); (3) cyclohexane extraction-ethyl acetate (6: 1); (4) petroleum ether (60～90 ℃)-ethyl acetate (4: 1); (5) petroleum ether (30～60 ℃)-ethyl acetate (3: 2).It is better that result (1), (2) launch effect, the clear spot rounding.
The comparison of Rhizoma Acori Graminei inspection knowledge condition: (1) 2% vanillin sulphuric acid liquid; (2) 10% sulphuric acid ethanol liquid; (3) iodine steam.Result (3) impurity is many, and (1) and (2) all can identify this speckle, and 2% vanillin sulphuric acid liquid colour developing color is more bright-coloured.
Three, Determination of Ephedrine Hydrochloride study on determination method in the Herba Ephedrae
1 instrument and reagent
Instrument: day island proper Tianjin LC-10ATvp high performance liquid chromatograph.
Reference substance: ephedrine hydrochloride derives from Nat'l Pharmaceutical & Biological Products Control Institute's (lot number: 0714-9903).Reagent:
Methanol (chromatographically pure), sodium hydroxide, hydrochloric acid, sodium dihydrogen phosphate, phosphoric acid and triethylamine are analytical pure.
Sample: DITONGBIYAN SHUIPENWUJI, lot number 021012.
2 need testing solution method for preparinies are selected
Method one is got these article, and the accurate 8.0ml that draws puts in the 250ml round-bottomed flask; Add sodium chloride 20g, 8% sodium hydroxide solution 150ml shakes up; With distillation (heating up in a steamer fast 0.5ml/min), receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 10ml, thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get.
The method two precision is got these article 8ml, regulates pH value to 8-9 with ammonia, with ether extraction 5 times, and each 10ml, the merging ether extracted liquid, evaporate to dryness, residue is used dissolve with methanol, and is transferred in the 100ml measuring bottle, adds methanol to scale.
Method three precisions are got these article 2ml, put in the 25ml measuring bottle, add 50% methanol 20ml, and supersound process 10 minutes adds 50% methanol to scale.
The method two ephedrine hydrochloride response rate is low as a result; Method three impurity are many, disturb greatly, are prone to pollute chromatographic column; Method one economy, environmental protection, impurity is few, and separating effect is good.
3. distillation speed is investigated
Heat up in a steamer speed and be unfavorable for that too soon ephedrine steams, too slow test period is long, is advisable with 0.2～1.0ml/min, and 0.5ml/min is more excellent.
4. measure the selection of wavelength
Precision takes by weighing ephedrine hydrochloride reference substance 10.1mg, puts in the 100ml measuring bottle, adds the mobile phase dissolving and is diluted to scale; Shake up; The accurate 1ml that draws puts in the 10ml measuring bottle, adds mobile phase and is diluted to scale, shakes up; In the interscan of 190～350nm scope, ephedrine hydrochloride has absorption maximum at the 210nm place as a result with ultraviolet spectrophotometer.
5 chromatographic conditions
Use octadecylsilane chemically bonded silica to be filler; Methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH2.5) (1: 10) is mobile phase; The detection wavelength is 210nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 4000.
Mobile phase compares: (1) methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH2.5) (1: 10); (2) methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH2.5) (0.8: 10); (3) methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH3) (1.2: 10); (4) methanol-0.02mol/L sodium dihydrogen phosphate (1: 10); (5) acetonitrile-potassium dihydrogen phosphate (6: 94); (6) acetonitrile-0.1% phosphoric acid solution (6: 94); (7) acetonitrile-0.1% phosphoric acid solution (9: 87); (8) methanol-0.02mol/L sodium dihydrogen phosphate (phosphoric acid is transferred pH2.5) (1: 10)
Mobile phase (1)-(3) separable ephedrine hydrochloride as a result, peak shape is better, is optimum with (1), and (4)-(8) have a separating effect not good, and the peak shape that has is bad.
6 negative samples are measured
Get the sample 8.0ml that does not contain Herba Ephedrae, put in the 250ml round-bottomed flask, add sodium chloride 20g; 8% sodium hydroxide solution 150ml shakes up, with distillation (heating up in a steamer fast 0.5ml/min); Receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 10ml, thin up shakes up to scale; Ultrasonic 10 minutes, promptly get.By above-mentioned chromatographic condition, precision is measured 20 μ l, injects chromatograph of liquid.The result does not have corresponding chromatographic peak in the ephedrine hydrochloride position, shows sample determination noiseless.
7 linear relationships are investigated
Precision takes by weighing ephedrine hydrochloride reference substance 10.1mg, puts in the 100ml measuring bottle, adds the mobile phase dissolving and is diluted to scale, shakes up; Accurate 1ml, 2ml, 3ml, 4ml, 5ml, the 6ml of drawing puts respectively in the 10ml measuring bottle, adds mobile phase and is diluted to scale, shakes up; Promptly get, by above-mentioned chromatographic condition, difference sample introduction 20 μ l; Measure, ephedrine hydrochloride sample size C (μ g) is carried out regression analysis, get regression equation: A=1.0538 * 10 with peak area integration A
6C-4.963 * 10
3, r=0.9999.Show that the ephedrine hydrochloride sample size is good in 0.202～1.212 μ g scope internal linear relation, and the straight-line pass initial point, available single-point external standard method is measured and is calculated, and linear relationship data is seen table 1.
Table 1 linear relationship is investigated data
The test of 8 precision
Precision is measured ephedrine hydrochloride reference substance solution (30.3 μ g/ml) 20 μ l, by above-mentioned chromatographic condition, injects chromatograph of liquid, repeats sample introduction 5 times, calculates the relative standard deviation of ephedrine hydrochloride peak area A, and the result sees table 2, shows that precision is better.
The test of table 2 precision
9 stability tests
Get need testing solution (lot number 021012), preparing back placement 0,2,4,6,8h and 24,48,72h respectively, 20 μ l take a sample; By above-mentioned chromatographic condition; Inject chromatograph of liquid, measure ephedrine hydrochloride peak area A, calculate relative standard deviation; The result sees table 3, table 4, shows that stability better.
Table 3 stability test (day interpolation)
Table 4 stability test (poor in the daytime)
10 replica tests
(lot number: 021012), the accurate 8.0ml that draws puts in the 250ml round-bottomed flask to get same sample; Add sodium chloride 20g, 8% sodium hydroxide solution 150ml shakes up; With distillation (heating up in a steamer fast 0.5ml/min), receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 10ml, thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get need testing solution.Be equipped with 5 parts of need testing solutions with legal system, the 20 μ l that take a sample by above-mentioned chromatographic condition, inject chromatograph of liquid, measure, and calculate Determination of Ephedrine Hydrochloride and relative standard deviation, and the result sees table 5, shows method repeatability better.
Table 5 method replica test
The test of 11 average recoveries
Accurate sample (the lot number 021012 content 0.492mg/ml) 4.0ml that has predicted content that draws puts in the 250ml round-bottomed flask, accurate respectively again ephedrine hydrochloride reference substance solution (1.02mg/ml) 1.7ml or the 2ml of adding; Add sodium chloride 20g, 8% sodium hydroxide solution 150ml shakes up; With distillation (heating up in a steamer fast 0.5ml/min), receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 10ml, thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get.By above-mentioned chromatographic condition, inject chromatograph of liquid, measure, calculate recovery rate, the result sees table 6, shows that the method response rate is better.
The test of table 6 average recovery
Four, microbial limit test checking
1. trial test
According to Chinese Pharmacopoeia microbial limit test conventional method DITONGBIYAN SHUIPENWUJI has been carried out trial test; Visible these article of result all do not have the biocidal property influence to the growth of escherichia coli, staphylococcus aureus, bacillus subtilis, Candida albicans, aspergillus niger, therefore can adopt conventional method that bacterial population, mycete and yeast count are verified.
DITONGBIYAN SHUIPENWUJI limit test of microbe method (conventional method) checking trial test result
2 count of bacteria method validations
According to the trial test result; Make the recovery test of each representative strain of conventional method; The response rate of three lot sample article all is higher than 70% as a result, shows that the test liquid of dilution does not have the biocidal property influence to the growth of escherichia coli, staphylococcus aureus, bacillus subtilis, therefore can adopt the conventional method check.
The response rate result of each representative strain of count of bacteria method validation test
3. mycete and yeast method of counting checking
According to the trial test result, do the recovery test of each representative strain with conventional method, the response rate of three lot sample article all is higher than 70% as a result, shows that these article do not have the biocidal property influence to the growth of Candida albicans, aspergillus niger, therefore can adopt the conventional method check.
The response rate result of mycete and each representative strain of yeast method of counting demonstration test
4 control bacterium (escherichia coli, staphylococcus aureus, Pseudomonas aeruginosa) inspection method checkings
Make the demonstration test of escherichia coli inspection method of conventional method.(lot number: demonstration test result 060115,060116,060117): test group all detects escherichia coli to three lot sample article, and the negative bacterium matched group does not all detect staphylococcus aureus.Show that these article do not have the biocidal property influence to the growth of escherichia coli, can adopt the conventional method check.
Make the demonstration test of staphylococcus aureus inspection method of conventional method.(lot number: demonstration test result 060115,060116,060117): test group all detects staphylococcus aureus to three lot sample article, and the negative bacterium matched group does not all detect escherichia coli.Show that these article do not have the biocidal property influence to the growth of staphylococcus aureus, can adopt the conventional method check.
Make the demonstration test of Pseudomonas aeruginosa inspection method of conventional method.(lot number: demonstration test result 060115,060116,060117): test group all detects Pseudomonas aeruginosa to three lot sample article, and the negative bacterium matched group does not all detect escherichia coli.Show that these article do not have the biocidal property influence to the growth of Pseudomonas aeruginosa, can adopt the conventional method check.
Above-mentioned steps need not carried out according to sequencing, and can also carry out conventional sense simultaneously, and like character, pH value, this finished product character is brown supernatant liquid; Gas fragrance, mildly bitter flavor; PH value should be 5.0～7.0.This finished product also should meet each item regulation relevant under an appendix I of Chinese Pharmacopoeia version in 2005 the X nasal formulations item.
Embodiment 1:
The quality control step of DITONGBIYAN SHUIPENWUJI:
1. with thin layer chromatography the Flos Magnoliae in the medicine is carried out qualitative identification:
Get DITONGBIYAN SHUIPENWUJI 50ml, put in the separatory funnel, with ethyl acetate extraction 2 times; Each 15ml, merge extractive liquid,, evaporate to dryness; Residue adds ethyl acetate 1ml makes dissolving, and the neutral alumina post that is added on internal diameter 9mm is (on 2～4g), with methanol 15ml eluting; Collect eluent, be concentrated into about 1ml, as need testing solution.Other gets Flos Magnoliae control medicinal material 1g, adds ethyl acetate 15ml, and supersound process 30 minutes filters, and filtrating is concentrated into about 1ml, and the neutral alumina post that is added on internal diameter 9mm (on 2～4g), with methanol 15ml eluting, is collected eluent, is concentrated into about 1ml, as need testing solution.Get the magnelin reference substance again, add methanol and process the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With 5: 1 chloroform-ether was developing solvent, launched, and took out; Dry, spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing in 105 ℃.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.
2. with thin layer chromatography the Rhizoma Acori Graminei in the medicine is carried out qualitative identification:
Get DITONGBIYAN SHUIPENWUJI 20ml, put in the separatory funnel, with ether extraction 2 times, each 20ml, merge extractive liquid, adds anhydrous sodium sulfate 2g, and jolting filters, the filtrating evaporate to dryness, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution.Other gets Rhizoma Acori Graminei control medicinal material 0.5g, adds dehydrated alcohol 25ml, and reflux 20 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution.According to thin layer chromatography test, draw need testing solution 5 μ l, control medicinal material solution 2 μ l, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With cyclohexane extraction-chloroform-ethyl acetate (8: 2: 1) is developing solvent; Launch, take out, dry; Spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing in 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
3. with the Determination of Ephedrine Hydrochloride in the high effective liquid chromatography for measuring DITONGBIYAN SHUIPENWUJI:
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filler; Methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH2.5) (1: 10) is mobile phase; The detection wavelength is 210nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 4000.
It is an amount of that the ephedrine hydrochloride reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds mobile phase and processes the solution that contains 30 μ g among every 1ml, promptly gets.
These article are got in the preparation of need testing solution, and the accurate 8.0ml that draws puts in the 250ml round-bottomed flask; Add sodium chloride 20g, 8% sodium hydroxide solution 150ml shakes up; With distillation (heating up in a steamer fast 0.5ml/min), receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 10ml, thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure.The every 1ml of these article contains Herba Ephedrae with ephedrine hydrochloride (C
10H
15NOHCl) meter must not be less than 0.30mg.
4. microbial check
These article of getting 10ml adds the aseptic sodium chloride-peptone buffer agent of pH7.0 to 100ml, and mixing was as 1: 10 test liquid.
Bacterial population is got 1: 10 test liquid 1ml of these article and is annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Mycete and yeast count are got 1: 10 test liquid 1ml of these article and are annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Escherichia coli is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
Staphylococcus aureus is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml nutrient broth medium, in accordance with the law inspection.
Pseudomonas aeruginosa is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
The every 1ml of these article bacterial population must not cross 100, and the every 1ml of mycete and yeast count must not cross 10, and the every 1ml of escherichia coli must not detect, and staphylococcus aureus, the every 1ml of Pseudomonas aeruginosa must not detect.
Above-mentioned steps need not carried out according to sequencing, and can also carry out conventional sense simultaneously, and like character, pH value, this finished product character is brown supernatant liquid; Gas fragrance, mildly bitter flavor; PH value should be 5.0～7.0.This finished product also should meet each item regulation relevant under an appendix I of Chinese Pharmacopoeia version in 2005 the X nasal formulations item.
Embodiment 2: the method for quality control of DITONGBIYAN SHUIDIJI comprises the following steps:
1. with thin layer chromatography the Flos Magnoliae in the medicine is carried out qualitative identification:
Get DITONGBIYAN SHUIDIJI 50ml, put in the separatory funnel, with ether extraction 2 times; Each 30ml, merge extractive liquid,, evaporate to dryness; Residue adds methanol 1ml makes dissolving, and the neutral alumina post that is added on internal diameter 12mm is (on 2～4g), with ethanol 30ml eluting; Collect eluent, be concentrated into about 1ml, as need testing solution.Other gets Flos Magnoliae control medicinal material 1g, the 30ml that adds diethyl ether, and reflux 40 minutes filters, and filtrating is concentrated into about 1ml, and the neutral alumina post that is added on internal diameter 12mm (on 2～4g), with ethanol 30ml eluting, is collected eluent, is concentrated into about 1ml, as need testing solution.Get the magnelin reference substance again, add ethyl acetate and process the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With 6: 1 chloroform-ether was developing solvent, launched, and took out; Dry, spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing in 105 ℃.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.
2. with thin layer chromatography the Rhizoma Acori Graminei in the medicine is carried out qualitative identification:
Get DITONGBIYAN SHUIDIJI 20ml, put in the separatory funnel, with ethyl acetate extraction 2 times, each 30ml, merge extractive liquid, adds anhydrous sodium sulfate 2g, and jolting filters, the filtrating evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution.Other gets Rhizoma Acori Graminei control medicinal material 0.5g, adds methanol 15ml, and supersound process 20 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution.According to thin layer chromatography test, draw need testing solution 5 μ l, control medicinal material solution 2 μ l, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With normal hexane-dichloromethane-ethyl acetate (8: 2: 1) is developing solvent; Launch, take out, dry; Spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing in 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
3. with the Determination of Ephedrine Hydrochloride in the high effective liquid chromatography for measuring DITONGBIYAN SHUIDIJI:
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filler; Methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH3) (1.1: 10) is mobile phase; The detection wavelength is 208nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 4000.
It is an amount of that the ephedrine hydrochloride reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and processes the solution that contains 30 μ g among every 1ml, promptly gets.
These article are got in the preparation of need testing solution, and the accurate 6.0ml that draws puts in the 250ml round-bottomed flask; Add sodium chloride 20g, 8% sodium hydroxide solution 130ml shakes up; With distillation (heating up in a steamer fast 0.3ml/min), receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 8ml, thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure.The every 1ml of these article contains Herba Ephedrae with ephedrine hydrochloride (C
10H
15NOHCl) meter must not be less than 0.30mg.
4. microbial check
These article of getting 10ml adds the aseptic sodium chloride-peptone buffer agent of pH7.0 to 100ml, and mixing was as 1: 10 test liquid.
Bacterial population is got 1: 10 test liquid 1ml of these article and is annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Mycete and yeast count are got 1: 10 test liquid 1ml of these article and are annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Escherichia coli is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
Staphylococcus aureus is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml nutrient broth medium, in accordance with the law inspection.
Pseudomonas aeruginosa is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
The every 1ml of these article bacterial population must not cross 100, and the every 1ml of mycete and yeast count must not cross 10, and the every 1ml of escherichia coli must not detect, and staphylococcus aureus, the every 1ml of Pseudomonas aeruginosa must not detect.
Above-mentioned steps need not carried out according to sequencing, and can also carry out conventional sense simultaneously, and like character, pH value, this finished product character is brown supernatant liquid; Gas fragrance, mildly bitter flavor; PH value should be 5.0～7.0.This finished product also should meet each item regulation relevant under an appendix I of Chinese Pharmacopoeia version in 2005 the X nasal formulations item.
Embodiment 3: the method for quality control of DITONGBIYAN SHUIPENWUJI comprises the following steps:
1. with thin layer chromatography the Flos Magnoliae in the medicine is carried out qualitative identification:
Get DITONGBIYAN SHUIPENWUJI 50ml, put in the separatory funnel, extract 1 time the extracting solution evaporate to dryness with dichloromethane 30ml; Residue adds ethanol 1ml makes dissolving, and the neutral alumina post that is added on internal diameter 5mm is (on 2～4g), with methanol 30ml eluting; Collect eluent, be concentrated into about 1ml, as need testing solution.Other gets Flos Magnoliae control medicinal material 1g, the 20ml that adds methylene chloride, and supersound process 20 minutes filters, and filtrating is concentrated into about 1ml, and the neutral alumina post that is added on internal diameter 5mm (on 2～4g), with methanol 30ml eluting, is collected eluent, is concentrated into about 1ml, as need testing solution.Get the magnelin reference substance again, add ethanol and process the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With 4: 1 chloroform-ether was developing solvent, launched, and took out; Dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing in 105 ℃.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.
2. with thin layer chromatography the Rhizoma Acori Graminei in the medicine is carried out qualitative identification:
Get DITONGBIYAN SHUIPENWUJI 20ml, put in the separatory funnel, with chloroform extraction 3 times, each 25ml, merge extractive liquid, adds anhydrous sodium sulfate 2g, and jolting filters, the filtrating evaporate to dryness, residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Other gets Rhizoma Acori Graminei control medicinal material 0.5g, adds ethanol 30ml, and reflux 40 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution.According to thin layer chromatography test, draw need testing solution 5 μ l, control medicinal material solution 2 μ l, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With cyclohexane extraction-chloroform-ethyl acetate (8: 3: 1) is developing solvent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing in 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
3. with the Determination of Ephedrine Hydrochloride in the high effective liquid chromatography for measuring DITONGBIYAN SHUIPENWUJI:
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filler; Methanol-0.02mol/L sodium dihydrogen phosphate (contain 0.2% triethylamine, phosphoric acid is transferred pH2.6) (0.9: 10) is mobile phase; The detection wavelength is 212nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 4000.
It is an amount of that the ephedrine hydrochloride reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds mobile phase and processes the solution that contains 30 μ g among every 1ml, promptly gets.
These article are got in the preparation of need testing solution, and the accurate 10.0ml that draws puts in the 250ml round-bottomed flask; Add sodium chloride 20g, 8% sodium hydroxide solution 180ml shakes up; With distillation (heating up in a steamer fast 1.0ml/min), receive distillate to about 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 10ml, thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get.
Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure.The every 1ml of these article contains Herba Ephedrae with ephedrine hydrochloride (C
10H
15NOHCl) meter must not be less than 0.30mg.
4. microbial check
These article of getting 10ml adds the aseptic sodium chloride-peptone buffer agent of pH7.0 to 100ml, and mixing was as 1: 10 test liquid.Bacterial population is got 1: 10 test liquid 1ml of these article and is annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Mycete and yeast count are got 1: 10 test liquid 1ml of these article and are annotated ware, and 2 plates of parallel preparation are pressed Plating and measured.
Escherichia coli is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
Staphylococcus aureus is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml nutrient broth medium, in accordance with the law inspection.
Pseudomonas aeruginosa is got 1: 10 test liquid 10ml of these article and is seeded in the 100ml cholate lactose medium, in accordance with the law inspection.
The every 1ml of these article bacterial population must not cross 100, and the every 1ml of mycete and yeast count must not cross 10, and the every 1ml of escherichia coli must not detect, and staphylococcus aureus, the every 1ml of Pseudomonas aeruginosa must not detect.
Claims (2)
1. the quality determining method of DITONGBIYAN SHUIDIJI and spray; Wherein said pharmaceutical formulation is made up of Chinese crude drug Herba Taraxaci 120g, Radix Scutellariae 60g, Herba Ephedrae 50g, Fructus Xanthii 50g, Flos Magnoliae 25g, Radix Angelicae Dahuricae 25g, Herba Asari 5g, Rhizoma Acori Graminei 60g; Above-mentioned raw materials is processed 1000ml altogether, it is characterized in that this method comprises following detection:
(1) gets these article 50ml, put in the separatory funnel, with dichloromethane, ethyl acetate or ether extraction 1～3 time; Each 10～30ml, merge extractive liquid,, evaporate to dryness; Residue solubilizer methanol, ethanol or ethyl acetate 1ml make dissolving, and the neutral alumina post 2～4g that is added on internal diameter 5～15mm is last, with methanol or ethanol 10～30ml eluting; Collect eluent, be concentrated into 1ml, as need testing solution; Other gets Flos Magnoliae control medicinal material 1g, adds methylene chloride, ethyl acetate or ether 10～30ml supersound process or reflux 20～60 minutes; Filter, filtrating is concentrated into 1ml, and the neutral alumina post 2～4g that is added on internal diameter 5～15mm is last; With methanol or ethanol 10～40ml eluting; Collect eluent, be concentrated into 1ml, as need testing solution; Get the magnelin reference substance again, add methanol, ethanol or ethyl acetate and process the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; Chloroform-ether with 10-4: 3-1 is developing solvent; Launch, take out, dry; Spray is with 2% vanillin sulphuric acid or 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing in 105 ℃; In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;
(2) get these article 20ml, put in the separatory funnel, with chloroform, ether or ethyl acetate extraction 1～3 time; Each 10～40ml, merge extractive liquid, adds anhydrous sodium sulfate 2g; Jolting filters, the filtrating evaporate to dryness; Residue adds methanol, ethanol, dehydrated alcohol or ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Rhizoma Acori Graminei control medicinal material 0.5g, adds methanol, ethanol or dehydrated alcohol 10～50ml, and reflux or supersound process 10～60 minutes filter, and filtrating is concentrated into 2ml, as control medicinal material solution; According to thin layer chromatography test, draw need testing solution 5 μ l, control medicinal material solution 2 μ l, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With 16～8: 4～1: 4～0.5 normal hexane or cyclohexane extraction dichloromethane or chloroform-ethyl acetate are developing solvent; Launch, take out, dry; Spray is with 2% vanillin sulphuric acid or 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing in 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;
(3) according to the high effective liquid chromatography for measuring Determination of Ephedrine Hydrochloride:
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filler; Containing 0.2% triethylamine, phosphoric acid, to transfer pH2-3, ratio be the mobile phase of methanol-0.02mol/L sodium dihydrogen phosphate 0.8～1.2: 10; The detection wavelength is 210 ± 2nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the ephedrine hydrochloride reference substance, and accurate the title decides, and adds mobile phase or methanol and processes the solution that contains 30 μ g among every 1ml, promptly gets;
The preparation of need testing solution: get these article, the accurate 4.0～12.0ml that draws puts in the 250ml round-bottomed flask; Add sodium chloride 20g, 8% sodium hydroxide solution, 120～180ml shakes up; To heat up in a steamer speed is 0.2～1.0ml/min distillation, receives distillate to 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 5～10ml, and thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get;
Algoscopy: accurate respectively reference substance solution and each 10～20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure; The every 1ml of these article contains Herba Ephedrae in ephedrine hydrochloride, must not be less than 0.30mg;
(4) shine the microorganism that microbial limit test is checked these article:
These article of getting 10ml adds the aseptic sodium chloride-peptone buffer agent of pH7.0 to 100ml, and mixing was as 1: 10 test liquid;
Bacterial population: get 1: 10 test liquid 1ml of these article and annotate ware, 2 plates of parallel preparation are pressed Plating and are measured;
Mycete and yeast count: get 1: 10 test liquid 1ml of these article and annotate ware, 2 plates of parallel preparation are pressed Plating and are measured;
Escherichia coli: get 1: 10 test liquid 10ml of these article and be seeded in the 100ml cholate lactose medium, in accordance with the law inspection;
Staphylococcus aureus: get 1: 10 test liquid 10ml of these article and be seeded in the 100ml nutrient broth medium, in accordance with the law inspection;
Pseudomonas aeruginosa: get 1: 10 test liquid 10ml of these article and be seeded in the 100ml cholate lactose medium, in accordance with the law inspection;
The every 1ml of these article bacterial population must not cross 100, and the every 1ml of mycete and yeast count must not cross 10, and the every 1ml of escherichia coli must not detect, and staphylococcus aureus, the every 1ml of Pseudomonas aeruginosa must not detect.
2. quality determining method as claimed in claim 1 is characterized in that this method comprises following detection:
(1) get these article 50ml, put in the separatory funnel, with ethyl acetate extraction 2 times, each 15ml; Merge extractive liquid,, evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, and the neutral alumina post 2～4g that is added on internal diameter 9mm is last; With methanol 15ml eluting, collect eluent, be concentrated into 1ml, as need testing solution; Other gets Flos Magnoliae control medicinal material 1g, adds ethyl acetate 15ml, and supersound process 30 minutes filters, and filtrating is concentrated into 1ml, and the neutral alumina post 2～4g that is added on internal diameter 9mm is last, with methanol 15ml eluting, collects eluent, is concentrated into 1ml, as need testing solution; Get the magnelin reference substance again, add methanol and process the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With chloroform-ether of 5: 1 was developing solvent, launched, and took out; Dry, spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing in 105 ℃; In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;
(2) get these article 20ml, put in the separatory funnel, with ether extraction 2 times, each 20ml, merge extractive liquid, adds anhydrous sodium sulfate 2g, and jolting filters, the filtrating evaporate to dryness, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Acori Graminei control medicinal material 0.5g, adds dehydrated alcohol 25ml, and reflux 20 minutes filters, and filtrating is concentrated into 2ml, as control medicinal material solution; According to thin layer chromatography test, draw need testing solution 5 μ l, control medicinal material solution 2 μ l, put in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With cyclohexane extraction-chloroforms of 8: 2: 1-ethyl acetate is developing solvent; Launch, take out, dry; Spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing in 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;
(3) according to high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filler; Containing 0.2% triethylamine, phosphoric acid, to transfer pH2.5, ratio be the mobile phase of methanol-0.02mol/L sodium dihydrogen phosphate of 1: 10; The detection wavelength is 210nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the ephedrine hydrochloride reference substance, and accurate the title decides, and adds mobile phase and processes the solution that contains 30 μ g among every 1ml, promptly gets;
The preparation of need testing solution: get these article, the accurate 8.0ml that draws puts in the 250ml round-bottomed flask; Add sodium chloride 20g, 8% sodium hydroxide solution 150ml shakes up; To heat up in a steamer fast 0.5ml/min distillation, receive distillate to 95ml with the 100ml measuring bottle that fills 1mol/L hydrochloric acid 10ml, thin up is to scale; Shake up, ultrasonic 10 minutes, promptly get;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure; The every 1ml of these article contains Herba Ephedrae in ephedrine hydrochloride, must not be less than 0.30mg.
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| CN105902645A (en) * | 2016-04-26 | 2016-08-31 | 于龙 | External-use ointment for treating rhinitis |
| CN107496504A (en) * | 2017-09-04 | 2017-12-22 | 大连大学 | A kind of Chinese medicine compound prescription liniment for treating allergic rhinitis and preparation method thereof |
| CN108362796A (en) * | 2018-02-23 | 2018-08-03 | 贵州省科晖制药厂 | A kind of detection method of CHENXIANGLU BAILU PIAN traditional Chinese medicine ingredients |
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| CN1762465A (en) * | 2005-10-01 | 2006-04-26 | 合肥立方制药有限公司 | Rhinitis gel, its preparation method and quality control technology |
| CN101002909A (en) * | 2005-12-05 | 2007-07-25 | 北京奇源益德药物研究所 | Antiviral traditional Chinese medicine, its preparing method and quality control method |
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| CN1762465A (en) * | 2005-10-01 | 2006-04-26 | 合肥立方制药有限公司 | Rhinitis gel, its preparation method and quality control technology |
| CN101002909A (en) * | 2005-12-05 | 2007-07-25 | 北京奇源益德药物研究所 | Antiviral traditional Chinese medicine, its preparing method and quality control method |
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| Title |
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| 叶庆棠等.滴通鼻炎水的质量标准研究.《现代医院》.2005,第5卷(第12期), * |
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