CN101961430A - Quality analysis method of compound Ganmaoling tablets - Google Patents
Quality analysis method of compound Ganmaoling tablets Download PDFInfo
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Abstract
The invention discloses a quality analysis method of compound Ganmaoling tablets. In the method, high-performance liquid chromatography (HLPC) is used for measure the chlorogenic acid content, acetaminophen content and caffeine content of the compound Ganmaoling tablets, test product solution is prepared and reference product solution is prepared and measured. The quality analysis method of the invention can detect the acetaminophen, chlorogenic acid and caffeine components under the same color spectrum condition at the same time, so the test time and cost are saved and the working efficiency is improved; the test method has high sensitivity, high separating degree; the basic line is stable; a negative reference is not affected; and the accuracy, repeatability, linearity and stability of the test method meet scientific research and production requirements, and the detection method is suitable for promotion and application.
Description
Technical field
The invention belongs to Chinese medicine and western medicine in conjunction with the pharmaceutical technology field, be specifically related to a kind of mass analysis method of fu fang gan mao ling tablets.
Background technology
Fu fang gan mao ling tablets is to add the Chinese medicine and western medicine compound preparation that acetaminophen, caffeine, three chemical medicine compositions of chlorphenamine maleate are made by Six-element Chinese medicines such as Flos Lonicerae, Folium vilicis Negundo, Flos Chrysanthemi Indici, Foliumet Ramulus Evodiae, Rhizoma Et Radix Baphicacanthis Cusiae, Flos Ilicis Asprellae.Have relieving the exterior syndrome with drugs of pungent in flavor and cool in nature, the effect of heat-clearing and toxic substances removing.Be widely used in the heating of anemopyretic cold clinically, micro evil wind is cold, a general pain, and xerostomia is and thirsty, and the nasal obstruction tears are turbid, red swelling and pain of throat, cough, the yellow thickness of expectorant.This product standard now is a tentative standard, and standard No. WS-11191 (ZD-1191)-2002 has the assay project of chlorogenic acid and acetaminophen in the standard.
Accuracy for the quality that guarantees this product and chemical medicine composition feed intake simultaneously also for patient's drug safety, is necessary to increase chemical medicine caffeine, content for Chlorphenamine Maleate is measured project.If increase this two assays, then this kind has chlorogenic acid, acetaminophen, caffeine, 4 assay projects of chlorphenamine maleate, adopts different chromatographic conditions to measure respectively, and detection time is long, detects the cost height, and workload is big.Not about the bibliographical information of the assay of measuring acetaminophen, chlorogenic acid, three compositions of caffeine in the compound recipe cold drug simultaneously, the existing literature report is only at the assay of one of them or two compositions in the prior art; When all being not suitable for above-mentioned three compositions, the experimental verification literature method detects.
Given this, be necessary content assaying method is studied,, reduce and detect cost to shorten Check-Out Time.
Summary of the invention
The object of the present invention is to provide a kind of mass analysis method of fu fang gan mao ling tablets, this mass analysis method can be measured acetaminophen in the fu fang gan mao ling tablets, chlorogenic acid and three kinds of content of effective of caffeine simultaneously.
For achieving the above object, the present invention has taked following technical scheme:
A kind of mass analysis method of fu fang gan mao ling tablets adopts HLPC to measure the content of chlorogenic acid, acetaminophen and caffeine in the fu fang gan mao ling tablets, may further comprise the steps:
(1) preparation need testing solution
Get 10 of fu fang gan mao ling tabletses, remove coating, porphyrize, precision takes by weighing the about 0.67g of fine powder, and adding volume ratio is the phosphoric acid solution 20~30ml of 12: 88 methanol-0.4%, and precision is weighed, after sonic oscillation 15-30 minute, taking-up is put cold, and weight decided in accurate again title, with volume ratio is that the phosphoric acid solution of 12: 88 methanol-0.4% is supplied and subtracted weight loss, shakes up, and filters, the accurate filtrate 2ml that draws, the phosphoric acid solution that adds volume ratio and be 12: 88 methanol-0.4% is diluted to 10ml, shakes up, filter, promptly;
(2) preparation reference substance solution
Precision takes by weighing acetaminophen, chlorogenic acid and caffeine reference substance, be that 12: 88 the phosphoric acid solution dissolving of methanol-0.4% and dilution are made and contained the solution that acetaminophen, chlorogenic acid and caffeine are respectively 0.65mg/ml, 0.03mg/ml and 0.04mg/ml with volume ratio, shake up, filter, promptly;
(3) measure
Chromatographic condition: with the octadecylsilane chemically bonded silica is filler, and the detection wavelength is 290nm; As mobile phase, carry out gradient elution with the phosphoric acid solution of methanol-0.4%; Be 2~4 column volumes of phosphoric acid solution eluting of 12: 88 methanol-0.4% with volume ratio earlier, be 1~3 column volume of phosphoric acid solution eluting of 15: 85 methanol-0.4% then with volume ratio, be 2~4 column volumes of phosphoric acid solution eluting of 20: 80 methanol-0.4% then with volume ratio, 2~4 column volumes of the phosphoric acid solution eluting of reuse volume ratio 12: 88 methanol-0.4%; Flow velocity is 0.5~1.5ml/min; 33~37 ℃ of column temperatures; Theoretical cam curve is pressed acetaminophen, chlorogenic acid and caffeine peak and is calculated, and should be not less than 1200 respectively;
Accurate reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate according to the fu fang gan mao ling tablets finished product, every finished product contains acetaminophen, caffeine and chlorogenic acid and is no less than setting value for qualified.
Preferably, flow velocity is 1ml/min in the described step (3).
Preferably, calculate according to the fu fang gan mao ling tablets finished product, every finished product contains the 90.0%-110.0% that acetaminophen and caffeine should be labelled amount, and chlorogenic acid must not be less than 1.0mg.
Compared with prior art, the present invention has following beneficial effect:
Mass analysis method of the present invention can detect acetaminophen, chlorogenic acid, three compositions of caffeine simultaneously under same chromatographic condition, Check-Out Time and cost have been saved, improved work efficiency, its detection method is highly sensitive, has good separating degree, baseline stability, negative control is noiseless; The accuracy of detection method, repeatability, linear relationship, stability all can reach the requirement of research and production, are suitable for applying.
The specific embodiment
Below describe the present invention in detail by specific embodiment.
The mass analysis method of 1 one kinds of fu fang gan mao ling tabletses of embodiment
Adopt HLPC to measure the content of chlorogenic acid, acetaminophen, caffeine in the fu fang gan mao ling tablets, may further comprise the steps:
(1) preparation need testing solution
Get 10 of fu fang gan mao ling tabletses, remove coating, the accurate title, decided porphyrize, precision takes by weighing the about 0.67g of fine powder, place tool plug triangular flask, adding volume ratio is the phosphoric acid solution 25ml of 12: 88 methanol-0.4%, and precision is weighed, ultrasonic jolting is after 20 minutes, taking-up is put cold, accurately again claims to decide weight, is that the phosphoric acid solution of 12: 88 methanol-0.4% is supplied and subtracted weight loss with volume ratio, shake up, filter, precision is drawn subsequent filtrate 2ml, places the brown measuring bottle of 10ml, the phosphoric acid solution that adds volume ratio and be 12: 88 methanol-0.4% is diluted to scale, shake up, filter, promptly.
(2) preparation reference substance solution
Precision takes by weighing acetaminophen, chlorogenic acid and caffeine reference substance and places brown measuring bottle in right amount, be that 12: 88 the phosphoric acid solution dissolving of methanol-0.4% and dilution are made and contained the solution that acetaminophen, chlorogenic acid and caffeine are respectively 0.65mg/ml, 0.03mg/ml and 0.04mg/ml with volume ratio, shake up, filter, promptly;
(3) measure
Chromatographic condition: with the octadecylsilane chemically bonded silica is filler, and the detection wavelength is 290nm; Mobile phase: A is methanol mutually, and B is 0.4% phosphoric acid solution mutually, carries out gradient elution; Earlier with percent by volume be 12% A phase with 88% B as 3 column volumes of mobile phase eluting, then with percent by volume be 15% A mutually with 85% B as 2 column volumes of mobile phase eluting, then with percent by volume be 20% A mutually with 80% B as 3 column volumes of mobile phase eluting, the A of reuse percent by volume 12% phase and 88% B are as 3 column volumes of mobile phase eluting; Flow velocity is 1.0ml/min; 35 ℃ of column temperatures; Theoretical cam curve is pressed acetaminophen, chlorogenic acid and caffeine peak and is calculated, and should be not less than 1200 respectively;
Accurate reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly.
Get 3 batch samples, carry out acetaminophen, chlorogenic acid and content of caffeine according to the method for this embodiment and measure, with the content of acetaminophen, chlorogenic acid, caffeine in the external standard method calculation sample, measurement result such as table 1.
Table 1
The conditional filtering of the mass analysis method of 2 one kinds of fu fang gan mao ling tabletses of embodiment and methodology checking
1. test sample preparation
With reference to data of literatures, adopt the water supersound extraction, impurity is more.Adopt methanol to make solvent, chromatograph peak-to-peak type is relatively poor.Behind an amount of dissolve with methanol, reuse phosphate buffer dilution standardize solution, the separating degree of acetaminophen, chlorogenic acid and caffeine chromatographic peak and adjacent peak greater than 1.5, purity is better, content is the highest.So select this preparation method for use.
2. mensuration wavelength
Acetaminophen, chlorogenic acid and the caffeine reference substance solution of preparation debita spissitudo, the accurate reference substance solution 20 μ l that draw inject chromatograph of liquid, check acetaminophen, chlorogenic acid and caffeine chromatographic peak spectrogram, the result shows that the acetaminophen spectrogram has absorption maximum under 200nm, 249nm wavelength, cutoff wavelength is 305nm, the chlorogenic acid spectrogram has absorption maximum under 220nm, 327nm wavelength, cutoff wavelength is 375nm, the caffeine spectrogram has absorption maximum under 205nm, 275nm wavelength, cutoff wavelength is 305nm.Because acetaminophen content is big in this product, the content of chlorogenic acid and caffeine is little, if select the 275nm wavelength for detecting wavelength, the chromatographic peak of acetaminophen is higher in chromatogram, and the chlorogenic acid chromatographic peak is little, if select 300nm for detecting wavelength, the chromatograph peak-to-peak of acetaminophen and caffeine is little in chromatogram, if select 290nm for detecting wavelength, acetaminophen, chlorogenic acid and caffeine chromatograph peak-to-peak type are better, the purity height of chromatographic peak is so select 290nm for detecting wavelength.
3. methodology checking
Because this product composition complexity, it is more to detect composition, once adopts the mode of isocratic elution, and the separating degree of target chromatographic peak and adjacent chromatographic peak is relatively poor.The inventor adopts the mode of gradient elution through a large amount of research, and the chromatographic peak separating degree is good, the collection of illustrative plates baseline stability.
(1) linear relationship
Precision takes by weighing the brown measuring bottle that acetaminophen reference substance 54.7mg, chlorogenic acid reference substance 12.1mg and caffeine reference substance 15.8mg place 25ml, the phosphoric acid solution (12: 88) that adds methanol-0.4% dissolves and is diluted to scale, draw 2 respectively again, 1,1,2ml places the brown measuring bottle of 5ml, 5ml, 10ml, 50ml respectively, the phosphoric acid solution (12: 88) that adds methanol-0.4% is diluted to scale, shake up, filter, each 10 μ l of sample introduction carry out linear regression with peak area (Y) to concentration (X) respectively, get regression equation.Regression equation sees Table 2.
Table 2
The material title | Regression equation | R | The range of linearity |
Acetaminophen | Y=4303.9x+40.266 | 0.9999 | 2.188~0.08752mg |
Chlorogenic acid | Y=20661x-73.818 | 0.9999 | 0.484~0.01936mg |
Caffeine | Y=9735.2x-9.6338 | 1 | 0.632~0.02528mg |
(2) specificity test
Form and ratio in prescription, take by weighing raw material that removes acetaminophen, caffeine and the medical material that contains chlorogenic acid, make sheet according to method for preparing tablet thereof, and make negative controls by the need testing solution preparation method, according to (1) method sample introduction, the negative control chromatogram with mix contrast acetaminophen, chlorogenic acid and caffeine place and do not have absworption peak, show under experiment condition, in the prescription in other medical material composition noiseless to measurement result.
(3) precision test
Accurate same sample solution 10 μ l, continuous sample introduction 6 times, the record chromatogram peak area, and the meansigma methods and the RSD% of calculating acetaminophen, chlorogenic acid and caffeine peak area drawn.Acetyl aminophenol peak area RSD% is 0.85, and chlorogenic acid peak area RSD% is 0.67, and caffeine peak area RSD% is 0.67, and the result shows that precision is good.
(4) stability test
Accurate draw same sample solution 10 μ l, respectively 0,2,4,6,8, the 12h sample introduction, record chromatogram peak area, and calculate the meansigma methods and the RSD% of acetaminophen, chlorogenic acid and caffeine peak area.Acetaminophen peak area RSD% is 1.6, and chlorogenic acid peak area RSD% is 1.53, and caffeine peak area RSD% is 1.46, and the result shows that sample solution is stable in 12 hours.
(5) repeatability test
Get the fu fang gan mao ling tablets of same lot number, press the operation of sample determination method, measure content, acetaminophen RSD% is 1.05, and chlorogenic acid RSD% is 1.06, and caffeine RSD% is 1.21, and the result shows: this method repeatability is good.
(6) recovery test
Adopt the application of sample absorption method, precision takes by weighing 6 parts in same sample, and each is an amount of to add acetaminophen, chlorogenic acid and caffeine respectively in every part, presses method operation under the assay item, measures, and calculates average recovery rate and RSD% value.The result: the chlorogenic acid average recovery rate is 98.5%, and RSD is 1.03%, and the acetaminophen average recovery rate is 98.4%, and RSD is 1.20%, and the caffeine average recovery rate is 99.04%, and RSD is 1.04%.
More than be at the specifying of possible embodiments of the present invention, but this embodiment is not in order to limiting claim of the present invention, does not allly break away from equivalence of the present invention and implement or change, all should be contained in the claim of the present invention.
Claims (3)
1. the mass analysis method of a fu fang gan mao ling tablets is characterized in that, adopts HLPC to measure the content of chlorogenic acid, acetaminophen and caffeine in the fu fang gan mao ling tablets, may further comprise the steps:
(1) preparation need testing solution
Get 10 of fu fang gan mao ling tabletses, remove coating, porphyrize, precision takes by weighing the about 0.67g of fine powder, and adding volume ratio is the phosphoric acid solution 20~30ml of 12: 88 methanol-0.4%, and precision is weighed, behind the sonic oscillation 15~30 minutes, taking-up is put cold, and weight decided in accurate again title, with volume ratio is that the phosphoric acid solution of 12: 88 methanol-0.4% is supplied and subtracted weight loss, shakes up, and filters, the accurate filtrate 2ml that draws, the phosphoric acid solution that adds volume ratio and be 12: 88 methanol-0.4% is diluted to 10ml, shakes up, filter, promptly;
(2) preparation reference substance solution
Precision takes by weighing acetaminophen, chlorogenic acid and caffeine reference substance, be that 12: 88 the phosphoric acid solution dissolving of methanol-0.4% and dilution are made and contained the solution that acetaminophen, chlorogenic acid and caffeine are respectively 0.65mg/ml, 0.03mg/ml and 0.04mg/ml with volume ratio, shake up, filter, promptly;
(3) measure
Chromatographic condition: with the octadecylsilane chemically bonded silica is filler, and the detection wavelength is 290nm; As mobile phase, carry out gradient elution with the phosphoric acid solution of methanol-0.4%; Be 2~4 column volumes of phosphoric acid solution eluting of 12: 88 methanol-0.4% with volume ratio earlier, be 1~3 column volume of phosphoric acid solution eluting of 15: 85 methanol-0.4% then with volume ratio, be 2~4 column volumes of phosphoric acid solution eluting of 20: 80 methanol-0.4% then with volume ratio, 2~4 column volumes of the phosphoric acid solution eluting of reuse volume ratio 12: 88 methanol-0.4%; Flow velocity 0.5~1.5ml/min; 33~37 ℃ of column temperatures; Theoretical cam curve is pressed acetaminophen, chlorogenic acid and caffeine peak and is calculated, and should be not less than 1200 respectively;
Accurate reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate according to the fu fang gan mao ling tablets finished product, every finished product contains acetaminophen, chlorogenic acid and caffeine and is no less than setting value for qualified.
2. the mass analysis method of fu fang gan mao ling tablets according to claim 1 is characterized in that, flow velocity is 1ml/min in the described step (3).
3. the mass analysis method of fu fang gan mao ling tablets according to claim 1, it is characterized in that, calculate according to the fu fang gan mao ling tablets finished product, every finished product contains acetaminophen and caffeine should be 90.0%~110.0% of labelled amount, and chlorogenic acid must not be less than 1.0mg.
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