Study On Quality Control,Material Basis Of Efficacy And Pharmacokinetics Of Ganmaoqing Tablet

Ganmaoqing tablet,a kind of Chinese-Western combination preparation for treating flu,was collected in nineteenth volunmes of Chinese Materia Medica preparation?WS3-B-3716-98?,which made of Radix Isatidis,folium isatidis,B.biternata,Radix et Caulis Ilicis asprellae,Radix Helicteris,Andrographis paniculata leaf,p-Acetaminophen,morpholinium hydrochloride and chlorphenamine maleate.At present,there are few studies on the quality control and no studies on material basis of efficacy as well as pharmacokinetics of the Ganmaoqing tablet.Therefore,in this paper,there are several aspects about Ganmaoqing tablet were studied.1.Chemical consitituents of Ganmaoqing tablet based on UPLC-Q-TOF/MSThe seperation was performed on Thermo C18 column?100 mm×4.6 mm,2.6?m?,using acetonitrile-0.1%formic acid as mobile phase.Detected with both positive and negative ion modes in ESI sourse of Q-TOF/MS.A total of 32 chemical consitituents were identified,which include alkaloids,organic acid,diterpenoids and flavone.This provides an accurate and rapid method for identification of chemical consitituents and a foundation for further studying quality control and material basis of efficacy of Ganmaoqing tablet.2.HPLC fingerprint and chemical pattern recognition method of Ganmaoqing tabletThe seperation was performed on Thermo Hypersil GOLD C18?250 mm×4.6 mm,5?m?,using acetonitril-0.2%phosphoric acid as mobile phase.The detection wavelengths were set at 220 nm.Establish fingerprint chromatogram of 22 batches from three companies and 20 public peaks were shown in the HPLC fingerprint,whose similarity was 0.885⁰.967.There were three groups by hierarchical cluster analysis,and 4 main makers were found through principal component analysis and orthogonal partial least squares discriminant analysis.This can provide an accurate and reliable method for quality control for Ganmaoqing tablet.3.Simultaneous determination of three chemical consitituents,content uniformity and stability of Ganmaoqing tablet by HPLCTo develop an HPLC method for simultanously determination the content of p-acetamidophenol,moroxydine hydrochloride,chlorphenamine maleate and its content uniformity as well as its stability in Ganmaoqing tablet.The seperation was carried out on Agilent TC-C₁₈?2??250 mm×4.6 mm,5?m?,using acetonitril-5 mmol·L^-11 Sodium heptane sulfonic acid?adjust pH to 3.0 with phosphoric acid?as mobile phase.The detection wavelengths were set at 249 nm and 262 nm.The three chemical consitituents with adjacent peaks reaching the baseline separation by chromatography.The regression equation showed good liner relationship between the peak area and the concentration of the three chemical consitituents?r=0.9990¹.0000?.The repeatability was good?RSD?0.90%,n=6?.The average recoveries were 101.4%¹03.7%?RSD?0.72%,n=6?.The test solution was stable at room temperature within 24 h?RSD?1.66%?.There were some batches of Ganmanqing tablet unqualified in both content and uniformity.Moroxydine hydrochloride and chlorphenamine maleate were unstable at some conditions.The method was exclusive,accurate,simple and rapid,which can be applied to the quality control of Ganmaoqing tablet.4.Major bioactive components and their metabolites in rats'plasma,urine and feces after oral administration of Ganmaoqing tablet based on UPLC-Q-TOF/MSMajor bioactive components and metabolites in plasma,urine and feces of SD rat were analyzed by UPLC-Q-TOF/MS.The seperation was performed on Thermo C18 column?100 mm×4.6 mm,2.6?m?,using acetonitrile-0.1%formic acid as mobile phase.Detected with both positive and negative ion modes in ESI sourse of Q-TOF/MS.As a result,9 parent compounds and 17 metabolites were detected in plasma,urine and feces samples of SD rats.The main metabolism pathways of these compounds in rats might include glucuronidation,sulfating,methylation,demethylation,isomerization,and so on.The method was accuracy and rapid,which can be used to identify parent compounds and metabolites of Ganmaoqing tablet in SD rats,thus provided scientific basis for pharmacological basis of Ganmaoqing tablet.5.Study on pharmacokinetics of three chemical consitituents from Ganmaoqing tabletTo develop a LC-MS method simultanous determination the plasma-drug content of p-acetamidophenol,moroxydine hydrochloride,chlorphenamine maleate,and study the effect from traditional Chinese medicine on their pharmacokinetics by setting up a control group.The seperation was performed on phenomen C18 column?75 mm×2.1 mm,2.6?m?,using acetonitrile-5 mmol·L^-1 ammonium acetate?containing 0.1%formic acid?as mobile phase.Detected with multiple reaction monitoring mode in ESI sourse.The regression equation showed good liner relationship between the peak area and the concentration of the three chemical consitituents?r=0.9992⁰.9998?.Intra and inner precision were both less 15.0%,and their extract recoveries were 86.6%¹13.5%.The absorption half-life of Ganmaoqing group was greater than that of the chemotherapy group in pharmacokinetics parameter.This method was simple and accuracy,which can be used to simultanously determination the plasma-drug content of p-acetamidophenol,moroxydine hydrochloride,chlorphenamine maleate,and traditional Chinese medicine affects the absorption half-life of chemical drugs.