Pharmacokinetic Studies Of Major Bioactive Compounds From Traditional Chinese Medicines In Rat Plasma By UHPLC-MS/MS

The curative effects of traditional Chinese medicines(TCM)are the results of complex interactions between various active ingredients,so pharmacokinetic studies are indispensable for elucidating their mechanism of action in vivo,and have been paid more and more attention in recent years.The mature UHPLC-MS/MS has the advantages of simple operation,high sensitivity,high efficiency and strong specificity,so it plays an irreplaceable role in the research of TCM's pharmacokinetics.In this study,UHPLC-MS/MS was used to qualitative and quantify the active components of Solanum lyratum,Erigeron breviscapus and Salvia chinensis benth in rat plasma and to explore the pharmacokinetic characteristics of rats following oral administration.The methods provide scientific basis for the mechanism of drug efficacy,the development of new drugs and the clinical references on rational administration.1.A simple and selective UHPLC-MS/MS method was developed and utilized to pharmacokinetic study of eight major absorbed compounds of Solanum lyratum,including protocatechuic acid,chlorogenic acid,caffeic acid,scopoletin,scutellarin,rutin,isochlorogenic acid C,and diosgenin.The plasma samples were acidified by formic acid and pretreated with 600?L ethyl acetate-acetonitrile(3:1,v/v).A 15 min linear gradient elution was used at a flow rate of 0.3 mL/min with a mobile phase of 0.1%formic acid in water and methanol.Intra-run and inter-run precision were in the range of 1.1%?11.4%and 1.407%?12.8%.Matrix effect was found to be minimal.Extraction recoveries of all the analytes were all more than 84.8%.The comprehensive studies provided essential data for further quality control,pharmacological as well as toxicological researches of Solanum lyratum.2.A sensitive and accurate UHPLC-MS/MS analytical method for simultaneous quantitation of twelve major bioactive ingredients(neochlorogenic acid,chlorogenic acid,caffeic acid,cynarin,scopoletin,scutellarin,isochlorogenic acid A,apigenin-7-o-glucuronide,isochlorogenic acid C,scutellarein,luteolin,and apigenin)in rat plasma was developed and fully validated.The plasma samples were pretreated and extracted by liquid-liquid extraction and protein precipitation with ethyl acetate-acetonitrile(95:5,v/v).The 0.1%formic acid aqueous solution and acetonitrile were used as mobile phase under gradient conditions at a flow rate of 0.3 mL/min.The correlation coefficients of all components were higher than 0.9990 within the linear ranges,which showed excellent correlation.Within-and between-day accuracy values were over the range of-10.43%to 9.76%and-10.14%to 10.33%.Mean extraction recovery ranged from 85.0%?111.9%in plasma,matrix effect and stability were found to be the required limits.This method could provide pharmacokinetic evidence for in vivo study and action mechanism of Erigeron breviscapus.3.Reliable,rapid analytical method was applied to simultaneously determine danshensu,protocatechuic acid,neochlorogenic acid,caffeic acid,rutin,isoquercitrin,vincetoxicoside B,apigenin-7-O-glucronide,rosmarinic acid,and methylrosmarinate in plasma at different time following administration the extract of Salvia chinensis benth by UHPLC-MS/MS.Gallic acid(IS1)and wogonoside(IS2)were selected as internal standards.Chromatography was performed with a gradient elution of 0.2%formic acid and acetonitrile at 0.3 mL/min.Detection and quantitation were performed by multiple reaction monitoring(MRM)scanning using electrospray ionization(ESI)in positive and negative mode.The correlation coefficients of all components were greater than 0.9990.The LLOQ of ten ingredients were 100,3.0,7.0,500,8.0,0.5,0.2,8.0,6.0 and 1.5 ng·mL-1,respectively.The intraday precision was<9.6%and the interday precision was?12.1%.This method was simple,sensitive and stable,so it was appropriate for plasma concentration monitoring and pharmacokinetic studies.