Pharmacokinetics Study Of Multiple Components Of Traditional Chinese Medicine In Rat Plasma By Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

The composition of traditional Chinese medicine is complex,and the effect of treatment is the result of interaction of various components.Therefore,it is very important to study the dynamic change process and rule of active components of traditional Chinese medicines in the living body and elucidate the pharmacokinetic characteristics of drug absorption and distribution.Ultra performance liquid chromatography tandem mass spectrometry has the characteristics of high selectivity and high sensitivity.It is widely applied in pharmacokinetics research of traditional Chinese medicine,and it is an effective way of drug metabolism research.In this paper,a rapid,sensitive and selective ultra performance liquid chromatography tandem mass spectrometry method was developed and validated for simultaneous determination of embelin and various active substances of Saururus chinensis(Lour.)Baill and Viola yedoensis Makino in rat plasma.And then establish the corresponding plasma concentration-time curve,calculate the pharmacokinetic parameters and explore pharmacokinetic characteristics for promoting the development of traditional Chinese medicine,elucidating the mechanism and designing of reasonable dosing regimen.1.A simple and selective ultra-performance liquid chromatography-tandem mass spectrometry method was developed for simultaneous determination of seven bioactive constituents(protocatechuic acid,rutin,hyperin,isoquercitrin,quercitrin,luteolin and sauchinone)of Saururus chinensis(Lour.)Baill and investigate the pharmacokinetics characteristics.The sample preparation was accomplished through a deproteinization procedure using puerarin and daidzein as internal standards.Chromatographic separation was carried out on a C18 column(2.1 × 50 mm,1.8 ?m)using water containing 0.10%(v/v)acetic acid and acetonitrile with gradient elution.Detection was performed under the multiple reaction-monitoring mode.The method was validated over the linear range of 15.0-600 ng·mL-1,25.0-400 ng·mL-1,15.0-600 ng·mL-1,15.0-300 ng·mL-1,12.5-250 ng·mL-1,0.25-20 ng·mL-1 and 15.0-400 ng·mL-1 for protocatechuic acid,rutin,hyperin,isoquercitrin,quercitrin,luteolin and sauchinone in rat plasma,respectively.The intra-and inter-day precisions were less than 10.8%,and intra-and inter-day accuracies ranged from-9.8%to 10.7%.The extraction recoveries of the seven compounds ranged from 86.3%to 104.8%.The accurate and rapid method was fully validated and successfully applied to the pharmacokinetic study of seven ingredients in rat plasma.2.In this paper,a method for the determination of embelin in rat plasma using emodin as internal standard was established.Plasma samples were processed by protein precipitation with 200 ?L acetone and 200 ?L ethyl acetate.The separation was achieved by 0.10%ammonia and methanol with a gradient elution at a flow rate of 0.3mL·min-1.The results showed that embelin has a good linearity in the range of 10-1200 ng·mL-1 with a correlation coefficient of 0.9990.The intra-and inter-day precisions were less than 8.7%,and intra-and inter-day accuracies ranged from-8.8%to 14.0%.The extraction recoveries of embelin were greater than 89.2%.The simple,rapid,accurate and selective method was fully validated and successfully applied to the pharmacokinetic study of embelin in rat plasma.3.A rapid,sensitive and selective ultra performance liquid chromatography tandem mass spectrometry method was developed and validated for simultaneous determination of ten constituents(esculetin,caffeic acid,isoscopoletin,scopoletin,orientin,isoorientin,rutin,luteolin,apigenin and ?-sitosterol)in rat plasma using daidzein as internal standard.Plasma samples were processed with a protein precipitation.Chromatographic separation was achieved by using 0.1%formic acid water and methanol as mobile phase with a gradient elution within a runtime of 25 min.The linear ranges of esculetin,caffeic acid,isoscopoletin,scopoletin,orientin,isoorientin,rutin,luteolin,apigenin and P-sitosterol were from 25-7000 ng·mL-1,25-800 ng mL-1,0.5-3000 ng·mL-1,0.5-4000 ng·mL-1,20-500 ng·L-1,25-4000 ng/mL,50-3000 ng·mL-1,25-1000 ng·mL-1,0.025-3 ng·mL-1 and 50-3000 ng ·mL-1,respectively,with the correlation coefficients greater than 0.9977.Mean recovery were ranged from 80.5%-117.8%.The intra-and inter-day precisions were less than 14.5%and accuracies were ranged from-13.8%to 14.6%.The simple and rapid method was successfully applied to investigate the pharmacokinetics study of ten compounds in rat plasma.