| UHPLC-MS/MS is one of the most widely used method in the separation and determination of complex compounds.It has the advantages of high sensitivity,high selectivity and high separation ability,which plays an important role in the field of drug analysis.The composition of Traditional Chinese Medicine?TCM?is complex,and its efficacy will be affected by the interaction of multiple components.Therefore,it is necessary to understand the pharmacokinetics and tissue distribution characteristics of the bioactive components of TCM.In this paper,UHPLC-MS/MS method was established to simultaneous determine the concentration of multiple bioactive components in SD rats,and the characteristics of pharmacokinetics and distribution of bioactive components in tissues were studiedA rapid and sensitive UHPLC-MS/MS method was developed for simultaneous determination of thirteen bioactive compounds?gallic acid,protocatechuic acid,puerarin,p-hydroxycinnamic acid,daidzin,ononin,daidzein,naringenin,genistein,apigenin,formononetin,biochanin A,and?-sitosterol?of Radix Puerariae extract in rat plasma and tissues.The biological samples were pretreated by protein precipitation extraction,and umbelliferone?IS1?and rutin?IS2?were used as internal standards.The analytical method exhibited good linearity with correlation coefficient of analytes in the range of concentration r?0.9983.The lower limit of quantitation range for the thirteen analytes was 0.20-35 ngˇmL-1.The intra-and inter-day precision of all the analytes were less than 12%,with an accuracy ranging from-13%to 12%.The recovery and matrix effect ranged from 81%to 114%and 80%to 113%,respectively.There was no residue in the sample after injection,and the stability of the analytes in the sample was good under four different conditions.This method was successfully applied to pharmacokinetic and tissue distribution study of the thirteen compounds in rats after oral dosing of Radix Puerariae extract.It provides basis for further research on the pharmacology of Radix Puerariae and also provides useful information for the safety of this herb medicine in clinical useBased on the technology of UHPLC-MS/MS,a rapid and sensitive analytical method was established to simultaneous determination of ten bioactive components?gallic acid,protocatechuic acid,chlorogenic acid,caffeic acid,p-hydroxycinnamic acid,hyperoside,rutin,isochlorogenic acid C,quercetin and luteolin?of Senecio scandens extract in rat plasma and tissues Puerarin?IS1?and emodin?IS2?were used as double internal standards,and acetonitrile was selected as the protein precipitant for sample treatment.The flow phase consisting of methanol-water?containing 0.1%formic acid?.Ten bioactive components and two internal standards could be separated within 18 minutes.The analytes exhibited good linearity in the corresponding linear range.The lower limits of quantification were between 1.0-50 ngˇmL-1 for all analytes.The intra-and inter-day precision were less than 13%,the intra-and inter-day accuracy ranged from-13%to 12%and-11%to 12%,and the recovery and matrix effect were within 80%to 118%and 81%to 116%.The carry-over effect of the samples could be ignored,and the stability of the analytes in four different environments was good.The data shown that the analytical method was accurate and reliable,and it was successfully applied to pharmacokinetic and tissue distribution study of ten bioactive components in rats after oral administration of Senecio scandens extract.The results of the study make a contribution to provide reference for the pharmacology,toxicology,quality control,and clinical drug safety of Senecio scandens. |
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