摘要
Abstract
7–ethyl–14–Aminocamptothecin (EAC) is a novel derivate of camptothecin which exhibits favorable antitumor activity.To determine the concentrations of the parent drug and lactam form metabolite (EAC-M) in vivo after intravenous administration of EAC,an UPLC-MS/MS method was established to simultaneously quantify EAC and EAC-M in SD rat plasma.7–ethyl–14–Nitrocamp-tothecin (ENC) and Chimmitecan (CMT) were used as internal standards.After protein precipitation by adding solution of internal standards and acetonitrile,plasma samples were separated on a Phenyl-Hexyl colum (2.1mm 50 mm,1.8 μm) with the mobile phase con-sisting of 0.5 mmoL/L ammonium acetate solution (containing 0.1% formic acid)-acetonitrile.Gradient elution was employed at a flow rate of 0.5 mL/min.Eluted compounds were ionized by an electrospray ionization source (ESI) in positive mode.The parent to production transitions for both the parent drug (m/z 392→291) and the metabolite (m/z 392→290) were monitored on a triple quadrupole mass spectrometer,using the multiple reaction monitoring (MRM).Based on the results from method validation,no interference was observed in blank plasma samples.The linear ranges for EAC and EAC–M were 0.500~250 ng/mL,the limits of quantitation were 0.500 ng/mL.The inter- and intra-day precisions(RSD)were not more than 7.6%,the accuracies(RE)were–4.2%~5.9%.The extraction recovery values were 84.4%~98.7%,and the matrix effects were 94.8%~104%.Samples beyond the upper limits of quantification were analyzed after a 6–fold-dilution with ×blank plasma,the precisions (RSD)were 1.8%,and the accuracies(RE) were –10%~–3.0%.In stability tests,the accuracies (RE) were–8.6%~11% and the precisions(RSD)were blow 7.2%.Data from pharmacokinetic tests indicated that EAC is moderately cleaned and is widely distributed in tissues.EAC-M was rapidly detected in plasma after intravenous administration which showed a 5% exposure of EAC.No accumula-tion of EAC or EAC-M was observed after an administration for 5 consecutive days.In summary,the provided method is convenient,sensitive,se-lective,and suitable for investigation of EAC and EAC-M in pharmacokinetic research.关键词
7–乙基–14–氨基喜树碱/内酰胺代谢产物/大鼠血浆/超高效液相色谱–串联质谱/同时定量检测Key words
7–ethyl–14–Aminocamptothecin/lactam metabolite/rat plasma/UPLC-MS/MS/simultaneous quantitative analysis分类
医药卫生
