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目的建立快速、灵敏、简便的超高效液相色谱-串联质谱(UPLC-MS/MS)法同时测定大鼠血浆中芍药苷及其代谢产物芍药内酯苷的暴露量,并研究单次ig给予大鼠低、中、高剂量(20、60、120 mg/kg)的芍药苷水溶液后,芍药苷、芍药内酯苷在大鼠体内的药动学特征。方法以栀子苷为内标,血浆样品经甲醇(含0.1%甲酸)沉淀蛋白后,通过ACQUITY UPLC BEH-C_(18)柱(50 mm×2.1 mm,1.7μm),以0.1%甲酸水溶液-乙腈为流动相梯度洗脱,色谱运行时间为4.5 min。采用电喷雾离子源(ESI),负离子扫描模式,以多反应监测方式(MRM)进行定量测定。结果芍药苷、芍药内酯苷的标准曲线的线性范围均为2.00~400 ng/m L,定量下限均为2.00 ng/m L,2者日内和日间精密度RSD均小于9.10%。芍药苷在低、中、高3个剂量给药组中的t_(max)分别为(1.56±0.62)、(1.13±0.35)、(1.28±1.92)h,Cmax分别为(85.45±47.49)、(390.75±139.26)、(1 223.5±420.15)μg/L;其中芍药苷水溶液低、中剂量组未检测到代谢产物芍药内酯苷,芍药苷水溶液高剂量组芍药内酯苷的tmax为(1.81±0.53)h,C_(max)为(19.81±8.98)μg/L。结论本方法简便、准确、专属性强,适用于芍药苷和芍药内酯苷在大鼠体内的药动学研究。
OBJECTIVE To establish a rapid, sensitive and simple method for the simultaneous determination of paeoniflorin and its metabolite of paeoniflorin in rat plasma by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) Pharmacokinetics of paeoniflorin and paeoniflorin in rats after low, medium and high dose (20, 60, 120 mg / kg) of paeoniflorin aqueous solution. Methods Geniposide was used as an internal standard. The plasma samples were precipitated with methanol (containing 0.1% formic acid) and eluted with ACQUITY UPLC BEH-C 18 column (50 mm × 2.1 mm, 1.7 μm) Acetonitrile gradient elution mobile phase, chromatographic run time was 4.5 min. The electrospray ionization source (ESI) and negative ion scanning mode were used for quantitative determination by multiple reaction monitoring (MRM). Results The calibration curves of paeoniflorin and paeoniflorin showed a linear range of 2.00-400 ng / mL with a lower limit of quantification of 2.00 ng / mL. Both RSD and RSD of intra-day and inter-day precision were less than 9.10%. The t max of paeoniflorin were (1.56 ± 0.62), (1.13 ± 0.35) and (1.28 ± 1.92) h in the low, medium and high dose groups, respectively, with Cmax of (85.45 ± 47.49) (390.75 ± 139.26) and (1223.5 ± 420.15) μg / L, respectively. The metabolites of paeoniflorin were not detected in the medium and low doses of paeoniflorin, and the tmax of the paeoniflorin in the high dose of paeoniflorin solution was (1.81 ± 0.53) h, C max (19.81 ± 8.98) μg / L. Conclusion The method is simple, accurate and specific, and is suitable for the pharmacokinetics of paeoniflorin and paeoniflorin in rats.