UPLC-MS/MS法测定抗菌活性成分DP413_在大鼠血浆中的浓度及药代动力学研究

目的: 建立超高效液相色谱-串联质谱法测定大鼠血浆中活性成分DP413浓度。方法: 血浆样品经蛋白沉淀后,采用Hypersil Gold C₁₈色谱柱(50 mm×2.1 mm,1.9 μm)分离,流动相为甲醇-水,梯度洗脱;选用电喷雾离子化电离源(ESI),负离子方式检测;扫描方式为选择反应监测,用于定量分析的离子反应为m/z 413.2→369.2(DP413)和m/z 331.2→303.3(内标脱水穿心莲内酯,DAG)。结果: 测定DP413的线性范围为5~1 000 ng·mL^-1,在该范围内DP413血药浓度呈良好的线性关系(r=0.999 6);RSD均小于6%,准确度在78.5%~91.1%。结论: 本法经方法学验证,适用于大鼠血浆中DP413浓度测定,可用于DP413大鼠体内药代动力学研究。

Objectives: To establish a UPLC-MS/MS method for simultaneous determination of DP413 in rat plasma,and to reveal corresponding pharmacokinetics. Methods: After a simple protein precipitation using methanol,andrographolide sodium bisulphite was chromatographed on a Hypersil Gold C₁₈ column (50 mm×2.1 mm,1.9 μm) using gradient mobile phase of methanol and water.The detection was performed by the selected reaction monitoring (SRM) mode via the electrospray ionization source (ESI) operating in the negative ionization mode.The precursor-product ion transitions used for quantification were m/z 413.2→369.2 for DP413 and m/z 331.2→303.3 for dehydroandrographolide(DAG). Results: The linear calibration curve was observed in a concentration range of 5-1 000 ng·mL^-1.RSD was less than 6%,and the accuracy was in the range of 78.5%-91.1%. Conclusion: The validation results showed that the established method can be applied to the determination of DP413 concentration in plasma of rats,and was suitable to the pharmacokinetics study of DP413.

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