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目的建立液质联用(HPLC-MS/MS)法同时测定人血浆中伊立替康(CPT-11)及主要活性代谢产物7-乙基-10-羟基喜树碱(SN-38)及无活性的葡萄糖醛酸化SN-38(SN-38G)的浓度。方法以氘代伊立替康、氘代SN-38、氘代SN-38G为内标,沉淀蛋白法进行提取。色谱柱:ZORBAX XDBC8(2.1 mm×50 mm,5μm),水相:0.1%甲酸,有机相:纯甲醇,进行梯度洗脱,流速:0.35 m L·min-1。通过电喷雾离子源,以正离子多反应监测模式(MRM)进行检测,考察该方法的专属性、标准曲线和定量下限、精密度和准确度、稳定性、基质效应和提取回收率。结果伊立替康标准曲线方程为y=1.32×10-3x-5.30×10-3(r=0.999 7),在10~2000 ng·m L-1线性关系良好,定量下限为10 ng·m L-1。SN-38标准曲线方程为y=8.50×10-3x+4.00×10-3(r=0.999 5),在2~400 ng·m L-1线性关系良好,定量下限为2 ng·m L-1。SN-38G标准曲线方程为y=6.53×10-3x-1.28×10-3(r=0.9997),在5~1000 ng·m L-1线性关系良好,定量下限为5 ng·m L-1。日内与日间RSD均小于15%,平均回收率>92%。结论该方法简便快速,准确度高,灵敏度好,专属性强,适用于同时测定人血浆中伊立替康、SN-38及SN-38G的浓度。
Objective To establish a HPLC-MS / MS method for the simultaneous determination of irinotecan (CPT-11) and 7-ethyl-10-hydroxycamptothecin (SN-38) Activity of glucuronidation of SN-38 (SN-38G). Methods The deuterated irinotecan, deuterated SN-38 and deuterated SN-38G were used as internal standard, and precipitated protein was extracted. Column: ZORBAX XDBC8 (2.1 mm × 50 mm, 5 μm), water phase: 0.1% formic acid, organic phase: pure methanol, gradient elution, flow rate: 0.35 m L · min-1. The electrospray ionization source was used for detection in positive ion multiple reaction monitoring mode (MRM). The specificity, standard curve and quantitation limit, precision and accuracy, stability, matrix effect and extraction recovery of this method were investigated. Results The standard curve of irinotecan was y = 1.32 × 10-3x-5.30 × 10-3 (r = 0.999 7). The linear range was 10-2000 ng · m L-1 with the lower limit of quantitation of 10 ng · m L -1. The calibration curve of SN-38 was y = 8.50 × 10-3x + 4.00 × 10-3 (r = 0.999 5). The linear relationship was good at 2 ~ 400 ng · m L -1 with the lower limit of quantitation of 2 ng · m L- 1. The calibration curve of SN-38G was y = 6.53 × 10-3x-1.28 × 10-3 (r = 0.9997), with good linearity at 5-1000 ng · m L-1 with a lower limit of quantitation of 5 ng · m L -1 . The RSDs were both less than 15% and the average recovery was> 92%. Conclusion The method is simple and rapid, accurate, sensitive and specific. It is suitable for the simultaneous determination of irinotecan, SN-38 and SN-38G in human plasma.