摘要
目的建立一种快捷、简便、灵敏的LC-MS/MS方法测定小鼠血浆中的厄他培南。方法以XZP-3408为内标,血浆样品经过甲醇沉淀蛋白,离心后取上清液进行分析。色谱柱:ZORBAX Eclipse plus C18(4. 6 mm×50. 0 mm,3. 5micron);流动相:乙腈含0. 2%甲酸-0. 2%甲酸水溶液含10 mmol·L~(-1)乙酸铵;梯度洗脱;流速:0. 4 m L·min~(-1);进样量:5μL。电喷雾正离子检测方式,多反应监测。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应、残留效应、稀释效应和稳定性。结果厄他培南在8~2000 ng·m L~(-1)内,线性关系良好,定量下限为8 ng·m L~(-1),批内及批间精密度分别为≤4. 20%和≤5. 81%,样品及内标回收率分别为96. 22%~100. 40%和88. 29%~96. 50%,日内及日间准确度均为-9. 30%~14. 00%,样品及内标基质效应分别为86. 10%~94. 87%和90. 01%~97. 32%,残留效应、稀释效应和稳定性等方法学考察的结果均符合要求。结论本文建立了一种灵敏、快速、稳定的LC-MS/MS方法用于测定小鼠血浆中厄他培南的浓度。
Objective To establish a rapid,sensitive method for the determination of ertapenem in mice plasma. Methods XZP-3408 was taken as the internal standard,the plasma samples were extracted by methanol to precipitate protein. After centrifugation,the supernatant was analyzed by LC-MS/MS. Chromatographic column of ZORBAX Eclipse plus C18( 4. 6 mm × 50. 0 mm,3. 5 micron),mobile phase composed of acetonitrile containing 0. 2% formic acid-water( 0. 2% formic acid)containing 10 mmol ·L~(-1) ammonium acetate in gradient elution mode,flow-rate of 0. 4 m L·min~(-1),injection volume of 5 μL. Electrospray positive ion detection method,multi-reaction monitoring. The specificity,standard curve and lower limit of quantification,precision and recovery,matrix effect,residual effect,dilution effect and stability of the method were investigated. Results Calibration curves were linear between 8 and 2000 ng·m L~(-1). The quantitation limit was 8 ng·m L~(-1).The intra-and inter-precision were ≤4. 20% and ≤5. 81%,the extraction recovery of ertapenem and internal standard were 96. 22%-100. 40%and 88. 29%-96. 50%,the intra-and inter-accuracy were-9. 30%-14. 00%,and the matrix effects of ertapenem and internal standard were 86. 10%-94. 87% and 90. 01%-97. 32%,respectively. All results of methodological study conformed to the requirements. Conclusion A reliable LC-MS/MS method with high sensitivity,simple sample preparation and short elution time was developed for the determination of ertapenem in mice plasma.
引文
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