HPLC/MS/MS法测定大鼠肝微粒体系统中非那西丁及其代谢物的浓度

目的:以高效液相色谱-串联质谱法测定大鼠肝微粒体孵育液中非那西丁及其代谢物对乙酰氨基酚的浓度。方法:色谱柱为XTerraMSC18,流动相为甲醇-0.1%甲酸不同比例梯度洗脱,流速为0.2mL·min-1;质谱仪为电喷雾-三重四极杆质谱,以多反应监测方式采集数据。结果:非那西丁和对乙酰氨基酚的检测浓度线性范围分别为45～9000(r=0.9998)、15.2～1520ng·mL-1(r=0.9996);平均回收率分别为(96.2±2.3)%～(98.3±2.4)%、(99.6±2.1)%～(100.2±2.6)%;日内及日间精密度均小于5%,最低检测浓度分别为9、10ng·mL-1。结论:本法简单、快速、灵敏,适宜于检测大鼠肝微粒体孵育液中非那西丁及其代谢物浓度。

Determination of Phenacetin and Paracetamol in Rat Liver Microsomal Incubation System with HPLC/MS/MS

OBJECTIVE: To develop a method of HPLC/MS/MS to determine phenacetin and paracetamol in rat liver microsomal incubation system. METHODS: Samples were separated on XTerra MS C18 column, different ratios of methanol- 0.1% formic acid were used as the gradient eluent, and the flow rate was 0.2mL·min-1. The electrospray ion-quadrupole mass spectrometry and multiple reaction monitor were adopted to detect the concentration of phenacetin and paracetamol. RESULTS: The calibration curves were linear in the ranges of 45～9 000ng·mL-1(r=0.999 8)and 15.2～1 520ng·mL-1(r=0.999 6) for phenacetin and paracetamol respectively; The lowest limits of assay were 9ng·mL-1 and 10ng·mL-1.The average recoveries at three concentrations of phenacetin were (96.2±2.3)%～(98.3±2.4)% and those of paracetamol were (99.6±2.1)%～(100.2±2.6)%; RSD of the intra-day and inter-day were less than 5%. CONCLUSION: The method is rapid, sensitive and suitable for determination of phenacetin and paracetamol in rat liver microsomal incubation system.