摘要
A rapid and sensitive high performance liquid chromatography-mass spectrometry （HPLC-MS） method for the quantification of nimesulide in human plasma was developed and validated. Sample aliquots of 100μL were extracted by one-step liquid-liquid extraction after addition of hydrochlorothiazide as the internal standard （IS）. Analytes were separated on a reverse phase C18 column using methanol-water （84：16, v/v） as the mobile phase and detected by a single quadrupole mass spectrometer in selected ion monitoring （SIM） negative mode. Monitored m/z values for nimesulide and IS were 307.00 and 295.90, respectively. The overall run time was 4.2 min. Validation experiments demonstrated good precision and accuracy over a wide concentration range of 20.0-7000 ng/mL with a lower limit of quantification （LLOQ） at 20.0 ng/mL. No interference by endogenous substances or matrix effect was observed. Average extraction recoveries for nimesulide and IS were all greater than 84.4%. The assay was successfully applied to a bioequivalence study of nimesulide dispersible tablets in Chinese male volunteers after oral administration.
建立和确证了适用于测定人血浆中尼美舒利含量的液相色谱-质谱(LC-MS)方法,并对尼美舒利参比和受试制剂进行生物等效性评价。人血浆样品(100μL)采用液液萃取法提取,内标为氢氯噻嗪。采用C_(18)柱,以甲醇-水(86:14,v/v)为流动相,选择离子监测负离子模式检测,尼美舒利m/z为307.00,内标氢氯噻嗪m/z为295.90。分析时间4.2分钟。尼美舒利在20.0-7000.0ng/mL浓度范围内线性关系良好,定量下限20.0ng/mL。内源性物质不干扰尼美舒利和内标的检测,无基质效应。尼美舒利和内标氢氯噻嗪的平均提取回收率均大于84.4%。所建立的方法已成功应用于尼美舒利生物等效性研究。
