中国临床药学杂志

目的采用高效液相色谱-串联质谱(LC-MS/MS)法建立用于检测人血浆中利奈唑胺浓度的方法。方法以CAPCELL PAK C₁₈(100 mm×2.0 mm,TYPE:MGⅡ,5μm)为色谱分析柱,流动相为乙腈-水-甲酸(30∶70∶0.1),流速为0.40 m L·min^-1。以利奈唑胺-d₃为内标,采用蛋白沉淀法进行样本处理。采用多反应监测(MRM),三重四极杆串联质谱仪正离子模式分析利奈唑胺(m/z 338.2→296.2)。本方法学考察了选择性、标准曲线、精密度及准确度、提取回收率及基质效应、残留、稀释可靠性和稳定性。结果利奈唑胺在0.020 00~20.00 mg·L^-1范围内线性良好,回归方程为y=0.376x+0.002 69(r=0.999 2),最低定量下限为0.020 00 mg·L^-1。最低定量下限、低、中、高质控浓度的准确度在84.95%~106.2%,批内和批间精密度(RSD)均<15%。经内标校正后,低、中、高质控浓度的提取回收率分别为104.8%、98.49%和105.3%;低、中、高浓度的基质效应分别为98.50%、89.92%和96.30%。且经验证,在实际生物样本检测过程中各步骤均有足够的稳定性。结论本方法可简便、灵敏、高效测定人血浆中利奈唑胺的血药浓度,且选择性良好,人血浆内源性杂质不干扰利奈唑胺的测定,可以满足临床药物监测及药动学研究的需要。

AIM To develop an LC-MS/MS method to determine the concentration of linezolid in human plasma. METHODS The chromatographic separation was achieved on a CAPCELL PAK C₁₈ column( 100 mm ×2. 0 mm,TYPE: MGⅡ,5 μm) with the mobile phase,consisted of a mixture of acetonitrile∶ water∶ formic acid( 30∶ 70∶ 0. 1),in isocratic mode at a flow rate of 0. 40 m L·min^-1. Deuterium-labeled compound linezolid-d₃ was used as the internal standards. The analyte was extracted from human plasma through protein precipitation method.Data acquisition was carried out in multiple reactions monitoring( MRM) mode. Analyte was detected on a tandem mass spectrometer,equipped with an electrospray ionization source that was operated in the positive mode,using the transitions of m/z 338. 2→m/z 296. 2. The selectivity,standard curve,precision and accuracy,extraction recoveries and matrix effect,carry-over,dilution integrity and stability were investigated. RESULTS The linear range of linezolid was 0. 020 00-20. 00 mg·L^-1. The linear regression equation was y = 0. 376 x + 0. 002 69( r =0. 999 2) with the lower limit of quantitation( LLOQ) of 0. 020 00 mg·L^-1. The accuracy of lower limit of quantitation,low,medium and high concentrations of linezolid was 84. 95 ~ 106. 2 %. The intra-day and inter-day relative standard deviations( RSD) were both less than 15%. The extraction recoveries of lower quality control( LQC),middle quality control( MQC) and higher quality control( HQC) were 104. 8%,98. 49% and 105. 3%respectively. The matrix effects of LQC,MQC and HQC were 98. 50%,89. 92% and 96. 30% respectively. Also,the stability during each step of bioanalysis was validated. CONCLUSION The established method is rapid,sensitive,accurate,specific and reliable,and suitable for determination of linezolid in human plasma,which meets the needs of pharmacokinetic research.