液相色谱-质谱法测定替考拉宁在重症感染患者血浆中的浓度

Determination the concentration of teicoplanin in human plasma of patients with severe infenction by liquid chromatography tandem mass spectrometry

目的建立液相色谱-质谱联用法(LC-MS/MS)测定人血浆中替考拉宁的浓度。方法人血浆样品用甲醇沉淀蛋白后,选用Agilent Eclipse Plus C_18(2.1 mm×100 mm,3.5μm)色谱柱,以甲醇-10 mmol·L^(-1)醋酸铵溶液(均含0.1%甲酸)85∶15为流动相,流速为0.25 m L·min^(-1),柱温为25~℃,用电喷雾离子化源,正离子方式,多反应监测(MRM)扫描方式进行监测,用于定量分析的离子反应分别为m/z 1880.1→m/z 1563.0(替考拉宁)和m/z 748.8→m/z158.1(内标克拉霉素)。考察其专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。结果血浆中替考拉宁的标准曲线方程为y=1.63×10^(-2)x-2.95×10^(-3)(r=0.996 3),在0.10~20.00μg·m L^(-1)内线性关系良好,定量下限为0.10μg·m L^(-1);质控样品的准确度在93.56%~105.12%;日内、日间RSD均小于15%;提取回收率80.32%~88.28%。基质效应为91.78%~108.56%。结论该方法快速、灵敏、准确,专属性强,重复性好,适用于人血浆中替考拉宁浓度的测定,可应用于替考拉宁的血药浓度检测和药物动力学研究。

Objective To develop a liquid chromatography tandem mass spectrometry（ LC -MS/MS） method for determination of s teicoplanin in human plasma. Methods The plasma samples were precipitated by methnaol, then the supernatant were separated on Eclipse Plus C1scolumn （2. 1 mm× 100 mm, 3. 5 μm） and eluted with methnaol - 10 mmol·L -1 ammonium acetate（85： 15, v/v, containing 0. 1% formic acid） , and the column temperature was 25 ℃. Detection of the analyte was achieved using positive ion electrospray ionization （ESI） in the multiple reaction monitoring （MRM） mode. The MS/MS ion transitions monitored were m/z 1880. 1→m/z 1563.0, m/z→748.8 m/z 158.1 for teicoplaninand internal standard, respectively. The specificity, standard curve and lower limitation of quantitation, precision and recovery rate, stability and ma- trix effect were investigated. Results The linear regression equation was y = 1.63× 10-2x -2.95 × 10-3（r =0. 9963）. The linear range of teico- planin was 0. 10 - 20. 00μg·mL-1, with the lower limit of quantitation （LLOQ） of 0. 10 μg · mL-1. The accuracy was 93.56% - 105.12%. Intraday and inter - dayrelative standard deviations were both below 15%. The absolute recovery in plasma was 80. 32% -88.28%. Thematrix effect was 91.78% - 108.56%. Conclusion The established method is rapid, sensitive,accurate, specific and reliable, and it is suitable for the determination of teicoplanin in human plasma and pharmacokinetic study.