HPLC-MS/MS测定人血浆中妥舒沙星的浓度

Determination of Tosufloxacin in Plasma by HPLC-MS/MS

目的 建立快速、灵敏测定人体血浆中妥舒沙星浓度的高效液相色谱-串联质谱方法。方法 采用Agilent EclipsePlus C18色谱柱（4.6 mm×150 mm,3.5μm）,乙腈（含0.1%甲酸）-5 mmol.L 1甲酸铵水溶液（36∶64）为流动相等度洗脱,流速为0.5 mL.min 1,柱温40℃。采用多重反应监测（MRM）对妥舒沙星（m/z 405.2→387.2）和内标环丙沙星（m/z 332.2→314.2）的测定。结果 妥舒沙星的线性范围为6-2 000 ng.mL 1,回归方程为y=0.183 5x＋1.461 9×10 4,r=0.996 2,最低定量下限为6 ng.mL 1（S/N〉5）,日内、日间RSD均〈10%,高（1 800 ng.mL 1）、中（750 ng.mL 1）、低（15 ng.mL 1）浓度标准血样的平均回收率分别为97.5%,103.3%,105.9%。结论 该方法灵敏、准确、简单、快速,可用于妥舒沙星的临床血药浓度监测和药动学研究。

OBJECTIVE To establish a rapid and sensitive HPLC-MS/MS method for tosufloxacin in plasma. METHODS Tosufloxacin was extracted with acetonitrile and the residues were analyzed with a HPLC-MS/MS system used Agilent Eclipse Plus Cl8 column（4.6 mm× 150 mm, 3.5 min） with the mobile phase consisted of acetonitrile （0.1% formic acid） with 5 mmol＇L-1 ammonium formate（36 ： 64）, with a flow rate of 0.5 mL·min-1 and temperature of 40℃. Multiple reaction monitoring（MRM） using the precursor to production combinations of m/z 405.2→387.2 and m/z 332.2→314.2 was performed to detect tosufloxacin and the internal standard, respectively. RESULTS The calibration curves for tosufloxacin had good linearity within the range of 6-2 000 ng·mL-1, r=0.996 2. The limits of quantitation for tosufloxacin was 6 ng.mL 1. The intra-day and inter-day precision of variation was 〈10%. The average recoveries for high-, middle-, low-dose of tosufloxacin（1 800, 750, 15 ng·mL-1） were 97.5%, 103.3% and 105.9%, respectively. CONCLUSION The method provides a sensitive, accurate, precise and reliable analytical procedure for clinical monitoring oftosufloxacin plasma and its pharmacokinetic studies.