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

关键词:  妥舒沙星  血药浓度  高效液相色谱-串联质谱法

Key words:  tosufloxacin  plasma concentration  HPLC-MS/MS

摘要:

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 Eclipse Plus C₁₈色谱柱(4.6 mm×150 mm，3.5 μm)，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。

Abstract:

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 C₁₈ column(4.6 mm×150 mm, 3.5 μm) 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 of tosufloxacin plasma and its pharmacokinetic studies.