液相色谱-串联质谱法同时测定血清中5个三唑类抗真菌药物浓度及其治疗药物监测应用

目的：建立同时测定血清中5个三唑类抗真菌药物（氟康唑、伏立康唑、伊曲康唑、羟基伊曲康唑、泊沙康唑）浓度的液相色谱-串联质谱（LC-MS/MS）方法，并应用于临床三唑类抗真菌药物的治疗监测。方法： 50 μL血清样品加入同位素内标，采用乙腈直接沉淀法处理样品后稀释进样分析。采用XSELECT^TMCSH^TM C₁₈（2.1 mm&#215;100 mm，3.5 μm）色谱柱进行色谱分离，流动相为0.1%甲酸10 mmol&#183;L^-1乙酸铵水溶液-0.1%甲酸乙腈水溶液，梯度洗脱，流速为0.5 mL&#183;min^-1，进样量5 μL，色谱分析时间5.5 min；采用电喷雾离子源，反应监测模式，正离子扫描进行测定。结果：5个三唑类抗真菌化合物的线性范围为0.02~20μg&#183;mL-1，线性相关系数均大于0.998 6，各浓度的批内和批间相对标准差（RSD）<10.6%，提取回收率为80.5%~98.4%，方法准确度为87.4%~111.0%，收集临床ICU和血液科严重侵袭性真菌病20例接受伏立康唑治疗的患者，伏立康唑的血药浓度0.24~4.06 μg&#183;mL^-1，个体间差异较大。结论：本方法同时定量检测5个三唑类抗真菌药物的血药浓度，可应用于临床治疗药物监测。

Objective: To develop a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for simultaneous determination of five azole antifungals(fluconazole, voriconazole, itraconazole, hydroxitraconazole, posaconazole)in human serum and then to apply it to clinical therapeutic drug monitoring.Methods: The serum samples(50 μL)were precipitated with acetonitrile after the addition of isotope internal standards.The chromatographic separation was performed on XSELECT^TM CSH^TM C₁₈(2.1 mm&#215;100 mm, 3.5 μm)with the mobile phase of 10 mmol&#183;L^-1 ammonium acetate solution(containing 0.1% formic acid)-acetonitrile(containing 0.1% formic acid)at a flow rate of 0.5 mL&#183;min^-1.The injection volume was 5 μL and liquid chromatography duration was 5.5 min.The deprotonated ion of analyte was ionized by turbo ion spray and was detected in positive ionization by multiple reaction monitoring mode.Results: The linear concentration ranges of the calibration curves for five azole antifungals were 0.02-20 μg&#183;mL^-1.The coefficients of linearity were more than 0.998 6.Withinbatch and between-batch RSD were less than 10.6%.The average recovery ranged from 80.5% to 98.4%.The accuracy ranged from 87.4% to 111.0%.Twenty patients received voriconazole therapy were collected from ICU and hemotology department.Blood concentration of voriconazole ranged from 0.24 μg&#183;mL^-1 to 4.06 μg&#183;mL^-1.Significant variance was found among patients.Conclusion: The method is suitable for quantitative determination of five azole antifungals in clinical therapeutic drug monitoring practice.

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