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  

Quantitative determination of five azole antifungals in human serum by LC-MS/MS and its application to therapeutic drug monitoring

作　　者：张素洁[1] 孙贺伟[1] 郭君君[1] 陈梅[1] 袁想妹 李水军[2,3] 胡新华[2,3] 王艳煜[2,3] 余琛[2,3] 朱建民[2,3] 

机构地区：[1]上海大学生命科学学院,上海200444 [2]上海市徐汇区中心医院/复旦大学附属中山医院徐汇医院,上海200031 [3]中国科学院上海临床研究中心,上海200031

出　　处：《药物分析杂志》2017年第6期1038-1045,共8页Chinese Journal of Pharmaceutical Analysis

摘　　要：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:50μL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XSELECTTM CSHTM C_(18)(2.1 mm×100 mm,3.5μm)色谱柱进行色谱分离,流动相为0.1%甲酸10 mmol·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 m L·min^(-1),进样量5μL,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·m L^(-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0.24~4.06μg·m L^(-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 （ 50μL ） were precipitated with acetonitrile after the addition of isotope internal standards. The chromatographic separation was performed on XSELECTTM CSHTM C18（2. 1 mm 100 mm, 3.5 μm ）with the mobile phase of 10 mmol L-1 ammonium acetate solution （ containing 0. 1% formic acid ） - acetonitrile （ containing O. 1% formic acid ）at a flow rate of 0. 5 mL 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 O. 02-20 μg mL-1. The coefficients of linearity were more than 0. 998 6. Within- batch 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 mL-1 to 4.06μg 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.

关 键 词：氟康唑 伏立康唑 伊曲康唑 羟基伊曲康唑 泊沙康唑 临床药物监测 抗真菌治疗 液相色谱-串联质谱法 

分 类 号：R917[医药卫生—药物分析学]