超高效液相色谱-串联质谱法测定伏立康唑血药浓度及其临床应用

Determination of Voriconazole Blood Concentration by Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry and Its Clinical Application

目的:建立快速准确高通量测定人血清伏立康唑(VRC)血药浓度的超高效液相色谱-串联质谱(UPLC-MS/MS)法,阐明不同年龄段患者VRC血药浓度监测的必要性。方法:以VRC同位素(VRC-d3)为内标,建立UPLC-MS/MS法测定人血清VRC血药浓度,并通过美国病理学家协会(CAP)室间质评活动验证方法准确性。对临床应用结果进行回顾性分析,探讨不同年龄段患者VRC血药浓度监测的重要性。结果:LC-MS/MS运行时间为1.5 min,在0.280.0μg/mL质量浓度范围内与待测物与内标峰面积比值的线性关系良好(R>0.999)。准确度103.5%107.3%,日内、日间精密度相对标准偏差(RSD)<10%。VRC提取回收率为95.20%98.27%,VRC-d3提取回收率为97.12%99.73%,RSD均<10%。CAP室间质评检测结果与均值的偏差为-7.74%-0.85%。50%临床样本检测结果不在治疗范围内,个体差异较大,尤其是儿童患者。结论:本研究建立的UPLC-MS/MS法灵敏度和准确度高,且简便快速,极大缩短了报告周期,适用于儿童和成人VRC血药浓度的快速检测。不同年龄段患者的VRC血药浓度个体差异较大,尤其是02岁低龄儿童急需VRC血药浓度监测。

Objective:To establish the rapid,accurate and high-throughput method for the determination of voriconazole(VRC)concentration in human serum by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),and to clarify the necessity of drug concentration monitoring for VRC in different age groups.Methods:VRC isotope(VRC-d3)was set as the internal standard,UPLC-MS/MS method was established to detect the concentration of VRC in human serum.The accuracy of the method was verified by College of American Pathologists(CAP)external quality assessment.The clinical application results were retrospectively analyzed to discuss the importance of drug concentration monitoring of VRC in different age groups.Results:The LC-MS/MS running time was 1.5 min and showed good linearity(R>0.999)in the mass concentration range from 0.2 to 80.0μg/mL with respect to the peak area ratio of the analyte to the internal standard.The accuracy ranged from 103.5%to 107.3%,and the intra-day and inter-day precision relative standard deviation(RSD)were<10%.The extraction recoveries of VRC were from 95.20%to 98.27%with RSD<10%.The extraction recoveries of VRC-d3 were from 97.12%to 99.73%with RSD<10%.The deviation of CAP results from the mean value ranged from-7.74%~-0.85%.Nearly 50%of the clinical samples tested were out of the therapeutic range,with large individual differences,especially in pediatric patients.Conclusion:The UPLC-MS/MS method established in this study is sensitive and accurate,simple and rapid,and greatly shortens the reporting period,which is suitable for the rapid detection of VRC blood concentration in children and adults.Individual differences in VRC blood concentration in patients of different age groups are large,and VRC blood concentration monitoring is urgently needed,especially in younger children from 0 to 2 years.