液相色谱串联质谱同时检测伏立康唑、泊沙康唑和利奈唑胺血清药物浓度方法学建立

Validation of a fast and reliable liquid chromatography tandem mass spectrometry for simultaneous quantitation of Voriconazole,Posaconazole and Linezolid in human serum

目的建立基于液相色谱串联质谱(LC-MS/MS)同时检测伏立康唑(VRC)、泊沙康唑(PCZ)和利奈唑胺(LNZ)血清药物浓度方法。方法方法学验证。收集2022年6月至12月浙江大学医学院附属第二医院服用伏立康唑、泊沙康唑和利奈唑胺的重症感染患者标本分别为155、44和59例。以利奈唑胺-[2H3]为内标,准确称量和配制伏立康唑、泊沙康唑和利奈唑胺工作液,采用ACE PFP色谱柱,梯度洗脱,进行液相色谱质谱分析。质谱分析采用正离子电喷雾离子化多离子反应监测模式(MRM)。根据CLSI中C62-A文件指南和液相色谱质谱临床应用建议指南评价该方法的线性、精密度、最低检测限和回收率等。结果VRC和PCZ线性范围均为0.10~10.00 mg/L,最低检测限均为0.01 mg/L;LNZ线性范围为0.50~50.00 mg/L,最低检测限为0.05 mg/L,范围内线性良好(r均>0.99)。VRC批内变异系数(CV)≤8.30%,批间CV≤6.99%;PCZ批内CV≤9.78%,批间CV≤7.98%;LNZ批内CV≤6.14%,批间CV≤7.14%。VRC、PCZ和LNZ回收率分别为90.96%~103.18%、91.84%~99.17%和97.04%~100.41%。血药浓度VRC155例、PCZ44例和LNZ59例。结论本研究成功建立LC-MS/MS法同时检测VRC、PCZ和LNZ血清药物浓度,该方法检测可靠实用,满足临床治疗药物监测(TDM)需求。

Objective To establish a high-performance liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the determination of voliconazole(VRC),posaconazole(PCZ),and linazolam(LNZ)in human serum.Methods This study is a methodological validation by LC-MS/MS.The blood concentration results of VRC,PCZ,and LNZ in our hospital′s anti-infection patients were collected.Voriconazole,Posaconazole,and Linezolid were accurately weighed and prepared.Linezolid-[2H3]was used as the internal standard.After gradient elution on the ACE PFP column,the residuals were analyzed by LC-MS/MS in the positive electrospray ionization mode and multiple reaction monitor(MRM)mode.The method′s linearity,precision,lower limit of detection,and recovery rate were validated according to standard guidelines.Results The linear correlation coefficient(r)of the standard curve was above 0.99(r>0.99).The linear range of VRC and PCZ were 0.10 mg/L~10.00 mg/L,and the lower limit of detection were 0.01 mg/L.The linear range of LNZ was 0.50 mg/L~50.00 mg/L,and the lower limit of detection was 0.05 mg/L.The recoveries of VRC,PCZ and LNZ were 90.96%-103.18%,91.84%-99.17%,and 97.04%-100.41%,respectively.Intra-and inter-batch precision(%CV)for VRC were less than 8.30%.Intra-and inter-batch precision(%CV)for PCZ was less than 9.78%.Intra-and inter-batch precision(%CV)for LNZ was less than 7.14%.Drug concentrations in 155 cases of VRC,44 cases of PCZ,and 59 cases of LNZ were detected.Conclusion We have established an LC-MS/MS method for the rapid,accurate,highly specific determination of VRC,PCZ,and LNZ concentrations in human serum.This method is suitable for analyzing large clinical sample sets.