LC-MS/MS法同时检测人血浆中氟尿嘧啶、尿嘧啶和二氢尿嘧啶的浓度及其临床应用  

Simultaneous determination of the concentrations of fluorouracil,uracil and dihydrouracil in human plasma by LC-MS/MS method and its clinical application

作　　者：范先煜 汪硕闻[1] 范国荣[1] FAN Xianyu;WANG Shuowen;FAN Guorong(Department of Clinical Pharmacy,Shanghai General Hospital,Shanghai Jiao Tong University,Shanghai 200080,China)

机构地区：[1]上海交通大学附属第一人民医院临床药学科,上海200080

出　　处：《药学服务与研究》2021年第1期12-18,共7页Pharmaceutical Care and Research

基　　金：上海市卫生计生系统重要薄弱学科建设计划资助项目(2016ZB0302)。

摘　　要：目的:建立同时测定人血浆中氟尿嘧啶、尿嘧啶和二氢尿嘧啶浓度的方法,并应用于临床。方法:建立LC-MS/MS法,以溴尿嘧啶为内标,采用Waters Atlantis T3色谱柱(100 mm×3.0 mm,3μm);0.1%甲酸水溶液(V∶V,A相)-甲醇(B相)为流动相,梯度洗脱(0~2 min,5%B;2.0~2.1 min,5%~40%B;2.1~5 min 40%B;5.0~5.1 min,40%~5%B;5.1~7.0 min,5%B),流速0.35 ml/min。采用电喷雾离子源,多反应监测模式(MRM)进行质谱定量分析。检测离子对:氟尿嘧啶m/z 129→42,尿嘧啶m/z 113→70,二氢尿嘧啶m/z 115→55,内标溴尿嘧啶m/z 189→42。以乙酸乙酯-异丙醇(85∶15,V/V)作为液-液萃取提取剂处理血浆样品。将该方法应用于消化道肿瘤患者的治疗药物监测。结果:氟尿嘧啶和二氢尿嘧啶在10~1000 ng/ml,尿嘧啶在5~500 ng/ml范围内线性良好。日内与日间准确度在88.3%~111.9%,精密度RSD均<8%,基质效应在100.0%~106.9%。9名患者氟尿嘧啶的稳态血药浓度为174.66~634.95 ng/ml,且给药后二氢尿嘧啶与尿嘧啶血浆浓度比值较给药前明显下降。结论:本研究建立的测定方法专属性强,灵敏度高,能快速、准确地对人血浆中氟尿嘧啶、尿嘧啶和二氢尿嘧啶的浓度进行同时检测,满足治疗药物监测的要求,为临床化疗方案的优化提供依据。Objective:To establish an analytical method for simultaneous determination of the concentrations of fluorouracil,uracil and dihydrouracil in human plasma,and to apply it clinically.Methods:The LC-MS/MS method was established with bromouracil as the internal standard.Chromatographic separation was performed on Waters Atlantis T3 column(100 mm×3.0 mm,3μm).0.1%formic acid aqueous solution(A)and methanol(B)were used as the mobile phase by gradient elution(0-2 min,5%B,2.0-2.1 min,5%-40%B,2.1-5 min 40%B,5.0-5.1 min,40%-5%B,5.1-7.0 min,5%B),and flow rate was 0.35 ml/min.Electrospray ion source and multiple reaction monitoring mode(MRM)were used for MS analysis.The detected ion pairs were m/z 129→42(fluorouracil),m/z 113→70(uracil),m/z 115→55(dihydrouracil)and m/z 189→42(bromouracil).Ethyl acetate isopropanol(85∶15,V/V)was used as the liquid-liquid extraction solvent.The established method was used for therapeutic drug monitoring of the patients with gastrointestinal cancer.Results:Good linearity was shown when fluorouracil and dihydrouracil were within the range of 10-1000 ng/ml and when uracil was within the range of 5-500 ng/ml.The intra-day and inter-day accuracy were in the range of 88.3%-111.9%,and relative standad deviation(RSD)were less than 8%.The matrix effect was in the range of 100%-106.9%.The steady state blood concentration of fluorouracil in the 9 patients was 174.66-634.95 ng/ml,and the ratio of plasma concentration of dihydrouracil to uracil after administration was significantly lower than that prior to administration.Conclusion:The established method had high specificity and high sensitivity,and could quickly and accurately determine the concentrations of fluorouracil,uracil and dihydrouracil in human plasma.For this reason,it could meet the requirements of drug concentration monitoring and help to provide good evidence for the optimization of clinical chemotherapy regimens.

关 键 词：氟尿嘧啶 二氢尿嘧啶 尿嘧啶 血药浓度 色谱法 液相 质谱法 联用 治疗药物监测 

分 类 号：R969[医药卫生—药理学] R979.1[医药卫生—药学]