UPLC-MS/MS测定人血浆中诺氟沙星的含量及其应用  

UPLC-MS/MS determination of norfloxacin content in human plasma and its application

作　　者：刘海姣 廖音娟[1] 唐智 LIU Hai-jiao;LIAO Yin-juan;TANG Zhi(The Fourth Hospital of Changsha,Changsha 410006;Changsha Duzheng Biotechnology Co.,Ltd,Changsha 410000)

机构地区：[1]长沙市第四医院,长沙410006 [2]长沙都正生物科技有限责任公司,长沙410000

出　　处：《中南药学》2021年第9期1859-1864,共6页Central South Pharmacy

基　　金：复杂基质样本生物分析湖南省重点试验室(No.2017TP1037);药物临床评价关键技术创新及应用示范(No.2019SK2241);长沙市科技计划项目(No.kq1907011)。

摘　　要：目的建立超高效液相色谱-串联质谱(UPLC-MS/MS)测定人血浆样品中诺氟沙星浓度并研究其生物等效性。方法血浆样品经过蛋白沉淀法处理,采用诺氟沙星-d8作为内标。色谱柱为Waters ACQUITY UPLC HSS T3(2.1 mm×50 mm,1.8μm),流动相为0.2%甲酸水-乙腈(85∶15),流速0.4 mL·min^(-1)。电喷雾离子源(ESI),正离子多反应监测模式进行检测。诺氟沙星与内标诺氟沙星-d8的离子对分别为m/z 320.11→m/z 276.13和m/z 328.17→m/z 284.20。对健康受试者口服相同剂量的诺氟沙星片(100 mg)受试制剂或参比制剂后的血药浓度进行定量分析。结果血浆中内源性物质不干扰待测物的测定,人血浆中诺氟沙星的定量范围分别为10~800 ng·mL^(-1)(r^(2)=0.9995),提取回收率为82.7%~86.0%;无明显基质效应,批内、批间精密度和准确度均满足要求。结论该方法准确可靠,且样品前处理和进样过程简单、快速,充分满足诺氟沙星人体生物等效性研究的定量分析需求。Objective To determine the concentration of norfloxacin in human plasma by ultraperformance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method and its human bioequivalence.Methods Plasma samples were processed by protein precipitation method,and with norfloxacin-d8 as the internal standard.The chromatographic column was Waters ACQUITY UPLC HSS T3(2.1 mm×50 mm,1.8μm),the mobile phase was 0.2%formylic acid in wateracetonitrile with a flow rate of 0.4 mL·min^(-1).Electrospray ion source was detected by the positive ion multiple reaction monitoring mode.The ion pairs of norfloxacin and the internal standard norfloxacin-d8 were m/z 320.11→m/z 276.13 and m/z 328.17→m/z 284.20,respectively.The plasma concentration of norfloxacin tablets(100 mg)as the test or reference preparation in healthy volunteers after the oral administration with the same dose were analyzed quantitatively.Results The determination of the substances to be measured was not interfered by the endogenous substances in the plasma,and the quantitative range of norfloxacin in human plasma was 10~800 ng·mL^(-1)with good linearity(r^(2)=0.9995).The recovery of norfloxacin was 82.7%~86.0%,without obvious matrix effect.The intra-and inter-batch accuracy and precision all met requirement.Conclusion The method is accurate and reliable.The sample pretreatment and injection is simple and fast,meeting the quantitative analysis of human bioequivalence study of norfloxacin.

关 键 词：诺氟沙星 生物等效性 超高效液相色谱-串联质谱 

分 类 号：R96[医药卫生—药理学]