HPLC-ECD与HPLC-MS联用测定人尿样中的罗红霉素及其代谢物

 目的:建立灵敏专一的分析方法,测定人尿样中的罗红霉素(E-Rox)及其活性代谢产物Z-Rox?方法:采用高效液相色谱-电化学检测法(HPLC-ECD)与高效液相色谱-质谱法(HPLC-MS),样品经液-液萃取后,通过HPLC-ECD定量分析E-Rox浓度;然后,采用HPLC-MS,通过选择性离子反应监测方式(SRM),测定Z-Rox与E-Rox的尿药浓度比值,SRM扫描方式进行定量分析的碎片离子为m/z679(母离子m/z837)?结果:E-Rox和Z-Rox的线性范围分别为0.1～10mg·L^-1和0.05～5mg·L^-1,最低检测限分别为0.1mg·L^-1和0.05mg·L^-1,日内、日间RSD均小于12.5%?两法联用,分析了4名健康受试者单剂量口服150mg E-Rox后的尿样?结论:为深入研究E-Rox在人体内的代谢规律提供了可靠的分析方法,首次测定了人尿样中的活性代谢产物Z-Rox?

Determination of roxithromycin and its active metabolite in human urine by liquid chromatography electrochemical detection and liquid chromatography mass spectrometry

OBJECTIVE:To establish a sensitive method for the determination of roxithromycin (E-Rox) and its active metabolite,(9Z)-epimeric isomer (Z-Rox).METHOD:E-Rox and Z-Rox were determined by using HPLC-ECD in combination with HPLC-MS.Quantitative analyse of E-Rox was performed with HPLC-ECD after liquid-liquid extraction.Afterwards,the isomeric ratios of Z-Rox and E-Rox were determined by using HPLC-MS in the selected reaction monitoring (SRM) mode via electrospray ionization (ESI).The SRM transition m/z 837 to m/z 679 was monitored for detection of Z-Rox and E-Rox isomers.RESULT:The calibration curves were over the concentration range of 0.1～10mg·L^-1 for E-Rox and 0.05～5mg·L^-1 for Z-Rox.The limits of quantification were found to be 0.1mg·L^-1 for E-Rox and 0.05mg·L^-1 for Z-Rox.The intra-and inter-day RSD were within 12.5% for each of analytes.The combined method was applied to the human urine samples from 4 healthy volunteers after oral administration of 150 mg E-Rox.CONCLUSION:The combined method proved to be suitable for studies on the metabolism of E-Rox in human.With the method,the concentrations of the active metabolite Z-Rox in human urine were for the first time investigated.