LC-MS/MS测定人血浆中罗红霉素的浓度及其在生物等效性研究中的应用

Determination of Roxithromycin in Plasma by LC-MS/MS and Its Application in the Bioequivalence Study

关键词:  罗红霉素  液相色谱-串联质谱法  生物等效性

Key words:  roxithromycin  LC-MS/MS  bioequivalence

摘要:

目的　建立LC-MS/MS测定人血浆中罗红霉素浓度的方法，并研究其制剂的生物等效性。方法　以克拉霉素为内标，色谱柱：Alltech Alltima(2.1 mm×100 mm，3.5 μm)；流动相：0.1%甲酸溶液-乙腈(45∶55)；流速：0.2 mL·min^-1；柱温：30 ℃。采用三重四极杆串联质谱，电喷雾离子源，正离子模式检测，以选择反应监测(SRM)方式进行检测，罗红霉素母离子[M+H]+m/z 837.5，子离子m/z 679.2，内标克拉霉素母离子[M+H]+m/z 748.5，子离子m/z 590.2。结果　罗红霉素线性范围为0.10~20.00 μg·mL^-1，最低检测限为0. 01 μg·mL^-1，3种浓度的相对回收率为100.8%~103.1%，日内、日间RSD均<5.9%(n=5)。生物等效性研究表明受试制剂和参比制剂生物等效，受试制剂相对生物利用度为(100.655±9.552)% (P=0.5%)。结论　该方法专属、灵敏、快速，适用于罗红霉素制剂的生物等效性研究。

Abstract:

OBJECTIVE To establish a LC-MS/MS method for determination of roxithromycin in plasma and to study its bioequivalence. METHODS Clarithromycin was used as internal standard. The method was performed on Alltech Alltima (2.1 mm×100 mm, 3.5 μm) at 30 ℃, and 0.1% formic acid-acetonitrile(45∶55) was used as mobile phase with the flow rate of 0.2 mL·min^-1. Electrospray ionization source was applied and operated in positive ion mode. Selected reaction monitoring(SRM) mode with the transitions of m/z 837.5→679.2 and m/z 748.5→590.2 was used to quantify roxithromycin and clarithromycin, respectively. RESULTS The assay was linear over the range of 0.10-20.00 μg·mL^-1, the limit of detection was 0.01 μg·mL^-1, relative recoveries at three level concentrations ranged from 100.8% to 103.1%, the intra- and inter-day precision(RSD) were below 5.9%. Bioequivalence study showed the test roxithromycin tablets was bioequivalent to the reference tablets. The relative bioavail-ability of the test sample was (100.655±9.552)%(P=0.5%). CONCLUSION The method is simple, rapid, accurate and can be applied to perform the bioequivalence research of roxithromycin tablets.