HPLC-FLD同时测定全血中加替沙星和莫西沙星含量

Determination of Gatifloxacin and Moxifloxacin in Whole Blood by HPLC Coupled with Fluorescence Detection

目的建立一种灵敏、可靠的全血中加替沙星和莫西沙星的高效液相色谱荧光检测方法。方法样品经磷酸盐缓冲液提取后,用Waters OasisMAX小柱进行净化,甲醇-乙腈-0.2%甲酸(15∶15∶70)为流动相,采用Cloversil-C18柱(3.0mm×150mm,5μm)分离,荧光检测波长:λex288nm和λem493nm,内标法定量。结果加替沙星和莫西沙星在15.0～240.0μg·L-1内呈良好线性,方法回收率在97.7%～101.3%之间,日内RSD<6.0%,其定量限为15.0μg·L-1。结论本方法简便、灵敏、干扰少,特异性好,能满足血液中加替沙星和莫西沙星的检测要求。

OBJECTIVE To develop a sensitive and accurate method for the determination of gatifloxacin and moxifloxacin in whole blood by high-performance liquid chromatography coupled with fluorescence detection（HPLC-FLD）. METHODS After gatifloxacin and moxifloxacin in whole blood were extracted by phosphate buffer, the sample was cleaned with solid-phase extraction （SPE） using Oasis MAX cartridges.The separation was performed on Cloversil - C18 column （3.0 mm × 150 mm, 5 μm） using the mobile phase consisting of methanol-acetonitrile-0.2% formic acid（15 ： 15 ： 70）.Detection was carried out on a fluorescence detection at λex 288 nm and λem 493 nm. RESULTS Calibration curves were linear within the range of 15.0-240.0 μg·L^-1, and the extraction recoveries were from 97.7% to 101.3%, the RSDs were less than 6.0%.The limits of quantification were found to be 15.0 μg ·L^-1.CONCLUSION This method is found to be simple,sensitive,little interferential and good specificity for the determination of gatifloxacin and moxifloxacin in whole blood.