荧光衍生化高效液相色谱法测定大鼠血浆中威替米星的含量

High performance liquid chromatography determination of vertilmicin in plasma using sensitive fluorometric derivatization

目的:建立一种高效液相色谱法(HPLC)用于测定血浆中的威替米星含量。方法:样品处理采用乙腈蛋白沉淀法,并以奈替米星作为内标,将威替米星与氯甲酸芴甲酯(FMOC Cl)衍生化后在反相C18柱上达到分离,以荧光检测器检测反应生成的衍生物。试验中对不同衍生化条件进行了考察。流动相为乙腈水(95∶5),荧光检测激发波长263nm,发射波长315nm。结果:威替米星线性范围是0.02～45mg·L-1,定量限为0.02mg·L-1。血浆中的相对平均回收率为99.84%。结论:本法重现性好,灵敏度高,可以用于治疗药物监控和药动学研究中测定血浆中威替米星的含量。

AIM: To develop a high-performance liquid chromatographic method with fluorescence detection for the determination of vertilmicin in plasma. METHODS: Derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by C_(18) reversed-phase chromatography allowed the fluorimetric detection of vertilmicin. Optimal conditions for the derivatization of vertilmicin are described.The mobile phase consisted of a mixture acetonitrile∶water (95∶5). A fluorescence was set as λ_(ex) 263 nm, λ_(em) 315 nm. RESULTS: The linear range of this method for determination of vertilmicin was 0.02-45 mg·L^(-1), the limit of quantification was 0.02 mg·L^(-1). The relative average recovery rates in plasma was 99.84 %. CONCLUSION: The method is sensitive and reproductive and this method can be used to determine vertilmicin in plasma.