人体体液中环丙沙星及其某些代谢物的液相色谱测定法。

Liquid chromatographic determination of ciprofloxacin and some metabolites in human body fluids.

作者信息

Borner K, Lode H, Höffken G, Prinzing C, Glatzel P, Wiley R

出版信息

J Clin Chem Clin Biochem. 1986 May;24(5):325-31. doi: 10.1515/cclm.1986.24.5.325.

DOI:10.1515/cclm.1986.24.5.325

Abstract

Two column liquid chromatographic (HPLC) methods for the determination of ciprofloxacin and three metabolites are described. Both use reversed phase chromatography, the stationary phase being Nucleosil 5C18. Method A separates ciprofloxacin, metabolite M1 and another metabolite of unknown structure using fluorometric detection. Method B allows the determinations of metabolite M3 (oxo-ciprofloxacin) in urine by UV absorption. Serum was deproteinised with acetonitrile. Urine was diluted with buffer solution. The detection limit of ciprofloxacin was 0.010 mg/l serum and 0.2 mg/l urine and for the metabolite M3, 1 mg/l urine. Within-batch precision (coefficient of variation) for ciprofloxacin in serum was 0.8 to 2.4% and between-batch precision 4.8 to 9.3%. In urine within-batch precision was 1.7 to 2.1% and between-batch precision 2.4 to 7.2%. Recovery rates of ciprofloxacin from three groups of spiked sera was 94.5 +/- 2.6%, 97.2 +/- 1.1% and 95.0 +/- 1.8% and from urine 99.6%. Results obtained by HPLC (method A) were compared with those from a standard microbiological assay by means of bivariate regression analysis. In 12 subsets of data the slope of the regression line varied from 1.042 to 1.556. Significantly higher results from the microbiological assay were probably due to the presence of microbiologically active metabolites. We conclude that HPLC is the more specific method of determination. The described methods were applied for pharmacokinetic studies and therapeutic drug monitoring.

摘要

本文描述了两种用于测定环丙沙星及其三种代谢物的高效液相色谱（HPLC）方法。两种方法均采用反相色谱法，固定相为Nucleosil 5C18。方法A使用荧光检测法分离环丙沙星、代谢物M1和另一种结构未知的代谢物。方法B通过紫外吸收法测定尿液中的代谢物M3（氧代环丙沙星）。血清用乙腈进行脱蛋白处理。尿液用缓冲溶液稀释。环丙沙星在血清中的检测限为0.010 mg/l，在尿液中的检测限为0.2 mg/l；代谢物M3在尿液中的检测限为1 mg/l。血清中环丙沙星的批内精密度（变异系数）为0.8%至2.4%，批间精密度为4.8%至9.3%。尿液中的批内精密度为1.7%至2.1%，批间精密度为2.4%至7.2%。环丙沙星在三组加标血清中的回收率分别为94.5±2.6%、97.2±1.1%和95.0±1.8%，在尿液中的回收率为99.6%。通过双变量回归分析，将HPLC（方法A）获得的结果与标准微生物测定法的结果进行了比较。在12个数据子集中，回归线的斜率在1.042至1.556之间变化。微生物测定法得到的结果明显更高，可能是由于存在具有微生物活性的代谢物。我们得出结论，HPLC是更具特异性的测定方法。所描述的方法应用于药代动力学研究和治疗药物监测。