Schaefer H G
Pharma Research Center, Bayer AG, Wuppertal, Germany.
J Chromatogr. 1993 Jun 23;616(1):87-93. doi: 10.1016/0378-4347(93)80475-j.
A reversed-phase high-performance liquid chromatographic (HPLC) method that allows the sensitive and selective quantification of a novel 4-quinolone (BAY y 3118, I) in biological fluids is described. After sample dilution with 0.05 M phosphoric acid (plasma) or 0.1 M phosphate buffer pH 7.5 (urine), samples can be directly injected into the HPLC system. Prior to fluorescence detection, I is decomposed to fluorescence compound(s) by post-column derivatization utilizing either photolysis (Beam Boost reaction unit) or a combination of thermolysis and photolysis (laboratory-made post-column reactor). Compared with fluorescence detection alone, derivatization increases the signal intensity (about 80-fold) and the selectivity of the detection significantly. Concentrations down to 0.01 mg/l could be quantified in biological fluids. Only thermolysis was not able to decompose I to fluorescence products. Investigations on the stability of I in plasma and urine demonstrate good stability under the different conditions tested. The method was applied to human plasma and urine samples from a subject after a single oral dose of 100 mg of I.
本文描述了一种反相高效液相色谱（HPLC）方法，该方法可对生物流体中的新型4-喹诺酮（BAY y 3118，I）进行灵敏且选择性的定量分析。用0.05 M磷酸（血浆）或0.1 M pH 7.5的磷酸盐缓冲液（尿液）稀释样品后，可将样品直接注入HPLC系统。在荧光检测之前，通过柱后衍生化利用光解（Beam Boost反应单元）或热解与光解相结合的方式（自制柱后反应器）将I分解为荧光化合物。与单独的荧光检测相比，衍生化显著提高了信号强度（约80倍）和检测的选择性。生物流体中低至0.01 mg/l的浓度也可进行定量分析。仅热解无法将I分解为荧光产物。对I在血浆和尿液中的稳定性研究表明，在测试的不同条件下其稳定性良好。该方法应用于一名受试者单次口服100 mg I后的人体血浆和尿液样本。
