Mukherjee P S, Karnes H T
Department of Pharmacy and Pharmaceutics, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0533, USA.
Anal Chem. 1996 Jan 15;68(2):327-32. doi: 10.1021/ac950737k.
This report describes the first application of 5-(bromo-methyl)fluorescein (5-BMF) for the quantitation of a pharmaceutically relevant carboxyl-containing analyte in a biological matrix. An analytical method for quantitation of gamma-(cholesteryloxy)butyric acid (CBA), a relatively new antitumor agent, in different tissues of Sprague-Dawley rats was developed. 5-BMF was employed to form a stable and spectrally well-characterized conjugate of CBA. The derivatization yield was maximized by optimizing several reaction variables. The conjugate was separated by high-performance liquid chromatography and quantitated by a laboratory-constructed argon ion laser fluorometer. The detection limits for CBA were 4.6 x 10(-9) and 6.34 x 10(-11) M by conventional and laser-induced fluorescence (LIF), respectively. A derivatization limit of detection of 1.85 x 10(-9) M was achieved by LIF for the conjugate The analytical method was useful for quantitation of CBA in various tissues in the picogram per milliliter range.
本报告描述了5-（溴甲基）荧光素（5-BMF）首次应用于生物基质中与药物相关的含羧基分析物的定量分析。开发了一种用于定量分析γ-（胆固醇氧基）丁酸（CBA，一种相对较新的抗肿瘤剂）在Sprague-Dawley大鼠不同组织中的分析方法。采用5-BMF形成稳定且光谱特征良好的CBA共轭物。通过优化几个反应变量使衍生化产率最大化。共轭物通过高效液相色谱分离，并用实验室构建的氩离子激光荧光计进行定量分析。通过常规荧光和激光诱导荧光（LIF）检测CBA的检测限分别为4.6×10^(-9) M和6.34×10^(-11) M。通过LIF对共轭物实现了1.85×10^(-9) M的衍生化检测限。该分析方法可用于定量分析各种组织中皮克每毫升范围内的CBA。
