Fries Harvey E, Evans Christopher A, Ward Keith W
Preclinical Drug Discovery, Cardiovascular and Urogenital Center of Excellence in Drug Discovery, GlaxoSmithKline, King of Prussia, PA 19406, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 May 25;819(2):339-44. doi: 10.1016/j.jchromb.2005.02.009.
Development of a sensitive and specific technique for the quantitation of drug metabolites without the use of synthetic analytical standards or radiolabel would represent a major advance in preliminary route of metabolism screening in drug discovery. In this study, the ability of evaporative light-scattering detection (ELSD) to quantify metabolites of 7-ethoxycoumarin (EC) was evaluated. Because ELSD operates as a mass detector, the complex nature of in vitro-derived samples from hepatocyte incubations resulted in an inability to detect the analytes of interest in this matrix using ELSD. Additionally, the gradient nature of the analysis required to temporally separate ethoxycoumarin from its metabolites and matrix components interfered with the ELSD response. Furthermore, using less-complex contrived mixtures, ELSD demonstrated insufficient sensitivity (limit of detection of 1000-10,000 ng/mL) and an inconsistent inter-analyte response. Together, the limitations outlined in these experiments demonstrate that ELSD is at present an inadequate technique for generating semi-quantitative data on metabolites in drug discovery.
开发一种灵敏且特异的技术，用于在不使用合成分析标准品或放射性标记的情况下定量药物代谢物，这将代表药物发现中初步代谢途径筛选方面的一项重大进展。在本研究中，评估了蒸发光散射检测（ELSD）对7-乙氧基香豆素（EC）代谢物进行定量的能力。由于ELSD作为质量检测器运行，来自肝细胞孵育的体外衍生样品的复杂性质导致无法使用ELSD检测该基质中感兴趣的分析物。此外，为了在时间上分离乙氧基香豆素与其代谢物和基质成分而进行的分析的梯度性质干扰了ELSD响应。此外，使用不太复杂的人工合成混合物时，ELSD表现出灵敏度不足（检测限为1000 - 10000 ng/mL）以及分析物间响应不一致。总之，这些实验中概述的局限性表明，目前ELSD是一种不适合用于生成药物发现中代谢物半定量数据的技术。
