Fukushima Takeshi, Usui Noriko, Santa Tomofumi, Imai Kazuhiro
Laboratory of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
J Pharm Biomed Anal. 2003 Jan 15;30(6):1655-87. doi: 10.1016/s0731-7085(02)00511-3.
The derivatization procedure with a suitable fluorescence or chemiluminescence reagent is performed for the purpose of increasing the detection sensitivity and selectivity, in high-performance liquid chromatography (HPLC) and/or capillary electrophoresis (CE). In this article, recent derivatization methods and their applications to biosamples are described. In HPLC, femto mol order of mass detection limits are obtained by derivatization. Regarding the fluorescence reagents, the use of water-soluble reagents has been effective to avoid an undesired adsorption in the process of determination of peptides. In CE, the advantages of having extremely low mass detection limits (ranging from atto to yocto mol level) and requiring only a very short analysis time (less than a few minutes) are made possible by using laser-induced fluorescence or near infra-red detections.
在高效液相色谱(HPLC)和/或毛细管电泳(CE)中,为提高检测灵敏度和选择性,需使用合适的荧光或化学发光试剂进行衍生化处理。本文介绍了近期的衍生化方法及其在生物样品中的应用。在HPLC中,通过衍生化可实现飞摩尔级别的质量检测限。对于荧光试剂,使用水溶性试剂可有效避免在肽测定过程中出现不必要的吸附。在CE中,利用激光诱导荧光或近红外检测,可实现极低的质量检测限(范围从阿托摩尔到幺摩尔级别)且仅需极短的分析时间(少于几分钟)。