Wise Dana D, Shear Jason B
Department of Chemistry & Biochemistry, The University of Texas, Austin, 78712-0165, USA.
J Chromatogr A. 2006 Apr 14;1111(2):153-8. doi: 10.1016/j.chroma.2005.07.067. Epub 2005 Aug 22.
Capillary electrophoresis (CE) with multiphoton-excited fluorescence detection (CE-MPE) allows low-background analysis of spectrally distinct fluorophores using a single long-wavelength laser. Extracts were prepared from immortalized rat raphe nuclei neurons, and were analyzed by CE-MPE. Native fluorescence was detected from reduced nicotinamide adenine dinucleotide (NADH) and its phosphorylated form (NADPH), flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN), riboflavin, serotonin, and 5-hydroxytryptophan (5HTrp). Quantitation of exogenous serotonin (taken up by cells) and endogenous NADH and 5HTrp was possible using internal standards or standard addition. This system should be useful to study monamine oxidase inhibitors (MAOIs) and selective serotonin reuptake inhibitors (SSRIs).
采用多光子激发荧光检测的毛细管电泳（CE-MPE）能够使用单一长波长激光对光谱上不同的荧光团进行低背景分析。从永生化大鼠中缝核神经元制备提取物，并通过CE-MPE进行分析。检测到了还原型烟酰胺腺嘌呤二核苷酸（NADH）及其磷酸化形式（NADPH）、黄素腺嘌呤二核苷酸（FAD）、黄素单核苷酸（FMN）、核黄素、血清素和5-羟色氨酸（5HTrp）的天然荧光。使用内标或标准加入法可以对外源血清素（细胞摄取）以及内源性NADH和5HTrp进行定量。该系统对于研究单胺氧化酶抑制剂（MAOIs）和选择性5-羟色胺再摄取抑制剂（SSRIs）应该是有用的。
