Liu Bi-Feng, Hisamoto Hideaki, Terabe Shigeru
Graduate School of Science, Himeji Institute of Technology, Kamigori, Hyogo 678-1297, Japan.
J Chromatogr A. 2003 Dec 22;1021(1-2):201-7. doi: 10.1016/j.chroma.2003.09.012.
In this article, it was demonstrated that a subsecond separation of cellular metabolites such as riboflavin (RF), flavin mononucleotide (FMN), and flavin-adenine dinucleotide (FAD) was achieved using microchip capillary electrophoresis with laser-induced fluorescence detection. The influences of crucial parameters that governed analysis time (e.g., channel length and electric field for separation) and separation resolution (e.g., sample size) were investigated, both in theoretical aspects and experimental practice. Quantitative analyses were performed that exhibited linear dynamic range of two orders of magnitude, with calculated detection limits of 34, 201, and 127 nM for RF, FAD, and FMN, respectively. To test the validity of the method, it was successfully applied to characterize several recombinant flavin-binding domains in a human neuronal nitric oxide synthase.
在本文中，已证明使用带有激光诱导荧光检测的微芯片毛细管电泳可实现对细胞代谢物如核黄素（RF）、黄素单核苷酸（FMN）和黄素腺嘌呤二核苷酸（FAD）的亚秒级分离。从理论和实验实践两方面研究了控制分析时间（如分离通道长度和电场）和分离分辨率（如样品量）的关键参数的影响。进行了定量分析，其线性动态范围为两个数量级，计算得出的RF、FAD和FMN的检测限分别为34、201和127 nM。为检验该方法的有效性，已成功将其应用于表征人神经元型一氧化氮合酶中的几个重组黄素结合结构域。
