Capillary electrophoresis apparatus equipped with a bioluminescence detector using a batch- or flow-type detection cell.
Capillary electrophoresis apparatus equipped with a bioluminescence detector using a batch- or flow-type detection cell.
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DOI：
10.1016/j.chroma.2005.09.048
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发表时间：
2005-11
期刊：
影响因子：
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通讯作者：
K. Tsukagoshi;Masayuki Tahira;R. Nakajima
中科院分区：
文献类型：
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作者：
K. Tsukagoshi;Masayuki Tahira;R. Nakajima
我们开发了一种配备批量或流动型生物发光（BL）检测池的毛细管电泳（CE）装置。萤火虫荧光素-荧光素酶 BL 反应用于分析核苷酸样品，例如 ATP、dATP、ADP、GTP、UTP、CTP、ITP 和 TTP。在带有批式细胞的CE装置中，ATP在5-100μM浓度下被检测到，而其他核苷酸在浓度低于500μM时未检测到。 ATP的电泳图包括两个BL峰；后一个峰表现出特殊的加宽，一直持续到大约。 2.5小时。在带有流动型池的CE装置中，ATP、dATP和ADP均呈单峰检测，检测限分别为1、75和100μM。其他核苷酸（GTP、UTP、CTP、ITP 和 TTP）在浓度低于 0.5 mM 时未检测到。使用带有流式 BL 检测池的 CE 装置检查 10μM ATP 和 100μM dATP 的混合物。使用含有 1mM 苯基硼酸的 pH 10 运行缓冲液分离 ATP 和 dATP。 ATP与苯基硼酸之间的相互作用延迟了ATP的迁移时间。
We developed a capillary electrophoresis (CE) apparatus equipped with a batch- or flow-type bioluminescence (BL) detection cell. Firefly luciferin–luciferase BL reaction was used to analyze samples of nucleotides, such as ATP, dATP, ADP, GTP, UTP, CTP, ITP, and TTP. In the CE apparatus with the batch-type cell, ATP was detected at concentrations of 5–100μM, while the other nucleotides were not detected at concentrations less than 500μM. The electropherogram of ATP included two BL peaks; the latter peak showed peculiar broadening, which continued up to ca. 2.5h. In the CE apparatus with the flow-type cell, ATP, dATP, and ADP were detected with single peaks with detection limits of 1, 75, and 100μM, respectively. The other nucleotides, GTP, UTP, CTP, ITP, and TTP, were not detected at concentrations less than 0.5mM. A mixture of 10μM ATP and 100μM dATP was examined using the CE apparatus with the flow-type BL detection cell. ATP and dATP were separated using running buffer at pH 10 containing 1mM phenylboronic acid. The interaction between ATP and phenylboronic acid delayed the migration time of ATP.