通过毛细管电泳-激光诱导荧光检测法选择性测定含腺嘌呤化合物。

Selective determination of adenine-containing compounds by capillary electrophoresis with laser-induced fluorescence detection.

作者信息

Tseng H C, Dadoo R, Zare R N

机构信息

Department of Chemistry, Stanford University, California 94305.

出版信息

Anal Biochem. 1994 Oct;222(1):55-8. doi: 10.1006/abio.1994.1453.

DOI:10.1006/abio.1994.1453

PMID:7856871

Abstract

Capillary electrophoresis coupled with laser-induced fluorescence detection provides a selective analysis of mixtures of adenine-containing compounds (adenine, adenosine, cAMP, AMP, ADP, and ATP) that are derivatized using chloroacetaldehyde as a fluorogenic reagent. The components can be detected with linear response over the concentration range of 10(-4) to 10(-9) M, and the detection limit corresponds to 5 to 10 x 10(-19) mol for each nucleotide. Analysis is achieved in less than 10 min with high separation efficiencies (up to 400,000 theoretical plates) and shows good reproducibility for migration times (0.5 to 1.0%) and peak heights (2.8 to 7.7%). To demonstrate the rapid analysis of small-volume samples, the adenine-containing compounds present in an oocyte from Xenopus laevis are determined.

摘要

毛细管电泳结合激光诱导荧光检测可对含腺嘌呤化合物（腺嘌呤、腺苷、环磷酸腺苷、一磷酸腺苷、二磷酸腺苷和三磷酸腺苷）的混合物进行选择性分析，这些化合物使用氯乙醛作为荧光试剂进行衍生化。各组分在10⁻⁴至10⁻⁹ M的浓度范围内呈线性响应，每种核苷酸的检测限对应于5至10×10⁻¹⁹ mol。分析可在不到10分钟内完成，分离效率高（高达400,000理论塔板数），迁移时间（0.5%至1.0%）和峰高（2.8%至7.7%）的重现性良好。为了证明对小体积样品的快速分析，测定了非洲爪蟾卵母细胞中存在的含腺嘌呤化合物。