高效液相色谱-电化学检测法测定鸟嘌呤：在DNA和RNA分析中的应用
Determination of guanine by high-performance liquid chromatography with electrochemical detection: application to DNA and RNA assays.
作者信息
Nakahara T, Shiraishi A, Hirano M, Matsumoto T, Kuroki T, Tatebayashi Y, Tsutsumi T, Nishiyama K, Ooboshi H, Nakamura K
机构信息
Department of Chemistry, Faculty of Science, Kyushu University, Fukuoka, Japan.
出版信息
Anal Biochem. 1989 Jul;180(1):38-42.
A highly sensitive assay for guanine was developed using high-performance liquid chromatography with electrochemical detection (ECD). Guanine was susceptible to the electrochemical oxidation, and ECD response was proportional to the amount of guanine in the range 0.25-4 pmol of guanine. The ECD of guanine was applicable to the analysis of nucleic acids. DNA and RNA were hydrolyzed in 0.03 and 3 M HCl, respectively, and guanine liberated from the nucleic acids was separated on a reverse-phase column and determined by ECD. The method allowed detection of 0.2 ng of calf thymus DNA or tRNA. An application of the method is shown for DNA and RNA assays in trichloroacetic acid extracts of rat adrenal and liver.
采用高效液相色谱-电化学检测（ECD）技术开发了一种高灵敏度的鸟嘌呤检测方法。鸟嘌呤易于发生电化学氧化，在0.25 - 4皮摩尔鸟嘌呤范围内，ECD响应与鸟嘌呤的量成正比。鸟嘌呤的ECD可用于核酸分析。DNA和RNA分别在0.03 M和3 M HCl中水解，从核酸中释放出的鸟嘌呤在反相柱上分离并用ECD测定。该方法可检测0.2 ng的小牛胸腺DNA或tRNA。展示了该方法在大鼠肾上腺和肝脏的三氯乙酸提取物中DNA和RNA检测的应用。
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