转移核糖核酸中甲基化嘌呤和嘧啶碱基的高效液相色谱分析

High-resolution liquid chromatographic analysis of methylated purine and pyrimidine bases in transfer RNA.

作者信息

Lakings D B, Waalkes T P, Mrochek J E

出版信息

J Chromatogr. 1976 Jan 7;116(1):83-92. doi: 10.1016/s0021-9673(00)83702-4.

Abstract

Methylated and major purine and pyrimidine bases were separated and quantified by high-resolution liquid chromatography after hydrolyzing transfer ribonucleic acids (tRNAs). Separation was accomplished by eluting the hydrolyzed samples from an anion-exchange column with a concentration gradient of ammonium acetate at pH 9.2. Isolated sample of tRNA were hydrolyzed to the free bases with a trifluoroacetic acid-formic acid mixture of 200 degrees. Detection limits of 100-200 ng/ml were measured for the methylated bases; analytical data are reported for ten methylated bases plus the four major bases of calf liver and rat liver tRNA.

摘要

在对转移核糖核酸（tRNA）进行水解后，通过高效液相色谱法分离并定量甲基化的以及主要的嘌呤和嘧啶碱基。通过用pH 9.2的乙酸铵浓度梯度从阴离子交换柱上洗脱水解后的样品来实现分离。将分离得到的tRNA样品用200℃的三氟乙酸 - 甲酸混合物水解为游离碱基。甲基化碱基的检测限为100 - 200 ng/ml；报告了小牛肝脏和大鼠肝脏tRNA的十种甲基化碱基以及四种主要碱基的分析数据。