转运核糖核酸在球形羟基磷灰石微珠上的高效液相色谱法。II. pH值和氯化钠对色谱的影响。

High-performance liquid chromatography of transfer ribonucleic acids on spherical hydroxyapatite beads. II. Effects of pH and sodium chloride on chromatography.

作者信息

Yamakawa Y, Miyasaka K, Ishikawa T, Yamada Y, Okuyama T

机构信息

Department of Applied Immunology, National Institute of Health, Tokyo, Japan.

出版信息

J Chromatogr. 1990 Aug 31;515:563-72. doi: 10.1016/s0021-9673(01)89355-9.

DOI:10.1016/s0021-9673(01)89355-9

PMID:2126546

Abstract

The effects of pH and sodium chloride on the high-performance liquid chromatography of transfer ribonucleic acids (tRNAs) on spherical hydroxyapatite beads were investigated. Binding of tRNAs on the beads was strengthened by increasing the pH of the mobile phase (phosphate buffer, pH 6.4-8.0), a phenomenon which is opposite to the retention of proteins on the beads. By using phosphate buffer of pH 8.0 instead of pH 6.8, the resolution of tRNAs was improved significantly; as many as ten times more theoretical plates were calculated with the use of the former buffer. Addition of sodium chloride to the phosphate buffer has a bifunctional effect on the retention of tRNAs: elution of tRNA(f-Met) and tRNA(Val) was retarded whereas that of tRNA(Phe) was facilitated.

摘要

研究了pH值和氯化钠对转运核糖核酸（tRNA）在球形羟基磷灰石珠上进行高效液相色谱分析的影响。通过提高流动相（pH 6.4 - 8.0的磷酸盐缓冲液）的pH值，tRNA与珠子的结合得到增强，这一现象与蛋白质在珠子上的保留情况相反。使用pH 8.0的磷酸盐缓冲液而非pH 6.8的缓冲液时，tRNA的分离度显著提高；使用前一种缓冲液计算得到的理论塔板数多达后者的十倍。向磷酸盐缓冲液中添加氯化钠对tRNA的保留具有双重作用：tRNA（f - Met）和tRNA（Val）的洗脱受到延迟，而tRNA（Phe）的洗脱则得到促进。