红细胞中腺苷和脱氧腺苷代谢物的快速放射分析

Rapid radioassay for metabolites of adenosine and deoxyadenosine in erythrocytes.

作者信息

Szabados E, Christopherson R I

机构信息

Department of Biochemistry, University of Sydney, NSW, Australia.

出版信息

J Chromatogr B Biomed Appl. 1995 Dec 1;674(1):132-7. doi: 10.1016/0378-4347(95)00288-x.

DOI:10.1016/0378-4347(95)00288-x

PMID:8749261

Abstract

A radioassay has been developed to quantify the uptake and initial metabolism of adenosine (Ado) or deoxyadenosine (dAdo) by human erythrocytes. Cell suspension and [3H]Ado are mixed at 3-s intervals with a novel dual-syringe apparatus, and uptake and metabolism of Ado is stopped by centrifuging the cells through a dibutylphthalate layer into perchloric acid. The neutralized cell extract is analyzed by two-dimensional chromatography on poly(ethyleneimine)-cellulose plates by two procedures using combinations of solvents optimised for the separation of nucleosides and nucleobases, and for nucleotides derived from the exogenous [3H]Ado.

摘要

已开发出一种放射分析方法，用于定量测定人红细胞对腺苷（Ado）或脱氧腺苷（dAdo）的摄取和初始代谢。细胞悬液与[³H]Ado通过一种新型双注射器装置以3秒的间隔混合，通过将细胞离心穿过邻苯二甲酸二丁酯层进入高氯酸来停止Ado的摄取和代谢。用针对核苷、核碱基以及源自外源性[³H]Ado的核苷酸分离进行优化的溶剂组合，通过两种方法在聚（乙烯亚胺） - 纤维素板上进行二维色谱分析中和后的细胞提取物。