通过高效液相色谱法筛查人类红细胞中的腺嘌呤和次黄嘌呤磷酸核糖基转移酶缺陷。

Screening for adenine and hypoxanthine phosphoribosyltransferase deficiencies in human erythrocytes by high-performance liquid chromatography.

作者信息

Sakuma R, Nishina T, Kitamura M, Yamanaka H, Kamatani N, Nishioka K

机构信息

Department of Clinical Chemistry, Toranomon Hospital, Tokyo, Japan.

出版信息

Clin Chim Acta. 1987 Dec;170(2-3):281-9. doi: 10.1016/0009-8981(87)90138-0.

DOI:10.1016/0009-8981(87)90138-0

PMID:3436062

Abstract

A screening method using high-performance liquid chromatography (HPLC) for the simultaneous detection of deficiencies of adenine phosphoribosyltransferase (APRT) and hypoxanthine phosphoribosyltransferase (HPRT) activities in human erythrocytes is described. Both enzyme reactions of APRT and HPRT in lysates treated with a charcoal-dextran were simultaneously carried out in the same reaction tube and the enzyme activities were determined by measuring the increases in absorbance at 260 nm of adenosine and inosine converted from adenosine-5'-monophosphate and inosine-5'-monophosphate with alkaline phosphatase. Adenosine and inosine were separated from adenine and hypoxanthine by a reversed-phase column. The method could detect 1% of normal APRT activity and 0.3% of normal HPRT activity. The within-run coefficients of variation for APRT and HPRT activities were 3.2 and 3.4%, respectively.

摘要

描述了一种使用高效液相色谱（HPLC）同时检测人红细胞中腺嘌呤磷酸核糖转移酶（APRT）和次黄嘌呤磷酸核糖转移酶（HPRT）活性缺陷的筛查方法。APRT和HPRT在经活性炭-葡聚糖处理的裂解物中的两种酶反应在同一反应管中同时进行，通过测量碱性磷酸酶将5'-单磷酸腺苷和5'-单磷酸肌苷转化为腺苷和肌苷后在260 nm处吸光度的增加来测定酶活性。腺苷和肌苷通过反相柱与腺嘌呤和次黄嘌呤分离。该方法可检测到正常APRT活性的1%和正常HPRT活性的0.3%。APRT和HPRT活性的批内变异系数分别为3.2%和3.4%。