用于改进酶促糖化白蛋白测量方法的糖化氨基酸消除反应研究

Study of glycated amino acid elimination reaction for an improved enzymatic glycated albumin measurement method.

作者信息

Kouzuma Takuji, Uemastu Yumiko, Usami Tomomi, Imamura Shigeyuki

机构信息

Diagnostics R&D Department, Fine Chemicals and Diagnostics Division, Asahi Kasei Pharma Corporation, 632-1, Mifuku, Ohito-cho, Tagata-gun, Shizuoka-ken 410-2321, Japan.

出版信息

Clin Chim Acta. 2004 Aug 16;346(2):135-43. doi: 10.1016/j.cccn.2004.02.019.

DOI:10.1016/j.cccn.2004.02.019

PMID:15256314

Abstract

BACKGROUND

In order to improve enzymatic glycated albumin measurement, we studied the endogenous glycated amino acid elimination reaction and the new bromocresolpurple (BCP) method for albumin measurement.

METHODS

In the assay, endogenous glycated amino acids are first eliminated by oxidation by ketoamine oxidase. Second, glycated albumin is hydrolyzed to glycated amino acids by proteinase digestion, and glycated amino acids are oxidized to produce hydrogen peroxide, which is quantitatively measured. Third, albumin is measured by the new BCP method. Finally, glycated albumin value is calculated as the percentage of glycated albumin in total albumin.

RESULTS

Glycated amino acid concentrations in prepared total parenteral nutrition products were increased in direct proportion to storage time and temperature. The glycated amino acid elimination reaction using ketoamine oxidase may be able to eliminate more than 15 mmol/l glycated amino acids. The glycated albumin values of samples calculated from the albumin concentrations using the new BCP method accorded with those calculated with the HPLC method. Fundamental performances (linearity, dilution test, analytical recovery, within-run and between-run CVs, interference study) of the present method were good. Detection of glycated albumin by the present method was significantly correlated with detection of glycated albumin by the high-performance liquid chromatography method (r(p) = 0.995).

CONCLUSIONS

This new improved method is free of interference by endogenous glycated amino acids and is unaffected by albumin concentration, and enables more accurate analysis of glycated albumin.

摘要

背景

为了改进酶法糖化白蛋白的检测，我们研究了内源性糖化氨基酸消除反应以及用于白蛋白检测的新型溴甲酚紫（BCP）法。

方法

在该检测中，首先通过酮胺氧化酶氧化消除内源性糖化氨基酸。其次，糖化白蛋白经蛋白酶消化水解为糖化氨基酸，糖化氨基酸被氧化产生过氧化氢，对其进行定量测定。第三，采用新型BCP法检测白蛋白。最后，计算糖化白蛋白值占总白蛋白中糖化白蛋白的百分比。

结果

在配制的全胃肠外营养产品中，糖化氨基酸浓度与储存时间和温度成正比增加。使用酮胺氧化酶的糖化氨基酸消除反应可能能够消除超过15 mmol/l的糖化氨基酸。使用新型BCP法根据白蛋白浓度计算的样品糖化白蛋白值与使用高效液相色谱法计算的值一致。本方法的基本性能（线性、稀释试验、分析回收率、批内和批间变异系数、干扰研究）良好。本方法检测糖化白蛋白与高效液相色谱法检测糖化白蛋白显著相关（r(p) = 0.995）。