摘要
目的:建立一种快速测定血浆中乙醇浓度的新方法.方法:选用三(羟甲基)氨基甲烷-盐酸(Tris-HCL)作为缓冲体系,在碱性条件下,乙醇脱氢酶(ADH)催化乙醇转化成乙醛,同时生成还原型辅酶I (NADH).在340 nm波长处检测吸光度的变化,对照标准计算乙醇的浓度.结果:ADH最适用量为753 KU/L,辅酶I(NAD+)最适浓度为60.0 mmol/L,检测过程仅需90 s,线性范围可达0～68.60 mmol/L,变异系数(CV)为2.31%～3.25%,回收率为98.4%～101%,与美国DADE试剂盒比较具有良好的相关,γ=0.985,y=0.987x+0.024. 结论:本法测定血浆中乙醇无需除蛋白,具有快速、简便等优点,可用于自动生化分析仪及手工操作,适于临床常规运用.
Objective：To establish a new method for rapid measuring alcohol （ALC） in plasma. Methods： rlft （hydroxymethyl） aminomethane- hydrochloride（Tris- HCL） buffer was used in this study. In the condition of alkalescence, alcohol dehydrogenase （ADH） catalyzed the oxidation of ALC to acetaldehyde , with the simultaneous production of deoxidized nicotinamide adenine dinucleotide（NADH）. The variance of absorbance was determined by a filter of 340 nm. The concentration of ALC was calculated according to the standard. Results： The most adapt quantity of ADH was 753 KU/L, and the best concentration of the NAD ^＋ was 60.0 mmoL/L. It cost only 90 s in the whole reaction. The range of linearity was from zero to 68.60mmoL/L. The coefficient of variation （CV） in different concentrations was from 2.31% to 3.25%. The recovery, rate ranged from 98.4% to 101%. Compared with DADE （America） reagents method, the regression equation was obtained. Conclusion： This ADH method for measuring the concentration of ALC in plasma is rapid and simple without removing protein. It could be used both automatically and manually and suitable for clinical application.
出处
《西北国防医学杂志》
CAS
2005年第5期345-347,共3页
Medical Journal of National Defending Forces in Northwest China
关键词
实验室诊断
乙醇
乙醇脱氢酶
检测
临床应用
Laboratory diagnosis
Alcohol
Alcohol dehydrogenase
Testing
Clinical application
