酶促电化学发光法测定血清总胆汁酸  

Determination of Total Bile Acids in Serum by Enzymatic Electrochemiluminescence

作　　者：丁敏[1] 王箭[1] 张晓清[1] 邓文平[1] 

机构地区：[1]重庆医科大学医学检验系 临床检验诊断学教育部重点实验室,重庆400016

出　　处：《分析化学》2010年第12期1793-1796,共4页Chinese Journal of Analytical Chemistry

摘　　要：利用胆汁酸在氧化型辅酶I(NAD+)共存下被3α-类固醇脱氢酶(3α-HSD)催化脱氢,同时NAD+被还原为还原型辅酶I(NADH)可增强三联吡啶钌(Ru(bpy)32+)的电化学发光(ECL)信号,实现电化学发光法间接检测胆汁酸。检测池中含2.5mmol/LRu(bpy)23+,833U/L3α-HSD,70nmol/LNAD+,50mmol/L磷酸盐缓冲液(pH8.0)。ECL强度与胆汁酸的浓度在1.0～100fmol/L范围内呈良好的线性关系,线性方程y=0.7707x+12.04(r=0.9975);检出限为0.02fmol/L;平均回收率为97.5%。本方法与临床采用的酶循环放大法有较好的相关性。Bile acids are dehydrogenated to 3-oxo bile acids by 3α-hydroxysteroid dehydrogenase(3α-HSD) with concomitant reduction of nicotinamide adenine dinucleotide(NAD +) to reduced nicotinamide adenine dinucleotide(NADH),which could enhance the electrochemiluminescence intensity of tris(2,2’-bipyridine) ruthenium (Ⅱ)(Ru(bpy) 32 +).Based on the principle above mentioned,bile acids could be detected by electrochemiluminescence method indirectly.The detection cell was filled with 2.5 mmol/L Ru(bpy) 23 +,833 U/L 3α-HSD,70 nmol/L NAD + and 50 mmol/L phosphate buffer saline(pH 8.0).The ECL intensity was linear with the concentration of bile acids in the range from 1.0 fmol/L to 100 fmol/L and the linear equation was y = 0.7707x + 12.04(r = 0.9975).The detection limit was 0.02 fmol/L and the average recovery was 97.5%.The ECL method established in this experiment was closely related to the enzymatic cycle method in clinical analysis.

关 键 词：胆汁酸 电化学发光 酶 血清 

分 类 号：R446.1[医药卫生—诊断学]