Indirect electrochemical detection for total bile acids in human serum
Indirect electrochemical detection for total bile acids in human serum
复制标题
人血清中总胆汁酸的间接电化学检测
DOI：
10.1016/j.bios.2016.05.055
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发表时间：
2016
影响因子：
12.6
通讯作者：
Min Ding
中科院分区：
文献类型：
--
作者：
Xiaoqing Zhang;Mingsong Zhu;Biao Xu;Yue Cui;Gang Tian;Zhenghu Shi;Min Ding
血清胆汁酸水平是肝胆疾病筛查和诊断的重要指标。胆汁酸浓度与肝胆疾病的严重程度密切相关，检测胆汁酸浓度是了解疾病分期的重要因素。胆汁酸的测定目前普遍采用酶循环法，该方法灵敏度低，试剂消耗大。因此，需要开发一种成本低、灵敏度高的新方法。采用丝网印刷碳电极（SPCE）建立了人血清胆汁的间接电化学检测方法acids.in由于胆汁酸不显示电化学信号，因此在烟酰胺腺嘌呤二核苷酸（NADPH）存在下，胆汁酸通过3-α-羟基类固醇脱氢酶（3α-HSD）转化为3-酮类固醇，后者还原为NADH。然后，NADH可以在SPCE的表面上被氧化，产生用于计算总胆汁酸（TBA）浓度的信号。TBA在5.00 ~ 400 μM浓度范围内线性关系良好。精密度和回收率均足以用于临床环境。35份人血清中TBA的含量经配对t检验，IED法与酶循环法无显著性差异。此外，我们的IED方法节省试剂，灵敏度和成本效益。
Bile acids level in serum is a useful index for screening and diagnosis of hepatobiliary diseases. As bile acids concentration is closely related to the degree of hepatobiliary diseases, detecting it is a vital factor to understand the stage of the diseases. The prevalent determination for bile acids is the enzymatic cycling method which has low sensitivity while reagent-consuming. It is desirable to develop a new method with lower cost and higher sensitivity. An indirect electrochemical detection (IED) for bile acids.in human serum was established using the screen printed carbon electrode (SPCE). Since bile acids do not show electrochemical signals, they were converted to 3-ketosteroids by 3-α-hydroxysteroid dehydrogenase (3α-HSD) in the presence of nicotinamide adenine dinucleotide (NADþ), which was reduced to NADH. NADH could then be oxidized on the surface of SPCE, generating a signal that was used to calculate the total bile acids (TBA) concentration. A good linear calibration for TBA was obtained at the concentration range from 5.00 μM to 400 μM in human serum. Both the precisions and recoveries were sufficient to be used in a clinical setting. The TBA concentrations in 35 human serum samples by our IED method didn't show significant difference with the result by enzymatic cycling method, using the paired t-test. Moreover, our IED method is reagent-saving, sensitive and cost-effective.