Sequential Injection Analysis of Cholesterol Using an Oxidation-Reduction Electrode Detector.

IF 3.5 3区综合性期刊 Q2 CHEMISTRY, ANALYTICAL

Sensors Pub Date : 2025-09-19 DOI:10.3390/s25185863

Takato Imanaka, Takashi Masadome

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Abstract

A new automated method for the determination of cholesterol in serum was developed by combining sequential injection analysis (SIA) with potentiometric detection using a gold oxidation-reduction potential (ORP) electrode because serum cholesterol is an important indicator of abnormal lipid metabolism, arteriosclerosis, and hypertension in clinical diagnosis. The method is based on enzymatic hydrolysis of cholesterol esters by cholesterol esterase (CE) to yield free cholesterol, followed by oxidation with cholesterol oxidase (COD) to produce hydrogen peroxide. In the presence of horseradish peroxidase (HRP) and potassium ferrocyanide (K₄[Fe(CN)₆]), hydrogen peroxide oxidizes ferrocyanide to ferricyanide (K₃[Fe(CN)₆]), and the concentration ratio of ferri-/ferrocyanide is determined potentiometrically. Experimental conditions were optimized as follows: 5.0 mM K₄[Fe(CN)₆], 2 min reaction time, 0.5 units/mL HRP, 0.75 units/mL COD for free cholesterol, 1.5 units/mL COD and 10.0 units/mL CE for total cholesterol, and 5.0% (w/v) Triton X-100 with 5.0% (v/v) isopropanol as solubilizing agents. Under these conditions, the calibration curve for total cholesterol exhibited a Nernstian slope of 47.6 mV/decade over the range of 1.0 × 10^-5-1.0 × 10^-3 M, with no significant interference from common serum constituents. This method offers low reagent consumption, high automation, and simple operation, making it promising for clinical cholesterol analysis.

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查看原文本刊更多论文

使用氧化还原电极检测器对胆固醇进行序贯注射分析。

鉴于血清胆固醇在临床诊断中是脂质代谢异常、动脉硬化和高血压的重要指标，本文建立了一种将序次注射分析（SIA）与金氧化还原电位（ORP）电极电位检测相结合的血清胆固醇自动检测方法。该方法是基于胆固醇酯酶（CE）酶解胆固醇酯生成游离胆固醇，然后与胆固醇氧化酶（COD）氧化生成过氧化氢。在辣根过氧化物酶（HRP）和亚铁氰化钾（K4[Fe(CN)6]）存在下，过氧化氢氧化亚铁氰化物（K3[Fe(CN)6]），用电位法测定了铁/亚铁氰化钾的浓度比。优化实验条件为：5.0 mM K4[Fe(CN)6]，反应时间2 min， HRP 0.5单位/mL，游离胆固醇COD 0.75单位/mL，总胆固醇COD 1.5单位/mL， CE 10.0单位/mL， 5.0% (w/v) Triton X-100加5.0% （v/v）异丙醇为增溶剂。在此条件下，总胆固醇的校正曲线在1.0 × 10-5-1.0 × 10-3 M范围内的纳氏斜率为47.6 mV/ 10年，无血清中常见成分的显著干扰。该方法具有试剂用量少、自动化程度高、操作简单等优点，在临床胆固醇分析中具有广阔的应用前景。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Sensors 工程技术-电化学

CiteScore

7.30

自引率

12.80%

发文量

8430

审稿时长

1.7 months

期刊介绍： Sensors (ISSN 1424-8220) provides an advanced forum for the science and technology of sensors and biosensors. It publishes reviews (including comprehensive reviews on the complete sensors products), regular research papers and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced.