College of Public Health, Zhengzhou University, Zhengzhou, 450001, China.
Luminescence. 2013 Nov-Dec;28(6):905-9. doi: 10.1002/bio.2455. Epub 2013 Feb 14.
Acetaminophen, also called paracetamol, is found in Tylenol, Excedrin and other products as over-the-counter medicines. In this study, acetaminophen as a luminol signal enhancer was used in the chemiluminescence (CL) substrate solution of horseradish peroxidase (HRP) for the first time. The use of acetaminophen in the luminol-HRP-H2O2 system affected not only the intensity of the obtained signal, but also its kinetics. It was shown that acetaminophen was to be a potent enhancer of the luminol-HRP-H2O2 system. A putative enhancement mechanism for the luminol-H2O2-HRP-acetaminophen system is presented. The resonance of the nucleophilic amide group and the benzene ring of acetaminophen structure have a great effect on O-H bond dissociation energy of the phenol group and therefore on phenoxyl radical stabilization. These radicals act as mediators between HRP and luminol in an electron transfer reaction that generates luminol radicals and subsequently light emission, in which the intensity of CL is enhanced in the presence of acetaminophen. In addition, a simple method was developed to detect acetaminophen by static injection CL based on the enhanced CL system of luminol-H2O2-HRP by acetaminophen. Experimental conditions, such as pH and concentrations of substrates, have been examined and optimized. The proposed method exhibited good performance, the linear range was from 0.30 to 7.5 mM, the relative standard deviation was 1.86% (n = 10), limit of detection was 0.16 mM and recovery was 99 ± 4%.
对乙酰氨基酚，也称为扑热息痛，在泰诺林、雅维、其他产品中作为非处方药。在这项研究中，首次将对乙酰氨基酚作为一种鲁米诺信号增强剂应用于辣根过氧化物酶（HRP）的化学发光（CL）底物溶液中。在鲁米诺-HRP-H2O2 体系中使用对乙酰氨基酚不仅影响了获得信号的强度，而且影响了其动力学。结果表明，对乙酰氨基酚是鲁米诺-HRP-H2O2 体系的有效增强剂。提出了鲁米诺-H2O2-HRP-对乙酰氨基酚体系的增强机制。对乙酰氨基酚结构中亲核酰胺基团和苯环的共振对苯酚基团的 O-H 键离解能有很大影响，因此对苯氧自由基的稳定有很大影响。这些自由基在电子转移反应中作为 HRP 和鲁米诺之间的介体，生成鲁米诺自由基，随后发光，其中在存在对乙酰氨基酚的情况下增强 CL 强度。此外，还基于对乙酰氨基酚增强的鲁米诺-H2O2-HRP CL 体系，建立了一种通过静态注射 CL 检测对乙酰氨基酚的简单方法。考察并优化了实验条件，如 pH 值和底物浓度。该方法具有良好的性能，线性范围为 0.30-7.5 mM，相对标准偏差为 1.86%（n=10），检出限为 0.16 mM，回收率为 99±4%。
