Arakawa Hidetoshi, Nakabayashi Shigeo, Ohno Ken-Ichi, Maeda Masako
School of Pharmacy, Showa University, Shinagawa-ku, Tokyo 142-8555, Japan.
J Pharm Anal. 2012 Apr;2(2):156-159. doi: 10.1016/j.jpha.2012.01.004. Epub 2012 Jan 24.
Horseradish peroxidase (HRP) is generally used as a label enzyme in enzyme immunoassay (EIA). The procedure used for HRP detection in EIA is critical for sensitivity and precision. This paper describes a novel fluorimetric assay for horseradish peroxidase (HRP) using sesamol as substrate. The principle of the assay is as follow: sesamol (3,4-methylenedioxy phenol) is reacted enzymatically in the presence of hydrogen peroxide to produce dimeric sesamol. The dimer is fluorescent and can be detected sensitively at ex. 347 nm, em. 427 nm. The measurable range of HRP was 1.0×10 to 1.0×10 mol/assay, with a detection limit of 1.0×10 mol/assay. The coefficient of variation (CV, =8) was examined at each point on the standard curve, with a mean CV percentage of 3.8%. This assay system was applied to thyroid stimulating hormone (TSH) EIA using HRP as the label enzyme.
辣根过氧化物酶（HRP）通常用作酶免疫分析（EIA）中的标记酶。在EIA中用于检测HRP的程序对于灵敏度和精密度至关重要。本文描述了一种以芝麻酚为底物检测辣根过氧化物酶（HRP）的新型荧光测定法。该测定法的原理如下：芝麻酚（3,4-亚甲二氧基苯酚）在过氧化氢存在下发生酶促反应生成二聚体芝麻酚。该二聚体具有荧光，可在激发波长347nm、发射波长427nm处进行灵敏检测。HRP的可测量范围为1.0×10至1.0×10摩尔/测定，检测限为1.0×10摩尔/测定。在标准曲线上的每个点检测变异系数（CV，n = 8），平均CV百分比为3.8%。该测定系统应用于以HRP作为标记酶的促甲状腺激素（TSH）EIA。
