An enzymatic amplification Flow Injection Analysis (FIA) system for the sensitive determination of phenol

被引:23
作者
Fuhrmann, B [1 ]
Spohn, U [1 ]
机构
[1] Univ Halle Wittenberg, Inst Biotechnol, D-06120 Halle, Germany
关键词
phenol detection; chemoenzymatic substrate recycling; flow injection analysis; derivatization of o-benzoquinone; fluorescence;
D O I
10.1016/S0956-5663(98)00061-X
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
A highly sensitive, enzymatic FIA procedure was developed to determine phenol fluorimetrically. Tyrosinase was immobilized in a packed bed flow reactor. Upon contact with tyrosinase, phenol is oxidized to o-benzoquinone, which oxidizes ascorbic acid to dehydroascorbic acid producing catechol, which is enzymatically reoxidized to o-benzoquinone. Dehydroascorbic acid reacts with o-phenylenediamine forming a highly fluorescent product, which is excited at lambda(exc) = 345 nm and detected at lambda(em) = 410 nm. Dehydroascorbic acid was detected in the range between 0.5 and 100 mu M. The chemoenzymatic substrate recycling enhances the sensitivity of the detection of phenol and catechol with a detection limit of around 0.02 mu M in both cases. Amplification factors between 8 and 12 were estimated. Phenol and catechol can be determined in the approximately linear ranges between 0.1 and 2 mu M and between 0.02 and 2 mu M, respectively. It is possible to perform 20 phenol detections per hour by the automatic FIA procedure with high operational stability. (C) 1998 Elsevier Science S.A. All rights reserved.
引用
收藏
页码:895 / 902
页数:8
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