Continuous Flow Analyses of Nadh Using Bacterial Bioluminescent Enzymes Immobilized on Nylon enzymes, flow analysis
作者:
S. Girotti,
A. Roda,
S. Ghini,
B. Grigolo,
C. Carrea,
R. Bovara,
期刊:
Analytical Letters
(Taylor Available online 1984)
卷期:
Volume 17,
issue 1
页码: 1-12
ISSN:0003-2719
年代: 1984
DOI:10.1080/00032718408065261
出版商: Taylor & Francis Group
关键词: Bioluminescence;NADH determination;immobilized enzymes;flow analysis.
数据来源: Taylor
摘要:
A continuous flow bioluminescent method for NADH analysis has been developed using nylon 6 as a solid support. Bacterial luciferase and NAD:FMN oxidoreductase are covalently co-immobilized to a nylon coil (1 m × 1.0 mm ID). The reactor (nylon coil) is placed in front of a photomultiplier tube inside a luminometer linked to an air segmented continuous flow system. Response is linear from 1 to 2500 pmol/tube allowing determination of NADH at picomolar levels. Inter and intra assay precision of the method is satisfactory (5-10%). Range of sample volume injected is 5-100 μl. More than 20 samples per hour can be analyzed and no carryover was observed. The stability of the nylon immobilized enzymes is high (over 2 months) and over 500 samples can be analyzed with a few mg of enzymes.
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