Chemiluminescent Determination of Xanthine Oxidase Activity Using A Sensitive Low-Light Detection System
作者:
Stefano Girotti,
Patrizia Pasini,
Elida Ferri,
Severino Ghini,
Fabiana Fini,
Rolando Budini,
Aldo Roda,
期刊:
Analytical Letters
(Taylor Available online 1996)
卷期:
Volume 29,
issue 12
页码: 2097-2114
ISSN:0003-2719
年代: 1996
DOI:10.1080/00032719608002234
出版商: Taylor & Francis Group
关键词: Xanthine oxidase;Chemiluminescence;Luminol;Enzymatic Analysis;Milk
数据来源: Taylor
摘要:
A highly sensitive and rapid chemiluminescent assay for the determination of the activity of xanthine oxidase (XOD) was developed. The chemiluminescent signal was obtained from the catalyzed oxidation of hypoxanthine, accelerated and amplified using a Fe-EDTA complex and perborate, which acts on luminol. The same luminescent mixture was previously used as detection system for immunoassays. Two different mixtures were used, which differ in their luminol and perborate content, with (CLMrho) or without (CLMb) addition of 0.1 μM rhodamine fluorophor. The response obtained from XOD standard solutions in buffer was linear from 5 to 500 U L−1and from 0.7 to 250 U L−1for CLMrho and CLMb respectively, at 25°C. 5 and 0.7 U L−1were the detection limits at 1 standard deviation level. The intra- and inter-assay relative standard deviations ranged from 6 to 12 % for both CLM. Measurements were made using the high performance, low-light level imaging Berthold luminograph LB-980 which allows simultaneous determination of several samples distributed on a microtiterplate. Various kinds of milk were analyzed for XOD content, which in pasteurized milk depends on the fat content and in the UHT milk disappears owing to the heat treatment.
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