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Free fatty acid determination by peroxidase catalysed luminol chemiluminescence

✍ Scribed by Näslund, Birgitta M. A. ;Bernström, Kerstin ;Lundin, Arne ;Arner, Peter

Publisher
John Wiley and Sons
Year
1989
Weight
612 KB
Volume
3
Category
Article
ISSN
0884-3996

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✦ Synopsis

A sensitive, specific, and partly automatic method for the analysis of free fatty acids is described. The assay involves activation of free fatty acids by acyl-CoA synthetase (EC 6.2.1.3) followed by oxidation of the thioesters by acyl-CoA oxidase. The H202 formed is determined in a reaction catalysed by horseradish peroxidase (EC 1.11.1.7) using luminol as electron donor. The assay has a linear range of 0.05 t o 5 nmol of different free fatty acids (C,o-Ct~) in the original sample. The efficiency of the method toward capric, lauric, myristic, palmitic, palmitoleic, stearic, oleic, and linoleic acid measured as recovery of light emission compared t o that of HzOz standards, was over 90%. AffiGel 501 was used t o covalently bind the free thiol group in CoASH eliminating interference of this substance in the peroxidase-luminol reaction.

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