Alcohol oxidase- and formaldehyde dehydrogenase-based enzymatic methods for formaldehyde assay in fish food products

Abstract

For Gadoid fishes, formaldehyde can be generated in tissues in huge amounts during endogenous enzymatic degradation of natural osmoprotectant trimethylamine-N-oxide. This paper describes two enzymatic methods for assay of formaldehyde in fish food products using alcohol oxidase (AOX) and formaldehyde dehydrogenase (FdDH) isolated from the thermotolerant methylotrophic yeast Hansenula polymorpha. AOX-based method exploits an ability of the enzyme to oxidise a hydrated form of formaldehyde to formic acid and hydrogen peroxide monitored in peroxidase-catalysed colorimetric reaction. In FdDH-based method, a monitored coloured formazane is formed from nitrotetrazolium salt during reduction by NADH, produced in formaldehyde-dependent reaction. It was demonstrated an applicability of both methods for assay of formaldehyde in fish products. The optimal protocols for analysis procedures have been elaborated and analytical parameters of both enzymatic methods have been established. The both methods were demonstrated that some fish products (hake and cod) contain high formaldehyde concentrations (up to 100mg/kg wet weight).