Use of β-hydroxyacyl-CoA-dehydrogenase (HADH) activity to differentiate frozen from unfrozen fish and shellfish

Abstract

 Further work on an enzymic method to differentiate frozen from unfrozen fish and shellfish is reported. The method is based on the release of the β-hydroxyacyl-CoA-dehydrogenase (HADH) from mitochondria during freezing. Enzymic activity was evaluated in fresh and frozen thawed samples from sole (Solea solea), sea bream (Pagellus centrodontus), hake (Merluccius merluccius), gilt headed bream (Sparus aurata), sea bass (Dicentrarchus labrax), salmon (Salmo salar), prawn (Penaeus japonicus) and Norwegian lobster (Nephrops norvegicus). Changes in the HADH activity of fresh and frozen thawed samples were compared after freezing at –196  °C for 15 min. Two values were obtained: U (by dividing: HADH activity of samples frozen at –196  °C, then thawed/HADH activity of unfrozen samples) and F (by dividing: HADH activity of samples frozen at –18  °C, thawed, then frozen at –196  °C /HADH activity of samples frozen at –18  °C, then thawed). Statistical analysis showed significant differences (P≤0.05) between both quotients for gilt headed bream, salmon, sea bream, sole and prawn, and an arbitrary limit was set at 2 to differentiate frozen thawed from unfrozen samples. The application of this limit made it possible to discriminate the unfrozen from the frozen thawed state of around 90% of the total samples analysed. Best results were obtained for prawn (100% of samples differentiated). In the present paper, a laboratory routine is proposed based on the comparison of the HADH activity of a sample analysed straight away and that of a sample frozen at –196  °C and then thawed. The reported method is simple and fast. The entire laboratory procedure can be performed in 45 min.