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Food Control
Volume 150, 2023, 109784

Development of a simple method of unravelling catalytic activity of some mitochondrial and cytosolic enzymes in meat express juice and it's application in differentiation of fresh and frozen-thawed meat for authentication

A.K. Biswas, A. Arsalan, S. Valecha, A. Jangir, S. Swami, F. Rahman, S. Talukder, R.K. Agrawal, S. Chand, S.K. Mendiratta

Division of Livestock Products Technology, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, 243 122, U.P., India.

Abstract

The aim of this study was to develop a simple, low cost and robust analytical method for unraveling activity of catalytically active enzymes released from frozen-thawed meat samples for authentication. The conventionally used G-Clamp method requires additional equipment for the extraction of meat express juice and it is often subjected to handling errors since the manually applied pressure on the G-Clamp apparatus varies from person to person. The method was developed by standardization of extraction buffer, purification of enzymes on DEAE sephacel column, and identification on Biospectrophotometer. For confirmation, it was run on vertical gel electrophoresis and performed enzyme-linked immunosorbent assay (ELISA). Further, the method was validated for Inter-day and Intra-day assay precision as determined by coefficient of variation, repeatability, and reproducibility study. The standardized method was also compared with the standard clamp method. Results of this study suggest that the developed method is much more effective than the currently used clamp method and efficient in extracting cellular enzymes with less error percentages. The activities of all enzymes exhibited clear differences between chilled and frozen-thawed meat regardless of animal species or meat types. Thus, the immersion/dipping method developed is more reliable than the clamp method approach and may find its place in application of meat authentication.

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