Development of superoxide dismutase based visual and spectrophotometric method for rapid differentiation of fresh and frozen-thawed buffalo meat

Study aimed to develop biomarker-based assay for rapid detection of fresh and frozen -thawed buffalo meat in the supply chain. The method is based on development of a solvent system and identification of suitable substrate and developer for screening of biomarkers. For the confirmation column chromatography, gel electrophoresis and Western Blotting were carried out. Validation was done by intra- and inter -day validation, storability study, and determination of thermal history. Best results were shown with pH 8.0 Tris-HCl; extraction buffer, 205 mu M nicotinamide adenine dinucleotide hydrogen; substrate, 184 mu M Nitroblue tetrazolium, and 1.9 mu M phenazine methosulfate; developer. The thermal history ranged from 0.14 to 0.17 during storage at -20 C-degrees. The intra- and inter -day assay precision (CV %) ranged from 5.3 to 6.5 %; in chilled and 14.1 - 9.2 % in frozen -thawed samples. The study confirmed SOD as a viable biomarker. Developed method using SOD has significant potential for rapidly differentiating chilled or frozen -thawed meat.