Unveiling citrate synthase enzyme as potential biomarker in developing a rapid method for monitoring of chilled and defrosted buffalo meat in supply chain

Aim of this study was to develop a rapid spectrometric method for monitoring chilled (CM) and frozen-thawed meat (FTM) in supply chain to find out the regulatory compliance. The method was developed with the standardization of extraction buffer, substrate and developer. SDS-PAGE clearly displayed presence of citrate synthase (CS) corresponding to MWt. of 42.5 KDa while Western blotting and ELISA data confirmed the presence of this enzyme. Validation data revealed very good precision of the method with coefficient of variation (CV, %) less than 10. To ascertain the thermal history, a threshold value (R1 value) less than 0.2 was established for samples subjected to freezing and thawing cycles, while meat samples exhibiting an R1 value of 0.9 or higher are classified as chilled meat which made clear demarcation in between the two types of meat. The CS enzyme showed very good stability on repeated freezing-thawing, and thus, CS has greater potential to acts a biomarker for distinguishing chilled and defrosted meat to meet the regulatory compliance.