Vol. 9, Special Issue 5, Part D (2025)

Mislabeling of defrosted meat as fresh or chilled meat is a serious concern in the meat supply chain. There are various methods used for the identification of such malpractices. However, an approach based on enzyme biomarkers can be considered as most suitable. A mitochondrial enzyme citrate synthase (CS) was identified as a potential biomarker enzyme for the differentiation of fresh and frozen thawed meat was done in this study. The meat express juice (MEJ) was prepared and purified using anion exchange chromatography. SDS-PAGE and Western blot analysis of the eluted fractions showed thicker bands at 47 kDa in the MEJ prepared from frozen thawed chicken samples compared to fresh/chilled. CS enzyme activity assay revealed a 6-fold increase in CS activity of frozen thawed chicken meat samples as compared to fresh samples. Thus, this study identifies CS as a potent biomarker enzyme for the differentiation of fresh and defrosted chicken meat.