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Analysis of adulterant meat species in raw pork samples by heme protein staining of isoelectric focusing gels

Analysis of adulterant meat species in raw pork samples by heme protein staining of isoelectric focusing gels

Japanese Journal of Swine Science 30(3): 215-219

In the previous paper, authors made clear by polyacrylamide gel isoelectric focusing (PAG-IEF) that the specific staining for heme protein by peroxidase reaction, in which o-toluidine-one of the bendizine derivatives- was used as substrate, was available for meat species identification (T-Hp method). Fluorene, naphthol and carbazole derivatives are used for peroxidase reaction as the substrates besides bendizine derivatives. In this paper, the peroxidase reaction by SHAW and PRASAD, in which 3-amino-9-ethylcarbazole was used as substrate, was applied to stain for heme protein on isoelectric focusing gels (C-Hp method) and the availability for identification of raw cattle, horse, pig, goat and sheep meats and the minimum level of detection in mixtures of pig meat with cattle, horse and sheep meat were compared with T-Hp method in previous paper. The supernatants from fresh meats and binary mixture meats were separated by PAG-IEF using 1.0 mm thick PAG plates (120 times 230 mm) containing 3.0% ampholytes in the pH range 5.0 apprx 8.0. Gels were stained for heme protein by C-Hp method. The results obtained in this experiments showed that C-Hp method was able to distinguish between fresh meat from cattle, horse and pig, however, goat and sheep meat could not be differentiated by this method. In detecting different meat species in binary mixture meats of pig with cattle, horse and sheep, approximately 1 to 5% of the contaminating species could be detected by C-Hp method, respectively. From above results, it was appeared that the heme protein staining of isoelectric focusing gels by C-Hp method for identification of meat species was not at all inferior to T-Hp method.

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