ポリアクリルアミドゲル等電点電気泳動法による生および加熱肉の畜種鑑別

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  • Species Identification of Raw and Cooked Meats by Polyacrylamide Gel Isoelectric Focusing
  • ポリアクリルアミドゲル トウデンテン デンキ エイドウホウ ニ ヨル セイ オ

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In the previous paper, the authors made clear by using starch gel electrophoresis (SGE) that sarcoplasmic protein (Sp), tetrazorium oxidase (To) and esterase (Es) were available for meat species identification, and that heme protein (Hp), creatin kinase (CK), adenylate kinase (AK) and phosphoglucomutase (PGM) were stained stronger compared with other 17 proteins and enzymes but could identify only a few animal meat species. In the present experiments, in order to identify raw and cooked cattle, horse, pig, goat and sheep meats, and in order to detect small proportions of contaminating species in fresh binary mixture meats of the different species, proteins and enzymes mentioned above were analyzed by polyacrylamide gel isoelectric focusing (PAG-IEF).<br>The soluble proteins of supernatants from fresh meats and meats cooked for 30 min. at 65°C, 75°C and 100°C were separated by PAG-IEF using 1.0mm thick PAG plates (120×230mm) containing 3.0% ampholytes in the pH range 5.0-8.0 and 3.0-10.0. Gels were stained for enzymes in each substrate, for Sp in Coomassie Brilliant Blue R 250 and for Hp in o-tolidin dihydrochloride solution.<br>The five animal species used in this experiment could be distinguished individually byeither To or Es. Within-species variabilities of Es were confirmed in goats, sheep and pigs, however those of To was not. The results obtained by staining for Sp, Hp and CK showed that these proteins and enzyme were capable of distinguishing between fresh meat from cattle, horse and pig. However, sheep and goat could not be identified. Only a few animal meat species could be identified by staining for AK and PGM.<br>In detecting different meat species in binary mixture meats, approximately 1 to 5% of the contaminating species could each be detected from distinguishable species by staining for Sp, Hp, Ak and CK. PAG-IEF detected lower proportion of contaminants and reduced the experimental time.<br>Although the Hp of each animal species decreased in the intensity of staining after heating for 30min. at 65°C, the electrophoretic differences of the meat species were still detectable even after heating at 75°C or 100°C for 30min. except pork. PGM withstood heating at 100°C for 30min. Therefore, the detectable animal species by Hp and PGM were the same as the above-mentioned results for the fresh meat species.

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