Determination of K Value for Fish Freshness by Filter Paper Electrophoresis
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Abstract:
In order to monitor the freshness level of fish, a simple filter paper electrophoresis was developed for the analysis of K value in fish meat. ATP and its breakdown products were extracted and homogenized from fish meat samples by 1N HCl. Each homogenate was centrifuged at 8000 xg for 15 min, then the paper disc (6 mm in diameter) containing 10 ?L supernatant was set at the electrophoresis paper which immersed in the pyridine–acetic acid buffer solution (acetic acid:pyridine:water﹦1:10:289, pH 3.7), ATP and its breakdown products were separated by electrophoresis paper and analyzed under UV light at 254 nm. The visualized sample colored spots was digitized and calculated by image processing software. Successful separation of HxR and Hx from ATP and other ATP breakdown products were thus achieved And the linearities of the mixture of ATP, ADP and IMP, AMP, and the mixture of HxR and Hx were within 0.001~0.01 ??mol (R2＝0.99). The average recoveries of ATP and its breakdown products were 98%, 96% and 103%, respectively, with the relative standard deviations (RSD) being of 0.75~5.8%. The method was simple, effective and accurate that could be applied to determinate K value for fish freshness.
