滤纸电泳分析检测鱼的鲜度K值

Determination of K Value for Fish Freshness by Filter Paper Electrophoresis

为了监控鱼类的鲜度水平，本文建立了简单的鱼肉鲜度K值的滤纸电泳检测方法。样品经1N HCl提取、混匀后，经高速离心机（8000 xg）离心15 min，取上清液10 μL于直径6 mm的滤纸片上，然后把含样品液的滤纸片放置在用醋酸吡啶缓冲液浸润的（醋酸：啶：＝10：1：289，pH＝3.7）电泳用滤纸上，经800 V 20 in的滤纸电泳分离后，在254 nm的紫外光视觉化处理下，观察到ATP＋ADP＋IMP, AMP, HxR＋Hx有效的分离，样品经数据处理软件处理后进行定量分析。标准品ATP＋ADP＋IMP, AMP, HxR＋Hx在0.001 umol至0.01 μmol浓度之间，有良好的线性关系线，线性关系系数均为0.99。鱼肉样品分别添加0.3 μmol ATP及其分解产物，其平均回收率为98%、96%、103%，相对标准偏差为0.75-5.8%。结果表明该方法具有快速、简便、准确的优点，并且不需要昂贵的设备，适合鱼肉鲜度K值的现场测定。

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.