HPLC法同时测定肉鸡各组织中游离棉酚和棉酚酮的含量

Simultaneous determination of free gossypol and gossypolone in chicken broiler tissues by HPLC

建立同时定量测定肉鸡各组织中游离棉酚及其代谢产物棉酚酮的高效液相色谱(High performance liquid chromatography,HPLC)法。以鸡血浆为基质,甘草次酸作为内标物,乙腈/1%甲酸(75∶25,体积比)为流动相,经ZORBA Extend-C18反相色谱柱分离棉酚、棉酚酮及甘草次酸,在254 nm下进行定性定量分析。棉酚和棉酚酮在0.5μg/mL^20μg/mL时线性关系良好,相关系数分别为0.9989和0.9992;两者的日内精密度RSD均小于10%,平均回收率均大于70%。方法学再验证试验表明该方法适用于肉鸡体内各组织器官如心、肝、脾、肺中游离棉酚及棉酚酮的定量分析。应用该方法对市场上销售的17份鸡肝样品和10份鸡肉样品进行分析,结果仅在15份鸡肝中检测出了游离棉酚,棉酚酮在样品中均未检出。该方法专属性强,准确度高、实用性强,为动物源性食品中游离棉酚及其代谢产物棉酚酮含量检测标准的建立提供一定的参考。

A high performance liquid chromatography（HPLC）method for the determination of free gossypol and its metabolite gossypolone in chicken broiler tissues was established. Gossypol,gossypolone,and internal standard Glycyrrhetic acid were separated on C_（18） column and detected at 254 nm. The mobile phase consisted of acetonitrile/1% formic acid（75 ∶ 25,volume ratio）. The calibration curves for the determination of gossypol and gossypolone were linear in the range of 0.5 μg/mL ~ 20 μg/mL with good correlation. Their correlation coefficient was 0.9989 and 0.9992,respectively. The mean recoveries of gossypol and gossypolone were all higher than 70%,and RSD of intraday precision were all below 10%（n=5）. Method validation test showed that this method was suitable for the quantitative analysis of free gossypol and gossypolone in broiler tissues,such as heart,liver,spleen,and lung. Total of 17 chicken liver samples and 10 chicken samples were analyzed. As a result,only free gossypol was detected in 15 chicken livers,and gossypolone was not detected in all samples. The method had high specificity,good accuracy and strong practicality,and it can provide a reference for establishing a standard of detecging free gossypol and its metabolite gossypolone in animal derived food.