高效液相色谱法检测鮰鱼体内丁香酚的残留

Determination of eugenol residues in channel catfish by high performance liquid chromatography

目的建立测定斑点叉尾鮰中丁香酚残留量的高效液相色谱法,幵探究高、中、低3个不同丁香酚浓度在鱼体内的残留规律。方法样品先经过冷冻干燥法将鱼肉冻干,磨成粉状过筛,称0.5g冻干鱼粉经5mL乙腈超声重复提取3次, 280 nm条件下,高效液相色谱技术进样检测,采用外标法定量。结果丁香酚标样在0.15～20.0 mg/L浓度范围内,线性关系良好,线性相关系数达到0.99996。在添加浓度为0.40-3.20 mg/kg时,加标回收率在78.67%～90.54%,相对标准偏差为3.37%～5.54%,方法的检出限为0.045 mg/kg。同时収现麻醉时所用的丁香酚溶液浓度越高,丁香酚在鱼体内积累越多,用60 mg/L丁香酚溶液麻醉过的鮰鱼,体内丁香酚完全代谢需6～8 d,而用40、16 mg/L丁香酚溶液麻醉过的鮰鱼,仅需4～6 d。结论本方法具有操作简便、快速、准确、所用试剂价格低廉的特点,适用于斑点叉尾鮰中丁香酚残留量批量检测。

Objective To establish a method for the determination of eugenol residues in channel catfish by high performance liquid chromatography(HPLC),and investigate the residues of eugenol in fish at 3 different concentrations.Methods The flesh of fish was frozen-dried by freeze dryer,grinded into powder and sifted.The 0.5 g frozen-dried sample was extracted by 5 mL acetonitrile ultrasound for 3 times.Under 280 nm conditions,the samples were detected by high performance liquid chromatography and quantified by external standard method.Results Eugenol standard samples had a good linear relationship within the concentration range of 0.15-20.0 mg/L,and the linear correlation coefficient reached 0.99996.When the added concentration was 0.40-3.20 mg/kg,the recoveries were 78.67%-90.54%,with the relative standard deviation were 3.37%-5.54%,and the limit of detection was 0.045 mg/kg.At the same time,it was also found that the higher the concentration of eugenol solution used during anesthesia,the more eugenol accumulated in the fish.With 60 mg/L of eugenol solution anesthetizing the fish,the metabolism of eugenol totally takes 6-8 d,and with 40,16 mg/L of eugenol solution anesthetizing the fish,only4-6 d.Conclusion This method is simple,rapid,accurate and cheap,which is suitable for the batch detection of eugenol residues in catfish.