柱前衍生-高效液相色谱法测定鱼类中组胺

Determination of histamine in fish by high performance liquid chromatography with pre-column derivatization

建立测定鱼类中组胺含量的柱前衍生-高效液相色谱(HPLC)联合紫外检测分析方法。样品均质后先用三氯乙酸水溶液震荡超声提取,再用丹磺酰氯衍生,采用HPLC-紫外检测器在254 nm处对组胺衍生物进行检测,以色谱峰面积外标法定量。组胺的质量浓度在1.0~50.0μg/mL范围内与色谱峰面积线性关系良好,相关系数为0.9997,方法检出限为7.2μg/kg,定量限为24μg/kg。样品加标回收率为96.8%~99.2%,测定结果的相对标准偏差为1.6%~3.7%(n=6)。该方法快速准确、灵敏度高、重复性好,可用于鱼类产品中组胺的定量分析。

A method for the determination of histamine in fish by pre-column derivatization and high performance liquid chromatography(HPLC)combined with ultraviolet detection was established.After homogenization,the samples were extracted by ultrasonic oscillation with trichloroacetic acid aqueous solution,and then derived by dansulfonyl chloride.Histamine derivatives were detected at 254 nm by HPLC-UV detector,and quantified by external standard method of chromatographic peak area.The mass concentration of histamine had a good linear relationship with the chromatographic peak area in the range of 1.0-50.0μg/mL with the correlation coefficient of 0.9997.The detection limit of the method was 7.2μg/kg,and the quantitation limit was 24μg/kg.The average recoveries of sample spiking were 96.8%-99.2%,and the relative standard deviations of the determination results were 1.6%-3.7%(n=6).The method is rapid,accurate,sensitive and reproducible,which can be used for the quantitative analysis of histamine in fish products.