HPLC法测定食品中组胺的研究

Study of Determination of Histamine in Food by HPLC

食品中组胺含量的检测是食品安全的研究热点。以国标为基础,对测定波长、柱温和流速进行了单因素优化,并用优化条件对基围虾和料酒样品进行检测。样品首先经过0.4 mol/L的高氯酸超声提取,正己烷除脂,饱和NaHCO3溶液缓冲后,用10 mg/mL丹磺酰氯溶液衍生,在40℃水浴振荡仪中水浴振荡35 min,乙腈定容过0.22μm滤膜。在流速为1.0 mL/min,检测波长为252 nm,柱温为40℃,进样体积为20μL的色谱条件下,通过Agilent C18(4.6 mm×250 mm,5μm)色谱柱,使用按一定比例混合后的甲基氰溶液和含0.1%乙酸的0.01 mol/L的乙酸铵水溶液进行梯度洗脱。结果表明:在该条件下,组胺标准曲线在1.0~50.0μg/mL浓度范围内呈现良好的线性趋势,R2≥0.999 9,其样品平均回收率为92.7%~101%,符合GB 27404—2008的相关要求,RSD(n=6)≤3.72%,仪器精密度较好,能够快速、简便、准确测定水产品及液态调味料食品中组胺的含量。

The detection of histamine content in food is a research hotspot in the field of food safety.This article opti‐mizes the determination wavelength,column temperature,and flow rate based on national standards,and detects samples of shrimp and cooking wine.The sample was first ultrasonically extracted with 0.4 mol/L perchloric acid,degreased with n-hexane,buffered with saturated NaHCO_(3) solution,and derivatized with 10 mg/mL dansyl chloride solution.The sam‐ple was shaken in a 40℃water bath shaker for 35 min,and acetonitrile was passed through a 0.22μm filter membrane at constant volume.At a flow rate of 1.0 mL/min,the detection wavelength is 252 nm,column temperature is 40℃.Un‐der chromatographic conditions with a sample volume of 20μL,gradient elution is carried out on Agilent C18(4.6 mm×250 mm,5μm)column with mixed methyl cyanide solution and 0.01 mol/L ammonium acetate aqueous solution con‐taining 0.1%acetic acid.Results:Under these conditions,the histamine standard curve showed a good linear trend in the concentration range of 1.0~50.0μg/mL,R2≥0.9999,the measured detection limit level was lower than the theoreti‐cal level specified in the national standard,the average recovery rate of samples was 92.7%~101%,which met the rele‐vant requirements of GB 27404-2008,RSD(n=6)≤3.72%.The method can quickly,simply and accurately determine the histamine content in aquatic products and liquid seasoning foods.