高效液相色谱法测定腊肉中生物胺含量

Determination of Biogenic Amines in Bacon by HPLC

本文建立了测定腊肉中生物胺含量的高效液相色谱方法,并对样品中8种生物胺进行了测定。采用0.4 mol·L^(-1)的高氯酸作为提取溶剂,提取腊肉样品中的生物胺,提取液经丹磺酰氯衍生后,采用Phenomenex Luna C18色谱柱(250 mm×4.6 mm,5μm),乙腈和超纯水作为流动相进行梯度洗脱,流速为1.0 mL·min^(-1),紫外检测器波长为254 nm。结果表明,色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺于25 min内能得到很好分离,各生物胺质量浓度在0.1~50 mg·L^(-1)时与相应峰面积有良好的线性关系;以信噪比大于3计,色胺、腐胺、亚精胺的检测限为0.03μg·mL^(-1),苯乙胺、组胺、酪胺、精胺的检测限为0.05μg·mL^(-1),尸胺的检测限为0.10μg·mL^(-1),8种生物胺的回收率在98.50%~104.05%;精密度相对标准偏差均小于4.0%。经检测的15个样品中生物胺总量在106.22~455.85 mg·kg^(-1)。本研究表明HPLC方法的灵敏度和精密度较高,是检测腊肉中生物胺含量的可靠方法。

In this paper,a high performance liquid chromatography method for the determination of biogenic amines in bacon was established,and 8 biogenic amines in the samples were determined.0.4 mol·L^(-1)perchloric acid was used as the extraction solvent to extract the biological amines from the bacon sample.After the extract was derivatized with dansyl chloride,the Phenomenex Luna C18 column(250 mm×4.6 mm,5μm),acetonitrile and ultrapure water was used as the mobile phase for gradient elution,the flow rate was 1.0 mL·min^(-1),and the UV detector wavelength was 254 nm.The results show that tryptamine,phenethylamine,putrescine,cadaverine,histamine,tyramine,spermidine and spermine can be separated well within 25 minutes,and the mass concentration of each biogenic amine is 0.1~50 mg·L^(-1)has a good linear relationship with the corresponding peak area;based on the signal-to-noise ratio greater than 3,the detection limit of tryptamine,putrescine and spermidine is 0.03μg·mL^(-1),the detection limit of phenethylamine,histamine,tyramine and spermine is 0.05μg·mL^(-1),and the detection limit of cadaverine is 0.10μg·mL^(-1).The recoveries of the 8 biogenic amines are between 98.50%and 104.05%;the relative standard deviations of precision are all less than 4.0%.The total amount of biogenic amines in the 15 samples tested was 106.22~455.85 mg·kg^(-1).This study shows that the HPLC method has high sensitivity and precision,and is a reliable method for detecting the content of biogenic amines in bacon.