食品科学 ›› 2019, Vol. 40 ›› Issue (16): 343-349.doi: 10.7506/spkx1002-6630-20180626-486

• 安全检测 • 上一篇    

高效液相色谱法测定东北农家酱中的5种生物胺

郝 宇,孙 波,张 宇,马艺荧,解双瑜,李天一   

  1. 东北农业大学食品学院,黑龙江 哈尔滨 150030
  • 出版日期:2019-08-25 发布日期:2019-08-26

Determination of Five Biogenic Amines in Northeast Farmhouse Soybean Paste by High Performance Liquid Chromatography

HAO Yu, SUN Bo, ZHANG Yu, MA Yiying, XIE Shuangyu, LI Tianyi   

  1. College of Food Science, Northeast Agricultural University, Harbin 150030, China
  • Online:2019-08-25 Published:2019-08-26

摘要: 建立柱前衍生-高效液相色谱法检测东北农家酱中的生物胺。采用5%的三氯乙酸提取东北农家酱中生物胺,正己烷除脂,丹磺酰氯作为柱前衍生试剂。采用C18(250 mm×4.6 mm,5 μm)色谱柱,以甲醇和水混合液进行梯度洗脱,流速为0.8 mL/min,在254 nm紫外检测波长处进行生物胺含量测定。结果表明,东北农家酱中的5 种生物胺(组胺、酪胺、β-苯乙胺、腐胺、亚精胺)在40 min内能很好地被分离。方法中5 种生物胺的线性范围为1~50 mg/kg,相关系数R2不小于0.99;检出限(RSN=3)为0.02~0.06 mg/kg,相对标准偏差低于5%,在添加量为5.0、10.0 mg/L和15.0 mg/L时,样品的平均回收率在82%~105%之间。本方法线性范围较广、重复性好、准确性高,能快速简便地对东北农家酱中的生物胺进行检测分析。

关键词: 生物胺, 东北农家酱, 高效液相色谱, 提取, 衍生物萃取

Abstract: In this study, we established a precolumn derivatization-high performance liquid chromatography (HPLC) method and applied it to determine biogenic amines in farmhouse soybean paste from northeastern China. Biogenic amines from samples were extracted with 5% trichloroacetic acid and were precolumn derivatized with dansyl chloride after defatting with n-hexane. A C18 (250 mm×4.6 mm, 5 μm) column was used for the chromatographic separation. Gradient elution was carried out with a mixture of methanol and water at a flow rate of 0.8 mL/min. The biogenic amine content was measured at a wavelength of 254 nm. The results showed that five biogenic amines (histamine, tyramine, β -phenylethylamine, putrescine, and spermidine) from the soybean paste were well separated within 40 min. The linear range for the analytes was 1–50 mg/kg, with correlation coefficients (R2) ≥ 0.99. The detection limits (LODs, RSN = 3) were 0.02–0.06 mg/kg, and the relative standard deviation (RSD) was less than 5%. The average recoveries of the biogenic amines from spiked samples at three concentration levels of 5.0, 10.0 and 15.0 mg/L were between 82% and 105%. The method had a wide linear range, good repeatability and high accuracy, and could allow for quick and simple detection of biogenic amines in northeast farmhouse soybean paste.

Key words: biogenic amines, northeast farmhouse soybean paste, high performance liquid chromatography, extraction, derivative extraction

中图分类号: