高效液相色谱法测定东北农家酱中的5种生物胺

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,5μm)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。

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

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.