高效液相色谱法分析发酵肉制品中生物胺

Determination of biogenic amines in fermented meats by HPLC

建立了高效液相色谱法(high performance liquid chromatography,HPLC)检测发酵肉制品中常见8种生物胺(色胺、β-苯乙胺、腐胺、尸胺、盐酸吡哆胺、组胺、酪胺和精精胺)的柱前衍生-HPLC分析方法,并对其在实际检测中的适用性进行验证。采用5%三氯乙酸作为提取溶剂,提取发酵肉制品中的生物胺,以丹磺酰氯作为衍生试剂,采用Waters Symmetry C_(18)色谱柱(250 mm×4. 6 mm,5μm),乙腈和水(含0. 1%甲酸的0. 01 mol/L乙酸铵溶液)作为流动相进行梯度进样洗脱,流速为0. 8 mL/min,紫外检测器波长为254 nm。结果表明,发酵肉制品中的8种生物胺在37 min内能够进行分离,在0. 05～50μg/mL,8种生物胺均呈现良好的线性相关性(R2>0. 998)。各生物胺3个水平(1. 0、5. 0、10μg/g)加标回收率为83. 34%～111. 4%,相对标准偏差(RSD)均小于4. 85%,8种生物胺检出限为0. 03～0. 21μg/g,定量限为0. 10～0. 69μg/g。

A precolumn derivatization-HPLC method for determining eight biogenic amines (tryptamine,β-phenylethylamine, putrescine, cadaverine, pyridoxamine hydrochloride, histamine, tyramine, and spermine) in fermented meats was established. The applicability of this method in practical was verified and discussed. Biogenic amines in fermented meats were extracted with 5% trichloroacetic acid, and tanshinoyl chloride was used as a derivative reagent. The column used was Waters Symmetry C 18 (250 mm×4.6 mm,5 μm), acetonitrile and 0.01 mol/L ammonium acetate solution that contained 0.1% acetic acid were used as the mobile phase for gradient injection elution. The flow rate was 0.8 mL/min, and the wavelength of the ultraviolet detector was 254 nm. The results showed that the eight biogenic amines in fermented meats could be separated within 37 min. In the range of 0.05-50 ug/mL, the eight biogenic amines showed good linear correlations ( R^2>0.998). The recoveries of biogenic amines were 83.34%- 111.4% at three concentrations (1.0, 5.0, and 10 μg/g), and the relative standard deviations (RSD) were less than 4.85%. The detection limits of these eight biogenic amines were 0.03-0.21 μg/g, and their quantitative limits were 0.10-0.69 μg/g.