高效液相色谱-串联质谱法测定食品中的5种生物胺

Determination of 5 biogenic amines in food by high performance liquid chromatography-tandem mass spectrometry

目的建立一种运用高效液相色谱-串联质谱法(HPLC-MS/MS)测定动物源性食品和腌制品食物中尸胺、腐胺、酪胺、组胺和色胺5种生物胺的分析方法。方法采用5%高氯酸冰浴超声提取试样中的生物胺,调节pH值为2~3后正己烷净化,以ACQUITY UPLC^(■)BEH HILIC(100 mm×2.1 mm,1.7μm)为色谱柱,0.5%甲酸的水溶液和0.5%甲酸的乙腈溶液为流动相梯度洗脱,在多反应监测(MRM)模式下测定,基质匹配标准溶液外标法定量。结果5种生物胺在8 min内得到有效分离,尸胺、腐胺和酪胺在10 ng/ml~1000 ng/ml浓度内线性系数(r)≥0.993,检出限为2.0 mg/kg。组胺和色胺在1 ng/ml~100 ng/ml浓度内线性系数(r)≥0.996,检出限为0.2 mg/kg。向鱿鱼样品中分别添加生物胺标准品8μg、80μg,2个含量水平5种生物胺平均回收率为85.9%~103.7%,相对标准偏差(RSD)为3.2%~8.9%。结论该方法前处理简单、快速,且具有灵敏度高,回收率和重复性好等优点,能满足日常检测工作的需求。

Objective To establish a method to determinate cadavetive(CAD),putrescine(PUT),tyramine(TYR),histamine(HIS),tryptamine(TYP)in animal derived food and preserved food by high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Methods To ultrasonic-extract the biogenic amines in ice bath from samples by 5%trichloroacetic acid aqueous solution,purify withn-Hexane after adjusting pH to the range of 2-3 and analyze the solution by HPLC-MS/MS in multiple reaction monitoring(MRM)mode.The chromatographic analysis is carried out on the ACQUITY UPLC^(■)BEH HILIC(100 mm×2.1 mm,1.7μm)column with 0.5%formic acid in water and 0.5%formic acid in acetonitrile as the mobile phases in gradient program,measured in multiple response monitoring(MRM)mode,and quantified by matrix-matched external standard method.Results Five bioamines were effectively isolated within 8 min.The linear coefficients of cadaverine,putrescine and tyramine ranged from 10 ng/ml to 1000 ng/ml(r≥0.993),and the detection limit was 2.0 mg/kg.The linear coefficients of histamine and tryptamine ranged from 1 ng/ml to 100 ng/ml(r≥0.996),and the detection limit was 0.2 mg/kg.The average recoveries of 5 bioamines were 85.9%-103.7%and the relative standard deviations(RSDs)were 3.2%-8.9%when adding 8μg and 80μg bioamines,respectively.Conclusion The method is simple,with high sensitivity,recoveries and good reproducibility,and can be used for daily inspection work.