摘要
该研究通过考察色谱条件、前处理方法及基质效应建立同时测定发酵酒中15种生物胺的通过式固相萃取-超高效液相色谱-串联质谱(UPLC-MS/MS)技术,并进行方法学考察。结果表明,样品加入同位素内标后经PRiME HLB柱净化,以0.1%甲酸乙腈-0.1%甲酸水为流动相,采用Waters ACQUITY UPLC^(®)BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm)能对15种生物胺进行有效分离,通过多反应监测(MRM)模式采集数据。其中,腐胺、N1-乙酰精胺、亚精胺、3-甲基组胺4种生物胺无基质效应,采用外标法进行定量,其余11种生物胺采用内标法进行定量。15种生物胺均在1~100μg/L的质量浓度范围内线性关系良好,相关系数R2均>0.99,检出限和定量限均分别为0.1 mg/L和0.3 mg/L;在3种发酵酒基质中添加3个不同质量浓度的生物胺(0.3 mg/L、1.0 mg/L、10.0 mg/L),平均加标回收率为75.6%~109.5%,精密度实验结果的相对标准偏差(RSD)为0.4%~6.4%,说明该方法适用于发酵酒中15种生物胺的同时检测。采用该方法从黄酒、葡萄酒、啤酒实际样品中均检出腐胺、三甲胺、酪胺。
A method was established for simultaneous determination of 15 biogenic amines in fermented alcoholic drink using pass-through solid phase extraction combined with ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),and the methodology was investigated.The results showed that the sample was purified by PRiME HLB column after adding isotopic internal standards,and 15 biogenic amines could be effectively separated by Waters ACQUITY UPLC^(®)BEH C_(18)chromatographic column(100 mm×2.1 mm,1.7μm)with 0.1%formic acid acetonitrile-0.1%formic acid water as the mobile phase.Data was collected by multiple reaction monitoring(MRM)mode.Four biogenic amines(putrescine,N1-acetylspermine,spermidine and 3-methylhistamine)without matrix effect were quantified using external standard method,but the remaining 11 biogenic amines were quantified using internal standard method.The linear relationship of 15 biogenic amines was good within the mass concentration range of 1-100μg/L,and the correlation coefficients R2 were all greater than 0.99.The detection limit and quantification limit were 0.1 mg/L and 0.3 mg/L,respectively.Three different mass concentrations of biogenic amines(0.3 mg/L,1.0 mg/L,10.0 mg/L)were added to three types of fermented alcoholic substrates,the average standard recovery rates were 75.6%-109.5%,and the relative standard deviation(RSD)of precision test results was 0.4%-6.4%,indicating that the method was suitable for simultaneous detection of 15 biogenic amines in fermented alcoholic drink.The putrescine,trimethylamine,and tyramine were detected in actual samples of Huangjiu,wine and beer using this method.
作者
徐潇颖
张一涵
梁晶晶
XU Xiaoying;ZHANG Yihan;LIANG Jingjing(Zhejiang Provincial Key Laboratory for Market Regulation and Health Food Quality and Safety,National Key Laboratory of Market Supervision(Quality and Safety of Functional Food),Zhejiang Institute for Food and Drug Control,Hangzhou 310052,China;College of Medical Technology and Information Engineering,Zhejiang Chinese Medical University,Hangzhou 310000,China)
出处
《中国酿造》
北大核心
2025年第8期263-269,共7页
China Brewing
基金
浙江省市场监督管理局检验监测技术装备项目(ZB2022466)
浙江省科技计划项目(2025C02126)
浙江省基础公益研究计划项目(LTGY23H260007)。
关键词
发酵酒
生物胺
通过式固相萃取
超高效液相色谱-串联质谱
fermented alcoholic drink
biogenic amines
pass-through solid phase extraction
ultra high performance liquid chromatography-tandem mass spectrometry
