白兰地原酒及成品酒中九种生物胺含量分析

Characteristic analysis of nine kinds of biogenic amine content inbrandy raw wine and finished wine

该研究建立了高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(liquid chromatography-quadrupole/electrostatic field orbit trap high-resolution mass spectrometry, HPLC-Q-Exactive-Orbitrap-HRMS)同时测定白兰地中9种生物胺的分析方法。白兰地样品经水相滤膜过滤后直接进样,经C18(100 mm×4.6 mm×2.6μm)色谱柱分离,乙腈和1%(体积分数)甲酸水溶液为流动相梯度洗脱,采用电喷雾正离子扫描,在tMS~2模式下提取目标化合物精确质量数,外标法定量。结果表明,质量浓度为0.01~10 mg/L时,各生物胺浓度与峰面积线性关系良好(r>0.995 2),检出限为0.02~2.26μg/L,定量限为0.05~7.55μg/L,样品平均回收率为80.8%~119.2%。原酒与成品酒中生物胺含量差异明显(分别为3.0~126.0μg/L和17.0~148.0μg/L)。不同档次成品酒样中主要含有的组胺、酪胺和盐酸吡哆胺含量差异较大。而不同产地和生产工艺的白兰地原酒中生物胺含量及种类同样显著不同。该定量检测方法简单、快速,具有良好的灵敏度和精密度,适用于白兰地中低浓度生物胺的测定。

A method for determining the content of 9 biogenic amines in brandy by liquid chromatography-quadrupole/electrostatic field orbit trap high-resolution mass spectrometry was established.The brandy samples were directly injected into HPLC after being filtrated by an aqueous filter membrane and separated by a C18(100 mm×4.6 mm×2.6μm)column.Acetonitrile and 1%formic acid aqueous solution were used as mobile phase for gradient elution.The mass spectrometry data was detected by electrospray ionization scanning in tMS 2 mode,and quantified by external standard method.Results showed that in the range of 0.01-10 mg/L,the concentration of biogenic amines had a good linear correlation with peak area(r>0.9952).The limits of detection were 0.02-2.26μg/L,and the limits of quantification were 0.05-7.55μg/L.The average recovery of samples was 80.8%-119.2%.The contents of bioamines in raw brandy(5.0-113μg/L)and finished brandy(21.0-106μg/L)were significantly different.Especially,histamine and tyramine were the main biogenic amines in hydrochloride in the low-end finished brandies,meanwhile,pyridoxamine hydrochloride had the highest content in high-end brandies.In addition,production processes and conditions could also affect the types and content of biogenic amines in brandy.The method is simple and rapid,has good sensitivity,recovery,and precision,and it is suitable for the detection of low-concentration bioamines in brandy.