Simultaneous determination of 13 biogenic amines and their metabolites in liquid fermented food by HPLC-QTRAP-MS without derivatization

Biogenic amines and their metabolites (BAs) are among the most frequently detected compounds in liquid fermented foods, raising significant concerns about the potential risks they pose. To rapidly and selectively determine BAs, this study proposed and validated a method for the simultaneous determination of 13 BAs in liquid fermented foods using high performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry (HPLC-QTRAP-MS) without derivatization. The simultaneous separation and determination of histamine (HIS), tyramine (TYR), putrescine (PUT), octopamine (OCT), 2-phenylethylamine (2-PHE), dopamine (DOP), spermidine (SPD), epinephrine (EPI), agmatine (AGM), 5-hydroxytryptamine (5-HYD), tryptamine (TRY), L-noradrenaline (L-NOR), cadaverine (CAD) were achieved using a Waters Atlantis Premier BEH Z-HILIC (1.7 mu m particle size, 100 mm x 2.1 mm id) column with a mobile phase consisting of water (containing 0.025 % formic acid) and acetonitrile (containing 0.1 % formic acid). The analysis of target compounds was performed with electrospray ionization (ESI) in positive ion mode, multiple reaction monitoring (MRM), information dependent acquisition (IDA), enhanced product ion (EPI), and multi-step mode analysis combined with EPI library searching. To enhance the extraction efficiency of BAs, factors such as extraction solvent, filter membranes, extraction methods, pH, oscillation speed, time and temperature were systematically investigated. The results demonstrated that the method exhibited low limits of detection (LOD) and quantification (LOQ), ranging from 0.05 to 2.00 mu g/L and 0.16 to 6.60 mu g/L, respectively. The method also showed satisfactory reproducibility, with intra-day and inter-day relative standard deviations ranging from 0.90 % to 3.80 % and 2.65 % to 8.27 %, respectively. Additionally, it had a broad linear range from 0.05 to 200.00 mu g/L with excellent linearity (R>0.99) and an average recovery rate between 65.68 % and 115.14 %. Overall, the results demonstrated that the established method was convenient to operate, offered a short analysis time, and achieved high accuracy and sensitivity. It met the qualitative and quantitative requirements for detecting 13 BAs in liquid fermented food, thus providing strong technical support for BAs detection.