Advancing Food Safety: A Rapid and High Throughput DART-MS/MS method for Biogenic Amines detection in foods

Biogenic amines (BAs) are low-molecular-weight compounds formed by the microbial decarboxylation of amino acids and are widely recognized as indicators of freshness and safety in different types of foods and in particular in fish and fishery products [1,2]. Scombroid dark-meat fish, such as tuna, are well-known sources of histamine intoxication, which arises from the microbial decarboxylation of histidine [1]. In addition to histamine, fish commonly contain other biogenic amines of concern—namely tyramine, putrescine, cadaverine, and spermidine [3,4]. BAs quantification in fish and fishery products is crucial for food safety monitoring. Traditional official methods primarily rely on chromatographic techniques such as High-Performance Liquid Chromatography (HPLC) with fluorescence or UV detection, offering high sensitivity and specificity [5]. These methods require derivatization and lengthy analysis times, motivating the development of faster, simpler alternatives. The present study aims to exploit a high throughput and streamlined workflow based on DART coupled to a EVOQ triple Quadrupole Mass Spectrometer for the rapid analysis of BAs in fish filets proposed as an alternative to HPLC methods. The method, environmentally friendly, requires a minimal sample prep and enables fast and efficient analysis. Preliminary analyses were carried out in Full-scan and Product Ion Scan mode on BAs standards and were directed to identify the characteristic markers for each investigated biogenic amine, then validated in food samples. After identification of the main fragment ions, MRM experiments were performed for targeted quantitative analysis. The ion transitions selected for each BA in the multi-target method are histamine (112.2 > 95.0 m/z), putrescine (89.0 > 72.1 m/z), spermidine (146.0 > 112.1 m/z), tyramine (138.1 > 121.1 m/z), and cadaverine (103.0 > 86.0 m/z). Calibration curves, generated spanning the concentration range from 200 to 800 mg/kg for each BA, exhibited strong linearity, supporting robust quantification aligned with international regulatory requirements (EU and FDA) [2—5]. Overall, the results confirm the establishment of a rapid, high-throughput approach suitable for routine quality and safety screening of biogenic amines in fish products.