Targeted and Rapid Determination of Key Biogenic Amines in Tuna by DART-TQ MS

The production of Biogenic Amines (BAs), important indicators of both quality and safety of fish and fishery products, is influenced by factors such as harvesting techniques, handling practices, and on-board processing operations. Scombroid dark-meat fish, such as tuna, are commonly associated with histamine poisoning, resulting from microbial decarboxylation of histidine [1]. Alongside histamine, other prevalent biogenic amines in fish include tyramine, putrescine, cadaverine, and spermidine. The study explores the potential of DART-MS combined with targeted Multiple Reaction Monitoring (MRM) on a DART-EVOQ triple quadrupole mass spectrometer for the detection and quantification of BAs in tuna samples, employing a straightforward sample preparation. Preliminary analyses focused on the selection of characteristic and specific ions for each biogenic amine under investigation, based on fragmentation patterns initially obtained from pure standard powders and subsequently confirmed in real tuna matrix. The tuna matrix was briefly extracted following a protocol adapted from Lioupi et al. (2024) [1] and analyzed using DART-MS. Full-scan and Product Ion Scan experiments enabled the identification of potential markers. Following the selection of the most suitable transitions for each compound and optimization of collision energy, MRM experiments were performed to enable targeted quantitative analysis. Specific markers and their main fragment ions were confirmed as suitable for quantification in a multi-target method, including: 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 for each BAs were created at crescent levels of 200, 400, 600, 800mg/kg for each BAs. Calibration curves showed a strong correlation, the method achieved reliable quantification in compliance with international regulatory limits (EU and FDA), demonstrating its effectiveness for routine safety monitoring [4] confirming the successful development of a rapid, high-throughput method for quality and safety screening.