Histamine is a biogene amine, derived from the decarboxylation of the amino acid histidine, as a result of microbial activity, mainly in protein-rich foods such as fish, cheese, meat and fermented products. Its analysis is important because its excessive content in organisms can cause serious poisoning. Reliable determination of histamine is crucial for ensuring consumer safety and compliance with regulatory limits. The purpose of the master's thesis was to optimize and validate a method for the determination of histamine in foods according to the ISO 19343:2017 standard, replacing environmentally harmful toluene with safer solvents and procedures. The method used is based on a combination of scientific literature and standard procedures, with special emphasis on the choice of mobile phase, flow rate, temperature and the use of derivatization. We found that increasing the proportion of methanol in the mobile phase shortens the retention time of histamine, and the optimal composition was at 35% methanol. A flow rate of 1.0 mL/min represented the best compromise between resolution and analysis duration, while a higher temperature (40 °C) improved histamine extraction without significantly affecting the symmetry of the peaks. Derivatization with dansyl chloride proved to be crucial for the sensitivity of the method, as histamine was not detected without this step. The calibration curve was linear (r=0.997), the limit of detection (LOD) was 1.0 mg/L, the limit of quantification (LOQ) was 4.0 mg/L, which is suitable for food quality control. Validation results (RSD < 15%) confirm the reliability of the method in the analysis of various matrices such as fish, cheeses and wines. Analyses of food samples revealed the presence of histamine in dried fish and at low concentrations in Parmigiano Reggiano cheese, with all levels remaining below the regulatory limits. The method enables fast, accurate and safe determination of histamine in complex food samples.
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