Dilute and shoot HPLC-fluorescence method for the quantification of free bioactive amines in dry-fermented sausage

Dry-fermented sausages are significant source of histamine and tyramine, both of which pose risks for healthy individuals, and, in special, for susceptible ones (histamine intolerants and those under mono aminoxidase inhibitor drugs). This study aimed to develop a dilute-and-shoot procedure for multi-amine analysis in sausages using HPLC-fluorescence. The proposed method is simple, rapid, and does not require fat extraction or exhibit a matrix effect. Amine recovery ranged from 86.1 % (phenylethylamine) to 109.9% (histamine). Analytical curves were linear (R2 >= 0.9811). The method was efficient and sensitive (LOQ-1.56-4.08 mg/kg). 54 dry-fermented sausages were analyzed. All 10 amines were detected in Italian, whereas Mini Hamburgues contained only three amines. Mean total amine levels were higher in Gourmet Italian (1587 mg/kg), and lower in Serrano (104.0 mg/kg). Tyramine was present in all types of sausage, accounting for 25.8-98.9 % of the total amine. Histamine was present in most types (77.8 % predominance), with higher levels in Gourmet Italian (650.5 mg/ kg). Principal component and hierarchical cluster analyses classified the nine sausage types into six clusters, distinguishing Serrano & Milano, and Brianza & Italian & Friolano, from the others. These findings highlight the need for industries to mitigate amine formation in dry-fermented sausage.