Pufa 3 menhaden oil

The preparation of the fatty acid methyl esters (FAME) was conducted on approximately 0.2 g of minced Coppa Piacentina samples belonging to different ripening times (i.e., T0, T60, T90, T180, and T240) according to the direct method described by O’Fallon et al. [39 (link)]. The FAME were analyzed using a gas chromatograph (model 2025, Shimadzu Corporation, Kyoto, Japan) equipped with an auto-sampler (model AOC-20s, Shimadzu), a flame ionization detector, and a CP-Select CB capillary column for FAME (100 m × 0.25 mm i.d.; 0.25 µm film thickness; Chrompack, Varian, Inc., Palo Alto, CA, USA). The injection volume was 1 mL in split mode (split ratio 30:1) and hydrogen was used as carrier gas at a constant flow of 1.5 mL/min. The injector and detector temperatures were set at 250 °C. The oven temperature program was set as follows: 60 °C for 2 min, from 60 to 170 °C at 10 °C/min for 35 min, and from 170 to 240 °C at 4 °C/min for 9.5 min. The FAME were identified by comparison with the retention times of external standards (Supelco 37 component FAME mix, PUFA-3 menhaden oil, conjugated octadecadienoic acid; Sigma Chemical Co, St. Louis, MO, USA). Data were expressed as a percentage of total fatty acids, calculated with peak areas corrected by factors according to the AOAC 963.22 method [35 ].