*2.6. Fatty Acid Composition*

A sub-sample of 60 steaks (*n* = 30 steaks closest to the mean marbling score of each treatment) were selected to evaluate fatty acid profile using direct FAME synthesis. Steaks were thawed slightly and external fat, epimysial connective tissue, and additional muscles were trimmed from the longissimus muscle. Samples were minced, immersed in liquid nitrogen, and powdered for 15 s using a Waring commercial blender (Waring Products Division, Model 51BL32, Landcaster, PA, USA). Duplicate 1 g samples were weighed and processed to generate FAMEs according to procedures of O'Fallon et al. [19]. Fatty acids were identified through comparison with retention times of an authentic fatty acid standard mixture (GLC-463, Nu-Check Prep Inc., Elysian, MN, USA). Quantities were computed as mg/g of raw wet tissue through an internal standard calibration method where C13:0 served as the internal standard. Final contents were then summed and %, g/100 g total fatty acids was produced after summing all fatty acids.
