Abstract Two different methods for single step transesterification from pig meat without fat extraction have been tested. Freeze-drying of the meat with and without anhydrous salt, followed by a base-catalyzed transmethylation (KOH/MeOH) was carried out. Both methods were compared with the standard Folch procedure of fat extraction followed by transmethylation. The methods were tested on a complete sample set of biceps femoris of pig thigh, used for the production of dry-cured ham. The set was divided in three subgroups according to total fat content. Both derivatization protocols on freeze-dried pork muscle were proven to be a valid alternative to the Folch procedure for FAME analysis. Freeze-drying method offered several advantages in comparison with the Folch procedure, including a lower solvent requirement, and process temperature, as well as considerable saving of time. In freeze-drying, the addition of an anhydrous salt (Na2SO4) gave more friable samples which resulted in higher yields for some fatty acids, particularly evident in the case of tissues with high lipid content. © 2017, Association of Food Scientists & Technologists (India).