In the Netherlands, butter is produced from milk originating from three different production systems: conventional, organic, and grass-fed cows. The aim of the current study was to characterize these types of butters, and pinpoint distinct compositional differences. Retail conventional (n = 28), organic (n = 14), and grass (n = 12) full-fat butters were collected during the winter and summer
seasons. Samples were analyzed for their fat content, free fatty acid (FFA) content, and triglyceride (TG) and fatty acid (FA) profiles. The fat content was significantly lower in conventional butters than in organic butters and the FFA content was significantly lower in conventional butters compared with grass butters. Also, organic butters differed significantly from their conventional counterparts with regard to their TG and FA profiles. The TG profiles of the organic and grass butters did not differ significantly. The FA profiles of grass butters were less distinct, since only a few FAs differed significantly from conventional (six FAs) and organic (eight FAs) butters.