The main aim of this study was to evaluate the volatile profile, sensory perception, and phytochemical content of bovine milk produced from cows fed on three distinct feeding systems, namely grass (GRS), grass/clover (CLV), and total mixed ration (TMR). Previous studies have identified that feed type can influence the sensory perception of milk directly via the transfer of volatile aromatic compounds, or indirectly by the transfer of non-volatile substrates that act as precursors for volatile compounds. In the present study, significant differences were observed in the phytochemical profile of the different feed and milk samples. The isoflavone formonoetin was significantly higher in CLV feed samples, but higher in raw GRS milk, while other smaller isoflavones, such as daidzein, genistein, and apigenin were highly correlated to raw CLV milk. This suggests that changes in isoflavone content and concentration in milk relate to diet, but also to metabolism in the rumen. This study also found unique potential volatile biomarkers in milk (dimethyl sulfone) related to feeding systems, or significant differences in the concentration of others (toluene, p-cresol, ethyl and methyl esters) based on feeding systems. TMR milk scored significantly higher for hay-like flavor and white color, while GRS and CLV milk scored significantly higher for a creamy color. Milk samples were easily distinguishable by their volatile profile based on feeding system, storage time, and pasteurization.Molecules 2020, 25, 26 2 of 28 of forage type on milk fat and composition, and highlighted the need to evaluate the impact of feeding systems on other aspects of milk fat quality, such as flavor and oxidative stability. Milk produced from many supplemented and altered diets have been investigated, including supplementation with flaxseed [8], lipid complex [9], crude protein [10], iodine [11] marine algae [12], oregano and caraway essential oils [13], hull-less barley [14] and sunflower/fish oil [15]. These studies focused mainly on animal production performance, milk composition, milk yield, milk fatty acid composition, and to a lesser extent on the flavor and sensory characteristics of milk. The study by O'Callaghan, et al. [16] investigated the influence of four supplemental feed choices for pasture-based cows on the fatty acid and volatile profile of milk. Some studies have also evaluated the effect of storage conditions on the microbiological quality of milk [17,18]. In the present study, the volatile profile and free fatty acid (FFA) content of the milk samples were evaluated over a 14-day storage period at 4 • C in order to ascertain the level of lipid oxidation occurring within the milk, and to track volatile compounds forming or changing during refrigerated storage. Free fatty acids (FFAs) in milk are produced by two mechanisms, namely incomplete esterification in the mammary gland before lipid excretion [19] or lipid hydrolysis after milking and during storage [20]. The FFAs influence product quality, flavor, nutrition, and texture, and...