Expressing human milk has become a more common alternative for mothers, as the average work demand has increased. As more mothers must work, bottle feeding trends are increasingly common. The handling and storage of human milk introduce the risk of degradation to expressed human milk and infant formula. In following a 20-minute simulated feeding, Vitamin C has been found to degrade. Vitamin C acts as an anti-oxidant and is responsible for shielding other nutrients from oxidation, such as retinol and alpha-tocopherol. By analyzing a 20-minute simulated feeding, retinol and alpha-tocopherol each displayed decreases over time significantly different than that of the Control, which was milk not exposed to bottle feeding. In human milk, retinol showed as high as a 9.5% decrease compared to the Control. Similar trends were seen with the infant formula samples. The correlation between degradation and bottle feeding systems was dependent upon the formation of bubbles in the milk as the milk was removed from the bottle. The analysis indicated a decrease of up to 12%, as seen in retinol, and 35%, as seen in alpha-tocopherol. These decreases in retinol and alpha-tocopherol should be considered when using a bottle feeding system to deliver either human milk or formula to an infant. More research is necessary to determine the effect of this decrease on the nutritional status of infants, particularly premature infants, who are at higher risk for nutrient deficiencies.