Vitamin consumption is essential to human health as they are not synthesized in the body, meaning they must be obtained from external sources, such as vegetables. The aim of this study was to design an efficient and robust method to extract vitamins, and quantify them using HPLC-MS/MS. These vitamins included: alpha-tocopherol, beta-carotene, phylloquinone, retinol, thiamine, pantothenic acid, niacin, choline, and folic acid. Further studies were performed to compare varying levels of pesticide exposure, and analyze concentration of vitamins over time. Ultimately the vitamin content remained relatively constant over time despite matrix degradation, with the exception of the statistically significant increase of phylloquinone. Further studies were performed with organic spinach to compare varying levels of pesticide exposure and it was found that the concentration of vitamins was not significantly affected by pesticides. However, concentrations of beta-carotene and choline were an anomaly as they increased with statistical significance in organic spinach samples.