Fluorotelomer alcohols (FTOHs) belong to a group of chemicals called per-and polyfluoroalkyl substances (PFAS). PFAS have been shown to cause toxicity in humans through exposure to household products, ingestion of dust, water, or inhalation of indoor air. Some studies have postulated the mechanism of action for FTOH metabolism, which is that certain metabolites can cause toxicity, such as hormonal discrepancies, immunotoxicity, and possible tumor aggregation. Two understudied groups of bioactive metabolites of FTOHs are the fluorotelomer saturated and unsaturated aldehydes (FTAL and FTUAL). As shown in Figure 1, these are reactive metabolites, due to their highly electrophilic α,β-unsaturated or saturated aldehyde moieties that will react with biological nucleophiles to potentially cause protein modification or inactivation. Yet, the proteins they covalently bind to are unknown. Determining specific proteins will demonstrate how biological mechanisms are affected in the presence of these metabolites. Some issues could include disruption of cellular events or cell signalling leading to toxicity. Therefore, this project focused on the specific proteins modified upon FTOH biotransformation. Our hypothesis was that 6:2 FTAL and/or 6:2 FTUAL would react immediately with the enzyme(s) that produce it, given their high reactivity. Enzymes of interest were cytochrome P450 (CYP) 2A6 and 2E1, which were determined as possible targets from previous studies as they were shown to be active for the metabolism of FTOHs. We also used CYP 3A4 as a negative control.