Abstract The goal of this project is to determine how mutations, elevated protein levels, and environmental conditions impact the function and dysfunction of Cytochrome P450 1B1 (CYP1B1). The addition of instrumentation is proposed to facilitate characterization of small molecule modulators of CYP1B1, and the metabolic products of this CYP. Selective inhibition of CYPs is an important goal in medicinal chemistry, but these enzymes can chemically modify their inhibitors, resulting in deactivation. The requested High Pressure Liquid Chromatography (HPLC) instrument, along with three orthogonal detection systems, will allow for accurate and rapid determination of chemical structures and enzymatic modifications. This will be accomplished with small amounts of sample, as the analytes will come from cells grown in tissue culture. As CYPs generally increase the polarity of hydrophobic substrates, the instrumentation includes specific modifications that are required for analysis of aqueous media solutions. Such solutions are incompatible with most LC mass spectrometers. This equipment will be also used to evaluate if mutations, protein:protein association, and environmental conditions alter CYP metabolic products, with implications for tissue-selective metabolism under hypoxia or due to the presence of signaling molecules.