Lipids are vital for cellular functions, acting as structural components, signaling molecules, and energy sources. Their metabolism, particularly through cytochrome P450s and other oxidizing enzymes, plays a key role in inflammation and disease. This study uses biochemical, analytical, and biophysical methods to examine the metabolism of endocannabinoids (a class of signaling lipids) and minor cannabinoids by membrane-bound cytochrome P450s, stabilized in nanoscale lipid bilayers (Nanodiscs). The pharmacological properties of lipid metabolites are evaluated, focusing on their impact on inflammation and pain receptor modulation. This integrated approach enhances our understanding of lipid metabolism and cannabinoid pharmacology, supporting the development of therapeutic strategies for pain and inflammatory diseases.