High plasma levels of saturated fatty acids (SFAs) are associated with lifestyle diseases such as atherosclerosis and diabetes, whereas the accumulation of very long-chain fatty acids (VLCFAs) is believed to be responsible for neuropathy in certain types of peroxisomal disorders. Despite their clinical relevance, the toxicity mechanisms of fatty acids (FAs) remain poorly understood, largely because of the challenges in solubilizing them for in vitro experiments. We recently developed a method to form stable complexes of FAs with bovine serum albumin (BSA) using isopropanol as the solvent. Here, we demonstrate the stability and concentration range of FA/BSA and lysophosphatidylcholine (LysoPtdCho)/BSA complexes prepared using this method. These complexes exhibit enhanced solubility, retain their biological activity in cellular uptake assays, and remain stable for up to 12 months. We believe that our method will contribute to a better understanding of the toxicity and metabolism of SFAs and SFA-LysoPtdChos, and offer new insights into their roles in metabolic diseases and peroxisomal disorders.