This chapter describes a thin layer chromatography (TLC) method for the quantitation of various lipids (such as phospholipids, sphingolipids, acylglycerols, and fatty acids) in lipid-based nanoparticle formulations such as liposomes and nanoemulsions. We illustrate this technique to quantify C6-ceramide (N -hexanoyl-d -erythro-sphingosine) in a nanoemulsion formulation. C6-ceramide is a powerful chemotherapeutic that is poorly soluble in aqueous buffers.