Solid-phase extraction (SPE) was used to demonstrate how various lipids and lipid classes could be separated in a biochemistry laboratory setting. Three different SPE methods were chosen on their ability to separate a lipid mixture, consisting of a combination of a either a fatty acid, a triacylglycerol, a mono- or diacylglycerol, phospholipid, cholesterol, or cholesteryl ester into distinct lipid fractions. These mini-scale SPE methods used aminopropyl bonded silica columns or silica Sep-Pak cartridges suitable for completion in a 2- to 3-h time block. Thin-layer chromatography of the isolated lipid fractions was used to show separation characteristics and to evaluate the utility and reproducibility of each SPE method.