This document provides instructions for using column chromatography and thin layer chromatography (TLC) to separate pigments from a spinach extract. Students will first use column chromatography with a hexanes/acetone solvent system to separate the spinach pigments into fractions on an alumina column. They will then use TLC to identify the separated pigments by comparing their retention factors (Rf) to standards. The principles of green chemistry are applied by using a renewable plant feedstock, safer solvents like alcohols and acetone that can be recycled, and reusable alumina.