Coumarins are, nowadays, an important group of organic compounds from natural and synthetic sources, possessing a wide range of pharmacological activities. Coumarins comprise a large group of phenolic substances, which occur in plants as secondary metabolites. On account of the significance of this group of compounds, the development of effective methods for the extraction, separation, qualitative and quantitative analyses, and purification remains of high importance. The purpose of the chapter is to present information about methods of analysis and separation of this important group of naturally occurring compounds.