The study course reviews the main techniques for isolation and purification of organic substances in preparative and semi-preparative scale. It also covers the commonly used analytical techniques for evaluation of the purity of organic compounds. In the study course, we will discuss the purification methods of organic compounds based on the presence of various functional groups. The study course gives a deeper insight into the use of specific laboratory equipment in order to solve the everyday as well as non-ordinary practical problems of organic chemistry. Two main sources of organic compounds: organic synthesis and isolation from nature are described. In this context, the main methods of fractionation are discussed. On the equal level, the classical analysis methods still in use (colour tests, titrimetric and gravimetric methods) and moderns ones (mass spectrometry, chromatography, NMR and IR spectrometry, circular dichroism, X-ray analysis etc.) are illustrated. Special attention is drawn to the determination of absolute configuration and enantiomeric excess of chiral substances.