Two powerful approaches are frequently used to trace incorporation and degradation of plant derived C in soil: 14C labelling/chasing and analysis of lipid composition. In this study, we coupled these approaches in order to trace short term incorporation of plant derived lipids into rhizosphere and non-rhizosphere soil. Methodological optimization was required and implied 14C liquid scintillation counting improvement for plant lipid extracts taking into account organic solvents, solvent-to-scintillation cocktail ratio, and amount of lipids. Following method optimization, 14C data of fatty acids indicated a notable contribution of root derived lipids to rhizosphere and non- rhizosphere soil. Coupling of 14C labelling/chasing with lipid analysis is a powerful and cheap approach for tracing of root derived C in soil allowing for estimation of C budget, for determination of C formation and translocation within plants and from plant to soil, as well as for identi- fication of short term dynamics of specific compound classes within soil.