Abstract: The lipophilic composition of honeybee pollen from Cistus ladanifer, Castanea sativa and Rubus sp. was analysed by GC-MS. The extracts are mainly composed by saturated and unsaturated fatty acids, sterols, long chain aliphatic alcohols, alkanes and alkenes. The profiles of chemical composition allow the chemical differentiation of pollen species. The biological activity of the lipophilic fractions against several Gram positive bacteria was demonstrated and related to the resistance of beehives to certain diseases when feed with such pollens.