Flavonoids present in the extracts from leaves of Pseudotsuga menziesii (Pinaceae), Ginkgo biloba (Ginkgoaceae) and Podocarpus dacrydioides (Podocarpaceae) were separated by use of the reversed phase HPLC method. The analysed compounds belong to different groups of flavonoids – biflavones (amentoflavone, bilobetin, 5’−methoxybilobetin, podocarpusflavone A, sequoiaflavone, podocarpusflavone B, ginkgetin, isoginkgetin, sciadopitysin, kayaflavone, hinokiflavone, 2,3−dihydrosciadopitysin, 2,3−dihydroisoginkgetin), O−acylated flavonol glycosides (daglesiosides I, II, III, IV, trans−tiliroside, trans−ditiliroside), flavonol O−glycosides (astragalin, isoquercetin) and flavonol aglycones (kaempferol, quercetin, isorhamnetin). The conditions for flavonoid separation were optimized using various RP−18 columns. The chromatographic resolution was performed with isocratic or gradient elution – optimized by Drylab program or by traditional “trial-and-error” method, depending on the composition of flavonoid complex.