The polar extracts of Potamogeton pectinatus, P. lucens, P. perfoliatus and P. crispus (Potamogetonaceae) were analyzed by HPLC-UV-MS and their chromatographic profiles were very similar. The polar constituents of P. pectinatus were more exhaustively investigated by HPLC-UV with post-column derivatization, HPLC-MS(n) and HPLC-NMR, which allowed the on-line identification of various known flavones (dereplication). One of these compounds, luteolin 3'-O-glucoside, has never been characterized in the Potamogeton genus. The HPLC-UV-MS and HPLC-NMR analyses revealed also the presence of ent-labdane diterpene glycosides in the polar extracts of P. pectinatus and P. lucens and led to the isolation of a new ent-labdane diglycoside from P. pectinatus, beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl-15,16-epoxy-12-oxo-8(17),13(16),14-ent-labdatrien-19-oate.