The fruit essential oils of two populations of Astrantia major L. (Apiaceae, subfamily Saniculoideae) were analyzed in detail by GC and GC/MS analyses. Seventy-six constituents identified accounted for 92.7-94.0% of the oils. The two oils differed significantly: the wild-growing population from Serbia contained zingiberene (47.9%), β-bisabolene (9.7%), and β-sesquiphellandrene (7.9%), while the one from Poland (botanical gardens) was sesquiterpene-poor with the major contributors oleic acid (38.6%), nonacosane (15.4%), and linoleic acid (5.1%). Motivated by the unresolved taxonomical relations between the Saniculoideae and Apioideae subfamilies, we performed multivariate statistical analyses on the compositional data of these A. major samples, and additional 14 Saniculoideae and 31 Apioideae taxa. This allowed us to assess the chemotaxonomical usefulness of such chemical data in differentiating taxa from these two Apiaceae subfamilies and to corroborate the existence of at least two A. major chemotypes. Diethyl ether extracts of the two samples of A. major fruits yielded seven diaryltetrahydrofurofurano lignans. Except for eudesmin that has been found for the first time in a Saniculoideae taxon, all other lignans (magnolin, epimagnolins A and B, epieudesmin, yangambin, and epiyangambin) are new for the entire plant family Apiaceae. The lignan profiles also supported the existence of two separate A. major chemotypes.