Aim. The genus Aconitum is represented in the Carpathian region by a significant number of endemic species. Some of these species, in particular A. pseudanthora and A. jacquinii, are included in the Red List of Ukraine. However, their taxonomic status and consequently the importance of conserving their natural resources remain uncertain. DNA barcoding techniques, e.g., the application of molecular markers based on variable regions of the chloroplast genome can help clarify these questions. Methods. DNA isolation, PCR amplification, sequencing of PCR products, phylogenetic analysis. Results. The psbA-trnH intergenic spacer region of A. pseudanthora and A. jacquinii specimens from Western Ukraine was amplified and sequenced. It is shown that the spacer sequences for these two species are identical, but differ in short indels and nucleotide substitutions from the psbA-trnH sequences of A. anthora s.l. specimens from Western Europe and China. Phylogenetic analyzes showed the phylogenetic affinity of the members of the A. anthora complex to the subgenus Lycoctonum. Conclusions. The use of the psbA-trnH intergenic spacer region for DNA barcoding makes it possible to unambiguously distinguish samples of A. jacquinii and A. pseudanthora from western Ukraine from samples of A. anthora s.l. of other geographical origins. The results obtained confirm the reticular nature of the evolution in the genus Aconitum.