Analysis of non-volatile constituents in Dracocephalum species by HPLC and GC-MS

In this paper we identify/determine the composition of the extracts of Dracocephalum moldavica L. and D. ruyschiana L. with special emphasis on their flavonoids, aliphatic, aromatic carboxylic acids and sugars. The plant materials were extracted using methanol-acetone 1 : 1 (v/v) acidified with HCI (0.05%). Composition of extract's most red fractions was identified by HPLC, while the constituents of the entire extract were identified and quantified as their trimethylsilyl-(oxime) ether/ ester derivatives by GC-MS. On the basis of HPLC analyses - (performed on Phenomenex Luna 5 pm C18 column, 250x 4.6 mm I.D.; eluent: acetonitrile - trifluoroacetic acid (0.1%); isocratic/5 mu m C18 column, 250x 4.6 mm I.D.; eluent: acetonitrile - trifluoroacetic acid (0.1%); isocratic/ gradient conditions) - delphinidin, cyanidin and pelargonidin (in traces) were identified by spectral analysis and on the basis of authentic standard's addition. GC-MS analyses were carried out immediately with extracts, as well as subsequently to extract's hydrolysis (trifluoroacetic acid: 2M, 2h, 4h). Constituents were identified and quantified as their trimethylsilyl-(oxime) ether/ester derivatives, from a single run, on the basis of their total (TIC) and selective fragment ion (SIM) responses. Calculations were related to the dry matter content of extracts. As main constituents monosaccharides, sugar alcohols di- and trisaccharides, aliphatic (phosphoric, succinic, levulinic, malic, tartaric, fatty acids) and aromatic (quinic, chlorogenic, caffeic, ferulic, 3,4-dihydroxyphenyllactic, rosmarinic) carboxylic acids and their corresponding esters; apigenin, luteolin flavon aglycons and tocopherol, in total 33 constituents were quantitated partly by their TIC, partly by their SIM responses. Identification/quantification proved to be in total of 35-69% (expressed in the dry matter content of extracts).