An HPLC method for quantitative determination of cynaropicrin in Centaurea scabiosa L. dry extract obtained using 40% aqueous ethanol has been developed and its performance has been validated. The proposed method is specific; ensures high accuracy, reproducibility, and linearity in an analytical range of ±30%; and can be used for reliable quality control of C. scabiosa L. dry extract and related preparations.

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Centaurea scabiosa L. (Asteraceae) is of great scientific and practical interest among the flora of Siberia. Its chemical composition includes various groups of compounds, of which sesquiterpene lactones (SL) have special significance. The guaiane-type SLs grosshemin and cynaropicrin were previously isolated for the first time from C. scabiosa and identified [1,2].Herein we report the isolation from the aerial part of C. scabiosa of another guaianolide, repin. The total SLs were separated over a column of silica gel (sorbent:compound ratio 30:1) with elution by petroleum ether and a gradient of petroleum ether:EtOAc with increasing fraction of the latter.Elution of the column by petroleum ether:EtOAc (90:10) afforded a brownish-yellow oil that was a mixture of eight compounds according to TLC. It was rechromatographed over a column of silica gel (sorbent:compound ratio 50:1) with elution by hexane and hexane:EtOAc.Removal of solvent from fractions eluted from the column by hexane afforded a colorless oil with crystalline inclusions that was soluble in benzene and CHCl 3 ; partly soluble in EtOH, EtOAc, Et 2 O, and hexane; and practically insoluble in H 2 O.Purification of the fraction with hexane followed by work up of the residue with EtOH (80%) isolated colorless crystals, mp 153-155°C (EtOH). IR spectrum (KBr, Q, cm -1 ): 3500 (OH), 1767 (J-lactone CO), and 1730 (ester).The PMR spectrum of the isolated compound (Table 1) showed doublets at 2.77 and 3.21 ppm with SSCC 6.0 Hz in addition to a singlet for methyl protons CH 3 (19) that were characteristic of epoxymetacrylic acid in the C-8 position of the guaiane skeleton.Resonances of exocyclic methylene protons conjugated to a J-lactone ring carbonyl were also found as a pair of doublets at 6.15 and 5.55 ppm. Resonances of protons of a second exocyclic methylene were observed as broad singlets at 5.12 and 4.96 ppm. A resonance at 4.90 ppm belonged to a hemiacyl proton. Table 1 presents the PMR and 13 C NMR spectra.The compound was identified as the guaiane-type SL repin based on the physicochemical properties and spectral data [3]. It was isolated from the first time from C. scabiosa.Thus, column chromatography over silica gel isolated for the first time the guaianolide repin from the aerial part of C. scabiosa. Repin was characterized before as having pronounced antibacterial and antifungal activity against Entamoeba histolytica and Trichomonas vaginalis [4].

Nowadays, the question about the negative impacts of liver fluke on the host organism and mechanisms of this damage is open. The bio-concentration of some heavy and toxic metals in tissue of adult forms of Opisthorchis felineus and its accumulation in liver tissue and cardiac muscle in the Syrian Golden Hamster (Mesocricetus auratus) model was analyzed. It is outlined, that Opisthorchis felineus infection leads to accumulating aluminum in a liver tissue of the host. It was also found, that adult fluke bio-concentrates iron and aluminum in their tissue. The shortage of such essential elements as Mn, Zn and Сu in infected organism is discussed.