Gentiopicroside (1) is the major secoiridoid glucoside constituent of Cephalaria kotschyi roots. The mutagenicity, DNAdamaging capacities, and clastogenicity of this molecule were evaluated by the Salmonella typhimurium mutagenicity assay (Ames test) on tester strains TA97a, TA98, TA100, and TA102, the alkaline comet assay, and the micronucleus assay on CHO cells. All tests were performed with and without the metabolization mixture, S9 mix. In the Ames test, the mutagenicity of 1 was limited to TA102 without S9 mix (2.3 rev µg -1 ). The genotoxicity was more evident without S9 mix (0.78 OTMχ 2 units µg -1 mL) than with the metabolic mixture (0.16 OTMχ 2 units µg -1 mL) with the comet assay. Similarly, the clastogenicity without S9 mix was 0.99 MNC µg -1 mL and 0.38 MNC µg -1 mL with S9 mix in the micronucleus assay. The interaction of 1 with DNA is probably through the involvement of oxidative DNA lesions.

Cephalaria kotschyi Boiss. et Hoh. (Dipsacaceae) is an endemic plant of the Caucasus region growing in Azerbaijan. Earlier phytochemical studies reported the isolation of triterpene saponins, flavonoids, and gentianine type alkaloids from the roots and flowers of C. kotschyi [1][2][3][4][5][6][7]. In our previous study we described the isolation of the iridoid glycosides loganin and gentiopicroside from the roots of this plant [8].Herein we report, together with the gentiopicroside and loganin described previously, the isolation of three iridoid and secoiridoid glycosides: loganic acid [9], swertiamarin [10], and cantleyoside [11]; two furofuranic lignans: prinsepiol glycoside [12] and 8-hydroxypinoresinol-4c-O-E-D-glucoside [13]; and two triterpene saponins: giganteoside Jc [14] and dipsacoside B [15], from an investigation of the chemical constituents of the roots of C. kotschyi. The plant was collected in the Lerik region of Azerbaijan in September 2005. A voucher specimen (C 8131005) was deposited at the herbarium of the Botanical Institute, Academy of Science, Azerbaijan.Air-dried roots (100 g) were extracted by percolation at room temperature with MeOH (1 L for 24 h), and the obtained solution was concentrated under vacuum. The MeOH extract (5 g) was submitted to flash column chromatography on polyamide MN SC 6 Macherey Nagel® (70 Pm), using a step gradient of H 2 O-MeOH of 90:10, 50:50, and 0:100, to give 3 main fractions (F1-F3). Fraction F1 was further purified by low-pressure liquid chromatography on Prepamatic Chromatospac 10 (Jobin Yvon) with Lichroprep RP18 Merck® (25-40 Pm). A step gradient of H 2 O-MeOH and 0:100, 1 L for each) allowed us to obtain 20 fractions, Sp1-Sp20. Fractions Sp2, Sp5, Sp7, Sp9, and Sp16 afforded respectively loganic acid (18 mg), swertiamarin (73 mg), gentiopicroside (250 mg), loganin (75 mg), and cantleyoside (230 mg). Purification of fractions Sp12 and Sp13 by means of preparative high-performance liquid chromatography on a Symmetry C18 column (5 Pm, 250 u 4.6, waters®), using H 2 O-MeOH (75:25), yielded prinsepiol glycoside (20 mg) and 8-hydroxypinoresinol glycoside (18 mg) respectively. Fraction F2 (1 g) was fractionated by silica gel CC using CH 2 Cl 2 -MeOH-H 2 O (50:16:3) and (30:13:3) successively as eluent to give dipsacoside B (30 mg) and giganteoside Jc (30 mg). The structural elucidations of the obtained compounds were performed by spectroscopic analysis, including 2D NMR data, and confirmed by comparison with the previously reported data. NMR spectra were recorded in CD 3 OD (G ppm) on a Bruker DRX 500 spectrometer operating at 500.13 MHz and 125.13 MHz.All the isolated compounds, except loganin and gentiopicroside, have been reported for the first time from C. kotschyi. The iridoid glycosides loganin, loganic acid, and cantleyoside of Cephalaria and other genus of the Dipsacaceae family [16][17][18][19][20] have been previously reported. These compounds are used as chemosystematic markers for plants of the Dipsacaceae family. The secoiridoid swertiamarin has b...