Małgorzata Kozyra

Solid-phase extraction and reversed-phase high-performance liquid chromatography of free phenolic acids in some Echinacea species

Abstract A new method is described combining solid-phase extraction (SPE) and reversed-phase high-performance liquid chromatography (RP-HPLC) for the isolation, purification as well as qualitative and quantitative determination of free phenolic acids in six Echinacea species. Plant extracts were purified and phenolic acids isolated on octadecyl and quaternary amine Bakerbond SPE columns; final eluates were analysed by RP-HPLC. Significant differences in the composition and amount of phenolic acids within Echinacea genus have been shown. The method can be used for quick screening analysis of the content of phenolic acids in plant material.

Establishment of a co-culture of Ammi majus L. and Ruta graveolens L. for the synthesis of furanocoumarins

Abstract Ammi majus and Ruta graveolens plants were chosen to establish a co-culture due to their abilities to produce secondary metabolites from the furanocoumarin (FC) family. The conditions for cultivating A. majus hairy roots and R. graveolens shoots separately were established earlier. In continuation, the conditions of the co-culture of A. majus and R. graveolens were determined in this work. Two systems of co-cultivation were tested: A. majus hairy roots with R. graveolens cell suspension and A. majus hairy roots with R. graveolens shoots. The second model was chosen for further experiments, namely to test different tissues inoculation ratios as well as the influence of the photoperiod and darkness conditions on A. majus and R. graveolens tissues growth and the accumulation of secondary metabolites. The growth index ( t 25 / t 1 ) of R. graveolens shoots was higher in the co-cultures; moreover, 2.5 times higher concentration of xanthotoxin was observed in the co-culture in light conditions.

Chemical composition and variability of the volatile components from inflorescences of Cirsium species

The present study aimed to investigate the chemical composition of the essential oils of inflorescences Cirsium spp. (Asteraceae) by GC/MS method. Essential oils were extracted from the inflorescences of Cirsium pannonicum (Link), Cirsium ligulare Boiss., Cirsium heterophyllum (L.) Hill., Cirsium acaule (L.) Scop., Cirsium oleraceum (L.) Scop., Cirsium dissectum (L.) Hill., Cirsium decussatum (Janka) and Cirsium eriophorum (L.) Scop., using the steam distillation method. A gas chromatography–mass spectrometry method was employed for the analysis of essential oils. Our study shows the differences in chemical composition of volatile oils in the inflorescences of Cirsium spp. The main components of the essential oil were ketones and aldehydes with a long carbon side-chain. Volatile oils also contained small amounts of terpenes: thymol, β-linalool, eugenol, carvacrol and fatty acids with odd number of carbon atoms–waxes. The compounds in the essential oils obtained from inflorescences Cirsium L. species have been identified for the first time.

Column chromatography and preparative TLC for isolation and purification of coumarins from Peucedanum verticillare L. Koch ex DC

Preparative separation and isolation of coumarins from petroleum ether and methanol extracts of the fruits and roots of Peucedanum verticillare L. Koch ex DC (Umbelliferae = Apiaceae) have been achieved by column chromatography and normal- and reversedphase TLC with gradient elution. Isolated coumarins were identified by comparison with standards in analytical TLC and in RP HPLC with gradient elution, by melting point (m.p.) measurement, by UV spectroscopy, by MS, and by 1H and 13C NMR spectroscopy. Esculin, umbeliferone, bergapten, xanthotoxin, isoimperatorin, imperatorin, psoralen, and cis-kellactone were isolated for the first time from P. verticillare L. Koch ex DC. Two angular-type pyranocoumarins isolated from the roots and fruits of Peucedanum verticillare L. Koch ex DC. were identified as pteryxin and epoxypteryxin by MS and 1H and 13C NMR.

Chemical compositions and antibacterial activity of extracts obtained from the inflorescences of Cirsium canum (L.) all

The aim of this study was to investigate phenolic acids and flavonoids in methanolic, dichloromethane, acetone and ethyl acetate extracts and fractions from inflorescences of Cirsium canum (L.). RP-HPLC analysis enabled identification of the following: chlorogenic acid, caffeic acid, p-coumaric acid, protocatechuic acid, p-hydroxybenzoic acid, vanillic acid, syringic acid, trans-cinnamic acid, luteolin-7-glucoside, apigenin-7-glucoside, kaempferol-3-glucoside, linarin, apigenin, rutoside, luteolin and kaempferol. The antimicrobial activity of tested extracts was determined in vitro against reference microorganisms, including bacteria or fungi, belonging to yeasts. Our data showed that the tested extracts had no influence on the growth of the reference strains of Gram-negative bacteria and yeasts belonging to Candida spp. Among them, the fractions possessed the highest activity against Gram-positive bacteria, especially Streptococcus aureus and Streptococcus pneumoniae belonging to pathogens and Streptococcus epidermidis, Bacillus cereus and Bacillus subtilis belonging to opportunistic microorganisms.

Phenolic acids in Peucedanum verticillare L. Koch ex Dc

Phenolic acids in the leaves, roots, and fruits of Peucedanum verticillare L. Koch ex Dc have been qualitatively determined for the first time by the method of one- and two-dimensional TLC on cellulose layers. Phenolic acids in P. verticillare have not previously been studied.