The anticonvulsant and anti-plasmid conjugation potential of Thymus vulgaris chemistry: An in vivo murine and in vitro study.

The high-performance counter-current chromatography was used for the efficient purification of single constituents from Thymus vulgaris essential oil. Mixtures of n-heptane, ethyl acetate, methanol, and water (5:2:5:2 and 4:1:4:1 v/v), allowed purification of eugenol, 1-octen-3- ol, borneol, thymol, terpinen-4-ol, and camphor, while n-hexane, acetonitrile, and tert-butyl methyl ether (1:1:0.1 v/v) yielded carvacrol, borneol, linalyl acetate, caryophyllene oxide, p-cymene, and eucalyptol. The anticonvulsant activities were evaluated in the maximal electroshock-induced seizure test in mice model (systemic i. p. administration). The oil exerted protection against MES-induced seizures when administered 15 and 30 min before the tests (50 and 62.5%, respectively). Among the isolates, borneol, thymol, and eugenol exerted the strongest protection against seizures. Moreover, linalool had the ability to reduce the transfer of the pKM101 plasmid by 84%, what has the potential to reduce virulence and resistance spread in E. coli. No acute toxic effects towards the CNS were noticed either for the essential oil or for single compounds, in the chimney and grip-strength tests. The preclinical screening of Thymus vulgaris EO, as well as isolated terpenoids, provides evidence that the EO has partial protective activity against seizures and HPCCC technique is suitable for its large scale isolation.

Biological activity and safety profile of the essential oil from fruits of Heracleum mantegazzianum Sommier & Levier (Apiaceae).

A composition of essential oils obtained from Heracleum mantegazzianum (Apiaceae) was examined using a GC-MS method. n-Octyl acetate (19.92%), n-hexyl-2-methylbutanoate (10.84%), n-octanol (10.13%), n-octyl butanoate (8.88%), n-octyl-2-methylbutanoate (8.01%), n-hexyl acetate (7.11%), n-octyl isobutanoate (5.5%) and n-hexyl isobutanoate (5.43%) were the main compounds. The high-performance counter-current chromatography was applied for purification of aliphatic alcohols and esters. A mixture of n-hexane, acetonitrile and tetr-butyl methyl ether (1:1:0.1, v/v) allowed to obtain n-octanol, n-octyl acetate, n-hexyl-2- methylbutanoate, n-octyl isobutanoate and n-octyl-2-methylbutanoate, with the purity range of 94-99%, in one single 74 min run. The antimicrobial activity was also determined against plant and foodborne pathogens. While n-octanol shares responsibility for the antibacterial activity of the essential oil, n-octyl acetate determines its antifungal action. The cytotoxic activity assessed on two normal kidney fibroblast cell lines: Vero (animal) and HEK-293 (human embryonic), and two human cancer cell lines: FaDu (squamous cell carcinoma of the pharynx) and SCC25 (squamous cell carcinoma of the tongue), showed a moderate cytotoxicity with CC50 values ranging from 262.3 to 567.8 µg/mL. Results indicate that normal cell lines were more sensitive to the tested essential oil than cancer cell lines. The antioxidant activity of oil and pure compounds was not significant.

Efficient Isolation of Dihydropyranocoumarins and Simple Coumarins from Mutellina purpurea Fruits.

For the first time, the separation of coumarin derivatives from the petroleum ether extract of the fruits of Mutellina purpurea through high-performance countercurrent chromatography (HPCCC) is described. Four compounds, pteryxin (1; 2.72 mg), hyuganin C (2; 7.94 mg), osthol (3; 4.30 mg), and hyuganin A (4; 3.09 mg), were obtained in a single run following the injection of crude extract (300 mg). Additionally, auraptenol (5) and hyuganin D (6) were identified using LC-ESI-(Q)TOF-MS. The structures of the isolated compounds were elucidated through spectroscopic (NMR and MS) methods. This is apparently the first report of the identification of dihydropyranocoumarins in this plant, and the first time that HPCCC was used to separate them.

Bioactivity-guided isolation of antimicrobial coumarins from Heracleum mantegazzianum Sommier & Levier (Apiaceae) fruits by high-performance counter-current chromatography.

An efficient strategy, based on bioassay-guided fractionation, high-performance liquid chromatography (HPLC), and high-performance counter-current chromatography (HPCCC), was established to purify and evaluate the bioactive compounds from the dichloromethane extract of the fruits of Heracleum mantegazzianum Sommier & Levier (Apiaceae). The quaternary solvent system n-heptane-ethyl acetate-methanol-water (6:5:6:5 v/v) was used in the reversed phase mode. Using this method, in a single run, seven fractions were isolated, among them three were the pure furanocoumarins: pimpinellin, imperatorin, and phellopterin. In order to purify xanthotoxin a more polar system (1:1:1:1 v/v) was further applied. The antimicrobial activity of extract, chromatographic fractions, and single compounds were in the range of MIC = 0.03-1 mg mL(-1). Xanthotoxin may have priority as a compound of further interest based on its antimicrobial activity. For the first time, an extensive antimicrobial study was performed for pimpinellin and phellopterin.

Isolation of terpenoids from Pimpinella anisum essential oil by high-performance counter-current chromatography.

High-performance counter-current chromatography was successfully used for the isolation and purification of terpenoid compounds from the essential oil of Pimpinella anisum L. A two-phase solvent system composed of n-heptane/methanol/ethyl acetate/water (5:2:5:2, v/v/v/v) was suitable for the purification of linalool, terpinen-4-ol, α-terpineol, p-anisaldehyde, while n-heptane/methanol (1:1, v/v) was used for the isolation of anethole and foeniculin. A scale-up process from analytical to preparative was developed. Additionally, a stepwise gradient elution was applied and instead of two different runs, 40 min each, one 80 min separation was performed; although the time of separation remains the same, it was possible to repeat the efficiency even if the water-containing mobile phase was changed to a nonaqueous system. The obtained essential oil, as well as purified compounds, was analyzed by GC. A total of 0.64 mg of linalool, 0.52 mg of terpinen-4-ol, 0.10 mg of α-terpineol, 0.62 mg of p-anisaldehyde, 15 mg of anethole, and 2.12 mg of foeniculin were obtained from 210 mg of the essential oil of P. anisum L. in a short time with purities of 99, 98, 94, 93.54, 93, and 93.6%, respectively.