ABSTRACT
A new lignan from the non-transformed root in vitro cultures of Phyllanthus amarus was isolated. The structure of the compound was established on the basis of one- and two-dimensional NMR, as well as mass spectrometry data, as 7'-oxocubebin dimethylether (1,4-bis(benzo[d][1,3]dioxol-5-yl)-2,3-bis(methoxymethyl)butan-1-on). The non-transformed root cultures of P. amarus showed to be a selective source of this compound. The lignan revealed strong cytotoxic activity against HeLa cell line with an IC50 value of 3.8 µg/mL.
Subject(s)
Lignans/chemistry , Lignans/pharmacology , Phyllanthus/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Roots/chemistry , Cell Line, Tumor , HeLa Cells , Humans , Magnetic Resonance Spectroscopy/methods , Phytotherapy/methodsABSTRACT
Four O-acylated flavonol glycosides, new in the plant kingdom, were isolated from the needles of Pseudotsuga menziesii. Their structures were established by 1D and 2D NMR and MS data as: daglesioside I [kaempferol 3-O-[2",5"-O-(4''',4(IV)-dihydroxy)-beta-truxinoyl]-alpha-L-arabinofuranoside] (1), daglesioside II [kaempferol 3-O-[2",5"-O-(4"'-hydroxy)-beta-truxinoyl]-alpha-L-arabinofuranoside] (2), daglesioside III [kaempferol 3-O-[2",5"-di-O-(E)-p-coumaroyl]-alpha-L-arabinofuranoside] (3), and daglesioside IV [kaempferol 3-O-[3",6"-di-O-(E)-cinnamoyl]-beta-D-glucopyranoside] (4). In addition, the known flavonoids (E)-tiliroside, (E)-ditiliroside, astragalin (kaempferol 3-O-beta-D-glucopyranoside), isorhamnetin, kaempferol, and quercetin were identified. The cytotoxic activity of compounds 1 and 3 was evaluated towards the HL-60, HeLa, and MDA-MB468 cell lines.
Subject(s)
Flavonoids/isolation & purification , Pseudotsuga/chemistry , Cell Line, Tumor , Chromatography, Thin Layer , Flavonoids/chemistry , Flavonoids/pharmacology , Glycosides/chemistry , Humans , Magnetic Resonance Spectroscopy , Mass SpectrometryABSTRACT
An HPTLC densitometric method was established for the determination of beta-sitosterol in in vitro cultures of some species of the genus Phyllanthus. Two derivatization reagents commonly used in TLC for the visualization and detection of sterols, namely, anisaldehyde reagent and 5% phosphomolybdic acid, were compared with vanillin reagent. The densitometric quantification of beta-sitosterol was carried out on HPTLC Si60 F254 plates with the mobile phase chloroform-hexane-methanol (65 + 30 + 5, vlv/v) at 525 nm. The method was validated for each derivatization reagent in terms of linearity, precision, repeatability, intra- and interday precision, LOD, and LOQ. The presence of beta-sitosterol was revealed in all analyzed plant material. The concentrations of beta-sitosterol determined ranged from 0.48 to 2.75 mg/g (dry weight). In addition, traces of beta-amyrin were detected in some plant samples.
Subject(s)
Chromatography, High Pressure Liquid/methods , Chromatography, Thin Layer/methods , Densitometry/methods , Phyllanthus/metabolism , Sitosterols/chemistry , Benzaldehydes/analysis , Chemistry Techniques, Analytical , Chemistry, Pharmaceutical/methods , Chloroform/chemistry , Hexanes/chemistry , Methanol/chemistry , Models, Chemical , Oleanolic Acid/analogs & derivatives , Oleanolic Acid/chemistry , Plants/metabolism , Reproducibility of ResultsABSTRACT
A novel analytical approach involving the addition of an ionic liquid into the mobile phase of the thin-layer chromatography (TLC) system during the optimization of chromatographic separation of peptides was demonstrated. Different behavior of peptides in the TLC sytem was observed after the addition of 1,3-dimethylimidazolium methyl sulfate to the eluent in comparison to the system without the ionic liquid. The objective of the work was to study the effect of the addition of different contents of ionic liquid to the mobile phase comprising mostly water and to observe the behavior of peptides' retention. The potential usefulness of environmentally friendly ionic liquids for the optimization of separation of peptides was demonstrated. An increase of R(f) values was observed with increasing the ionic liquid content in the mobile phase. The benefits of the used approach were related to the separation achieved. Finally, quantitative structure-retention relationships (QSRR) were used for the studies on the predictions of peptides' retention in the TLC systems with the addition of ionic liquid in terms of the predictions performed recently in HPLC systems.
Subject(s)
Peptides/isolation & purification , Solvents , Amino Acid Sequence , Angiotensin II/chemistry , Angiotensin II/isolation & purification , Chromatography, Thin Layer/methods , Molecular Sequence DataABSTRACT
The addition of an ionic liquid into the mobile phase appeared to be useful in optimization of chromatographic separation of peptides. Different behavior of peptides in thin-layer chromatography (TLC) was observed after addition of 1-ethyl-3-methylimidazolium tetra fluoroborate to the eluent in comparison to the system without the ionic liquid. Nonlinear dependence of the retention coefficient, R(M), of peptides on the volume percentage of acetonitrile in the eluent was found in normal-phase TLC with and without immidazolium tetra fluoroborate in the mobile phase. In general, R(M) increased with increasing concentration of acetonitrile. In TLC systems without the ionic liquid, R(M) can be described well with a quadratic function. On the other hand, in a TLC system with an ionic liquid as the additive to the mobile phase, the retention behavior is better described with a third-degree polynomial function. The potential usefulness of ionic liquids for optimization of separation of peptides was demonstrated. Optimization of the separation conditions was supported by a commercially available computer program.
