A comparison and column selection of Hydrophilic Interaction Liquid Chromatography and Reversed-Phase High-Performance Liquid Chromatography for detection of DNA methylation.

5-methylcytosine (5mC) is an epigenetic mark which has a profound effect on various fundamental processes in cells. In present study, at first Hydrophilic Interaction Liquid Chromatography (HILIC) was compared with Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) based on their selectivity (α), retention factor (k), and resolution (R) for cytosine (C) and 5mC nucleobases. We tried to justify the separation mechanism on the basis of mobile phase and solute polarity, structural characterization of solute and stationary phases, log Do/w, and pka under both modes. Then, these two modes were compared in order to select the best column for measurement of methylation level in two real samples with less analytical complexity (i.e. animal and bacteria) and a highly complex sample (i.e. plant), after chemical hydrolysis of DNA. In this favor, diol and cyano (CN) columns in HILIC mode as well as C8 and C18 in RP-HPLC were investigated. Optimum separation and the best validation parameters were obtained for CN column with Limit of Detection (LOD) of 1.4 pmol and Limit of Quantification (LOQ) of 4.8 pmol for 5mC. When the CN column was used in HILIC-UV procedure, separation of 5mC and C bases was achieved in all types of hydrolyzed DNA solutions.

Biochemical and biophysical properties of a novel homoisoflavonoid extracted from Scilla persica HAUSSKN.

In this study isolation and structural elucidation of a homoisoflavonoid, 3-(3',4'-dihydroxybenzyl)-8-hydroxy-5,7-dimethoxychroman-4-one (Scillapersicone 1), is reported from Scilla persica HAUSSKN. The structure was solved by a single crystal X-ray analysis. The unit cell parameters are a=11.7676 (2)Å, b=20.1174 (4)Å, c=7.8645 (9)Å, ß=93.544 (2)°, V=1858.23 (7)Å(3), monoclinic space group P21/c and four symmetry equivalent molecules in an unit cell. The structure was consistent with the UV, IR, 1D and 2D NMR, HRFAB-MS data. The optimized molecular geometry agrees closely that obtained from the single crystal X-ray crystallography. Furthermore, cytotoxicity of this compound was evaluated by MTT assay on AGS and WEHI-164 cancerous cell lines.

Chemical Constituents of Cymbocarpum erythraeum (DC.) Boiss., and Evaluation of Its Anti-Helicobacter pylori Activity.

OBJECTIVE: Cymbocarpum erythraeum (Apiaceae) is an endemic species in Iran. Up to now, there have been no phytochemical and biological investigations on this species. Therefore, isolation of the main secondary metabolites of the plant along with its anti-H. pylori activity have been considered in this paper. MATERIALS AND METHODS: The dried parts of the plant were extracted with different solvents using solvent percolation and the antibacterial activity of the extracts evaluated by the disk diffusion method. Four compounds were isolated using different column chromatography methods. RESULTS: The compounds were identified by proton nuclear magnetic resonance and carbon-13 nuclear magnetic resonance as isoquercetin (1), rutin (2), ß-sitosterol (3) and 2-decenol (4). CONCLUSION: Anti-H. pylori evaluation of the extracts and isolated compounds against three clinical isolates of H. pylori revealed that hexane extract of the plant inhibited all H. pylori strains.

Isolation and characterization of homoisoflavonoids from Scilla persica HAUSSKN

abstract Medicinal plants have many traditional claims including the treatment of ailments of infectious origin. In the evaluation of traditional claims, scientific research is extremely important. In this study, five homoisoflavonoids named 3-(4'-hydroxybenzylidene)-5,7-dihydroxy-6-methoxychroman-4-one(Autumnalin),3-(4'-hydroxybenzyl)-5,7-dihydroxy-6-methoxychroman-4-one (3,9-dihydro-autumnalin), 3-(3',4'-dihydroxybenzyl)-5,8-dihydroxy-7-methoxychroman-4-one,3-(3',4'-dihydroxybenzylidene)-5,8-dihydroxy-7-methoxychroman-4-one and 3-(3',4'-dihydroxybenzylidene)-5,7-dihydroxy-6-methoxychroman-4-one, were isolated from the bulbs of the plant Scilla persica HAUSSKN. Their structures were established on the basis of extensive spectroscopic analyses such as NMR, MS, IR and UV.

resumo Plantas medicinais apresentam muitas atribuições tradicionais, incluindo o tratamento de doenças de origem infecciosa. A pesquisa científica é extremamente importante na avaliação dos usos tradicionais. Neste estudo, cinco homoisoflavonóides: 3-(4'-hidroxibenzilideno)-5,7-diidroxi-6-metoxicroman-4-ona(autumnalina), 3-(4'-hidroxibenzil)-5,7-diidroxi-6- metoxicroman-4-ona (3,9-diidro-autumnalina), 3-(3',4'-diidroxibenzil)-5,8-diidroxi-7- metoxicroman-4-ona, 3-(3',4'-diidroxibenzilideno)-5,8-diidroxi-7- metoxicroman-4-ona e 3-(3',4'-diidroxibenzilideno)-5,7-diidroxi-6- metoxicroman-4-ona foram isolados dos bulbos da planta Scilla persica HAUSSKN. Suas estruturas foram estabelecidas com base na extensa análise espectroscópica, como RMN, EM, IV e UV.