Purification of Food Color Red No. 106 (acid red) using pH-zone-refining counter-current chromatography

pH-Zone-refining counter-current chromatography was successfully applied to the separation of the main components of Food Color Red No. 106 (R-106, acid red, Color Index No. 45100). A 300-mg quantity of sample was separated using the following two-phase solvent system: n-butanol-water, 40 mM sulfuric acid in organic stationary phase and 30 mM ammonia in aqueous mobile phase. The obtained fractions were analyzed by high-performance liquid chromatography and fast atom bombardment mass spectrometry. The separation yielded 261.9 mg of main component of acid red with purity of 99.9%.     

Direct monitoring of DNA cleavages catalyzed by an ATP-dependent deoxyribonuclease on a 27 MHz quartz-crystal microbalance

Kinetic studies of enzymatic DNA cleavage reactions (the enzyme binding, hydrolysis along DNA strands, and then release of the enzyme from the completely hydrolyzed ssDNA) were carried out on a 27 MHz quartz-crystal microbalance.     

A highly cis-selective synthesis of 2-ethynylaziridines by intramolecular amination of chiral bromoallenes: improvement of stereoselectivity based on the computational investigation

The base-mediated intramolecular amination of bromoallenes having an axial chirality is described. The treatment of (4S,aR)-4-alkyl-4-[N-(arylsulfonyl)amino]-1-bromobuta-1,2-dienes with NaH in DMF affords 2,3-cis-2-ethynylaziridines in good to excellent selectivity (2,3-cis:trans = 92:8-99:1). The reaction of (4S,aS)-bromoallenes with NaH/DMF also gives 2,3-cis-2-ethynylaziridines selectively (79:21-91:9). These experimental results have been rationalized by B3LYP density functional calculations together with the 6-31+G(d) basis set and the Onsager solvation model. The transition structures for cis-aziridine formation of both (4S,aR)- and (4S,aS)-bromoallenes in DMF are favored over the corresponding trans transition structures by 4.35 and 1.41 kcal/mol, respectively. Furthermore, the calculations predicted that a less polar solvent gives higher cis selectivity for (4S,aS)-bromoallenes. In fact, improvement of the cis selectivity to 99:1 has been realized by using a less polar solvent such as THF. The cyclization of bromoallenes bearing a beta- or gamma-amino group also affords four- and five-membered azacycles in a highly cis-selective manner.     

Cidep study of radical-ion pair systems: Photooxidation reactions of carbazoles by maleic anhydride in alcohol solution

Photooxidation (charge transfer) reactions of carbazole derivatives by maleic anhydride (MA) in alcoholic media are studied by a time-resolved cw-ESR (TRESR) and Fourier transform ESR (FTESR) techniques. The CIDEP spectra observed in the systems (methyl-, ethyl-, and phenyl-substituted carbazoles with MA) remarkably depend on the MA concentration. Under the high concentration conditions, an emissive TM (triplet mechanism) polarization with a slight A/E (absorption/emission) pattern of the RPM (radical pair mechanism) is observed. In the low concentration of MA, a TM-like absorptive polarization is superimposed on the A/E pattern of the RPM. Since this reaction takes place exclusively through the triplet state, this spectral phase reveals that the singlet state of the radical-ion pair (RIP) has an energy higher than that of the triplet state, in other words, the sign of the apparent exchange interaction of the present RIP systems is positive. The total absorptive polarization observed in the low concentration of MA is tentatively explained by the polarization transfer from the excited triplet state of carbazoles in thermal equilibrium.     

Adsorption factor and photocatalytic degradation of dye-constituent aromatics on the surface of TiO2 in the presence of phosphate anions

The photocatalytic degradation for some kinds of dye-constituent aromatics with TiO₂ in the presence of phosphate anions in aqueous dispersion was investigated under both visible light (λ>480 nm) and UV irradiation. The influences of phosphate anion upon the degradation of organics under these different conditions was revealed by the measurement of point of zero ξ-potential (P _ZC) of TiO₂, UV-VIS spectra, HPLC and LC-MS. The adsorption and photodegradation of some organics, which adsorb on the surface of TiO₂ by a dominating group bearing a positive charge, was enhanced, while that of others, which adsorb on the surface of TiO₂ by a dominating group bearing negative charge, was depressed by the presence of phosphate anions under UV irradiation at the experimental conditions (pH 4.3). It was confirmed that better adsorption of organics on the surface of TiO₂ had an advantage in their photocatalytic degradation under UV irradiation. On the other hand, although the adsorption of rhodamine B (RhB) and methylene Blue (MB) markedly increased, their degradation under visible light irradiation was depressed in the presence of phosphate anions. It is suggested that phosphate anion greatly blocked the electron transfer from excited RhB and MB molecules as RhB and MB molecules predominantly adsorbed on the surface of TiO₂ through the electrostatic interaction with surface adsorbed phosphate anions.     

Characterization of atenolol transformation products on light-activated TiO2 surface by high-performance liquid chromatography/high-resolution mass spectrometry

We have studied the photocatalytic transformation of atenolol, 4-[2-hydroxy-3-[(1-methyl)amino]propoxyl]benzeneacetamide, a cardioselective beta-blocking agent used to treat cardiac arrhythmias and hypertension, under simulated solar irradiation using titanium dioxide as photocatalyst. The investigation involved monitoring drug decomposition, identifying intermediate compounds, assessing mineralization, and evaluating toxicity. High-performance liquid chromatography (HPLC) coupled to high-resolution mass spectrometry (HRMS) via an electrospray ionization (ESI) interface was a powerful tool for the identification and measurement of the degradation products; 23 main species were identified. Intermediates were characterized through their chromatographic behavior and evolution kinetics, coupled with accurate mass information. Through the full analysis of MS and MS(n) spectra and a comparison with parent drug fragmentation pathways, the diverse isomers were characterized. Neither atenolol nor the intermediates formed exhibit acute toxicity. All intermediates are easily degraded and no compound identified could withstand 2 h irradiation. Photomineralization of the substrate in terms of carbon mineralization and nitrogen release was rapid and, within 4 h of irradiation, organic nitrogen and carbon were completely mineralized.     

Development of pyrrole-imidazole polyamide for specific regulation of human aurora kinase-A and -B gene expression

Pyrrole-imidazole polyamide (PIP) is a nuclease-resistant novel compound that inhibits gene expression through binding to the minor groove of DNA. Human aurora kinase-A (AURKA) and -B (AURKB) are important regulators in mitosis during the cell cycle. In this study, two specific PIPs (PIP-A and PIP-B) targeting AURKA and AURKB promoter regions were designed and synthesized, and their biological effects were investigated by several in vitro assays. PIP-A and PIP-B significantly inhibited the promoter activities, mRNA expression, and protein levels of AURKA and AURKB, respectively, in a concentration-dependent manner. Moreover, 1:1 combination treatment with both PIPs demonstrated prominent antiproliferative synergy (CI value [ED(50)] = 0.256) to HeLa cells as a result of inducing apoptosis-mediated severe catastrophe of cell-cycle progression. The novel synthesized PIP-A and PIP-B are potent and specific gene-silencing agents for AURKA and AURKB.