Enantioselective analysis of ketamine and its metabolites in equine plasma and urine by CE with multiple isomer sulfated beta-CD

CE with multiple isomer sulfated beta-CD as the chiral selector was assessed for the simultaneous analysis of the enantiomers of ketamine and metabolites in extracts of equine plasma and urine. Different lots of the commercial chiral selector provided significant changes in enantiomeric ketamine separability, a fact that can be related to the manufacturing variability. A mixture of two lots was found to provide high-resolution separations and interference-free detection of the enantiomers of ketamine, norketamine, dehydronorketamine, and an incompletely identified hydroxylated metabolite of norketamine in liquid/liquid extracts of the two body fluids. Ketamine, norketamine, and dehydronorketamine could be unambiguously identified via HPLC fractionation of urinary extracts and using LC-MS and LC-MS/MS with 1 mmu mass discrimination. The CE assay was used to characterize the stereoselectivity of the compounds' enantiomers in the samples of five ponies anesthetized with isoflurane in oxygen and treated with intravenous continuous infusion of racemic ketamine. The concentrations of the ketamine enantiomers in plasma are equal, whereas the urinary amount of R-ketamine is larger than that of S-ketamine. Plasma and urine contain higher S- than R-norketamine levels and the mean S-/R-enantiomer ratios of dehydronorketamine in plasma and urine are lower than unity and similar.     

Determination of the cation-chelating potential of C-methylthiolated beta-lactams and their sulfones by electrospray ionization mass spectrometry

The chelation potential of highly lipophilic C-dimethylthiolated monocyclic beta-lactams was examined using electrospray ionization mass spectrometry (ESI-MS). The metal salts NaCl, KCl, CaCl2, ZnCl2, Cu(NO3)2, CdSO4, MnCl2, and Mg(NO3)2 were used for the analysis. The K+ adducts of the compounds studied were more responsive in ESI analysis, compared to their Na+ adducts, regardless of the oxidation state of the sulfur (in the methylthio or the sulfone groups) and the type of the group adjacent to the lactam carbonyl. Opening of the beta-lactam ring, leading to formation of a chargeable N-atom, had little to no effect on the K+ adduct formation. Interactions of the methylthio group with the divalent zinc ion were also observed.     

Aggregates of naphthalene chromophores in poly(vinylalcohol)-graft-poly(vinylnaphthalene) pseudomicelles

A novel amphiphilic grafted copolymer, poly(vinylalcohol)-graft-poly(vinylnaphthalene), was synthesized and studied spectroscopically. The unusual photophysical properties of its aqueous solutions were observed for the first time and are attributed to the aggregation of the naphthalene chromophores in the bulk of polymer pseudomicelles. The enormous red shift of the fluorescence spectra is interpreted in terms of the confinement effect. The interpretation is supported by model calculations, taking into account the mixing of Frenkel excitons and charge-transfer states of chromophore clusters in densely packed polymer domains. Gradual funneling of the excitation energy between aggregates of different sizes is discussed in the context of the measured fluorescence depolarization spectra.     

Pressure-driven phase transitions in NaBH4: theory and experiments

We have investigated pressure-induced structural transitions in NaBH4 through density-functional theory calculations combined with X-ray and neutron diffraction experiments. Our calculations confirm that the cubic phase is stable up to 5.4 GPa and an orthorhombic phase occurs above 8.9 GPa, as observed in X-ray diffraction experiments. Both the calculations and X-ray diffraction measurements identify an intermediate tetragonal phase that appears between 6 and 8 GPa; that is, between the cubic and orthorhombic phases. This result is also confirmed by high-pressure neutron diffraction experiments performed on NaBD4. Our calculations and X-ray diffraction measurements show that the space group of the orthorhombic phase above 8.9 GPa is Pnma and the orthorhombic phase remains stable up to 30 GPa. The calculated equations of state are in excellent agreement with experiments.     

Electrostatic versus nonelectrostatic effects in DNA sequence discrimination by divalent ions Mg2+ and Mn2+

Mg2+ and Mn2+ ions are critical to the functioning of phosphoryl transfer enzymes, such as restriction endonucleases. Although these ions play similar roles in the chemical steps, they govern substrate specificity via modulating sequence discrimination by up to a factor of 10(5) with Mg2+ and only up to a factor of 10 with Mn2+. To explain whether such diversity originates in fundamental differences in the electronic structures of the nucleobase-hydrated-metal ion complexes, structures and interaction energies were determined at the density functional (DFT) and second-order M?ller-Plesset (MP2) levels of theory. Although both metal ions favor identical binding sites, Mn2+ complexes exhibit greater distortions from the ideal octahedral geometry and larger variability than the corresponding Mg2+ systems. In inner-shell complexes, with direct contact between the metal and the nucleobase, Mg2+ is preferred over Mn2+ in the gas phase, due primarily to nonelectrostatic effects. The interaction energies of the two metal ions are more similar in the outer-shell complexes, likely due to reduced charge transfer between the hydrated metal ion and the base moieties. Inclusion of solvation effects can amplify the relative nucleobase preferences of Mg2+ and Mn2+, indicating that bulk hydration modulates the balance between electrostatic and nonelectrostatic terms. In most cases, the base substitutions in solution are facilitated more by Mn2+ than by Mg2+. Electrostatic properties of the environment were demonstrated to have a major influence on the nucleobase preferences of the two metal ions. Overall, quantum chemical calculations suggest that the contrasting selectivity of Mg2+ and Mn2+ cofactors toward nucleobases derives from the larger flexibility of the Mn2+ complexes accompanied by the excessive polarization and charge-transfer effects as well as less favorable solvation.     

Interfacial tension of the lipid membrane formed from lipid-fatty acid and lipid-amine systems

Interfacial tension has been determined for phosphatidylcholine-stearic acid and phosphatidylcholine-stearylamine membranes. Phosphatidylcholine, stearic acid and stearylamine were used in the experimental. The interfacial tension values of the pure components are 1.62x10(-3) N/m, - 1.54x10(-2) N/m and 4.40x10(-3) N/m (hypothetical values), respectively. The 1:1 complexes were formed during formation of phosphatidylcholine-stearic acid and phosphatidylcholine-stearylamine membranes. The following parameters describing the complexes were determined: the surface concentrations of the lipid membranes formed from these complexes, A(3)(-1), the interfacial tensions of such membranes, gamma(3) and the stability constants of these complexes, K.     

Biosensor for total cholesterol estimation using N-(2-aminoethyl)-3-aminopropyltrimethoxysilane self-assembled monolayer

Cholesterol oxidase (ChOx), cholesterol esterase (ChEt), and horseradish peroxidase (HRP) have been co-immobilized covalently on a self-assembled monolayer (SAM) of N-(2-aminoethyl)-3-aminopropyltrimethoxysilane (AEAPTS) deposited on an indium–tin–oxide (ITO) glass surface. These enzyme-modified (ChOx-ChEt-HRP/AEAPTS/ITO) biosensing electrodes have been used to estimate cholesteryl oleate from 10 to 500 mg dL⁻¹. The sensitivity, K _m value, and shelf-life of these ChEt-ChOx-HRP/AEAPTS/ITO biosensing electrodes have been found to be 124 nA mg⁻¹ dL, 95.098 mg dL⁻¹ (1.46 mmol L⁻¹), and ten weeks, respectively. The ChEt-ChOx-HRP/AEAPTS/ITO bio-electrodes have been used to estimate total cholesterol in serum samples. Figure Covalent immobilization of enzymes onto AEAPTS/ITO surface using EDC/NHS chemistry Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.