Analysis of nucleotides by pressure-assisted capillary electrophoresis-mass spectrometry using silanol mask technique

A method for the determination of nucleotides based on pressure-assisted capillary electrophoresis-electrospray ionization mass spectrometry (PACE-MS) is described. To prevent multi-phosphorylated species from adsorbing onto the fused-silica capillary, silanol groups were masked with phosphate ions by preconditioning the capillary with the background electrolyte containing phosphate. During preconditioning, nebulizer gas was turned off to avoid contamination of MS detector with phosphate ions. To detect nucleotides using the CE positive mode at a pH 7.5, it was necessary to apply air pressure to the inlet capillary during electrophoresis to supplement the electroosmotic flow (EOF) toward the cathode. Moreover, we exchanged the running electrolyte every analysis using the buffer replenishment system to obtain the required reproducibility. Under the optimized conditions, 14 phosphorylated species such as nucleotides, nicotinamide-adenine dinucleotides and coenzyme A (CoA) compounds were well determined in less than 20 min. The relative standard deviations (n=6) of the method were better than 0.9% for migration times and between 1.7% and 8.1% for peak areas. The detection limits for these species were between 0.5 and 1.7 micromol/L with pressure injection of 50 mbar for 30 s (30 nL) at a signal-to-noise ratio of 3. This approach is robust and quantitative compared to the previous method, and its utility is demonstrated by the analysis of intracellular nucleotides and CoA compounds extracted from Escherichia coli wild type, pfkA and pfkB knockout mutants. The methodology was used to suggest that pfkA is the main functional enzyme.     

Metabolome analysis by capillary electrophoresis-mass spectrometry

Capillary electrophoresis (CE)-mass spectrometry (MS), as an analytical platform, has made significant contributions in advancing metabolomics research, if still limited up to this time. This review, covering reports published between 1998 and 2006, describes how CE-MS has been used thus far in this field, with the majority of the works dealing with targeted metabolite analyses and only a small fraction using it in the comprehensive context. It also discusses how some of the key features of CE-MS were exploited in selected metabolomic applications.     

Phase structure and critical behavior of multi-Higgs U(1) lattice gauge theory in three dimensions

We study the three-dimensional (3D) compact U(1) lattice gauge theory coupled with N-flavor Higgs fields by means of the Monte Carlo simulations. This model is relevant to multi-component superconductors, antiferromagnetic spin systems in easy plane, inflational cosmology, etc. It is known that there is no phase transition in the N = 1 model. For N = 2, we found that the system has a second-order phase transition line in the c₂ (gauge coupling)-c₁ (Higgs coupling) plane, which separates the confinement phase and the Higgs phase. Numerical results suggest that the phase transition belongs to the universality class of the 3D XY model as the previous works by Babaev et al. and Smiseth et al. suggested. For N = 3, we found that there exists a critical line similar to that in the N = 2 model, but the critical line is separated into two parts; one for c₂<c_2tc=2.4±0.1 with first-order transitions, and the other for c_2tc<c₂ with second-order transitions, indicating the existence of a tricritical point. We verified that similar phase diagram appears for the N = 4 and N = 5 systems. We also studied the case of anistropic Higgs coupling in the N = 3 model and found that there appear two second-order phase transitions or a single second-order transition and a crossover depending on the values of the anisotropic Higgs couplings. This result indicates that an “enhancement” of phase transition occurs when multiple phase transitions coincide at a certain point in the parameter space.     

10-Oxo-10H-5λ,10λ-thianthren-5-ylideneamine as a probe for stereochemistry in the formation and amination of fluoro-λ-sulfanenitriles

The fluorination of 10-oxo-10H-5λ⁴,10λ⁴-thianthren-5-ylideneamine (2) with Selectfluor™ affords 5-fluoro-10-oxo-5,10-dihydro-5λ⁶,10λ⁴-thianthren-5-nitrile (4). The amination of 4 with morpholine gives 5-morpholino-10-oxo-5,10-dihydro-5λ⁶,10λ⁴-thianthren-5-nitrile (5). The stereochemical course of both reactions has been studied, while the configurations of their products, cis-isomer 4 and trans-5-morpholino-10-oxo-5,10-dihydro-5λ⁶,10λ⁴-thianthren-5-nitrile (trans-5) are elucidated by the use of X-ray crystallographic analyses.     

Hydrogen evolution from aliphatic alcohols and 1,4-selective hydrogenation of NAD+ catalyzed by a [C,N] and a [C,C] cyclometalated organoiridium complex at room temperature in water

A [C,N] cyclometalated Ir complex, [Ir(III)(Cp*)(4-(1H-pyrazol-1-yl-κN(2))benzoic acid-κC(3))(H(2)O)](2)SO(4) [1](2)·SO(4), was reduced by aliphatic alcohols to produce the corresponding hydride complex [Ir(III)(Cp*)(4-(1H-pyrazol-1-yl-κN(2))-benzoate-κC(3))H](-)4 at room temperature in a basic aqueous solution (pH 13.6). Formation of the hydride complex 4 was confirmed by (1)H and (13)C NMR, ESI MS, and UV-vis spectra. The [C,N] cyclometalated Ir-hydride complex 4 reacts with proton to generate a stoichiometric amount of hydrogen when the pH was decreased to pH 0.8 by the addition of diluted sulfuric acid. Photoirradiation (λ > 330 nm) of an aqueous solution of the [C,N] cyclometalated Ir-hydride complex 4 resulted in the quantitative conversion to a unique [C,C] cyclometalated Ir-hydride complex 5 with no byproduct. The complex 5 catalyzed hydrogen evolution from ethanol in a basic aqueous solution (pH 11.9) under ambient conditions. The 1,4-selective catalytic hydrogenation of β-nicotinamide adenine dinucleotide (NAD(+)) by ethanol was also made possible by the complex 1 to produce 1,4-dihydro-β-nicotinamide adenine dinucleotide (1,4-NADH) at room temperature. The overall catalytic mechanism of hydrogenation of NAD(+), accompanied by the oxidation of ethanol, was revealed on the basis of the kinetic analysis and detection of the reaction intermediates.     

Reaction of α,β‐unsaturated ketones with cerium(IV) salts in alcohol

Reaction of α,β‐unsaturated ketones with cerium(IV) salts or lanthanide triflates in alcohols gave good yields of the corresponding β‐alkoxy compounds. In the case of 2‐cyclopentenone and 2‐cyclohexenone, the 1,1,3‐trialkoxy acetal derivatives were obtained preferentially accompanied by β‐alkoxyketone, except 2‐cycloheptenone. However, in the reaction of 2‐cycloheptenone with alcohol using cerium(IV) sulfate (CS)‐molecular sieve, 1,1,3‐trialkoxy derivatives were obtained. Also, in the cases of 1‐penten‐3‐one, 4‐hexen‐3‐one and 3‐hepten‐2‐one, 1,1,3‐trialkoxy derivatives were obtained preferentially. Copyright © 2006 John Wiley & Sons, Ltd.