Analytical method for determination of disaccharides derived from keratan sulfates in human serum and plasma by high-performance liquid chromatography/turbo-ionspray ionization tandem mass spectrometry

We established a highly sensitive LC/MS/MS method for the analysis of the disaccharides produced from keratan sulfates (KS). It was revealed that the disaccharides produced by keratanase II enzymatic digestion of KS could be determined with high sensitivity by negative ion mode of multiple reaction monitoring. Furthermore, monosulfated and disulfated disaccharides can be separated using a Hypercarb (2.0 mm i.d. x 150 mm, 5 microm) with a gradient elution of acetonitrile-0.01 m ammonium bicarbonate (pH 10). This method was applied to the determination of KS in serum and plasma of control subjects. The intra-day precision expressed as %CV was within 6.8% for five replicate analyses with three different control serum. The inter-day (overall, n = 15) precision was within 7.3% for three days. This method is sensitive, reproducible and would be useful for clinical analysis.     

Semiconductor laser fluorimetry for enzyme and enzymatic assays

Near-infrared semiconductor laser fluorimetry is applied to assays of xanthine and xanthine oxidase. The fluorescence of indocyanine green in the near-infrared region is quenched by hydrogen peroxide. Xanthine is converted to uric acid by xanthine oxidase, in a reaction which also produces hydrogen peroxide; xanthine can be determined by measuring the decrease in fluorescence intensity of the dye added to the sample solution. The calibration graph for xanthine is linear from 5 × 10^?5 M to 5 × 10^?7 M. The enzyme activity can also be determined.     

Additive-free nucleophilic addition of imidazolines and imidazoles to haloacetylenes

Nucleophilic addition of imidazolines to 1-halo-1-alkynes takes place by simple heating in DMF without any additives to give (Z)-N-(1-halo-1-alken-2-yl)imidazolines in good yield and in a highly regio- and stereoselective manner. These reaction conditions are also valid for the similar addition of imidazoles.     

Photoisomerization locking of azobenzene by formation of a self-assembled macrocycle

Reaction of an azobenzene-linked biscatecholate ligand with boron and titanium sources gave ring- and cage-shaped complexes in a self-assembly fashion, respectively. These complexes were inert to photoisomerization though the ligand itself was isomerized upon photoirradiation. The self-assembled macrocyclization caused inhibition of the photoisomerization.     

High-performance liquid chromatography of apolipoproteins in serum high-density lipoproteins

A simple and rapid method for apolipoprotein analysis in serum high-density lipoproteins (HDL) has been developed using high-performance liquid chromatography (HPLC) with sodium phosphate buffer (pH 7.0) containing 0.1% sodium dodecyl sulphate (SDS) as eluent. In contrast to the use of urea solution as an eluent, apolipoproteins can be analysed by applying an incubation mixture of HDL and the eluent buffer. A TSK-GEL column of G3000SW was found to be more profitable than G2000SW or G4000SW for analysis of HDL apolipoproteins. Elution patterns monitored by absorbance at 280 nm using a G3000SW column can give precise quantitative as well as qualitative information about apolipoproteins of molecular weight between 10(4) and 10(5). HPLC patterns of HDL apolipoproteins were compared between individual human subjects with various diseases. Elution profiles for lipid components in an incubation mixture were also examined.     

Room Temperature Solution ENDOR of some Super Stable Fluorocarbon Radicals

Room temperature fluorine electron nuclear double resonance (ENDOR) has been successfully observed for several superstable fluorocarbon radicals ·C(C₂F₄R)(i-C₃F₇)₂ in solution. Three radicals were employed in which CF₃, F, and O-c-C₆F₁₀SO₃C₂F₅ were introduced as R, and all the hyperfine couplings (hfcs) obtained by ENDOR were assigned with the help of ESR simulation and ab initio MO calculation. In case of ·C(i-C₃F₇)₃ large ¹³C and considerable β-fluorine couplings suggest the nonplaner arrangement for the central and three carbons at the β-position, in spite of the fact that all the methyl fluorine show the same hfc. Therefore, a rapid puckering motion along the C₃ axis together with the methyl rotation should average the hfc’s of the 18 fluorine nuclei to give the same value. When one of the CF₃ groups is substituted with an F nucleus, the five CF₃ groups give two hfc values, suggesting some dynamics still exists for the molecular frame. When a large group, O-c-C₆F₁₀SO₃C₂F₅, is substituted for CF₃, all the five CF₃ groups become nonequivalent and the ENDOR signal becomes intensive and sharp even at 290 K, indicating that the molecular frame becomes rigid. The relation between the ENDOR spectra of these systems and the intramolecular dynamics is discussed.     

Thermotropic liquid crystalline ionic stilbazoles and their miscible mixtures with non-ionic carbazolyl compounds

New thermotropic ionic liquid crystals were prepared from trans-4-nitro-4'-stilbazole and alkyl halides. trans-N-Alkyl-4-nitro-4'-stilbazolium bromides containing alkyl chains with 7 to 10 carbons and the chloride homologues containing alkyl chains with 5 to 10 carbons exhibited smectic phases. For example, trans-N-decyl-4-nitro-4'-stilbazolium bromide and trans-N-hexyl-4-nitro-4'-stilbazolium chloride showed smectic phases from 175 to 186°C and from 129 to 190°C, respectively. The temperature range of mesophases increased with decreasing size of the counteranions. The miscibility of trans-N-alkyl-4-nitro-4'-stilbazolium bromide in Schiff's base compounds having various electronegative groups was examined by differential scanning calorimetry and polarizing microscopy. Miscible binary mixtures were prepared from trans-N-alkyl-4-nitro-4'-stilbazolium bromides and 4-alkoxy-N-(9-methyl-2-carbozolyl-methylene)anilines. The 1:1 (mole ratio) binary mixture of trans-N-hexyl-4-nitro-4'-stilbazolium bromide with 4-hexyloxy-N-(9-methyl-2-carbazolylmethylene)aniline exhibited a stable smectic phase between 83 and 149°C, though a smectic phase is not exhibited by both individual components. The miscibility in the binary mixtures might be caused by a combination of ionic and electron donor-acceptor interactions.     

Photoluminescence and population analysis of single-walled carbon nanotubes produced by CVD and pulsed-laser vaporization methods

Band gap photoluminescence (PL) behaviors of single-walled carbon nanotubes (SWNTs) grown by the methods of chemical vapor deposition and pulsed-laser vaporization are investigated over the wide diameter range (≈0.8–1.4 nm). The peak intensity of the PL signals strongly depends on chirality and the ‘(2n + m) family type’ of SWNTs. Based on the PL results, a population analysis of these SWNTs is conducted by combining the calculated PL yields for each (n, m) tube. The results are directly compared with the histograms of diameter distributions estimated by the transmission electron microscope (TEM) observations to check the validity of the analysis.