The Effect of Alkane Chain Length on the Liquid–Liquid Critical Temperatures of Oligostyrene/Linear-Alkane Mixtures

Summary. Upper critical solution temperatures (UCSTs) for liquid–liquid demixing in a set of mixtures of linear alkanes (pentane (N ₁=5) to pentacontane (N ₁=50)) with an oligostyrene (1241amu, N ₂=12) are reported. We find strong correlation between the Hildebrand solubility parameters of the alkanes and the UCST. Correlations are developed which enable predictions concerning the miscibility of mixtures of compounds with longer chains.     

Vibrational analysis of chlorinated methylphosphonic acids: Part 2. Vibrational analysis of CHCl2PO3H2 and its anions

The vibrational spectra (IR and Raman) of CHCl₂PO₃H₂ and its anions in H₂O and D₂O solutions are reported. The IR spectra of the solid dibasic sodium and potassium salts, the solid normal and O-deuterated monobasic sodium and potassium salt and the solid normal and O-deuterated acid are discussed. The principal results of the normal coordinate analysis of the compounds are tabulated.     

Chemical studies of carbohydrates. Part I. Conversion of derivatives of glucose and allose into pyrazoles and pyridazines

On reaction of 1,2:5,6-di-O-isopropylidenc-3-O-(p-tolylsulfonyl)-α-D-glueofuranose ( 1 ) with hydrazine hydrate at 140° besides formation of 3-deoxy-3-hydrazino-1,2:5,6-di-O-isopropylidene-α-D-allofuranose ( 2 ) and 3-dcoxy-1,2:5,6-di-O-isopropylidene-α-D-erythro-hex-3-enofuranose ( 3 ), ring transformation into 3-[4′-(2′,2′-dimethyl-1′,3′-dioxolanyl)]pyridazine ( 4 ) takes place. At 170°, however, only 2 and 4 are formed, indicating that 3 is the precursor of 4. Treatment of 3 with hydrazine hydrate at 170° indeed gives a nearly quantitative ring expansion into 4. Treatment of 3-dcoxy-3-hydrazino-1,2:5,6-di-O-isopropylidenc-α-D-glucofuranose ( 8 ) as well as the stereoisomeric allofuranose 2 with concentrated hydrochloric acid gives a nearly quantitative ring interconversion into 3-(D-erythro-trihydroxypropyl)pyrazole ( 9 ).     

Studies on borate esters 1 : The ph dependence of the stability of esters of boric acid and borate in aqueous medium as studied by 11B NMR

The pH dependent stability of esters of boric acid and borate with glycol, glycolic acid, oxalic acid and glyceric acid as dihydroxy compounds has been studied. ¹¹B NMR provides a suitable analytical tool for the quantitative determination and structure elucidation of the various esters in aqueous medium. The pH dependent stability of esters of boric acid and borate is formulated in a general rule of thumb: esters of boric acid and borate with dihydroxy compounds in aqueous medium show the highest stability at that pH where the sum of the charges of the free esterifying species is equal to the charge of the ester.     

Application of the FCP-activation procedure to the synthesis of a biospecific adsorbent for trypsin

The synthesis of a biospecific adsorbent for trypsin was chosen as a model to investigate the applicability of FCP activation in affinity chromatography.p-Aminobenzamidine was chosen as a ligand, directly suitable for immobilization. The nonspecific binding properties of the first series of synthesized agarose derivatives were obviated either by FCP activation of the ligand instead of the matrix, or by modifying the initial FCP-activation procedure. The adsorbents prepared in this way, however, demonstrated no selectivity between trypsin and chymotrypsin. The introduction ofe-aminocaproic acid as a spacer was ineffectual. These problems were solved by the application of glycylglycine as a spacer. The final affinity matrices had a degree of substitution of approximately 4 μ.mol of ligand per gram gel (100 μmol ligand per gram dry adsorbent). The specific activity of a current trypsin preparation was increased by 58% in a single cycle. The biospecificity of these adsorbents was demonstrated.     

Alignment of molecules in pulsed resonant laser fields

We investigate by numerical simulations the dynamics of alignment of linear molecules in resonant pulsed laser fields and its dependence on pulse length, field strength, and molecular parameters. We propose an analytical short-time approximation for the time-dependent wave packets. We provide a theoretical basis for the occurrence of saturation in the rotational pumping. We present a formula to predict the time at which the maximum alignment occurs. We discuss the magnitude of the laser-induced alignment and we relate it to a theoretical upper limit.     

Development of a quality control method for the characterization of oligonucleotides by capillary zone electrophoresis-electrospray ionization-quadrupole time of flight-mass spectrometry

A capillary zone electrophoresis-negative electrospray ionization-quadrupole time of flight-mass spectrometric method was developed for the characterization of oligonucleotides after synthesis, using model compounds. The major difficulty is the adduction of metal cations to the polyanionic backbone of the oligonucleotide sample, resulting in complex spectra and decreased sensitivity. Several approaches were investigated to circumvent this problem. Separation was performed in an ammonium carbonate buffer. During separation, the interfering metal ions were exchanged for ammonium ions, which are less tightly bound to the oligonucleotide when ionized. The influence of the addition of piperidine and imidazole or trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA) to the running buffer for further reduction of cation adduction was investigated. Addition of CDTA to the buffer system resulted in a deconvoluted spectrum with very little adducts. On-line sample stacking proved vital to preconcentrate the samples. The pH and the concentration of the ammonium carbonate buffer as well as the electrophoresis voltage were optimized to achieve the best signal response for the oligonucleotides and a maximum reduction of the cation adducts as well as a short analysis time. Finally, the sheath liquid composition was examined for further improvement of the signal. The developed method was used to analyze different oligonucleotides (5000-9200 Da) in light of its use as a final quality control method for oligonucleotides in terms of purity and sequence homogeneity of the synthesized products. In all cases, very little adducts were observed in the deconvoluted spectra, and the relative errors of the measured molecular masses ranged from 3 to 35 ppm.     

Time-resolved fluorescence analysis of the mobile flavin cofactor in p -hydroxybenzoate hydroxylase

Conformational heterogeneity of the FAD cofactor in p-hydroxybenzoate hydroxylase (PHBH) was investigated with time-resolved polarized flavin fluorescence. For binary enzyme/substrate (analogue) complexes of wild-type PHBH and Tyr222 mutants, crystallographic studies have revealed two distinct flavin conformations; the ‘in’ conformation with the isoalloxazine ring located in the active site, and the ‘out’ conformation with the isoalloxazine ring disposed towards the protein surface. Fluorescence-lifetime analysis of these complexes revealed similar lifetime distributions for the ‘in’ and ‘out’ conformations. The reason for this is twofold. First, the active site of PHBH contains various potential fluorescence-quenching sites close to the flavin. Fluorescence analysis of uncomplexed PHBH Y222V and Y222A showed that Tyr222 is responsible for picosecond fluorescence quenching free enzyme. In addition, other potential quenching sites, including a tryptophan and two tyrosines involved in substrate binding, are located nearby. Since the shortest distance between these quenching sites and the isoalloxazine ring differs only little on average, these aromatic residues are likely to contribute to fluorescence quenching. Second, the effect of flavin conformation on the fluorescence lifetime distribution is blurred by binding of the aromatic substrates: saturation with aromatic substrates induces highly efficient fluorescence quenching. The flavin conformation is therefore only reflected in the small relative contributions of the longer lifetimes.     

Analysis of unknown compounds in azithromycin bulk samples with liquid chromatography coupled to ion trap mass spectrometry

A selective reversed-phase liquid chromatography/mass spectrometry (LC/MS(n)) method is described for the identification of azithromycin impurities and related substances in commercial azithromycin samples. Mass spectral data are acquired on an LCQ ion trap mass spectrometer equipped with an atmospheric pressure chemical ionization interface operated in positive ion mode. The LCQ provides on-line LC/MS(n) capability, making it ideally suited for identification purposes. In comparison with UV detection, this hyphenated technique provides as its main advantage efficient identification of novel substances without time-consuming isolation and purification procedures. Using this technique, six novel related substances detected in commercial azithromycin samples have been studied.