Assay for codeine, morphine and ten potential urinary metabolites by gas chromatography--mass fragmentography

A mass fragmentography (MF) assay is described for ten potential, minor urinary metabolites of codeine (C) and morphine (M). Samples were hydrolyzed, extracted, derivatized with Tri-Sil Z and analyzed by methane chemical ionization (CI)-MF. The method is sensitive to ca. 0.01 microgram/ml for all compounds with the exception of normorphine (NM) which was difficult to extract with chloroform. The sensitivity of the MF assay for NM was only ca. 0.10 microgram/ml. Various solvent systems were investigated for optimization of extraction efficiency of all metabolites. A separate method for the extraction of NM is reported which utilizes a solid buffer--solvent combination, i.e., potassium carbonate--isopropanol. This latter method provided the best overall recovery of NM (39.0 +/- 3.4%). Gas chromatographic (GC) retention times of C, M and metabolites are reported for three liquid phases (3%) on Gas-Chrom Q (100-120 mesh). Resolution of metabolites (as trisilyl derivatives) was best on Silar-5CP and this phase was used in metabolic studies of C and M. GC resolution was not complete for all compounds; however, selection of specific ions for monitoring by MF provided the required specificity for all compounds except the 6 alpha- and 6 beta-hydroxy isomers. CI spectra for all metabolites are reported. The MF assay was used for urinary analysis of samples from guinea pigs that received single doses of C (15 mg/kg) or M (8 mg/kg). Following C administration 6 alpha- and 6 beta-hydrocodol, 6 alpha, beta-hydromorphol (undifferentiated), HM and M were measured. Following M administration only 6 alpha, beta-hydromorphol was found. The amount of total metabolite as percent dose for each component was calculated as less than 1%.     

Poly(ethylene terephthalate) copolymers containing nitroterephthalic units. III. Methanolytic degradation

The methanolytic degradation of poly(ethylene terephthalate) (PET) copolymers containing nitroterephthalic units was investigated. Random poly(ethylene terephthalate‐co‐nitroterephthalate) copolyesters (PETNT) containing 15 and 30 mol % nitrated units were prepared from ethylene glycol and a mixture of dimethyl terephthalate and dimethyl nitroterephthalate. A detailed study of the influence of the nitro group on the methanolytic degradation rate of the nitrated bis(2‐hydroxyethyl) nitroterephthalate (BHENT) model compound in comparison with the nonnitrated bis(2‐hydroxyethyl) terephthalate (BHET) model compound was carried out. The kinetics of the methanolysis of BHENT and BHET were evaluated with high‐performance liquid chromatography and ¹H NMR spectroscopy. BHENT appeared to be much more reactive than BHET. The methanolytic degradation of PET and PETNT copolyesters at 80 °C was followed by changes in the weight and viscosity, gel permeation chromatography, differential scanning calorimetry, scanning electron microscopy, and ¹H and ¹³C NMR spectroscopy. The copolyesters degraded faster than PET, and the degradation increased with the content of nitrated units and occurred preferentially by cleavage of the ester groups placed at the meta position of the nitro group in the nitrated units. For both PET and PETNT copolyesters, an increase in crystallinity accompanied methanolysis. A surface degradation mechanism entailing solubilization of the fragmented polymer and consequent loss of mass was found to operate in the methanolysis of the copolyesters. © 2002 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 40: 2276–2285, 2002     

Poly(ethylene terephthalate) copolymers containing 5‐nitroisophthalic units. II. Crystallization studies

The microstructure and crystallization behavior of a set of poly(ethylene terephthalate‐co‐5‐nitroisophthalate) copolymers (PETNI) containing 5‐nitroisophthalic units in the 10–50 mol % range were examined and compared to those of poly(ethylene terephthalate) (PET) and poly(ethylene terephthalate‐co‐isophthalate) (PETI) copolymers. A ¹³C NMR analysis of PETNI copolymers in a trifluoroacetic acid solution indicates that they are random copolymers with average sequence lengths in accordance with ideal polycondensation statistics. Differential scanning calorimetry (DSC) studies show that PETNI containing 5‐nitroisophthalic units up to 20 mol % are able to crystallize and that crystallization takes place in these copolymers at much slower rates than in PET. Wide‐angle X‐ray diffraction from powder and fibers reveals that crystallizable PETNI adopts the same triclinic crystal structure as PET, with the nitroisophthalate units being excluded from crystallites. Fourier transform infrared in combination with cross‐polarization/magic‐angle spinning ¹³C NMR spectroscopy demonstrates the occurrence of a gauche–trans conversion encompassing the crystallization process. A correlation between DSC and spectroscopic data leads us to conclude that the content of trans conformer in the noncrystallized phase of PETNI is higher than in both PET and PETI copolymers and suggests that secondary crystallization in the homopolymer must proceed by a mechanism different than that in copolymers. © 2001 John Wiley & Sons, Inc. J Polym Sci Part B: Polym Phys 39: 1553–1564, 2001     

Across-ear interference from parametrically degraded synthetic speech signals in a dichotic cocktail-party listening task

Recent results have shown that listeners attending to the quieter of two speech signals in one ear (the target ear) are highly susceptible to interference from normal or time-reversed speech signals presented in the unattended ear. However, speech-shaped noise signals have little impact on the segregation of speech in the opposite ear. This suggests that there is a fundamental difference between the across-ear interference effects of speech and nonspeech signals. In this experiment, the intelligibility and contralateral-ear masking characteristics of three synthetic speech signals with parametrically adjustable speech-like properties were examined: (1) a modulated noise-band (MNB) speech signal composed of fixed-frequency bands of envelope-modulated noise; (2) a modulated sine-band (MSB) speech signal composed of fixed-frequency amplitude-modulated sinewaves; and (3) a "sinewave speech" signal composed of sine waves tracking the first four formants of speech. In all three cases, a systematic decrease in performance in the two-talker target-ear listening task was found as the number of bands in the contralateral speech-like masker increased. These results suggest that speech-like fluctuations in the spectral envelope of a signal play an important role in determining the amount of across-ear interference that a signal will produce in a dichotic cocktail-party listening task.     

Limits to the role of a common fundamental frequency in the fusion of two sounds with different spatial cues

Two experiments establish constraints on the ability of a common fundamental frequency (F0) to perceptually fuse low-pass filtered and complementary high-pass filtered speech presented to different ears. In experiment 1 the filter cut-off is set at 1 kHz. When the filters are sharp, giving little overlap in frequency between the two sounds, listeners report hearing two sounds even when the sounds at the two ears are on the same F0. Shallower filters give more fusion. In experiment 2, the filters' cut-off frequency is varied together with their slope. Fusion becomes more frequent when the signals at the two ears share low-frequency components. This constraint mirrors the natural filtering by head-shadow of sound sources presented to one side. The mechanisms underlying perceptual fusion may thus be similar to those underlying auditory localization.     

Effects of reverberation on spatial, prosodic, and vocal-tract size cues to selective attention

Three experiments explored the resistance to simulated reverberation of various cues for selective attention. Listeners decided which of two simultaneous target words belonged to an attended rather than to a simultaneous unattended sentence. Attended and unattended sentences were spatially separated using interaural time differences (ITDs) of 0, +/-45, +/-91 or +/-181 micros. Experiment 1 used sentences resynthesized on a monotone, with sentence pairs having F0 differences of 0, 1, 2, or 4 semitones. Listeners' weak preference for the target word with the same monotonous F0 as the attended sentence was eliminated by reverberation. Experiment 1 also showed that listeners' ability to use ITD differences was seriously impaired by reverberation although some ability remained for the longest ITD tested. In experiment 2 the sentences were spoken with natural prosody, with sentence stress in different places in the attended and unattended sentences. The overall F0 of each sentence was shifted by a constant amount on a log scale to bring the F0 trajectories of the target words either closer together or further apart. These prosodic manipulations were generally more resistant to reverberation than were the ITD differences. In experiment 3, adding a large difference in vocal-tract size (+/- 15%) to the prosodic cues produced a high level of performance which was very resistant to reverberation. The experiments show that the natural prosody and vocal-tract size differences between talkers that were used retain their efficacy in helping selective attention under conditions of reverberation better than do interaural time differences.     

On-chip photoactivation of heterologously expressed rhodopsin allows kinetic analysis of G-protein signaling by surface plasmon resonance spectroscopy

Surface plasmon resonance spectroscopy allows the study of protein interaction dynamics in real-time. Application of this technique to G-protein coupled receptors, the largest family of receptors involved in signal transduction, has been complicated by their low level of expression and the critical dependence of their native conformation on the hydrophobic transmembrane lipid environment. Here, we investigate and compare three different strategies to immobilize rhodopsin, a prototypical G-protein coupled receptor on a sensor chip surface using antibodies and a lectin for receptor capturing. By further probing of different experimental conditions (pH, detergent type) we identified the optimal factors to maintain rhodopsin in a functional conformation and extended this approach to recombinant rhodopsin that was heterologously expressed in COS cells. Functional operation of rhodopsin on the sensor chip surface was proven by its activation and subsequent light-stimulated G-protein coupling. The influence of these experimental parameters on the association and dissociation kinetics of G-protein receptor coupling was determined. Thereby, we found that the kinetics of G_t interaction were not changed by the strategy of immobilization or the type of detergent. Regeneration of opsin directly on a chip allowed recycling of the immobilized native and recombinant receptor. Thus, the approach provides an experimental framework for choosing the most suitable conditions for the solubilization, immobilization, and for functional tests of rhodopsin on a biosensor surface.     

The synthesis and X-ray structure of five co-ordinate imidodiphosphinate complexes of indium(III)

We report the synthesis and structural characterisation of three five co-ordinate indium(III) bis-chelate complexes of imidodiphosphinates with general formula InCl{N(PR₂X)₂}₂, (R=Ph or Prⁱ, X=S or Se), formed from the 1:2 reaction of indium(III) trichloride and the potassium or sodium salt of the ligand in methanol/THF. The complexes consist of discrete monomeric, molecules and show a trigonal–bipyramidal co-ordination pattern. These are the first examples of five co-ordinate complexes containing this type of ligand, in contrast to other indium(III) and main group element complexes of similar ligands in which only the six co-ordinate tris-chelates are known.