Validation of a high performance liquid chromatography analysis for the determination of noradrenaline and adrenaline in human urine with an on-line sample purification

A high performance liquid chromatography (HPLC) method with fluorescence detection including an on-line purification was established for determination of catecholamines in human urine. The method was evaluated using samples of pooled urine spiked with catecholamines and validated for measurements of catecholamines in urine of healthy individuals in a field study. The laboratory method evaluation study showed that the recovery of the method was 0.82 (confidence interval (CI): 0.79-0.86) and 0.92 (CI: 0.89-0.95) for noradrenaline and adrenaline, respectively. Thus, correction factors of 0.82(-1) and 0.92(-1) were applied to correct the measurement results for this systematic effect. Furthermore, an uncertainty budget was generated for the analytical method using the BIPM-approach recommended by the International Organization for Standardization. The relative uncertainty of the method was estimated to be 10-12%, which was consistent with the observed relative variability found in the method evaluation. The method was evaluated in accordance with EURACHEM Guidance Document No 1 concerning accreditation for chemical laboratories with respect to accuracy and precision. The field study showed that the standard deviation of the method was sufficiently low to distinguish between persons working with two different processes of garbage collection, i.e. collection using four wheeled containers versus collection using bins.     

Proton-ionizable crown compounds. 4. New macrocyclic polyether ligands containing a triazole subcyclic unit

A series of macrocyclic polyether (crown) ligands containing the proton-ionizable s-triazole subcyclic unit were prepared by reacting the 1-THP blocked 3,5-bis(chloromethyl)-1H-1,2,4-triazole with various oligoethylene glycols. The starting bis(chloromethyl)triazole is a vessicant and must be used with caution. Triazolo-18-crown-6 ( 5 ) formed stable complexes with barium, strontium, copper and benzylammonium cations but not with potassium or lithium. The crystal structure of 5 showed the triazole proton to be on nitrogen 3 which is outside the macroring cavity.     

Tetrathiafulvalene-acetylene scaffolding: new pi-electron systems for advanced materials

Novel extended tetrathiafulvalenes (TTFs) with hexa-2,4-diyne-1,6-diylidene spacers between the two 1,3-dithiole rings and laterally appended alkynyl moieties for one- and two-dimensional scaffolding were synthesised and investigated for their electronic properties.     

Improved pulse sequences for pure exchange solid-state NMR spectroscopy

Spin-exchange experiments are useful for improving the resolution and establishment of sequential assignments in solid-state NMR spectra of uniformly (15)N-labeled proteins oriented macroscopically in phospholipid bilayers. To exploit this advantage fully, it is crucial that the diagonal peaks in the two-dimensional exchange spectra are suppressed. This may be accomplished using the recent pure-exchange (PUREX) experiments, which, however, suffer from up to a threefold reduction of the cross-peak intensity relative to experiments without diagonal-peak suppression. This loss in sensitivity may severely hamper the applicability for the study of membrane proteins. In this paper, we present a two-dimensional exchange experiment (iPUREX) which improves the PUREX sensitivity by 50%. The performance of iPUREX is demonstrated experimentally by proton-mediated (15)N-(15)N spin-exchange experiments for a (15)N-labeled N-acetyl-L-valyl-L-leucine dipeptide. The relevance of exchange experiments with diagonal-peak suppression for large, uniformly (15)N-labeled membrane proteins in oriented phospholipid bilayers is demonstrated numerically for the G-protein coupled receptor rhodopsin.     

A ruler for determining the position of proteins in membranes

Both the oxygen diffusion rate and the oxygen solubility vary with depth into the interior of biological membranes. The product of these two gradients generates a single gradient, a permeability gradient, which is a smooth continuous function of the distance from the center of the membrane. Using electron paramagnetic resonance and the spin-probe method, the relaxation gradient of oxygen, which is directly proportional to the permeability gradient, is the quantity that can be directly measured in membranes under physiological conditions. The gradient obtained provides a calibrated ruler for determining the membrane depth of residues either from loop regions of membrane-binding proteins or from the membrane-exposed residues of transmembrane proteins. We have determined the relaxation gradient of oxygen in zwitterionic and anionic phospholipid membranes by attaching a single nitroxide probe to a transmembrane alpha-helical polypeptide at specific residues. The peptide ruler was used to determine the depth of penetration of the calcium-binding loops of the C2 domain of cytosolic phospholipase A(2). The positions of selected residues of this membrane-binding protein that penetrate into the membrane, determined using this ruler, compared favorably with previous determinations using more complex methods. The relaxation gradient constrains the possible values of the membrane-dependent oxygen concentration and the oxygen diffusion gradients. The average oxygen diffusion coefficient is estimated to be at least 2-fold smaller in the membrane than that in water.     

Positron studies of hydrogen-defect interactions in proton irradiated molybdenum

Molybdenum single crystals are irradiated at 20 K with 6 MeV protons. The radiation damage and lattice defect annealing is studied by positron lifetime spectroscopy in the temperature range from 15 to 720 K. Loss of vacancies due to recombination with mobile interstitials is observed at 40 K (Stage I) in agreement with resistivity measurements. This is the first time Stage I is observed by positrons below 77 K. The implanted hydrogen decorates the vacancies around 100 K, which is consistent with a hydrogen migration energy in molybdenum:E _M ^H = 0.3–0.4 eV. Clustering of spatially correlated vacancies takes place in a wide temperature region below the usual vacancy clustering stage (Stage III). Stage III is observed at rather low temperatures (400–480 K) due to the very high vacancy concentration. Hydrogen bound to vacancies and vacancy clusters is released above 540 K, which puts an upper limit to the hydrogen binding energy:E _B ^H 1.4 eV. The present work emphasizes the advantage of employing a vacancy sensitive technique to study hydrogen in metals, where its intrinsic solubility is low. In such metals (as molybdenum) both the effective solubility and the effective mobility of hydrogen are strongly influenced by the presence of vacancies.     

New phosphonic analogs of aspartic and glutamic acid by aminoalkylation of trivalent phosphorus chlorides with ethyl acetyloacetate or ethyl levulinate and benzyl carbamate

Preparation of the phosphonic analogs of -methylaspartic (4 a–d), glutamic (7 a–b) and -methylpyroglutamic (5 a–b) acids by aminoalkylation of trivalent phosphorus chlorides with ethyl esters of oxoalkyloacids and benzyl carbamate is described. The phosphonic analogs of pyroglutamic acid (8 a–b) was obtained by the cyclization of the corresponding esters (9 a–b). The stability of the phosphonic analogs of pyroglutamic acid in acidic and alkaline media was also studied.

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Die Herstellung von neuen Phosphonanalogen der Asparagin- und Glutaminsäure in der Reaktion von trivalenten Phosphorchloriden mit Ethyl-acetyloacetat oder Ethyl-lävulinat bzw. Benzylcarbaminat

Zusammenfassung Es wurde die Darstellung der Phosphonanalogen der -Methylasparaginsäure (4 a–d), Glutamin- (7 a–b) und -Methylpyroglutaminsäure (5 a–b) in der Reaktion der trivalenten Phosphorchloride und der Oxoalkansäureethylester sowie des Benzylcarbaminats beschrieben. In der Ringschlußreaktion der Ester (9 a–b) erhält man Phosphonanaloge der Pyroglutaminsäure. Die Stabilität der Analogen der Pyroglutaminsäure wurde in sauren und alkalischen Medien geprüft.

     

The vibrational spectra of maleimide and N-D maleimide

The infrared spectra of maleimide as a vapour (160°C), melt (100°C), oriented polycrystalline film, pellet and when dissolved in various solvents were recorded between 4000 and 400 cm^?1. Also certain spectra in the far infrared region 400-40 cm^?1 were obtained. Raman spectra of the crystalline solid, melt and as a saturated solution in acetonitrile were recorded and semiquantitative polarization measurements carried out. For N-D maleimide infrared and Raman spectra of the solid compound were recorded.The fundamental frequencies have been assigned in terms of C_2v, symmetry on the basis of infrared vapour contours and dichroism of the oriented film as well as on Raman polarization data. A force field was derived for maleimide, by initially transferring force constants from maleic anhydride and subsequent refinement of the force constants. The agreement between observed and calculated frequencies for the in-plane modes was satisfactory whereas certain large discrepancies remained for the out-of-plane vibrations.