Use of cation exchange chromatography for human C-peptide isotope dilution - mass spectrometric assay

An application of ion exchange chromatography for C-peptide analysis is described here. At the stage of C-peptide isolation, a strong cation exchanger (SP HP or MonoS) was used to purify the analyte from ballast proteins and peptides. The conditions of ion-exchange chromatographic separations were optimized using theoretical modeling of the net surface electric charge of the peptide as a function of pH. The purified and concentrated sample was further subjected to LC-MS/MS. In order to improve the reliability of analysis, two fragment ions were monitored simultaneously both for native C-peptide and internal standard, isotopically labeled C-peptides analogues (fragments with m/z of 927.7 and 147.2). Using ion-exchange chromatography, it became possible to process larger sample volumes, important for testing patients with very low C peptide levels, compared to currently used solid phase extraction methods.     

Nitrile imines: matrix isolation, IR spectra, structures, and rearrangement to carbodiimides

The structures and reactivities of nitrile imines are subjects of continuing debate. Several nitrile imines were generated photochemically or thermally and investigated by IR spectroscopy in Ar matrices at cryogenic temperatures (Ph-CNN-H 6, Ph-CNN-CH(3)17, Ph-CNN-SiMe(3)23, Ph-CNN-Ph 29, Ph(3)C-CNN-CPh(3)34, and the boryl-CNN-boryl derivative 39). The effect of substituents on the structures and IR absorptions of nitrile imines was investigated computationally at the B3LYP/6-31G* level. IR spectra were analyzed in terms of calculated anharmonic vibrational spectra and were generally in very good agreement with the calculated spectra. Infrared spectra were found to reflect the structures of nitrile imines accurately. Nitrile imines with IR absorptions above 2200 cm(-1) have essentially propargylic structures, possessing a CN triple bond (typically PhCNNSiMe(3)23, PhCNNPh 29, and boryl-CNN-boryl 39). Nitrile imines with IR absorptions below ca. 2200 cm(-1) are more likely to be allenic (e.g., HCNNH 1, PhCNNH 6, HCNNPh 43, PhCNNCH(3)17, and Ph(3)C-CNN-CPh(3)34). All nitrile imines isomerize to the corresponding carbodiimides both thermally and photochemically. Monosubstituted carbodiimides isomerize thermally to the corresponding cyanamides (e.g., Ph-N═C═N-H 5 → Ph-NH-CN 8), which are therefore the thermal end products for nitrile imines of the types RCNNH and HCNNR. This tautomerization is reversible under flash vacuum thermolysis conditions.     

A twelve‐analyzer detector system for high‐resolution powder diffraction

A dedicated high‐resolution high‐throughput X‐ray powder diffraction beamline has been constructed at the Advanced Photon Source (APS). In order to achieve the goals of both high resolution and high throughput in a powder instrument, a multi‐analyzer detector system is required. The design and performance of the 12‐analyzer detector system installed on the powder diffractometer at the 11‐BM beamline of APS are presented.     

Was Biofilme zusammenhält: Proteine,Polysaccharide ..

Extracellular polymeric substances (EPS) are biopolymers of microbial origin and consist mainly of polysaccharides, proteins, lipids and nucleic acids. The EPS mediate adhesion to surfaces and form a hydrogel matrix for biofilms and other microbial aggregates. This matrix can be considered as “house” of the microorganisms which allows for the formation of stable communities (“microconsortia”) of synergistic strains and enables them to degrade recalcitrant substances. EPS retain water and prevent desiccation. Cohesion and adhesion are provided by a network of fluctuating adhesion points. External pressure can change the structure from a gel to a highly viscous liquid. Due to their sorptive properties, dissolved nutrients from the water phase are accumulated and increase the survival chances of biofilm organisms in oligotrophic environments. The matrix facilitates gene exchange and regulation processes via signalling molecules. It provides a template for extracellular enzymes and prevents that they are washed out. Thus, it is of great importance for the degradation of solids and particles. Remnants of lysed cells are retained and can be utilized as food source. Thus, biofilms can be considered as a natural example for sustainable use of nutrients. Some EPS are biotechnologically employed as additives for food, drilling fluids and as biosurfactants.