NMR analysis of a series of substituted pyrazolo[3,4-d] pyrimidines-4-amines

A series of 21 substituted pyrazolo[3,4-d]pyrimidines-4-amines were studied by (1)H and (13)C NMR. The application of two-dimensional techniques, HMQC and HMBC, allowed the complete assignment of the spectra for all the compounds.     

Homoleptic Organoderivatives of High‐Valent Nickel(III)

Homoleptic perhalophenyl derivatives of divalent nickel complexes with the general formula [NBu₄]₂[Ni^II (C₆X₅)₄] [X=F ( 1 ), Cl ( 2 )] have been prepared by low‐temperature treatment of the halo‐complex precursor [NBu₄]₂[NiBr₄] with the corresponding organolithium reagent LiC₆X₅. Compounds 1 and 2 are electrochemically related by reversible one‐electron exchange processes with the corresponding organometallate(III) compounds [NBu₄][Ni^III (C₆X₅)₄] [X=F ( 3 ), Cl ( 4 )]. The potentials of the [Ni^III (C₆X₅)₄]^?/[Ni^II (C₆X₅)₄]^2? couples are +0.07 and ?0.11 V for X=F or Cl, respectively. Compounds 3 and 4 have also been prepared and isolated in good yield by chemical oxidation of 1 or 2 with bromine or the amminium salt [N(C₆H₄Br‐4)₃][SbCl₆]. The [Ni^III (C₆X₅)₄]^? species have SP‐4 structures in the salts 3 and 4 , as established by single‐crystal X‐ray diffraction methods. The [Ni^II (C₆F₅)₄]^2? ion in the parent compound 1 has also been found to exhibit a rather similar SP‐4 structure. According to their SP‐4 geometry, the Ni^III compounds (d⁷) behave as S=1/2 systems both at microscopic (EPR) and macroscopic levels (ac and dc magnetization measurements). The spin Hamiltonian parameters obtained from the analysis of the magnetic behavior of 3 and 4 within the framework of ligand field theory show that the unpaired electron is centered mainly on the metal atom, with >97 % estimated d contribution. Thermal decomposition of 3 and 4 proceeds with formation of the corresponding C₆X₅? C₆X₅ coupling compounds.     

Electrophoretic deposition as a tool for separation of protein inclusion bodies from host bacteria in suspension

This paper shows, for the first time, that the electrophoretic deposition technique is able to selectively collect protein inclusion bodies (PBs) from the host bacteria suspensions. In the first step, zeta potential as a function of pH is carefully determined for both species involved. Based on the obtained dependencies, the pH of the mixture of PBs and bacteria is precisely adjusted and the electrophoretic experiment is carried out. We show that the efficiency of separation and the yield depends not only on the electrokinetic properties of given species but also on the electrode composition and surface morphology. The deposited species are easily removed by forced washing or reverse electric field. As a whole, the selectivity and the yields are higher than in most alternative state-of-the art techniques.     

Bulk and interfacial behaviour of caseinoglycomacropeptide (GMP)

Caseinoglycomacropeptide (GMP) is a hydrophilic glycopeptide released from milk κ-casein by chymosin hydrolysis during cheese making. GMP is thought to be a potential ingredient for specific dietary applications with several health benefits. In this study GMP was characterized at the air–water interface and its behaviour was related with the self-assembly of GMP in solution as affected by pH. This GMP self-assembly was investigated by dynamic light scattering and the interfacial properties were determined by tensiometry and surface dilatational measurements at pH 4, 5 and 7. At pH 5 GMP exhibited higher surface pressure at equilibrium than at pH 7. At pH 4 the behaviour was more complex due to self-assembly close to GMP pI. Dynamic measurement showed that the adsorption/penetration rate constant (K_ads) is facilitated at higher GMP bulk concentrations, while the rate constant of rearrangement (K_r) decreased at higher GMP concentrations which could be attributed to the existence of a steric restriction due to the higher GMP load at the interface. K_r was higher at pH 5 because of lower electrostatic interactions close to the pI. The viscoelastic properties showed a complex behaviour due to the existence of protein–protein interactions depending on the GMP concentration, on the pH of the bulk and on the rates of diffusion, adsorption and rearrangement of GMP at the air–water interface.     

Characterization of sepia ink in ancient graphic documents by capillary electrophoresis

A simple and rapid capillary electrophoresis with diode array detection method was developed for sepia ink identification in ancient graphic documents.Separation was performed in a fused-silica capillary (64.5 cm length, 50 μm i.d.). The running buffer was 20 mM sodium tetraborate solution, pH 9.2. The applied potential was 25 kV, temperature 25 °C and detection was at 220 nm. An appropriate extraction procedure was applied for the take and treatment of sample from the reference substances and ancient graphic documents. This method was successfully applied to the collection of drawings and maps from the Royal Chancellery Archives of Granada (Spain).     

Heart-cutting two-dimensional (size exclusion × reversed phase) liquid chromatography–mass spectrometry analysis of flavonol glycosides from leaves of Maytenus ilicifolia

Two-dimensional liquid chromatography–electrospray ionization mass spectrometry was employed to analyze flavonol glycosides present in leaves of Maytenus ilicifolia, frequently used in traditional Brazilian medicine. Since they contain many flavonol glycosides, including isomers, one-dimensional liquid chromatography did not give complete separation and identification, yielding overlapping of compounds with different molecular weights. Thus, employing size exclusion chromatography in the first and reversed phase in the second dimension, a great number of flavonol glycosides could be identified and its relative abundances determined. The majority of glycosides contained kaempferol or quercetin as aglycones, and glycosides with previously unreported structures were also present and characterized.     

Characterization of O-trimethylsilyl oximes of trisaccharides by gas chromatography–mass spectrometry

Gas chromatography (GC) data (linear retention indices and relative areas) and mass spectra (most representative m/z fragments) of 12 reducing trisaccharides as trimethylsilyl oximes (TMSO) and four non-reducing as trimethylsilyl (TMS) ethers have been described for the first time and related to their structural features. Some trends have been observed: earlier elution of non-reducing compounds and fructotrioses; aldotrioses bearing the reducing end with link in position 6 showing the highest retention. Abundance of several fragment ions and their ratios were useful for trisaccharide characterization; some of these features seem to be useful for the characterization of new trisaccharides.     

Determination of sulfonamide residues in water samples by in-line solid-phase extraction-capillary electrophoresis

An in-line solid-phase extraction-capillary electrophoresis method with UV–vis detection was developed for the monitoring of residues of five sulfonamides (sulfadoxin, sulfadimethoxine, sulfamerazine, sulfachloropyridazine and sulfamethoxazole) in tap, bottled mineral and river waters. For this purpose an analyte concentrator was constructed, based on the introduction of a small portion of a solid-phase extraction sorbent into the electrophoretic capillary to carry out an in-line concentration step, improving sensitivity. A detailed study was carried out to optimize parameters affecting the in-line solid-phase extraction process, such as the design of the concentrator device, type of sorbent and conditions of elution and injection. The proposed method is simple for the environmental monitoring of these antibiotic residues in waters, allowing the direct injection of the samples without any off-line pretreatment and achieving limits of detection between 0.3 and 0.6 μg/L. Recoveries ranging 52.2–109.2% and relative standard deviations below 13.4% were obtained.     

Ultra-High-Pressure Liquid Chromatographic method for the analysis of tocopherols in human colostrum and milk

A rapid, highly sensitive and direct Ultra-High-Pressure Liquid Chromatographic method was developed and validated for quantifying δ-, β + γ-, and α-tocopherol in human colostrum and milk. Two reversed-phase chromatographic columns and two detectors (Fluorescence Detector or FD and Photodiode Detector Array or PDA) were used and both methods were independently validated. Two internal standards were selected according to the detector used. Recoveries ranged from 96.71% to 103.55% and the relative standard deviations for the within-day precision were below 6% (PDA) and 3% (FD). Both approaches enabled to achieve low detection limits, on the order of ng (PDA) or pg (FD). Only 300 μL of sample and a chromatographic run of less than 1.6 min were enough to efficiently quantify the isomers in the colostrum and milk of Spanish women.     

LC Method for Studies on the Stability of Sibutramine in Soft Gelatin Capsules

A sensitive and rapid liquid chromatographic method was successfully developed and validated for the determination of sibutramine hydrochloride in bulk and capsules. Sibutramine in the presence of its degradation products was analyzed using UV detection at 225 nm. Chromatography was performed on a reversed-phase C₈ (150 × 4.0 mm I.D., 5 μm) analytical column under isocratic conditions. The mobile phase was composed of acetonitrile:water (aqueous phase containing 0.3% triethylamine and pH adjusted to 7.0) (75:25, v/v) at a flow-rate of 1.1 mL min⁻¹. No chromatographic interference was found during the analysis. Light was the stress condition which most contributed to sibutramine degradation. The method showed a linear response (r > 0.999) from 30 to 90 μg mL⁻¹. The mean recovery for capsules was 101.2%. Inter-day assays showed relative standard deviations of 0.42 and 1.62% for bulk and capsules, respectively. The developed method is able to separate sibutramine from its major degradation products and it may be used in the quality control of this active pharmaceutical ingredient in both bulk and capsules.