Solvent and temperature gradients in separation of synthetic oxyethylene-oxypropylene block (co)polymers using high-temperature liquid chromatography

Chromatographic behavior of synthetic block (co)oligomer samples (EO)n(PO)m(EO)n and (PO)n(EO)m(PO)n with different distribution of propylene oxide (PO) and ethylene oxide (EO) monomer units was investigated on three types of stationary phases on zirconium dioxide support: Zr-PS (polystyrene), Zr-carbon, and Zr-carbon C18. The effects of the distribution and sequence of the oxyethylene and oxypropylene monomer units on the chromatographic retention depend on the type of the stationary phase, but are strongly affected by the organic modifier (methanol or ACN) in aqueous-organic mobile phase. Special attention was focused on the influence of the mobile-phase composition on the separation according to the EO and PO distribution. Zirconia-based columns are stable at elevated temperatures and can be used in high-temperature LC (HTLC); hence, we investigated the temperature effects on the chromatographic behavior up to 90 degrees C. The applications of solvent and temperature gradients were compared on the zirconia stationary phases in the RP mode.     

Acetylcholinesterase and butyrylcholinesterase inhibitory compounds from Chelidonium majus (Papaveraceae)

The roots and aerial parts of Chelidonium majus L. were extracted with EtOH and fractionated using CHCl3 and EtOH. Repeated column chromatography, preparative TLC and crystallization led to the isolation of five isoquinoline alkaloids, stylopine (3), chelidonine (4), homochelidonine (5), protopine (6), and allocryptopine (7), along with two isolation artifacts 6-ethoxydihydrosanguinarine (1) and 6-ethoxydihydrochelerythrine (2). All isolated compounds were tested for human blood acetylcholinesterase (HuAChE) and human plasma butyrylcholinesterase (HuBuChE) inhibitory activity. The isolation artifacts exhibited the highest activity against HuAChE and HuBuChE with IC50 values of 0.83 +/- 0.04 microM and 4.20 +/- 0.19 microM for 6-ethoxydihydrochelerythrine and 3.25 +/- 0.24 microM and 4.51 +/- 0.31 microM for 6-ethoxydihydrosanguinarine. The most active of the naturally-occurring alkaloids was chelidonine, which inhibited both HuAChE and HuBuChE in a dose-dependent manner with IC50 values of 26.8 +/- 1.2 microM and 31.9 +/- 1.4 microM, respectively.     

A new chemical probe for the detection of the cancer-linked galectin-3

A chemical probe was developed for the detection of the emerging cancer marker galectin-3. The probe contains a benzophenone moiety which covalently attaches itself to the protein upon binding and irradiation. Introduction of a fluorescent label via'click' chemistry allows the labelled proteins to be visualized in a gel. With the probe, selective visualization of galectin-3 in protein mixtures was shown and remarkably even in cell lysates.     

Synthesis of thiolate-protected silver nanocrystal superlattices from an organometallic precursor and formation of molecular di-<Emphasis Type="Italic">n</Emphasis>-alkyldisulfide lamellar phases

Abstract  

Silver nanocrystal superlattices (NCSs) stabilized by hexadecanethiol have been prepared through reduction of [Ag(hexadecanethiolate)] _n , formed in situ by reaction of the organometallic precursor [Ag(C₆F₅)] and hexadecanethiol. The nanostructures have been characterized by transmission electron microscopy (TEM) and X-ray diffractometry (XRD). Several reaction parameters such as solvent (anisole or toluene), reaction temperature (150 or 120 °C) or silver:thiol ratio (1:1 or 2:1) have been studied. The NCSs are formed by silver nanoparticles which sizes range from 3.7 to 5.1 nm, depending on the reaction conditions. The formation a of lamellar phase of di-n-hexadecyldisulfide by oxidation of the hexadecanethiolate ligands bonded to Ag(I) is detected by XRD.     

Numerical simulation of bubble dynamics in a Phan-Thien–Tanner liquid: Non-linear shape and size oscillatory response under periodic pressure

We study non-linear bubble oscillations driven by an acoustic pressure with the bubble being immersed in a viscoelastic, Phan-Thien–Tanner liquid. Solution is provided numerically through a method which is based on a finite element discretization of the Navier–Stokes flow equations. The proposed computational approach does not rely on the solution of the simplified Rayleigh–Plesset equation, is not limited in studying only spherically symmetric bubbles and provides coupled solutions for the velocity, stress fields and bubble interface. We present solutions for non-spherical bubbles, with asphericity being addressed by means of Legendre polynomials or associated Legendre functions. A parametric investigation of the bubble dynamical oscillatory response as a function of the fluid rheological properties shows that the amplitude of bubble oscillations drastically increases as liquid elasticity (quantified by the Deborah number) increases or as liquid viscosity decreases (quantified by the Reynolds number). Extensive numerical calculations demonstrate that increasing elasticity and/or viscosity of the surrounding liquid tend to stabilize the shape anisotropy of an initially non-spherical bubble. Results are shown for pressure amplitudes 0.2–2 MPa and Deborah, Reynolds numbers in the intervals of 1–8 and 0.094–1.256, respectively.     

Optimization of separation in two-dimensional high-performance liquid chromatography by adjusting phase system selectivity and using programmed elution techniques

The overall peak capacity in comprehensive two-dimensional liquid chromatographic (LC x LC) separation can be considerably increased using efficient columns and carefully optimized mobile phases providing large differences in the retention mechanisms and separation selectivity between the first and the second dimension. Gradient-elution operation and fraction-transfer modulation by matching the retention and the elution strength of the mobile phases in the two dimensions are useful means to suppress the band broadening in the second dimension and to increase the number of sample compounds separated in LC x LC. Matching parallel gradients in the first and second dimension eliminate the necessity of second-dimension column re-equilibration after the independent gradient runs for each fraction, increase the use of the available second-dimension separation time and can significantly improve the regularity of the coverage of the available retention space in LC x LC separations, especially with the first- and second-dimension systems showing partial selectivity correlations. Systematic development of an LC x LC method with parallel two-dimensional gradients was applied for separation of phenolic acids and flavone compounds. Several types of bonded C18, amide, phenyl, pentafluorophenyl and poly(ethylene glycol) columns were compared using the linear free energy relationship method to find suitable column combination with low correlation of retention of representative standards. The phase systems were optimized step-by-step to find the mobile phases and gradients providing best separation selectivity for phenolic compounds. The optimization of simultaneous parallel gradients in the first and second dimension resulted in significant improvement in the utilization of the available two-dimensional retention space.     

CdS-containing nano-assemblies of double hydrophilic block copolymers in water

Two samples of poly(sodium(sulfamate-carboxylate)isoprene)-block-poly(ethylene oxide) copolymer (SCIEO-1 and SCIEO-2) differing in molecular weight and relative length of polyelectrolyte blocks have been used as templates for the synthesis of cadmium sulfide (CdS) nanoparticles in aqueous media. The double-hydrophilic copolymer SCIEO has very high 1D charge density, and its polymer chain structure mimics that of polysaccharide heparin. It is soluble in aqueous media, but the addition of cadmium acetate (Cd(Ac)₂) to its aqueous solution causes the formation of micellar aggregates with Cd²⁺containing insoluble cores above the threshold Cd²⁺ concentration. The trapped Cd²⁺ ions can be chemically transformed to CdS nanoparticles. The stability of hybrid SCIEO/CdS micelles depends on the ratio of PEO-to-SCI lengths: it was found that the SCIEO-2 copolymer with sufficiently long PEO block behaves as an effective stabilizer for the synthesis of CdS nanoparticles embedded in micelles, while SCIEO-1 does not. The morphology of aggregates varies with the Cd-to-SCI ratios and ranges from spherical to mixture of spherical and necklace-like micellar aggregates. A number of experimental techniques including static and dynamic light scattering, fluorescence correlation spectroscopy, atomic force and transmission electron microscopy, UV-vis, and fluorescence spectroscopy were employed for the characterization of both CdS containing hybrid micelles and embedded CdS nanoparticles.     

Use of buffering and other means to improve results of problematic pesticides in a fast and easy method for residue analysis of fruits and vegetables

A modification that entails the use of buffering during extraction was made to further improve results for certain problematic pesticides (e.g., folpet, dichlofluanid, chlorothalonil, and pymetrozine) in a simple, fast, and inexpensive method for the determination of pesticides in produce. The method, known as the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method for pesticide residues in foods, now involves the extraction of the sample with acetonitrile (MeCN) containing 1% acetic acid (HAc) and simultaneous liquid-liquid partitioning formed by adding anhydrous MgSO4 plus sodium acetate (NaAc). The extraction method is carried out by shaking a centrifuge tube which contains 1 mL of 1% HAc in MeCN plus 0.4 g anhydrous MgSO4 and 0.1 g anhydrous NaAc per g sample. The tube is then centrifuged, and a portion of the extract is transferred to a tube containing 50 mg primary secondary amine sorbent plus 150 mg anhydrous MgSO4/mL of extract. After a mixing and centrifugation step, the extract is transferred to autosampler vials for concurrent analysis by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/tandem mass spectrometry. Independent of the original sample pH, the use of buffering during the extraction yields pH <4 in the MeCN extract and >5 in the water phase, which increases recoveries of both acid- and base-sensitive pesticides. The method was evaluated for 32 diverse pesticides in different matrixes, and typical percent recoveries were 95 +/- 10, even for some problematic pesticides. Optional solvent exchange to toluene prior to GC/MS analysis was also evaluated, showing equally good results with the benefit of lower detection limits, but at the cost of more time, material, labor, and expense.     

Evaluation of two fast and easy methods for pesticide residue analysis in fatty food matrixes

Two rapid methods of sample preparation and analysis of fatty foods (e.g., milk, eggs, and avocado) were evaluated and compared for 32 pesticide residues representing a wide range of physicochemical properties. One method, dubbed the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method for pesticide residue analysis, entailed extraction of 15 g sample with 15 mL acetonitrile (MeCN) containing 1% acetic acid followed by addition of 6 g anhydrous magnesium sulfate and 1.5 g sodium acetate. After centrifugation, 1 mL of the buffered MeCN extract underwent a cleanup step (in a technique known as dispersive solid-phase extraction) using 50 mg each of C18 and primary secondary amine sorbents plus 150 mg MgSO4. The second method incorporated a form of matrix solid-phase dispersion (MSPD), in which 0.5 g sample plus 2 g C18 and 2 g anhydrous sodium sulfate was mixed in a mortar and pestle and added above a 2 g Florisil column on a vacuum manifold. Then, 5 x 2 mL MeCN was used to elute the pesticide analytes from the sample into a collection tube, and the extract was concentrated to 0.5 mL by evaporation. Extracts in both methods were analyzed concurrently by gas chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry. The recoveries of semi-polar and polar pesticides were typically 100% in both methods (except that basic pesticides, such as thiabendazole and imazalil, were not recovered in the MSPD method), but recovery of nonpolar pesticides decreased as fat content of the sample increased. This trend was more pronounced in the QuEChERS method, in which case the most lipophilic analyte tested, hexachlorobenzene, gave 27 +/- 1% recovery (n=6) in avocado (15% fat) with a<10 ng/g limit of quantitation.     

Study of posttranslational non-enzymatic modifications of collagen using capillary electrophoresis/mass spectrometry and high performance liquid chromatography/mass spectrometry

The depository effects that occur in slowly metabolized proteins (typically glycation) are very difficult to assess, owing to their extremely low concentration in the protein matrix. Collagen accumulates reactive metabolites through reactions that are not regulated by enzymes. A typical example of these non-enzymatic changes is glycation (the Maillard reaction, the formation of advanced glycation end products), resulting from the reaction of the oxo-group of sugars with the epsilon-amino group of lysine and arginine. Collagen samples (type I) as a test protein were incubated separately with glucose, ribose and malondialdehyde. Collagen was fragmented with cyanogen bromide and then digested with trypsin. This peptide digest was separated by CE, CE-MS/MS, and HPLC-MS/MS. An ion trap MS was used and MS conditions were optimized for both methods. These on-line CE-MS/MS and HPLC-MS/MS couplings made it possible to discover specific modifications such as (N(epsilon)-(carboxymethyl)-lysine) in the precise location in the structure of collagen corresponding to posttranslational non-enzymatic modifications. A new CE-MS/MS technique for peptide analysis was developed, and applied in the identification of posttranslational modifications in slowly metabolized test proteins.