Pressurized electrochromatography coupled with electrospray ionization mass spectrometry for analysis of peptides and proteins

Pressurized capillary electrochromatography (pCEC) was coupled with electrospray ionization mass spectrometry (ESI-MS) using a coaxial sheath liquid interface. It was used for separation and analysis of peptides and proteins. The effects of organic modifier and applied voltage on separation were investigated, and the effects of pH value of the mobile phase and the concentration of the electrolyte on ESI-MS signal were investigated. The resolution and detection sensitivity with different separation methods (pCEC, capillary high-performance liquid chromatography) coupled on-line with mass spectrometry were compared for the separation of a peptide mixture. To evaluate the feasibility and reliability of the experimental setup of the system, tryptic digests of cytochrome c and modified protein as real samples were analyzed by using pCEC-ESI-MS.

Capillary electrochromatographic separation of ionizable compounds with a molecular imprinted monolithic cationic exchange column

A polymer-based monolithic capillary column imprinted with 4-aminopyridine (4-AP) was prepared by a thermally-initiated polymerization process; and its performance as a capillary electrochromatographic medium was evaluated in separating 4-AP and 2-AP isomers. The effects of experimental parameters, such as pH value and ionic strength of the buffer, the acetonitrile content in the mobile phase, and the applied voltage, on the resolution of these isomers had been carefully investigated. It was found that in the retention process there were interplays of multiple mechanisms of ion-exchange, molecular imprinting, and electrophoresis. These mechanisms allowed more sophisticated control of experimental parameters in the separation of ionizable compounds.

Separation of peptides on mixed mode of reversed-phase and ion-exchange capillary electrochromatography with a monolithic column

The mixed mode of reversed phase (RP) and strong canon-exchange (SCX) capillary electrochromatography (CEC) based on a monolithic capillary column has been developed. The capillary monolithic column was prepared by in situ copolymerization of 2-(sulfooxy)ethyl methacrylate (SEMA) and ethylene dimethacrylate (EDMA) in the presence of porogens. The sulfate group provided by the monomer SEMA on the monolithic bed is used for the generation of the electroosmotic flow (EOF) from the anode to the cathode, but at the same time serves as a SCX stationary phase. A mixed-mode (RP/SCX) mechanism for separation of peptides was observed in the monolithic column, comprising hydrophobic and electrostatic interaction as well as electrophoretic migration at a low pH value of mobile phase. A column efficiency of more than 280000 plates/m for the unretained compound has been obtained on the prepared monoliths. The relative standard deviations observed for to and retention factors of peptides were about 0.32% and less than 0.71% for ten consecutive runs, respectively. Effects of mobile phase compositions on the EOF of the monolithic column and on the separation of peptides were investigated. The selectivity on separation of peptides in the monolithic capillary column could be easily manipulated by varying the mobile phase composition.

Pressurized capillary electrochromatography separation of peptides with strong cation exchange and hydrophilic interaction

A pressurized capillary electrochromatography (pCEC) instrument with solvent gradient capability has been used for the separation of a peptide mixture. Retention mechanism and selectivity of the peptides were studied by pCEC using a strong cation exchange (SCX) column. The effects of applied voltage, supplementary pressure, organic modifier concentration, ionic strength,, and pH value on pCEC separation were investigated. It was found that the retention mechanism of the peptides in this system is based on a mixed mode of hydrophilic interaction, strong cation exchange, and electrophoresis. Compared with the separation results obtained by reverse phase pCEC and capillary electrophoresis (CE), this mixed-mode pCEC is more powerful for the separation of hydrophilic peptides with similar charge-to-mass ratio.

Capillary electrochromatography for separation of peptides driven with electrophoretic mobility on monolithic column

A mode of capillary electrochromatography for separation of ionic compounds driven by electrophoretic mobility on a neutrally hydrophobic monolithic column was developed. The monolithic column was prepared from the in situ copolymerization of lauryl methacrylate and ethylene dimethacrylate to form a C-12 hydrophobic stationary phase. It was found that EOF in this hydrophobic monolithic column was very poor, even the pH value of mobile phase at 8.0. The peptides at acidic buffer were separated on the basis of their differences in electrophoretic mobility and hydrophobic interaction with the stationary phase; therefore, different separation selectivity can be obtained in CEC from that in capillary zone electrophoresis (CZE). Separation of peptides has been realized with high column efficiency (up to 150 000 plates/meter) and good reproducibility (migration time with RSD < 0.5%), and all of the peptides, including some basic peptides, showed good peak symmetry. Effects of the mobile phase compositions on the retention of peptides at low pH have been investigated in a hydrophobic capillary monolithic column. The significant difference in selectivity of peptides in CZE and CEC has been observed. Some peptide isomers that cannot be separated by CZE have been successfully separated on the capillary monolithic column in this mode with the same buffer used.

Direct spectroscopic evidence for the specific property of aqueous core phase in black soap film

The microenvironment of the aqueous core phase in the black soap film of cationic surfactant cetyltrimethylammonium bromide with the anionic dye Brilliant Yellow as spectral probe has been studied by UV-vis spectroscopy. Under neutral and basic conditions, the dye aggregates in the films exist as both the acid and base forms in contrast to a preference of the base form in the bulk solutions. The specific property of black soap film, that the intrinsic pH value of the aqueous core phase insensitively responds to pH changes of the bulk solution, is directly observed through UV-vis spectra.

Controllable synthesis of VSB-5 micirospheres and microrods: Growth mechanism and selective hydrogenation catalysis

Nanoporous VSB-5 nickel phosphate molecular sieves with relatively well controllable sizes and morphology of microspheres assembled from nanorods were synthesized at 140 degrees C over a short time in the presence of hexamethylenetetramine (HMT) by a facile hydrothermal method. The pH value, reaction time, and ratio of HMT to NaHPO2-H2O crucially influence the morphology and quality of the final products.

Analysis of amphetamines in urine with liquid-liquid extraction by capillary electrophoresis with simultaneous electrochemical and electrochemiluminescence detection

Amphetamines including methamphetamine, 3,4-methylenedioxyamphetamine and 3,4-methylenedioxymethamphetamine were separated and detected by CE using simultaneous electrochemical (EC) and electrochemiluminescence (ECL) detection (CE-EC/ ECL). Factors that influenced the separation and detection performance, such as the detection potential, the pH value and concentration of the running buffer, the separation voltage and the pH of the detection buffer, were investigated.

Microchip Micellar Electrokinetic Chromatography Based on One Functionalized Ionic Liquid and Its Excellent Performance on Proteins Separation

Herein, one water-soluble functionalized ionic liquid (IL), 1-butyl-3-methylimidazolium dodecanesulfonate (BAS), was designed, investigated and successfully applied to microchip micellar electrokinetic chromatography (MEKC) construction. It possessed the properties of both IL and surfactant. A fairly stable pH value similar to 7.4, which was fit to pH values of general biological buffers, was nicely placed at the optimum concentration of 20 mM BAS solution. While applying BAS solution as running buffer in poly(dimethylsiloxane) (PDMS) microfluidic systems, significantly enhanced electroosmotic flow (8-fold) and resolutions between analytes were obtained than that using other supporting electrolytes or surfactants.

Investigation of some critical parameters of buffer conditions for the development of quantum dots-based optical sensors

The unique surface-sensitive properties make quantum dots (QDs) great potential in the development of sensors for various analytes. However, quantum dots are not only sensitive to a certain analyte, but also to the surrounding conditions. The controlled response to analyte may be the first step in the designing of functional quantum dots sensors. In this study, taking the quenching effect of benzoquinone (BQ) on CdTe QDs as model, several critical parameters of buffer solution conditions with potential effect on the sensors were investigated. The pH value and the concentration of sodium citrate in the buffer solution critically influenced the quenching effects of BQ.

Phosphonate-Functionalized Polyfluorene as a Highly Water-Soluble Iron(III) Chemosensor

An anionic, phosphonate-functionalized polyfluorene, i.e., poly(9,9-bis(3'-phosphatepropyl)fluorene-alt-1,4-phenylene) sodium salt (PFPNa), has been synthesized by copolymerization of phosphonic acid-substituted 2,7-dibromofluorene and phenyldiboronic ester via direct Suzuki polycondensation reaction in DMF/water. Polymer PFPNa is highly soluble and emissive in water with a solubility of 60 mg/mL and a photoluminescence quantum yield of 75%. The absorption and fluorescence spectra of PFPNa are strongly dependent on pH value owing to the partial protonation of phosphate groups and the aggregation of the polymer chains.

Rare Earth Pyrophosphates: Effective Catalysts for the Production of Acrolein from Vapor-phase Dehydration of Glycerol

Vapor-phase dehydration of glycerol to produce acrolein was investigated at 320 A degrees C over rare earth (including La, Ce, Nd, Sm, Eu, Gd, Tb, Ho, Er, Tm, Yb, Lu) pyrophosphates, which were prepared by precipitation method. The most promising catalysts were characterized by means of XRD, FT-IR, TG-DTA, BET and NH3-TPD measurements. The excellent catalytic performance of rare earth pyrophosphate depends on the appropriate surface acidity which can be obtained by the control of pH value in the precipitation and the calcination temperature, e.g. Nd-4(P2O7)(3) precipitated at pH = 6 and calcined at 500 A degrees C in the catalyst preparation.

Synthesis of different gold nanostructures by solar radiation and their SERS spectroscopy

In this article, a simple and novel photochemical synthesis of different gold nanostructures is proposed using solar radiation. This method is rapid, convenient and of low cost, and can be performed under ambient conditions. By adjusting the concentration of sodium acetate (NaAc), different morphologies of the products can be easily obtained. Without NaAc, the products obtained are mainly polyhedral gold particles; lower concentration of NaAc (0.05 and 0.1 M) accelerates the formation of flowerlike gold nanostructures; while higher concentration of NaAc (0.5 M) facilitates the formation of a variety of gold nanowires and nanobelts. It is found that the morphology change of gold nanaostructures is the result of the synergistic effect of poly(diallyl dimethylammonium) chloride (PDDA), Ac- ions, and the pH value. In addition, the different gold nanostructures thus obtained were used as substrates for surface-enhanced Raman scattering (SERS) with p-aminothiophenol (p-ATP) as the probe molecule.

Effect of block sequence and block length on the stimuli-responsive behavior of polyampholyte brushes: hydrogen bonding and electrostatic interaction as the driving force for surface rearrangement

Diblock polyampholyte brushes with different block sequences (Si/SiO2/poly(acrylic acid)-b-poly (2-vinylpyridine) (PAA-b-P2VP) brushes and Si/SiO2/P2VP-b-PAA brushes) and different block lengths were synthesized by sequent surface-initiated atom transfer radical polymerization (ATRP). The PAA block was obtained through hydrolysis from the corresponding poly(tert-butyl acrylate). The polyampholyte brushes demonstrated unique pH-responsive behavior. In the intermediate pH region, the brushes exhibited a less hydrophilic wetting behavior and a rougher surface morphology due to the formation of polyelectrolyte complex through electrostatic interaction between oppositely charged blocks. In the low pH and high pH regions, the rearrangement of polyampholyte brushes showed great dependence on the block sequence and block length. The polyampholyte brushes with P2VP-b-PAA sequence underwent rearrangement during alternative treatment by acidic aqueous solution (low pH value) and basic aqueous solution (high pH value).

Size-tailored synthesis and luminescent properties of one-dimensional Gd2O3 : Eu3+ nanorods and microrods

Nearly monodisperse and well-defined one-dimensional (1D) Gd2O3:Eu3+ nanorods and microrods were successfully prepared through a large-scale and facile hydrothermal method followed by a subsequent heat treatment process, without using any catalyst or template. X-ray diffraction (XRD), thermogravimetric analysis and differential scanning calorimetry (TGA-DSC), scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), selected area electron diffraction (SAED), photoluminescence (PL) and cathodoluminescence (CL) spectra as well as kinetic decays were used to characterize the samples. The size of the Gd2O3:Eu3+ rods could be modulated from micro- to nanoscale with the increase of pH value using ammonia solution. The as-formed product via the hydrothermal process, Gd(OH)(3):Eu3+, could transform to cubic Gd2O3:Eu3+ with the same morphology and a slight shrinking in size after a postannealing process.