Nuclear magnetic resonance of mass-limited samples using small RF coils

Figure Schematic diagram of a typical arrangement used for hyphenating chemical microseparations (e.g. capillary HPLC, CE, or CEC) with microcoil NMR detection     

Biosensor for total cholesterol estimation using N-(2-aminoethyl)-3-aminopropyltrimethoxysilane self-assembled monolayer

Cholesterol oxidase (ChOx), cholesterol esterase (ChEt), and horseradish peroxidase (HRP) have been co-immobilized covalently on a self-assembled monolayer (SAM) of N-(2-aminoethyl)-3-aminopropyltrimethoxysilane (AEAPTS) deposited on an indium–tin–oxide (ITO) glass surface. These enzyme-modified (ChOx-ChEt-HRP/AEAPTS/ITO) biosensing electrodes have been used to estimate cholesteryl oleate from 10 to 500 mg dL⁻¹. The sensitivity, K _m value, and shelf-life of these ChEt-ChOx-HRP/AEAPTS/ITO biosensing electrodes have been found to be 124 nA mg⁻¹ dL, 95.098 mg dL⁻¹ (1.46 mmol L⁻¹), and ten weeks, respectively. The ChEt-ChOx-HRP/AEAPTS/ITO bio-electrodes have been used to estimate total cholesterol in serum samples. Figure Covalent immobilization of enzymes onto AEAPTS/ITO surface using EDC/NHS chemistry Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Amplification of noble gas ion lines in the afterglow of a pulsed hollow cathode discharge and possible benefit for analytical glow discharge mass spectrometry

A high-current pulsed hollow cathode discharge was used to study the role of atomic and ionic metastables involved in ionization plasma processes. We observed the enhancement of the spectral emission lines of noble gas ions in the afterglow. A study of the processes that involve atomic and ionic metastables is of great interest since it should lead to a better understanding of and enhanced control over the ionization mechanisms crucial to analytical glow discharge mass spectrometry (GDMS) analysis. Figure Time profile of Ti, Ti⁺, and Ne⁺ spectral lines     

Analysis of glycans in glycoproteins by diffusion-ordered nuclear magnetic resonance spectroscopy

Enzymatically cleaved glycans from sub-milligram quantities of erythropoietin (EPO) and ovalbumin have been analyzed, without further purification, by two-dimensional diffusion-ordered nuclear magnetic resonance spectroscopy. At NMR sample concentrations below 50 μmol L⁻¹ the major components of the oligosaccharide fractions could be distinguished by their anomeric proton chemical shift and their size-dependent diffusion coefficients. Figure ¹H NMR diffusion decay curves of anomeric protons in the EPO glycan fraction     

Prediction of retention times of polycyclic aromatic hydrocarbons and <Emphasis Type="Italic">n</Emphasis>-alkanes in temperature-programmed gas chromatography

We have developed an iterative procedure for predicting the retention times of polycyclic aromatic hydrocarbons (PAHs) and n-alkanes during separations by temperature-programmed gas chromatography. The procedure is based on estimates of two thermodynamic properties for each analyte (the differences in enthalpy and entropy associated with movements between the stationary and mobile phases) derived from data acquired experimentally in separations under isothermal conditions at temperatures spanning the range covered by the temperature programs in ten-degree increments. The columns used for this purpose were capillary columns containing polydimethylsiloxane-based stationary phases with three degrees of phenyl substitution (0%, 5%, and 50%). Predicted values were mostly within 1% of experimentally determined values, implying that the method is stable and precise. Figure Predicted values were mostly within 1 % of experimentally determined values, thus implying that the method is stable and precise     

31P NMR study on the autophosphorylation of insulin receptors in the plasma membrane

A nonradioactive ³¹P nuclear magnetic resonance (NMR) spectroscopy protocol has been developed and used to investigate in vitro autophosphorylation of insulin receptors. Optimum experimental conditions have been explored, and the effects of Mn²⁺ and phosphocreatine (PCr) on the determination of the phosphorylation reaction have been assayed. The method was used to monitor the time courses of the phosphorylation reaction in solution. The results from this NMR study were in agreement with observations of insulin receptor phosphorylation made by using Western blotting.      

Headspace mass spectrometry methodology: application to oil spill identification in soils

In the present work we report the results obtained with a methodology based on direct coupling of a headspace generator to a mass spectrometer for the identification of different types of petroleum crudes in polluted soils. With no prior treatment, the samples are subjected to the headspace generation process and the volatiles generated are introduced directly into the mass spectrometer, thereby obtaining a fingerprint of volatiles in the sample analysed. The mass spectrum corresponding to the mass/charge ratios (m/z) contains the information related to the composition of the headspace and is used as the analytical signal for the characterization of the samples. The signals obtained for the different samples were treated by chemometric techniques to obtain the desired information. The main advantage of the proposed methodology is that no prior chromatographic separation and no sample manipulation are required. The method is rapid, simple and, in view of the results, highly promising for the implementation of a new approach for oil spill identification in soils. Figure PCA score plots illustrate clear discrimination of types of crude oil in polluted soil samples (e.g. results are shown for vertisol)     

Spectrophotometric determination of total protein in serum using a novel near-infrared cyanine dye, 5,5′-dicarboxy-1,1′-disulfobutyl-3,3,3′,3′-tetramethylindotricarbocyanine

The application of near-infrared (NIR) dyes (λ _em > 750 nm) to the analysis of biological samples shows much promise, because the long emission wavelengths of such dyes allow interferences from biomolecule matrices to be minimized. In this paper, a novel NIR dye, 5,5′-dicarboxy-1,1′-disulfobutyl-3,3,3′,3′-tetramethylindotricarbocyanine (DCDSTCY) has been developed for the spectrophotometric determination of total protein in serum. Under acidic conditions, the binding of DCDSTCY to proteins caused a new peak at 878 nm, the height of which was proportional to the concentration of protein. The linear range of the method was found to be 0.04–0.5 μg mL⁻¹ for bovine serum albumin (BSA) and human serum albumin (HSA), and detection limits of 5 ng mL⁻¹ were obtained for these substances. The maximum binding number of BSA with DCDSTCY was measured to be 133. The method proposed here has been applied to the quantitation of total protein in serum, and recoveries of 96.6–104% were achieved. Figure Near-infrared probe for protein determination     

Wax components of larval cocoon silk of the hornet Vespa analis Fabricius

Wax, 85% of which consists of orthorhombic crystals, has been found in the cocoon of the hornet Vespa analis Fabricius by means of high-resolution ¹³C solid-state nuclear magnetic resonance (NMR). GC–MS analysis revealed the major components of the wax in the cocoon were linear alkenes and alkanes with a total of 23 or 34 carbon atoms. At 40.7 °C a DSC absorption peak and a ¹³C NMR chemical shift change were observed and interpreted as the result of a crystal transition from the orthorhombic to rotator phase of the wax molecules. At 55.5 °C melting of the wax was observed. The amount of crystalline wax deposition varied with the part of the cocoon—crystalline wax was concentrated in the silk sleeve lining the inner wall of each comb cell but there was very little in the silk cap projecting from the end of each cell. Because the wax components of the larval cocoon were almost identical to those of the larval cuticle, despite a slight difference in the profiles, they might have come from the larval cuticle via direct body contact with the cocoon. Figure Cocoon of the hornet Vespa analis Fabricius     

Computer simulations of a dielectric barrier discharge used for analytical spectrometry

A model developed for a dielectric barrier discharge (DBD) in helium, used as a new spectroscopic source in analytical chemistry, is presented in this paper. The model is based on the fluid approach and describes the behavior of electrons, He⁺ and ions, He metastable atoms, He atoms in higher excited levels, and He₂ dimers. The He ground-state atoms are regarded as background gas. The characteristic effect of charging/discharging of the dielectrics which cover both electrodes is also simulated. Typical results of the model include the distribution of potential inside the plasma (and the potential drop across the dielectrics), the electric current and gap voltage as a function of time for a given applied potential profile, the spatial and temporal number-density profiles of the different plasma species, and the relative contributions of the mechanisms of their production and loss. Figure Schematic diagram of the dielectric barrier discharge (left) and typical temporal profiles of voltage and current, as obtained from the simulations (right)     

Rapid response behavior,at room temperature,of a nanofiber-structured TiO<Subscript>2</Subscript> sensor to selected simulant chemical-warfare agents

A chemical prototype sensor was constructed based on nanofiber-structured TiO₂ and highly sensitive quartz resonators. The gas-sensing behavior of this new sensor to selected simulant warfare agents was investigated at room temperature. Results showed rapid response and good reversibility of this sensor when used with high-purity nitrogen. This provides a simple approach to preparation of materials needed as chemical sensors for selected organic volatiles or warfare agents. Figure Sensing behavior of TiO₂ nanofiber sensor to chemical vapors     

Size prediction of recombinant human growth hormone nanoparticles produced by supercritical fluid precipitation

Solution-enhanced dispersion by supercritical fluids (SEDS) was applied to produce nano-sized recombinant human growth hormone (hGH) particles. Ethanol was used to help the supercritical carbon dioxide to extract water from the aqueous protein solution. Various sizes of hGH nanoparticles were successfully prepared with a narrow particle size distribution from aqueous ethanol solution without using any additive. The theoretical particle sizes were deduced from the calculated droplet sizes based on a modified Jasuja’s equation. The calculated mean particle sizes and the experimentally obtained ones were compared and the results showed an excellent correlation coefficient (R ²) of 0.995. Figure Distribution of hGH Nano-particles     

Preparation and evaluation of a new synthetic polymeric chiral stationary phase for HPLC based on the trans-9,10-dihydro-9,10-ethanoanthracene-(11S,12S)-11,12-dicarboxylic acid bis-4-vinylphenylamide monomer

A new synthetic polymeric chiral stationary phase for liquid chromatography was prepared via free-radical-initiated polymerization of trans-9,10-dihydro-9,10-ethanoanthracene-(11S,12S)-11,12-dicarboxylic acid bis-4-vinylphenylamide. The new polymeric chiral stationary phase (CSP) showed enantioselectivity for many chiral compounds in multiple mobile phases. High stability and sample capacities were observed on this polymeric chiral stationary phase. Mobile phase components and additives affected chiral separation greatly. This new synthetic chiral stationary phase is complementary to two other related commercially available CSPs: the P-CAP and P-CAP-DP columns. Interactions between the chiral stationary phase and analytes that lead to retention and chiral recognition include hydrogen bonding, dipolar, and π–π interactions. Repulsive (steric) interactions also contribute to chiral recognition. Figure LC chromatograms showing the analytical (blue) and preparative (red) separations of N-(3,5-dinitrobenzoylleucine) enantiomers on a new synthetic polymeric chiral stationary phase     

Novel gel-based rapid test for non-instrumental detection of ochratoxin A in beer

A rapid easy-to-use immunoassay was optimised for the non-instrumental detection of ochratoxin A (OTA) in beer. The analytical method involves preconcentration on the immunoaffinity layer inside a column followed by direct competitive ELISA detection in the same layer. The visual cut-off value, i.e. the lowest OTA concentration resulting in no colour development, was 0.2 μg L^-1. Assay validation was performed using samples spiked with OTA. Thirty-seven naturally contaminated samples were screened with the gel-based method developed and no false-negative results were obtained. The method described offers a simple, rapid and cost-effective screening tool, thus contributing to better health protection of consumers. Figure Gel-based immunoassay of spiked beer samples.     

Investigating the post-chemiluminescence behavior of phenothiazine medications in the luminol–potassium ferricyanide system: molecular imprinting–post-chemiluminescence method for the determination of chlorpromazine hydrochloride

A new post-chemiluminescence (PCL) phenomenon was observed when phenothiazine medications were injected into the reaction mixture after the chemiluminescence (CL) reaction of luminol and potassium ferricyanide had finished. A possible reaction mechanism was proposed based on studies of the kinetic characteristics of the CL, CL spectra, fluorescence spectra, and on other experiments. The feasibility of determining various phenothiazine medications by utilizing these PCL reactions was examined. A molecular imprinting–post-chemiluminescence (MI-PCL) method was established for the determination of chlorpromazine hydrochloride using a chlorpromazine hydrochloride-imprinted polymer (MIP) as the recognition material. The method displayed high selectivity and high sensitivity. The linear range of the method was 1.0×10⁻⁸∼1.0×10⁻⁶, with a linear correlation coefficient of 0.9985. The detection limit was 3×10⁻⁹ g/ml chlorpromazine hydrochloride, and the relative standard deviation for a 1.0×10⁻⁷ g/ml chlorpromazine hydrochloride solution was 4.0% (n=11). The method has been applied to the determination of chlorpromazine hydrochloride in urine and animal drinking water with satisfactory results.      

Determination of norfloxacin in human urine by capillary electrophoresis with electrochemiluminescence detection

A fast and sensitive approach that can be used to detect norfloxacin in human urine using capillary electrophoresis with end-column electrochemiluminescence (ECL) detection of is described. The separation column was a 75-μm i.d. capillary. The running buffer was 15 mmol L⁻¹ sodium phosphate (pH 8.2). The solution in the detection cell was 50 mmol L⁻¹ sodium phosphate (pH 8.0) and 5 mmol L⁻¹ The ECL intensity varied linearly with norfloxacin concentration from 0.05 to 10 μmol L⁻¹. The detection limit (S/N=3) was 0.0048 μmol L⁻¹, and the relative standard deviations of the ECL intensity and the migration time for eleven consecutive injections of 1.0 μmol L⁻¹ norfloxacin (n=11) were 2.6% and 0.8%, respectively. The method was successfully applied to the determination of norfloxacin spiked in human urine without sample pretreatment. The recoveries were 92.7–97.9%.      

Electrochemical preparation of copper–dendrimer nanocomposites: picomolar detection of Cu<Superscript>2+</Superscript> ions

The present work describes, for the first time, in situ electrochemical preparation of dendrimer-encapsulated Cu nanoparticles using a self-assembled monolayer of fourth-generation amine-terminated polyamidoamine (PAMAM) dendrimer as the template. Atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) studies of the modified surface confirmed the presence of Cu nanoparticles entrapped in dendrimer film. Au electrode modified with a monolayer of the dendrimer enables preconcentration and subsequent voltammetric detection of Cu²⁺ at picomolar concentrations. Further, Cu nanoparticles in the dendrimer monolayer could be electrochemically derivatised to Cu hexacyanoferrate, which exhibits specific crystal planes, unlike the random distribution of crystal planes in bulk-formed Cu hexacyanoferrate, which is another catalytically active material for sensor applications. Figure Electrochemical preparation of copper–dendrimer nanocomposite     

Sequential preconcentration by coupling of field amplified sample injection with pseudo isotachophoresis–acid stacking for analysis of alkaloids in capillary electrophoresis

A novel on-column sequential preconcentration method based on the combination of field-amplified sample injection induced by acetonitrile and pseudo isotachophoresis (ITP)–acid stacking is developed for simply but efficiently concentrating alkaloid cations in a high-salt sample matrix in capillary electrophoresis. Acetonitrile (70%) added to a sample solution with a high-salt sample matrix not only induces field-amplified sample stacking by decreasing conductivity but also acts as a termination reagent in the succeeding pseudo ITP. After sample injection had been completed, a plug of H⁺ was injected electrokinetically and a neutralization reaction between H⁺ and tartrate from the buffer solution produced a low conductivity zone, in which the injected analyte cations were further concentrated. With the sequential preconcentration method, a 3 orders of magnitude detection sensitivity (1,400-fold) increase could be observed compared with the conventional electrokinetic injection method, without compromising separation efficiency and peak shape, and detection limits of 0.1 ng/mL for myosmine and 0.3 ng/mL for anabasine with the conditions selected were achieved. The calibration curves demonstrated good linearity in the concentration ranges 1.3–600 ng/mL for myosmine and 4.9–900 ng/mL for anabasine, respectively. The proposed method has been used to analyze successfully trace alkaloids in cigarette samples. Figure Sequential preconcentration processes: a sample injection; b introduction of HCl; c capillary zone electrophoresis separation. A ⁻ tartrate, white circles acetonitrile, black circles Na⁺, sample zone, myosmine, anabasine     

Rapid determination of p<Emphasis Type="Italic">K</Emphasis><Subscript>a</Subscript> values of 20 amino acids by CZE with UV and capacitively coupled contactless conductivity detections

A rapid and universal capillary zone electrophoresis (CZE) method was developed to determine the dissociation constants (pK _a) of the 20 standard proteogenic amino acids. Since some amino acids are poorly detected by UV, capacitively coupled contactless conductivity detection (C⁴D) was used as an additional detection mode. The C⁴D coupling proved to be very successful on a conventional CE-UV instrument, neither inducing supplementary analyses nor instrument modification. In order to reduce the analysis time for pK _a determination, two strategies were applied: (i) a short-end injection to reduce the effective length, and (ii) a dynamic coating procedure to generate a large electroosmotic flow (EOF), even at pH values as low as 1.5. As a result, the analysis time per amino acid was less than 2 h, using 22 optimized buffers covering a pH range from 1.5 to 12.0 at a constant ionic strength of 50 mM. pK _a values were calculated using an appropriate mathematical model describing the relationship between effective mobility and pH. The obtained pK _a values were in accordance with the literature. Figure a UV (1) and C⁴D (2) detectors placed on-line on the CE capillary. b Curve of effective mobility as a function of pH for histidine     

Profiling of 3,4-methylenedioxymethamphetamine by means of high-performance liquid chromatography

An impurity-profiling method for 3,4-methylenedioxymethamphetamine (MDMA) is presented. The impurities of interest were extracted by solid-phase extraction (SPE) on Bakerbond C₁₈ spe columns from a weakly alkaline solution (pH 8.5). Development of the extraction conditions covered selection of the buffer for dissolution of the sample and the volume of the eluent used to elute the impurities. An important part of the studies was to optimise the separation conditions, and the simplex method was used for this purpose. Cluster analysis was applied for comparison of samples and its grouping. The developed method was based on the areas of 33 selected peaks corresponding to MDMA impurities. All examined samples were correctly classified into clusters corresponding to the synthetic route.