Phase-changing sacrificial layers in microfluidic devices: adding another dimension to separations

The use of polymers in microchip fabrication affords new opportunities for the development of powerful, miniaturized separation techniques. One method in particular, the use of phase-changing sacrificial layers, allows for simplified designs and many additional features to the now standard fabrication of microchips. With the possibility of adding a third dimension to the design of separation devices, various means of enhancing analysis now become possible. The application of phase-changing sacrificial layers in microchip analysis systems is discussed, both in terms of current uses and future possibilities. Figure Phase-changing sacrificial materials enable multilayer microfluidic device layouts     

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)     

Chiral ligand-exchange chromatography of amino acids using porous graphitic carbon coated with a dinaphthyl derivative of neamine

In this paper, we describe the preparation and the evaluation of a porous graphitic carbon (PGC) column coated with a new dinaphthyl derivative of neamine for chiral ligand-exchange (LE) chromatography. It was shown that the graphitic surface/dinaphthyl anchor system efficiently (1.15 μmol/m²) and stably (three months of intensive use) adsorbs the neamine template onto the chromatographic support. The resulting coated PGC stationary phase showed appreciable LE-based enantioselective properties towards several native amino acids. Chromatographic separation of methionine enantiomers using a dinaphtyl neamine-based ligand-exchange chiral stationary phase     

Enhanced resonance light scattering properties of gold nanoparticles due to cooperative binding

The interaction of 11-mercaptoundecanoic acid capped gold nanoparticles (MUA-GNPs) with europium ions and aminoacids has been studied by UV-Vis spectrophotometry, fluorescence, confocal fluorescence microscopy, resonance light scattering and TEM. Results demonstrated that hyper-Rayleigh scattering emission occurs upon the addition of lysine to the MUA-GNPs–Eu(III) system, thus providing an inherently sensitive method for lysine determination. The effects of geometrical factors of the gold nanoparticles (aspect ratio, particle size, cluster formation) and the surrounding medium (pH) on this behavior are discussed. The cooperative binding interactions of Eu³⁺ and lysine with gold nanoparticles permitted the discrimination of lysine from other amino acids. The probable mechanism for the spectral changes and the enhanced resonance light scattering observed is outlined. Figure Gold nanoparticle resonance light scattering plasmon enhancement through cooperative binding with europium and lysine     

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     

Hydrophilic interaction liquid chromatography (HILIC) in proteomics

In proteomics, nanoflow multidimensional chromatography is now the gold standard for the separation of complex mixtures of peptides as generated by in-solution digestion of whole-cell lysates. Ideally, the different stationary phases used in multidimensional chromatography should provide orthogonal separation characteristics. For this reason, the combination of strong cation exchange chromatography (SCX) and reversed-phase (RP) chromatography is the most widely used combination for the separation of peptides. Here, we review the potential of hydrophilic interaction liquid chromatography (HILIC) as a separation tool in the multidimensional separation of peptides in proteomics applications. Recent work has revealed that HILIC may provide an excellent alternative to SCX, possessing several advantages in the area of separation power and targeted analysis of protein post-translational modifications. Figure Artistic impression of the HILIC separation mechanism     

A viscosity-tunable polymer for DNA separation by microchip electrophoresis

A thermo-responsive separation matrix, consisting of Pluronic F127 tri-block copolymers of poly(ethylene oxide) and poly(propylene oxide), was used to separate DNA fragments by microchip electrophoresis. At low temperature, the polymer matrix was low in viscosity and allowed rapid loading into a microchannel under low pressure. With increasing temperatures above 25°C, the Pluronic F127 solution forms a liquid crystalline phase consisting of spherical micelles with diameters of 17–19 nm. The solution can be used to separate DNA fragments from 100 bp to 1500 bp on poly(methyl methacrylate) (PMMA) chips. This temperature-sensitive and viscosity-tunable polymer provided excellent resolution over a wide range of DNA sizes. Separation is based on a different mechanism compared with conventional matrices such as methylcellulose. To illustrate the separation mechanism of DNA in a Pluronic F127 solution, DNA molecular imaging was performed by fluorescence microscopy with F127 polymer as the separation matrix in microchip electrophoresis. Figure Temperature dependence of the viscosity of 20% w/w Pluronic F127 solution in 1xTBE buffer. Dotted approximates resultant curve.     

Two-dimensional liquid chromatography of synthetic polymers

Two-dimensional liquid chromatography, 2D-LC of synthetic polymers is critically assessed. Similarities and differences of 2D-LC of low-molecular-mass and polymeric substances are reviewed. The rationale of application of 2D-LC to macromolecular substances is discussed. Basic information on retention mechanisms in liquid chromatography of synthetic polymers is furnished. The principles, reasons, and significance of coupling of retention mechanisms are explained. The resulting separation processes are elucidated, and the technical concepts of the corresponding experimental arrangements are described. The benefits of 2D-LC are demonstrated together with numerous problems and shortcomings of the method.      

Semi-quantitative determination of cationic surfactants in aqueous solutions using gold nanoparticles as reporter probes

Concentrations of cationic surfactants in aqueous solutions have been estimated on the basis of changes in the color of gold nanoparticles, used as reporter probes. We have shown that the colors of gold nanoparticles with anionic protective groups on their surfaces shift from red to indigo/purple and then back to red in a range of cationic surfactant solutions in which concentrations vary from very low to above the theoretical CMCs. The color changes occur near the theoretical CMCs, presumably because the presence of surfactant micelles in the solution prevents the gold nanoparticles from aggregating. We have used gold nanoparticles as reporter probes to determine the concentrations of cationic surfactants in products such as hair conditioners, which often contain large amounts of alkyltrimethylammonium halides. Although this approach can only provide an estimate, it can be performed simply by addition of a given amount of gold nanoparticles to a series of diluted solutions, without the need for instruments or labor-intensive procedures. Figure Photographs of a series of diluted hair conditioner solutions with added gold nanoparticles

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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     

Characterisation of historical organic dyestuffs by liquid chromatography–mass spectrometry

This review discusses the characterisation of natural organic dyestuffs of historical interest by liquid chromatography–mass spectrometry. The structures of the most important natural organic dyestuffs traditionally used are presented and discussed from the perspective of their analytical chemical determination. The practical aspects of the determination of this inhomogeneous range of compounds with different structures, such as anthraquinones, flavonoids, indigoids or tannins, are discussed with their implications for sample preparation, liquid chromatographic separation and mass spectrometric detection. The particular focus of this review is the discussion of the mass spectral fragmentation patterns of the different classes of natural organic dyestuffs, which in the ideal case allow the identification of the dyestuff actually used, and thereby provide a key to the better characterisation and understanding of historical objects dyed with natural organic dyestuffs. Figure LC-MS allows characterisation of natural dyestuff constituents: the MS spectrum of alizarin is superimposed over a photo of a textile coloured using this red dye     

Addition of gold nanoparticles to real-time PCR: effect on PCR profile and SYBR Green I fluorescence

Real-time quantitative polymerase chain reaction (qPCR) is the industry standard technique for the quantitative analysis of nucleic acids due to its unmatched sensitivity and specificity. Optimisation and improvements of this fundamental technique over the past decade have largely consisted of attempts to allow faster and more accurate ramping between critical temperatures by improving assay reagents and the thermal geometry of the PCR chamber. Small gold nanoparticles (Au-NPs) have been reported to improve PCR yield under fast cycling conditions. In this study, we investigated the effect of Au-NPs on optimised real-time qPCR assays by amplifying DNA sequences from genetically modified canola in the presence and absence of 0.9 nM Au-NPs of diameter 12 ± 2nm. Contrary to expectations, we found that Au-NPs altered the PCR amplification profile when using a SYBR Green I detection system due to fluorescence quenching; furthermore, high-resolution melt (HRM) analysis demonstrated that Au-NPs destabilised the double-stranded PCR product. The results indicate that effects on the assay detection system must be carefully evaluated before Au-NPs are included in any qPCR assay. Figure Raw amplification profiles in the presence and absence of gold nanoparticles     

Application of anodized aluminum in fluorescence detection of biological species

Thin nanoporous alumina obtained by anodization of aluminum films offers promising advantages for application in fluorescence-based biological sensors including convenient preparation, increased density of binding sites, and improved collection efficiency of fluorescence. These advantages are illustrated in the detection of streptavidin using biotin covalently bound to the surface of alumina nanopores. Fluorescence intensity enhancement as high as 7 times is observed in nanopores in comparison to flat glass surface.      

Nanoscale organic and polymeric field-effect transistors as chemical sensors

This article reviews recently published work concerning improved understanding of, and advancements in, organic and polymer semiconductor vapor-phase chemical sensing. Thin-film transistor sensors ranging in size from hundreds of microns down to a few nanometers are discussed, with comparisons made of sensing responses recorded at these different channel-length scales. The vapor-sensing behavior of nanoscale organic transistors is different from that of large-scale devices, because electrical transport in a nanoscale organic thin-film transistor depends on its morphological structure and interface properties (for example injection barrier) which could be modulated by delivery of analyte. Materials used in nanoscale devices, for example nanoparticles, nanotubes, and nanowires, are also briefly summarized in an attempt to introduce other relevant nano-transducers.      

Inorganic mass spectrometry as a tool for characterisation at the nanoscale

Inorganic mass spectrometry techniques may offer great potential for the characterisation at the nanoscale, because they provide unique elemental information of great value for a better understanding of processes occurring at nanometre-length dimensions. Two main groups of techniques are reviewed: those allowing direct solid analysis with spatial resolution capabilities, i.e. lateral (imaging) and/or in-depth profile, and those for the analysis of liquids containing colloids. In this context, the present capabilities of widespread elemental mass spectrometry techniques such as laser ablation coupled with inductively coupled plasma mass spectrometry (ICP-MS), glow discharge mass spectrometry and secondary ion/neutral mass spectrometry are described and compared through selected examples from various scientific fields. On the other hand, approaches for the characterisation (i.e. size, composition, presence of impurities, etc.) of colloidal solutions containing nanoparticles by the well-established ICP-MS technique are described. In this latter case, the capabilities derived from the on-line coupling of separation techniques such as field-flow fractionation and liquid chromatography with ICP-MS are also assessed. Finally, appealing trends using ICP-MS for bioassays with biomolecules labelled with nanoparticles are delineated.      

Aptamers as molecular recognition elements for electrical nanobiosensors

Recent advances in nanotechnology have enabled the development of nanoscale sensors that outperform conventional biosensors. This review summarizes the nanoscale biosensors that use aptamers as molecular recognition elements. The advantages of aptamers over antibodies as sensors are highlighted. These advantages are especially apparent with electrical sensors such as electrochemical sensors or those using field-effect transistors. Figure Feeling proteins with aptamer-functionalized carbon nanotubes     

Site-specific binding of chelerythrine and sanguinarine to single pyrimidine bulges in hairpin DNA

Spectrofluorometric titration, electrospray ionization time-of-flight mass spectrometric and UV melting methods were employed to study the binding of chelerythrine and sanguinarine to bulged DNA. The results showed that both alkaloids bind specifically to single pyrimidine (C, T) bulge sites. The ability of sanguinarine to bind to both regular and bulged hairpins was found to be stronger than that of chelerythrine, but the binding selectivity of chelerythrine toward single-base bulges was much larger than that of sanguinarine. Figure Association constants for chelerythrine and sanguinarine toward regular and single-base bulged hairpins obtained from fluorometric analysis     

The size distribution of 'gold standard' nanoparticles

The spherical gold nanoparticle reference materials RM 8011, RM 8012, and RM 8013, with a nominal radius of 5, 15, and 30 nm, respectively, have been available since 2008 from NIST. These materials are recommended as standards for nanoparticle size measurements and for the study of the biological effects of nanoparticles, e.g., in pre-clinical biomedical research. We report on determination of the size distributions of these gold nanoparticles using different small-angle X-ray scattering (SAXS) instruments. Measurements with a classical Kratky type SAXS instrument are compared with a synchrotron SAXS technique. Samples were investigated in situ, positioned in capillaries and in levitated droplets. The number-weighted size distributions were determined applying model scattering functions based on (a) Gaussian, (b) log-normal, and (c) Schulz distributions. The mean radii are 4.36 ± 0.04 nm (RM 8011), 12.20 ± 0.03 nm (RM 8012), and 25.74 ± 0.27 nm (RM 8013). Low polydispersities, defined as relative width of the distributions, were detected with values of 0.067 ± 0.006 (RM 8011), 0.103 ± 0.003, (RM 8012), and 0.10 ± 0.01 (RM 8013). The results are in agreement with integral values determined from classical evaluation procedures, such as the radius of gyration (Guinier) and particle volume (Kratky). No indications of particle aggregation and particle interactions—repulsive or attractive—were found. We recommend SAXS as a standard method for a fast and precise determination of size distributions of nanoparticles.