Evaluation of hydrodynamic chromatography coupled to inductively coupled plasma mass spectrometry for speciation of dissolved and nanoparticulate gold and silver.

In this study, hydrodynamic chromatography coupled to inductively coupled plasma mass spectrometry has been evaluated for the simultaneous determination of dissolved and nanoparticulate species of gold and silver. Optimization of mobile phase was carried out with special attention to the column recovery of the different species and the resolution between them. Addition of 0.05 mM penicillamine to the mobile phase allowed the quantitative recovery of ionic gold and gold nanoparticles up to 50 nm, whereas 1 mM penicillamine was necessary for quantitative recovery of ionic silver and silver nanoparticles up to 40 nm. The resolution achieved between ionic gold and 10-nm gold nanoparticles was 0.7, whereas it ranged between 0.31 and 0.93 for ionic silver and 10-nm silver nanoparticles, depending on the composition of mobile phase. Best-case mass concentration detection limits for gold and silver species were 0.05 and 0.75 µg L-1, respectively. The developed methods allowed the simultaneous detection of nanoparticulate and dissolved species of gold and silver in less than 10 min. Size determination and quantification of gold and silver species were carried out in different dietary supplements, showing good agreement with the results obtained by electron microscopy and total and ultrafiltrable contents, respectively. Due to the attainable resolution, the quality of the quantitative results is affected by the relative abundance of nanoparticulate and dissolved species of the element and the size of the nanoparticles if present.

Detection, size characterization and quantification of silver nanoparticles in consumer products by particle collision coulometry.

Silver nanoparticles (AgNPs) are widely used in industrial and consumer products owing to its antimicrobial nature and multiple applications. Consequently, their release into the environment is becoming a big concern because of their negative impacts on living organisms. In this work, AgNPs were detected at a potential of + 0.70 V vs. Ag/AgCl reference electrode, characterized, and quantified in consumer products by particle collision coulometry (PCC). The electrochemical results were compared with those measured with electron microscopy and single-particle inductively coupled plasma mass spectrometry. The theoretical and practical peculiarities of the application of PCC technique in the characterization of AgNPs were studied. Reproducible size distributions of the AgNPs were measured in a range 10-100 nm diameters. A power allometric function model was found between the frequency of the AgNPs collisions onto the electrode surface and the number concentration of nanoparticles up to a silver concentration of 1010 L-1 (ca. 25 ng L-1 for 10 nm AgNPs). A linear relationship between the number of collisions and the number concentration of silver nanoparticles was observed up to 5 × 107 L-1. The PCC method was applied to the quantification and size determination of the AgNPs in three-silver containing consumer products (a natural antibiotic and two food supplements). The mean of the size distributions (of the order 10-20 nm diameters) agrees with those measured by electron microscopy. The areas of current spikes from the chronoamperogram allow the rapid calculation of size distributions of AgNPs that impact onto the working electrode.

Voltammetric sensing of silver nanoparticles on electrodes modified with selective ligands by using covalent and electropolymerization procedures. Discrimination between silver(I) and metallic silver.

New electrochemical sensors are described for the rapid quantification of silver nanoparticles (AgNPs). They are based on the immobilization of L-cysteine on a glassy carbon electrode (GCE) and on the formation of electropolymerized oligomers on the GCE. Ligands with amino, sulfur and carboxy functional groups are used that are capable of selectively retaining AgNPs. The experimental conditions for electropolymerizations were optimized for each of four monomers studied: L-lysine, thiophene-3-carboxamide, thionin, and o-phenylene-diamine. The best retention capabilities and conditions for quantification of AgNPs were found for immobilized nanoparticle voltammetry. This method is more sensitive than others based on the sorption of AgNPs from dispersions. These ligands also retain ionic silver species. Hence, Ag(I) and Ag0 can be discriminated, and the total silver content can be quantified by stripping voltammetry. The best analytical properties (for dispersions of AgNPs of 40 nm diameter) were found with GCEs carrying electropolymerized L-lysine. Figures of merit include (a) sensitivity of 4.329 ± 0.031 µA µg-1 mL cm-2, (b) a detection limit of 0.010 µg mL-1, and (c) a relative standars deviation of about 7.2% (for n = 4). The poly-L-lysine sensors can also evaluate the size of the AgNPs in the range 20 to 80 nm diameter, owing to displacements of the maximum potential of the voltammetry peaks. Graphical abstractSchematic representation of the quantification of silver nanoparticles (AgNPs) with a glassy-carbon electrode (GCE) modified with electropolymerized poly(L-lysine). AgNPs (and Ag+ ions) are selectively preconcentrated in the polymer and determined by differential-pulse stripping voltammetry.

Evaluation of agarose gel electrophoresis for characterization of silver nanoparticles in industrial products.

Agarose gel electrophoresis (AGE) has been used extensively for characterization of pure nanomaterials or mixtures of pure nanomaterials. We have evaluated the use of AGE for characterization of Ag nanoparticles (NPs) in an industrial product (described as strong antiseptic). Influence of different stabilizing agents (PEG, SDS, and sodium dodecylbenzenesulfonate), buffers (TBE and Tris Glycine), and functionalizing agents (mercaptosuccinic acid (TMA) and proteins) has been investigated for the characterization of AgNPs in the industrial product using different sizes-AgNPs standards. The use of 1% SDS, 0.1% TMA, and Tris Glycine in gel, electrophoresis buffer and loading buffer led to the different sizes-AgNPs standards moved according to their size/charge ratio (obtaining a linear relationship between apparent mobility and mean diameter). After using SDS and TMA, the behavior of the AgNPs in the industrial product (containing a casein matrix) was completely different, being not possible their size characterization. However we demonstrated that AGE with LA-ICP-MS detection is an alternative method to confirm the protein corona formation between the industrial product and two proteins (BSA and transferrin) maintaining NPs-protein binding (what is not possible using SDS-PAGE).

Evaluation of number concentration quantification by single-particle inductively coupled plasma mass spectrometry: microsecond vs. millisecond dwell times.

The quality of the quantitative information in single-particle inductively coupled plasma mass spectrometry (SP-ICP-MS) depends directly on the number concentration of the nanoparticles in the sample analyzed, which is proportional to the flux of nanoparticles through the plasma. Particle number concentrations must be selected in accordance with the data acquisition frequency, to control the precision from counting statistics and the bias, which is produced by the occurrence of multiple-particle events recorded as single-particle events. With quadrupole mass spectrometers, the frequency of data acquisition is directly controlled by the dwell time. The effect of dwell times from milli- to microseconds (10 ms, 5 ms, 100 µs, and 50 µs) on the quality of the quantitative data has been studied. Working with dwell times in the millisecond range, precision figures about 5 % were achieved, whereas using microsecond dwell times, the suitable fluxes of nanoparticles are higher and precision was reduced down to 1 %; this was independent of the dwell time selected. Moreover, due to the lower occurrence of multiple-nanoparticle events, linear ranges are wider when dwell times equal to or shorter than 100 µs are used. A calculation tool is provided to determine the optimal concentration for any instrument or experimental conditions selected. On the other hand, the use of dwell times in the microsecond range reduces significantly the contribution of the background and/or the presence of dissolved species, in comparison with the use of millisecond dwell times. Although the use of dwell times equal to or shorter than 100 µs offers improved performance working in single-particle mode, the use of conventional dwell times (3-10 ms) should not be discarded, once their limitations are known.

Physical and chemical characterization of cerium(IV) oxide nanoparticles.

Chemical composition, size and structure of the nanoparticle are required to describe nanoceria. Nanoparticles of similar size and Ce(III) content might exhibit different chemical behaviour due to their differences in structure. A simple and direct procedure based on affordable techniques for all the laboratories is presented in this paper. The combination of Raman and UV-vis spectroscopy and particle impact coulometry (PIC) allows the characterization of nanoceria of small size from 4 to 65 nm at a concentration from micromolar to nanomolar, a concentration range suitable for the analysis of lab-prepared or commercial nanoparticle suspensions, but too high for most analytical purposes aimed at nanoparticle monitoring. While the PIC limits of size detection are too high to observe small nanoparticles unless catalytic amplification is used, the method provides a simple means to study aggregation of nanoparticles in the media they are needed to be dispersed for each application. Raman spectroscopy provided information about structure of the nanoparticle, and UV-vis about their chemical behaviour against some common reducing and oxidizing agents. Graphical Abstract To characterize nanoceria it is necessary to provide information about the shape, size and structure of the nanoparticles as well as the chemical composition.

Evaluation of gel electrophoresis techniques and laser ablation-inductively coupled plasma-mass spectrometry for screening analysis of Zn and Cu-binding proteins in plankton.

The determination of metal-binding proteins in plankton is important because of their involvement in photosynthesis, which is fundamental to the biogeochemical cycle of the oceans and other ecosystems. We have elaborated a new strategy for screening of Cu and Zn-containing proteins in plankton on the basis of separation of proteins by use of Blue-Native PAGE (BN-PAGE), which entails use of a non-denaturing Tris-tricine system and detection of metals in the proteins by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). For comparison, denaturing PAGE based on Tris-glycine and Tris-tricine systems and Anodic-Native PAGE have also been investigated. A large number of protein bands with MW between 20 and 75 kDa were obtained by use of Tris-glycine PAGE but detection of metals by LA-ICP-MS was unsuccessful because of loss of metals from the proteins during the separation process. Different protein extraction, purification, and preconcentration methods were evaluated, focussing on both issues-achieving the best extraction and characterization of the proteins while maintaining the integrity of metal-protein binding in the plankton sample. Use of 25 mmol L(-1) Tris-HCl and a protease inhibitor as extraction buffer with subsequent ultrafiltration and acetone precipitation was the most efficient means of sample preparation. Two Cu and Zn proteins were detected, a protein band corresponding to a MW of 60 kDa and another poorly resolved band with a MW between 15 and 35 kDa.

Applicability of a Material Constitutive Model Based on a Transversely Isotropic Behaviour for the Prediction of the Mechanical Performance of Multi Jet Fusion Printed Polyamide 12 Parts.

Multi Jet Fusion (MJF), an innovative additive manufacturing (AM) technique in the field of Powder Bed Fusion (PBF) developed by Hewlett-Packard (HP) Inc. (Palo Alto, CA, USA), has been designed to produce polymer parts using thermoplastic-based powders, primarily focusing on polyamide 12 (PA12). Employing a layer-by-layer approach, MJF enables the rapid production of intricate components, reportedly up to 10 times faster than other AM processes. While the mechanical properties of MJF-printed PA12 and the impact of build orientation on those properties have already been explored in various studies, less attention has been given to the mechanical performance of MJF-printed PA12 components under complex loads and accurate predictive models. This contribution aims to assess the applicability of a constitutive model based on a transversely isotropic behaviour under linear elastic deformation for predicting the mechanical response of MJF-printed PA12 parts through numerical simulations. Both uniaxial tensile and shear tests were carried out on printed samples to determine the elastic properties of MJF-printed PA12, with additional testing on printed complex handle-shaped parts. Finally, a numerical model was developed to simulate the mechanical tests of the handles. Results from tests on printed samples showed that MJF-printed PA12, to some extent, behaves as a transversely isotropic material. Furthermore, using a constitutive model that assumes a transversely isotropic behaviour under linear elastic deformation for predicting the mechanical response of MJF-printed PA12 parts in numerical simulations could be a reasonable approach, provided that the material stress levels remain within the linear range. However, the particularities of the stress-strain curve of MJF-printed PA12 complicate determining the elasticity-to-plasticity transition point.

An electrochemical immunosensor for ochratoxin A determination in wines based on a monoclonal antibody and paramagnetic microbeads.

We report a direct competitive immunosensor for the rapid determination of ochratoxin A (OTA) in wine samples. Magnetic beads (1 ± 0.5 µm diameter) covered with streptavidin were functionalized with a monoclonal antibody against OTA, and then left to incubate in a solution of tracer (ochratoxin conjugated to the enzyme peroxidase) and a range of OTA concentrations (10(-4) to 1,000 ng mL(-1)). After washing and separation steps helped with a magnetic field, a volume of the dispersion was put on screen-printed electrodes under a magnet, and after adding the substrate the p-benzoquinone generated enzymatically was detected by differential-pulse voltammetry. Wine samples (2 mL) were easily prepared simply by adjusting to pH = 7.5 with diluted NaOH and by adding polyvinylpyrrolidone for complexing polyphenols, without any other clean-up or preconcentration steps. The limit of detection for detecting OTA in wines was of 0.11 ± 0.01 ng L(-1), well below the permitted content of the mycotoxin by the European Union (<2 ng mL(-1)). Spiked wines were subjected to immunosensor calibrations to study the matrix effects. OTA concentrations measured with the immunosensor were compared with those obtained by high-performance liquid chromatography coupled to fluorescence detection (AOAC official method 2001.01). The OTA levels from two red wines of "Campo de Borja", Spain, ranged from about 0.027 to 0.033 ng mL(-1) of OTA.

How the use of a short channel can improve the separation efficiency of nanoparticles in asymmetrical flow field-flow fractionation.

The use of a commercially available short length channel (14 cm length) is proposed to improve the efficiency associated to the separation by asymmetrical flow field-flow fractionation of particles in the nanometer range respect to a standard channel (27 cm length). The effect of channel length on elution times, separation efficiency and resolution have been studied. Polystyrene particles between 50 and 500 nm in size have been used to compare the behavior of both channels. Theoretical aspects based on the different contributions on particle diffusion inside the channel during the separation process have been considered to justify the results obtained. Non-equilibrium diffusion contribution to the efficiency has shown to be the most relevant aspect to be controlled during the separation. The increment of the field strength applied through the cross-flow velocityallows the reduction of diffusion while keep elution times constant. The use of the same cross-flow in a channel with a smaller area is the key factor that justifies the better efficiencies observed along the whole size range studied (improvements that reach factors up to 4.7 in experimental efficiency respect to the standard channel were achieved). The separation of polystyrene particles of 100 and 200 nm was achieved with a resolution of 1.20, whereas a 0.66 value was obtained with the standard channel at the same elution times. Channel recoveries have been also compared under optimized conditions to ensure that no side effects are produced, including the separation of mixtures of TiO2 nanoparticles. Similar or even better values were obtained with the short length channel, with recoveries higher than 85% for all the polystyrene particles tested and 75% recovery for the TiO2 nanoparticle mixture, which justifies its use for the separation of nanoparticles, providing better resolutions without compromise elution times or recoveries.

Inorganic and organic characterization of Santa Lucía salt mine peloid for quality evaluations.

Santa Lucía peloid is a sediment used in pelotherapy in Cuban primary health care services. Therefore, in addition to physicochemical regulated parameters, other analyses are required to complement their physicochemical characterization and understand potential element mobility, radiological risk, and toxicity as well as likely bioactive compounds present in Santa Lucía peloid. For these purposes, inorganic and organic elements and compounds were considered to evaluate Santa Lucía peloid's quality. This was accomplished through an integral approach that included (1) determination of physicochemical parameters (pH, electrical conductivity, oxidation-reduction potential, temperature, dissolved oxygen, elemental C, H, and N analyses, organic matter, and hexane removable substances content); (2) determination of total concentration of elements with biological and toxicological importance (i.e., Na, K, Ca, Mg, Fe, Mn, Cr, Cu, Ni, Pb, and Zn), as well as their distribution in operationally defined solid phases, mineralogy, particle size distribution, and total content of radionuclides and radiological dose calculations; and (3) its organic characterization. Results from this study showed that Santa Lucía peloid was non-contaminated and showed low metal mobility and acceptable radiological dose levels, being safe for therapeutic uses. Additionally, these results contribute to the understanding of the organic composition of peloides, provide strong evidences to scientifically explain the therapeutic action of peloids in the treatment of inflammatory diseases, and set a new frame to improve peloid guidelines in Cuba and other countries.

Some pitfalls in PAGE-LA-ICP-MS for quantitative elemental speciation of dissolved organic matter and metalomics.

An experimental approach to evaluate the capabilities and limitations of polyacrylamide gel electrophoresis (PAGE) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) for quantitative elemental speciation is presented. Two metalloproteins (superoxide dismutase, containing Cu and Zn, and thyroglobulin, containing I) with high binding affinity for metals, and metal-dissolved organic matter (DOM) complexes (from a compost leachate sample) which show different types of metal binding are studied. Iodine can be quantitatively detected in thyroglobulin after PAGE-LA-ICP-MS using either sodium dodecyl sulfate (SDS) PAGE or native PAGE. However, detection of Cu and Zn in superoxide dismutase after PAGE-LA-ICP-MS depends on the conditions of the PAGE method because possible metal losses can occur (either with SDS-PAGE or with native PAGE). The use of PAGE-LA-ICP-MS to study the contribution of DOM to the mobilization of metals from environmental samples is possible, but it depends also on the PAGE separation conditions owing to disequilibrium effects of metal-DOM complexes.

A rapid magnetic particle-based enzyme immunoassay for human cytomegalovirus glycoprotein B quantification.

Human cytomegalovirus (HCMV) is a herpes virus that can cause severe infections. Still, the available methods for its diagnostic have the main disadvantage of requiring long time to be performed. In this work, a simple magnetic particle-based enzyme immunoassay (mpEIA) for the quantification of glycoprotein B of Human cytomegalovirus (gB-HCMV) in urine samples is proposed. The immunosensor scheme is based on the analyte protein gB-HCMV sandwiched between a primary monoclonal antibody, (MBs-PrG-mAb1), and a secondary anti-gB-HCMV antibody labelled with Horseradish peroxidase (Ab2-HRP) to allow spectrophotometric detection. The mpEIA analytical performance was tested in urine samples, showing a linear dependence between gB-HCMV concentration and the absorbance signal at 450 nm in a range of concentrations from 90 to 700 pg mL-1. The calculated detection limits for gB-HCMV were 90 ±â€¯2 pg mL-1 and the RSD was about 6.7% in urine samples. The immunosensor showed good selectivity against other viruses from Herpesviridae family, namely varicella zoster and Epstein Barr viruses. The recoveries of spiked human urine samples at 0.30-0.50 ng mL-1 concentration levels of gB-HCMV ranged between 91 to 105%. The proposed mpEIA method was validated following the guidelines of the European Medicines Agency (EMEA-2014), and allows rapid, successful and easy quantification of gB-HCMV in urine samples.

Mobilization and speciation of chromium in compost: a methodological approach.

An integral approach to study the mobility of chromium in compost is presented. The approach is based on batch pH dependence leaching tests and the analysis of the leachates for total chromium, chromium(VI) and complexes of chromium(III) with natural organic matter. As leachings are performed with no aggressive reagents (ultrapure water with added nitric acid or potassium hydroxide), the method can be considered a good approach to simulate natural scenarios. The analysis of leachates is complemented with the determination of total chromium and total Cr(VI) in the solid sample. Speciation analysis are done by high performance liquid chromatography with inductively coupled plasma mass spectrometry as detection technique; Cr(VI) is determined by ion chromatography, whereas Cr(III) complexes with natural organic matter by size exclusion chromatography. In the compost studied, Cr(VI) accounted for 6% of the total chromium in the solid, whereas no significant amounts of Cr(VI) were mobilized in the pH range studied (4-10). Chromium is mobilized as Cr(III) bound to organic matter, both to humic and fulvic acids, with an increasing contribution of humic acids at higher pHs.

Rapid determination of recent cocaine use with magnetic particles-based enzyme immunoassays in serum, saliva, and urine fluids.

Cocaine is one of the most worldwide used illicit drugs. We report a magnetic particles-based enzyme-linked immunoassay (mpEIA) method for the rapid and sensitive determination of cocaine (COC) in saliva, urine and serum samples. Under optimized conditions, the limits of detections were 0.09ngmL(-1) (urine), 0.15ngmL(-1) (saliva), and 0.06ngmL(-1) COC (human serum). Sensitivities were in the range EC50=0.6-2.5ngmL(-1) COC. The cross-reactivity with the principal metabolite benzoylecgonine (BZE) was only 1.6%. Recovering percentages of doped samples (0, 10, 50, and 100ngmL(-1) of COC) ranged from about 86-111%. Some advantages of the developed mpEIA over conventional ELISA kits are faster incubations, improved reproducibility, and consumption of lower amounts of antibody and enzyme conjugates due to the use of magnetic beads. The reported method was validated following the guidelines on bioanalytical methods of the European Medicines Agency (2011). Unmetabolized COC detection has a great interest in pharmacological, pharmacokinetics, and toxicokinetics studies, and can be used to detect a very recent COC use (1-6h).

Detection, characterization and quantification of inorganic engineered nanomaterials: A review of techniques and methodological approaches for the analysis of complex samples.

The increasing demand of analytical information related to inorganic engineered nanomaterials requires the adaptation of existing techniques and methods, or the development of new ones. The challenge for the analytical sciences has been to consider the nanoparticles as a new sort of analytes, involving both chemical (composition, mass and number concentration) and physical information (e.g. size, shape, aggregation). Moreover, information about the species derived from the nanoparticles themselves and their transformations must also be supplied. Whereas techniques commonly used for nanoparticle characterization, such as light scattering techniques, show serious limitations when applied to complex samples, other well-established techniques, like electron microscopy and atomic spectrometry, can provide useful information in most cases. Furthermore, separation techniques, including flow field flow fractionation, capillary electrophoresis and hydrodynamic chromatography, are moving to the nano domain, mostly hyphenated to inductively coupled plasma mass spectrometry as element specific detector. Emerging techniques based on the detection of single nanoparticles by using ICP-MS, but also coulometry, are in their way to gain a position. Chemical sensors selective to nanoparticles are in their early stages, but they are very promising considering their portability and simplicity. Although the field is in continuous evolution, at this moment it is moving from proofs-of-concept in simple matrices to methods dealing with matrices of higher complexity and relevant analyte concentrations. To achieve this goal, sample preparation methods are essential to manage such complex situations. Apart from size fractionation methods, matrix digestion, extraction and concentration methods capable of preserving the nature of the nanoparticles are being developed. This review presents and discusses the state-of-the-art analytical techniques and sample preparation methods suitable for dealing with complex samples. Single- and multi-method approaches applied to solve the nanometrological challenges posed by a variety of stakeholders are also presented.

Flow injection electrochemical hydride generation of hydrogen selenide on lead cathode: critical study of the influence of experimental parameters.

A flow injection system for the determination of selenium by electrochemical hydride generation and quartz tube atomic absorption spectrometry is described. The generator consists of an electrolytic flow-through cell with a concentric arrangement and a packed cathode made of particulated lead. The influences of sample flow rate, carrier gas flow rate and electrolysis current on the hydrogen selenide generation have been critically studied. Both sample flow rate and electrolysis current play important roles in the efficiency of the hydride generation process. A characteristic mass of 2.4 ng and a concentration detection limit of 17 microg l(-1) were obtained for a sample volume of 420 microl.

Development of a platinized and ferrocene-mediated cholesterol amperometric biosensor based on electropolymerization of polypyrrole in a flow system.

The preparation of a cholesterol amperometric biosensor using a platinized Pt electrode as a support for the electropolymerization of a polypyrrole film, in which cholesterol oxidase and ferrocene monocarboxylic acid (electron-transfer mediator) were co-entrapped, is described. All the biosensor preparation steps (platinization and electropolymerization) and the cholesterol determination take place in the same flow system. The presence of the mediator enhances the sensitivity and selectivity of the platinized biosensor without modifying the dynamic parameters of the response, and the platinized layer improves the operational lifetime of the mediated sensor. The sensitivity obtained was 88.51 nA mM(-1) and the limit of detection was 12.4 microM of cholesterol. The analytical properties of the biosensor for the flow-injection determination of cholesterol were studied and compared with those of other more simple amperometric biosensor configurations.