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Showing papers in "Analyst in 2014"


Journal ArticleDOI
26 Aug 2014-Analyst
TL;DR: A generalized table of extinction coefficient data for silver nanoparticles from 8 to 100 nm is presented, in good agreement with collated literature values measured by different authors with non-standardized methodology and each for a limited range of particle size.
Abstract: We present a generalized table of extinction coefficient data for silver nanoparticles from 8 to 100 nm. This table allows for easy and quick estimation of the concentration and size of modified and mono-dispersed silver nanoparticles from their optical spectra. We obtained data by determining the silver content of citrate-stabilised silver nanoparticles using sodium cyanide to dissolve the nanoparticles, and measuring solution conductivity with a pH meter and a cyanide-ion selective electrode. The quantification of the silver ion concentration enabled the calculation of extinction coefficients. Experimentally calculated extinction coefficients, in the current work, are in good agreement with collated literature values measured by different authors with non-standardized methodology and each for a limited range of particle size. They are also in good agreement with our theoretical calculations using Mie theory. Thus, we provide a highly standardized and comprehensive tabulated reference data-set.

526 citations


Journal ArticleDOI
06 May 2014-Analyst
TL;DR: This Critical Review aims to summarize progress that might enable practical applications of aptamers for biological samples that have been discussed in more detail since they are more likely to work in a complex sample matrix.
Abstract: Aptamers are single-stranded nucleic acids that selectively bind to target molecules. Most aptamers are obtained through a combinatorial biology technique called SELEX. Since aptamers can be isolated to bind to almost any molecule of choice, can be readily modified at arbitrary positions and they possess predictable secondary structures, this platform technology shows great promise in biosensor development. Over the past two decades, more than one thousand papers have been published on aptamer-based biosensors. Given this progress, the application of aptamer technology in biomedical diagnosis is still in a quite preliminary stage. Most previous work involves only a few model aptamers to demonstrate the sensing concept with limited biomedical impact. This Critical Review aims to summarize progress that might enable practical applications of aptamers for biological samples. First, general sensing strategies based on the unique properties of aptamers are summarized. Each strategy can be coupled to various signaling methods. Among these, a few detection methods including fluorescence lifetime, flow cytometry, upconverting nanoparticles, nanoflare technology, magnetic resonance imaging, electronic aptamer-based sensors, and lateral flow devices have been discussed in more detail since they are more likely to work in a complex sample matrix. The current limitations of this field include the lack of high quality aptamers for clinically important targets. In addition, the aptamer technology has to be extensively tested in a clinical sample matrix to establish reliability and accuracy. Future directions are also speculated to overcome these challenges.

428 citations


Journal ArticleDOI
03 Mar 2014-Analyst
TL;DR: The new mouthguard enzymatic biosensor, based on an immobilized lactate oxidase and a low potential detection of the peroxide product, exhibits high sensitivity, selectivity and stability using whole human saliva samples.
Abstract: The present work describes the first example of a wearable salivary metabolite biosensor based on the integration of a printable enzymatic electrode on a mouthguard. The new mouthguard enzymatic biosensor, based on an immobilized lactate oxidase and a low potential detection of the peroxide product, exhibits high sensitivity, selectivity and stability using whole human saliva samples. Such non-invasive mouthguard metabolite biosensors could tender useful real-time information regarding a wearer's health, performance and stress level, and thus hold considerable promise for diverse biomedical and fitness applications.

293 citations


Journal ArticleDOI
15 Apr 2014-Analyst
TL;DR: Fluorescent B-doped carbon quantum dots were prepared by a facile one-pot solvothermal route and can be used as a novel fluorescence sensing system for hydrogen peroxide and glucose detection.
Abstract: Fluorescent B-doped carbon quantum dots (BCQDs) were prepared by a facile one-pot solvothermal route. The BCQDs can be used as a novel fluorescence sensing system for hydrogen peroxide and glucose detection.

243 citations


Journal ArticleDOI
15 Apr 2014-Analyst
TL;DR: The most common problems encountered during sample preparation, ways to optimize established sample preparation techniques and important recent developments to reduce or eliminate major interferents from biofluids are described.
Abstract: Liquid chromatography-mass spectrometry analysis of small molecules from biofluids requires sensitive and robust assays. Because of the very complex nature of many biological samples, efficient sample preparation protocols to remove unwanted components and to selectively extract the compounds of interest are an essential part of almost every bioanalytical workflow. This review describes the most common problems encountered during sample preparation, ways to optimize established sample preparation techniques and important recent developments to reduce or eliminate major interferents from biofluids.

220 citations


Journal ArticleDOI
03 Feb 2014-Analyst
TL;DR: Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy is a technique capable of in situ characterization of the liquid-solid interface to probe surface adsorption on nanoparticle surfaces in environmentally and biologically relevant media and can directly measure ligand displacement reactions.
Abstract: Given the importance of nanoparticle surface composition in nanotoxicology, analytical tools that can probe nanoparticle surfaces in aqueous media are crucial but remain limited. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy is a technique capable of in situ characterization of the liquid–solid interface to probe surface adsorption on nanoparticle surfaces in environmentally and biologically relevant media. Furthermore, given that the interfacial region in these media is dynamic, ATR-FTIR spectroscopy facilitates monitoring these dynamics by interrogating a layer of immobilized nanoparticles coated on the ATR element while changing the overlying aqueous phase. The molecular information acquired from this technique allows for the determination of the adsorption mode, including conformational and structural changes of the coordinating ligand, and can directly measure ligand displacement reactions. Furthermore, in some cases, ATR-FTIR spectroscopy can be used as a quantitative surface analytical tool. In this article, we briefly review the fundamentals of the technique and then provide several examples of using ATR-FTIR spectroscopy to probe nanoparticle surfaces in general with respect to: (i) the adsorption of different environmentally and biologically relevant coordinating ligands; (ii) competitive ligand adsorption and; (iii) the determination of kinetic and thermodynamic parameters. We have also investigated surface adsorption of TiO2 nanoparticles in different biological media typically used for toxicity studies and show that the surface composition of TiO2 nanoparticles depends to a large extent on the composition of the medium due to surface adsorption. This result has important implications for the interpretation of toxicity data as well as inter-comparisons between toxicity studies.

201 citations


Journal ArticleDOI
Qun Zeng1, Yuzhi Wang1, Yanhua Huang1, Xueqin Ding1, Jing Chen1, Kaijia Xu1 
15 Apr 2014-Analyst
TL;DR: Results showed that hydrophobic interactions, hydrogen bonding interactions and the salting-out effect played important roles in the transfer process, and the aggregation and surrounding phenomenon were the main driving forces for the separation.
Abstract: Four kinds of green deep eutectic solvent (DES) were synthesized, including choline chloride (ChCl)–urea, tetramethylammonium chloride (TMACl)–urea, tetrapropylammonium bromide (TPMBr)–urea and ChCl–methylurea. An aqueous two-phase system (ATPS) based ChCl–urea DES was studied for the first time for the extraction of bovine serum albumin (BSA). Single factor experiments proved that the extraction efficiency of BSA was influenced by the mass of the DES, concentration of K2HPO4 solution, separation time and extraction temperature. The optimum conditions were determined through an orthogonal experiment with the four factors described above. The results showed that under the optimum conditions, the average extraction efficiency could reach up to 99.94%, 99.72%, 100.05% and 100.05% (each measured three times). The relative standard deviations (RSD) of extraction efficiencies in precision, repeatability and stability experiments were 0.5533% (n = 5), 0.8306% (n = 5) and 0.9829% (n = 5), respectively. UV-vis and FT-IR spectra confirmed that there were no chemical interactions between BSA and the DES in the extraction process, and the CD spectra proved that the conformation of BSA did not change after extraction. The conductivity, DLS and TEM were combined to investigate the microstructure of the top phase and the possible mechanism for the extraction. The results showed that hydrophobic interactions, hydrogen bonding interactions and the salting-out effect played important roles in the transfer process, and the aggregation and surrounding phenomenon were the main driving forces for the separation. All of these results proved that ionic liquid (IL)-based ATPSs could potentially be substituted with DES-based ATPSs to offer new possibilities in the extraction of proteins.

189 citations


Journal ArticleDOI
17 Mar 2014-Analyst
TL;DR: Results showed that a maximal mobility resolution can be achieved by optimizing the gas velocity, radial confinement (RF amplitude) and ramp speed (voltage range and ramp time) in a trapped ion mobility spectrometer.
Abstract: In the present paper, theoretical simulations and experimental observations are used to describe the ion dynamics in a trapped ion mobility spectrometer. In particular, the ion motion, ion transmission and mobility separation are discussed as a function of the bath gas velocity, radial confinement, analysis time and speed. Mobility analysis and calibration procedure are reported for the case of sphere-like molecules for positive and negative ion modes. Results showed that a maximal mobility resolution can be achieved by optimizing the gas velocity, radial confinement (RF amplitude) and ramp speed (voltage range and ramp time). The mobility resolution scales with the electric field and gas velocity and R = 100–250 can be routinely obtained at room temperature.

185 citations


Journal ArticleDOI
03 Jun 2014-Analyst
TL;DR: A simple yet reliable microfluidic device for the ultra-high-throughput, label-free, size-based isolation of CTCs from clinically relevant blood volumes that can be utilized for diverse biomarker studies or time-sensitive molecular assays such as RT-PCR.
Abstract: The detection and characterization of rare circulating tumor cells (CTCs) from the blood of cancer patients can potentially provide critical insights into tumor biology and hold great promise for cancer management. The ability to collect a large number of viable CTCs for various downstream assays such as quantitative measurements of specific biomarkers or targeted somatic mutation analysis is increasingly important in medical oncology. Here, we present a simple yet reliable microfluidic device for the ultra-high-throughput, label-free, size-based isolation of CTCs from clinically relevant blood volumes. The fast processing time of the technique (7.5 mL blood in less than 10 min) and the ability to collect more CTCs from larger blood volumes lends itself to a broad range of potential genomic and transcriptomic applications. A critical advantage of this protocol is the ability to return all fractions of blood (i.e., plasma (centrifugation), CTCs and white blood cells (WBCs) (size-based sorting)) that can be utilized for diverse biomarker studies or time-sensitive molecular assays such as RT-PCR. The clinical use of this biochip was demonstrated by detecting CTCs from 100% (10/10) of blood samples collected from patients with advanced-stage metastatic breast and lung cancers. The CTC recovery rate ranged from 20 to 135 CTCs mL−1 and obtained under high purity (of 1 CTC out of every 30–100 WBCs which gives ∼4 log depletion of WBCs). They were identified with immunofluorescence assays (pan-cytokeratin+/CD45−) and molecular probes such as HER2/neu.

173 citations


Journal ArticleDOI
21 Jan 2014-Analyst
TL;DR: A generic method to estimate the detection limit of biomolecular assays based on a step-by-step analysis of the assay procedure, using Enzyme-linked immunosorbent assay (ELISA) as an example.
Abstract: The detection limit is one of the most important performance parameters for bioanalytical techniques. Here we present a generic method to estimate the detection limit of biomolecular assays based on a step-by-step analysis of the assay procedure. Enzyme-linked immunosorbent assay (ELISA) is used here as an example; however, much of the information presented in this article may be applied to other types of biomolecular assays and analytical techniques. A clear understanding of what affects the detection limit can help researchers to evaluate different bio-analytical techniques properly, and to design better strategies to optimize and achieve the best analytical performance.

168 citations


Journal ArticleDOI
19 May 2014-Analyst
TL;DR: The fabricated AChE biosensor displayed high stability and excellent activity together with a fast response to organophosphorus pesticides and led to the rapid and sensitive detection of paraoxon-ethyl from 1.0 nM to 5 μM with a detection limit of 0.5 nM.
Abstract: A nanohybrid of gold nanoparticles, polypyrrole, and reduced graphene oxide sheets (named as Au–PPy–rGO) was achieved by electrochemical deposition of reduced graphene oxide with pyrrole and the introduction of gold nanoparticles. Acetylcholinesterase (AChE) was further encapsulated in a silica matrix and immobilized on the Au–PPy–rGO nanocomposite by co-deposition with (NH4)2SiF6. The presence of PPy helped to avoid the aggregation of rGO caused by van der Waals interactions between individual sheets and significantly increased the surface area of the modified electrode. The obtained Au–PPy–rGO nanocomposite not only showed excellent conductivity but also exhibited a high electrocatalytic activity and specific affinity for thiocholine, the hydrolysis product of the enzyme, and thus an improved detection sensitivity. Since AChE molecules were protected by the circumambient silica matrix, which provided a biocompatible environment and facilitated mass transport, the fabricated AChE biosensor displayed high stability and excellent activity together with a fast response to organophosphorus pesticides. Under optimum conditions, the biosensor led to the rapid and sensitive detection of paraoxon-ethyl from 1.0 nM to 5 μM with a detection limit of 0.5 nM.

Journal ArticleDOI
10 Nov 2014-Analyst
TL;DR: A smartphone-based device shows adequate analytical performance to offer a cost-effective alternative for non-invasive lactate measurement and could be used to evaluate lactate variation in relation to the anaerobic threshold in endurance sport and for monitoring lactic acidosis in critical-care patients.
Abstract: Increasingly, smartphones are used as portable personal computers, revolutionizing communication styles and entire lifestyles. Using 3D-printing technology we have made a disposable minicartridge that can be easily prototyped to turn any kind of smartphone or tablet into a portable luminometer to detect chemiluminescence derived from enzyme-coupled reactions. As proof-of-principle, lactate oxidase was coupled with horseradish peroxidase for lactate determination in oral fluid and sweat. Lactate can be quantified in less than five minutes with detection limits of 0.5 mmol L−1 (corresponding to 4.5 mg dL−1) and 0.1 mmol L−1 (corresponding to 0.9 mg dL−1) in oral fluid and sweat, respectively. A smartphone-based device shows adequate analytical performance to offer a cost-effective alternative for non-invasive lactate measurement. It could be used to evaluate lactate variation in relation to the anaerobic threshold in endurance sport and for monitoring lactic acidosis in critical-care patients.

Journal ArticleDOI
07 Jan 2014-Analyst
TL;DR: It was found that the electrochemical reduction of Cr(VI) at the Ti/TiO2NT/Au electrode exhibited an almost 23 fold improvement in activity as compared to a polycrystalline gold electrode, due to its nanoparticle/nanotubular heterojunction infrastructure.
Abstract: Owing to the severe toxicity and mobility of Cr(VI) in biological and environmental systems, it is of great importance to develop convenient and reliable methods for its detection. Here we report on a facile and effective electrochemical technique for monitoring Cr(VI) concentrations based on the utilization of Au nanoparticle-decorated titania nanotubes (TiO2NTs) grown on a titanium substrate. It was found that the electrochemical reduction of Cr(VI) at the Ti/TiO2NT/Au electrode exhibited an almost 23 fold improvement in activity as compared to a polycrystalline gold electrode, due to its nanoparticle/nanotubular heterojunction infrastructure. As a result, the Ti/TiO2NT/Au electrode demonstrated a wide linear concentration range from 0.10 μM to 105 μM, a low detection limit of 0.03 μM, and a high sensitivity of 6.91 μA μM−1 Cr(VI) via amperometry, satisfying the detection requirements of the World Health Organization (WHO). Moreover, the Ti/TiO2NT/Au electrode exhibited good resistance against interference from coexisting Cr(III) and other metal ions, and excellent recovery for Cr(VI) detection in both tap and lake water samples. These attributes suggest that this hybrid sensor has strong potential in applications for the selective detection of Cr(VI).

Journal ArticleDOI
17 Feb 2014-Analyst
TL;DR: This review critiques current protein-based microarray preparation techniques commonly used for analytical and function-based proteomics and their effects on array-based assay performance.
Abstract: Of the diverse analytical tools used in proteomics, protein microarrays possess the greatest potential for providing fundamental information on protein, ligand, analyte, receptor, and antibody affinity-based interactions, binding partners and high-throughput analysis. Microarrays have been used to develop tools for drug screening, disease diagnosis, biochemical pathway mapping, protein–protein interaction analysis, vaccine development, enzyme–substrate profiling, and immuno-profiling. While the promise of the technology is intriguing, it is yet to be realized. Many challenges remain to be addressed to allow these methods to meet technical and research expectations, provide reliable assay answers, and to reliably diversify their capabilities. Critical issues include: (1) inconsistent printed microspot morphologies and uniformities, (2) low signal-to-noise ratios due to factors such as complex surface capture protocols, contamination, and static or no-flow mass transport conditions, (3) inconsistent quantification of captured signal due to spot uniformity issues, (4) non-optimal protocol conditions such as pH, temperature, drying that promote variability in assay kinetics, and lastly (5) poor protein (e.g., antibody) printing, storage, or shelf-life compatibility with common microarray assay fabrication methods, directly related to microarray protocols. Conventional printing approaches, including contact (e.g., quill and solid pin), non-contact (e.g., piezo and inkjet), microfluidics-based, microstamping, lithography, and cell-free protein expression microarrays, have all been used with varying degrees of success with figures of merit often defined arbitrarily without comparisons to standards, or analytical or fiduciary controls. Many microarray performance reports use bench top analyte preparations lacking real-world relevance, akin to “fishing in a barrel”, for proof of concept and determinations of figures of merit. This review critiques current protein-based microarray preparation techniques commonly used for analytical and function-based proteomics and their effects on array-based assay performance.

Journal ArticleDOI
03 Feb 2014-Analyst
TL;DR: M mammalian oocytes were sensitive to silver containing nanoparticles, likely, the delicate process of completing meiosis in maternal gametes features high vulnerability towards nanomaterial derived toxicity.
Abstract: Metal and alloy nanoparticles are increasingly developed for biomedical applications, while a firm understanding of their biocompatibility is still missing. Various properties have been reported to influence the toxic potential of nanoparticles. This study aimed to assess the impact of nanoparticle size, surface ligands and chemical composition of gold, silver or gold–silver alloy nanoparticles on mammalian gametes. An in vitro assay for porcine gametes was developed, since these are delicate primary cells, for which well-established culture systems exist and functional parameters are defined. During coincubation with oocytes for 46 h neither any of the tested gold nanoparticles nor the gold–silver alloy particles with a silver molar fraction of up to 50% showed any impact on oocyte maturation. Alloy nanoparticles with 80% silver molar fraction and pure silver nanoparticles inhibited cumulus–oocyte maturation. Confocal microscopy revealed a selective uptake of gold nanoparticles by oocytes, while silver and alloy particles mainly accumulated in the cumulus cell layer surrounding the oocyte. Interestingly sperm vitality parameters (motility, membrane integrity and morphology) were not affected by any of the tested nanoparticles. Only sporadic association of nanoparticles with the sperm plasma membrane was found by transmission electron microscopy. In conclusion, mammalian oocytes were sensitive to silver containing nanoparticles. Likely, the delicate process of completing meiosis in maternal gametes features high vulnerability towards nanomaterial derived toxicity. The results imply that released Ag+-ions are responsible for the observed toxicity, but the compounding into an alloy seemed to alleviate the toxic effects to a certain extent.

Journal ArticleDOI
01 Apr 2014-Analyst
TL;DR: In this paper, the effect of paper type on the analytical performance of a series of microfluidic paper-based analytical devices (μPADs) fabricated using a CO2 laser engraver was investigated.
Abstract: A systematic investigation was conducted to study the effect of paper type on the analytical performance of a series of microfluidic paper-based analytical devices (μPADs) fabricated using a CO2 laser engraver. Samples included three different grades of Whatman chromatography paper, and three grades of Whatman filter paper. According to the data collected and the characterization performed, different papers offer a wide range of flow rate, thickness, and pore size. After optimizing the channel widths on the μPAD, the focus of this study was directed towards the color intensity and color uniformity formed during a colorimetric enzymatic reaction. According to the results herein described, the type of paper and the volume of reagents dispensed in each detection zone can determine the color intensity and uniformity. Therefore, the objective of this communication is to provide rational guidelines for the selection of paper substrates for the fabrication of μPADs.

Journal ArticleDOI
17 Feb 2014-Analyst
TL;DR: Cd(II) is chosen as a model of a small molecule with less sites, and a novel SELEX strategy of immobilizing ssDNA libraries rather than target molecules on a matrix is proposed, for selection of aptamers with high affinity to Cd( II).
Abstract: The demand for selection of aptamers against various small chemical molecules has substantially increased in recent years. To incubate and separate target-specific aptamers, the conventional SELEX procedures generally need to immobilize target molecules on a matrix, which may be impotent to screen aptamers toward small molecules without enough sites for immobilization. Herein we chose Cd(II) as a model of a small molecule with less sites, and proposed a novel SELEX strategy of immobilizing ssDNA libraries rather than target molecules on a matrix, for selection of aptamers with high affinity to Cd(II). After eleven rounds of positive and negative selection, twelve T and G-rich of nonrepeating ssDNA sequences were identified, of which the Cd-4 aptamer displayed the highest binding affinity to Cd(II). The secondary structures of these sequences revealed that a stem-loop structure folded by the domain of their 30-random sequence is critical for aptamers to bind targets. Then the interaction between the selected Cd-4 aptamer and Cd(II) was confirmed by CD analysis, and the binding specificity toward other competitive metal ions was also investigated. The dissociation constant (Kd) of Cd-4 aptamer was determined as 34.5 nM for Cd(II). Moreover, the Cd-4 aptamer was considered a recognition element for the colorimetric detection of Cd(II) based on the aggregation of AuNPs by cationic polymer. Through spectroscopic quantitative analysis, Cd(II) in aqueous solution can be detected as low as 4.6 nM. The selected Cd-4 aptamer will offer a new substitute for the detection of Cd(II) or other applications like recovery of cadmium from polluted samples.

Journal ArticleDOI
07 Jan 2014-Analyst
TL;DR: Positive charged graphene oxide was found to be more beneficial for neurite outgrowth and branching and could be implemented clinically, especially in cases wherein long-term presence of outgrowth modulation is necessary.
Abstract: Graphene oxides with different surface charges were fabricated from carboxylated graphene oxide by chemical modification with amino- (-NH2), poly-m-aminobenzene sulfonic acid- (-NH2/-SO3H), or methoxyl- (-OCH3) terminated functional groups. The chemically functionalized graphene oxides and the carboxylated graphene oxide were characterized by infrared spectroscopy, X-ray photoelectron spectroscopy, UV-Vis spectrometry, ζ potential measurements, field emission scanning electron microscopy, and contact angle analyses. Subsequently, the resulting graphene oxides were used as substrates for culturing primary rat hippocampal neurons to investigate neurite outgrowth and branching. The morphological features of neurons that directly reflect their potential capability in synaptic transmission were characterized. The results demonstrate that the chemical properties of graphene oxide can be systematically modified by attaching different functional groups that confer known characteristics to the substrate. By manipulating the charge carried by the functionalized graphene oxides, the outgrowth and branching of neuronal processes can be controlled. Compared with neutral, zwitterionic, or negatively charged graphene oxides, positively charged graphene oxide was found to be more beneficial for neurite outgrowth and branching. The ability to chemically modify graphene oxide to control neurite outgrowth could be implemented clinically, especially in cases wherein long-term presence of outgrowth modulation is necessary.

Journal ArticleDOI
03 Mar 2014-Analyst
TL;DR: Current trends suggest that biosensor-based techniques (BTs) for miRNA assay will complement MTs due to the advantages of amplification-free detection, LOD being femtomolar (fM)-attomolar (aM), short TTR, multiplexing capability, and minimal sample preparation requirement.
Abstract: The current state of biosensor-based techniques for amplification-free microRNA (miRNA) detection is critically reviewed. Comparison with non-sensor and amplification-based molecular techniques (MTs), such as polymerase-based methods, is made in terms of transduction mechanism, associated protocol, and sensitivity. Challenges associated with miRNA hybridization thermodynamics which affect assay selectivity and amplification bias are briefly discussed. Electrochemical, electromechanical, and optical classes of miRNA biosensors are reviewed in terms of transduction mechanism, limit of detection (LOD), time-to-results (TTR), multiplexing potential, and measurement robustness. Current trends suggest that biosensor-based techniques (BTs) for miRNA assay will complement MTs due to the advantages of amplification-free detection, LOD being femtomolar (fM)–attomolar (aM), short TTR, multiplexing capability, and minimal sample preparation requirement. Areas of future importance in miRNA BT development are presented which include focus on achieving high measurement confidence and multiplexing capabilities.

Journal ArticleDOI
28 Jul 2014-Analyst
TL;DR: A novel carbon paste electrode modified with ZnO nanorods and 5-(4'-amino-3'-hydroxy-biphenyl-4-yl)-acrylic acid was fabricated and used for the simultaneous determination of levodopa and carbidopa.
Abstract: A novel carbon paste electrode modified with ZnO nanorods and 5-(4'-amino-3'-hydroxy-biphenyl-4-yl)-acrylic acid (3,4'-AAZCPE) was fabricated. The electrochemical study of the modified electrode, as well as its efficiency for the electrocatalytic oxidation of levodopa, is described. The electrode was employed to study the electrocatalytic oxidation of levodopa, using cyclic voltammetry (CV), chronoamperometry (CHA), and square-wave voltammetry (SWV) as diagnostic techniques. It has been found that the oxidation of levodopa at the surface of the modified electrode occurs at a potential of about 370 mV less positive than that of an unmodified carbon paste electrode. The SWV results exhibit a linear dynamic range from 1.0 × 10(-7) M to 7.0 × 10(-5) M and a detection limit of 3.5 × 10(-8) M for levodopa. In addition, this modified electrode was used for the simultaneous determination of levodopa and carbidopa. Finally, the modified electrode was used for the determination of levodopa and carbidopa in some real samples.

Journal ArticleDOI
29 Sep 2014-Analyst
TL;DR: The potential of silica nanoparticles to avoid the washing away effect and improve the color uniformity and intensity in colorimetric bioassays performed on μPADs is demonstrated.
Abstract: This paper describes a silica nanoparticle-modified microfluidic paper-based analytical device (μPAD) with improved color intensity and uniformity for three different enzymatic reactions with clinical relevance (lactate, glucose, and glutamate). The μPADs were produced on a Whatman grade 1 filter paper and using a CO2 laser engraver. Silica nanoparticles modified with 3-aminopropyltriethoxysilane were then added to the paper devices to facilitate the adsorption of selected enzymes and prevent the washing away effect that creates color gradients in the colorimetric measurements. According to the results herein described, the addition of silica nanoparticles yielded significant improvements in color intensity and uniformity. The resulting μPADs allowed for the detection of the three analytes in clinically relevant concentration ranges with limits of detection (LODs) of 0.63 mM, 0.50 mM, and 0.25 mM for lactate, glucose, and glutamate, respectively. An example of an analytical application has been demonstrated for the semi-quantitative detection of all three analytes in artificial urine. The results demonstrate the potential of silica nanoparticles to avoid the washing away effect and improve the color uniformity and intensity in colorimetric bioassays performed on μPADs.

Journal ArticleDOI
17 Mar 2014-Analyst
TL;DR: The results show that the four pesticides can be captured and detected using principal component analysis based on their distinct fingerprint Raman peaks and demonstrated the super capacity of aptamer-based SERS in rapid detection and discrimination of multi-pesticides.
Abstract: The objective of this study was to develop a simple and rapid method that could detect and discriminate four specific pesticides (isocarbophos, omethoate, phorate, and profenofos) using a single aptamer-based capture procedure followed by Surface Enhanced Raman Spectroscopy (SERS). The aptamer is a single stranded DNA sequence that is specific to capture these four pesticides. The thiolated aptamer was conjugated onto silver (Ag) dendrites, a nanostructure that can enhance the Raman fingerprint of pesticides, through Ag–thiol bonds. It was then backfilled with 6-mercaptohexanol (MH) to prevent nonspecific binding. The modified SERS platform [Ag–(Ap + MH)] was then mixed with each pesticide solution (P) for 20 min. After capturing the pesticides, the Ag–(Ap + MH)–P complex was analyzed under a DXR Raman microscope and TQ Analyst software. The results show that the four pesticides can be captured and detected using principal component analysis based on their distinct fingerprint Raman peaks. The limits of detection (LODs) of isocarbophos, omethoate, phorate, and profenofos were 3.4 μM (1 ppm), 24 μM (5 ppm), 0.4 μM (0.1 ppm), and 14 μM (5 ppm) respectively. This method was also validated successfully in apple juice. These results demonstrated the super capacity of aptamer-based SERS in rapid detection and discrimination of multi-pesticides. This technique can be extended to detect a wide range of pesticides using specific aptamers.

Journal ArticleDOI
03 Feb 2014-Analyst
TL;DR: The comparison between heparin and EDTA anti-coagulant collection tubes, the former showed no interference of the polysaccharide, while conserving full spectral information, and the impact of long-term -80 °C storage was found almost negligible.
Abstract: This work contributes to fill in some existing gaps in the knowledge of human plasma degradability during handling and storage, a paramount issue in Nuclear Magnetic Resonance (NMR) metabolomics. Regarding the comparison between heparin and EDTA anti-coagulant collection tubes, the former showed no interference of the polysaccharide, while conserving full spectral information. In relation to time/temperature conditions, room temperature was seen to have a large impact on lipoproteins and choline compounds from 2.5 hours. In addition, short-term storage at −20 °C was found suitable up to 7 days but, for periods up to 1 month, −80 °C was recommended. Furthermore, in the case of reusing plasma samples, no more than 3 consecutive freeze–thaw cycles were found advisable. Finally, the impact of long-term −80 °C storage (up to 2.5 years) was found almost negligible, as evaluated on a partially matched non-fasting cohort (n = 49), after having investigated the possible confounding nature of the particular non-fasting conditions employed.

Journal ArticleDOI
03 Mar 2014-Analyst
TL;DR: As-prepared N-CDs displayed bright blue fluorescence under the irradiation of UV light, together with a high quantum yield and good biocompatibility as demonstrated by the cell viability assay, which opens a new way for the preparation of fluorescent carbon nanomaterials using small molecules as carbon sources.
Abstract: A simple, facile and green hydrothermal method was developed in the synthesis of water-soluble nitrogen-doped carbon dots (N-CDs) from streptomycin. The as-prepared N-CDs displayed bright blue fluorescence under the irradiation of UV light, together with a high quantum yield of 7.6% and good biocompatibility as demonstrated by the cell viability assay. Thus, the N-CDs can be used as fluorescent probes for cell imaging, which have potential applications in bioimaging and related fields. This strategy opens a new way for the preparation of fluorescent carbon nanomaterials using small molecules as carbon sources.

Journal ArticleDOI
10 Nov 2014-Analyst
TL;DR: A simple, low-cost, one-step fabrication method for microfluidic paper-based analytical devices (μPAD) using only polystyrene and a patterned screen is reported, verifying that the polymer screen-printing method can be used as an alternative method for μPAD fabrication.
Abstract: We report a simple, low-cost, one-step fabrication method for microfluidic paper-based analytical devices (μPAD) using only polystyrene and a patterned screen. The polystyrene solution applied through the screen penetrates through the paper, forming a three-dimensional hydrophobic barrier, defining a hydrophilic analysis zone. The optimal polystyrene concentration and paper types were first investigated. Adjusting polystyrene concentration allows for various types of paper to be used for successful device fabrication. Using an optimized polystyrene concentration with Whatman#4 filter paper, a linear relationship was found to exist between the design width and the printed width. The smallest hydrophilic channel and hydrophobic barrier that can be obtained are 670 ± 50 μm and 380 ± 40 μm, respectively. High device-to-device fabrication reproducibility was achieved yielding a relative standard deviation (%RSD) in the range of 1.12–2.54% (n = 64) of the measured diameter of the well-shaped fabricated test zones with a designed diameter of 5 and 7 mm. To demonstrate the significance of the fabricated μPAD, distance-based and well-based paper devices were constructed for the analysis of H2O2 and antioxidant activity, respectively. The analysis of H2O2 in real samples using distance-based measurement with CeO2 nanoparticles as the colorimetric agent produced the same results at 95% confidence level, as those obtained using KMnO4 titration. A proof-of-concept antioxidant activity determination based on the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was also demonstrated. The results verify that the polymer screen-printing method can be used as an alternative method for μPAD fabrication.

Journal ArticleDOI
03 Feb 2014-Analyst
TL;DR: It is demonstrated that Raman microscopy will open avenues as a non-invasive and label-free technique to investigate pharmacokinetics at the highest possible resolution in living cells.
Abstract: Targeted cancer therapies block cancer growth and spread using small molecules. Many molecular targets for an epidermal growth factor receptor (EGFR) selectively compete with the adenosine triphosphate-binding site of its tyrosine kinase domain. Detection of molecular targeted agents and their metabolites in cells/tissues by label-free imaging is attractive because dyes or fluorescent labels may be toxic or invasive. Here, label-free Raman microscopy is applied to show the spatial distribution of the molecular targeted drug erlotinib within the cell. The Raman images show that the drug is clustered at the EGFR protein at the membrane and induces receptor internalization. The changes within the Raman spectrum of erlotinib measured in cells as compared to the free-erlotinib spectrum indicate that erlotinib is metabolized within cells to its demethylated derivative. This study provides detailed insights into the drug targeting mechanism at the atomic level in cells. It demonstrates that Raman microscopy will open avenues as a non-invasive and label-free technique to investigate pharmacokinetics at the highest possible resolution in living cells.

Journal ArticleDOI
07 Jan 2014-Analyst
TL;DR: In this paper, the collision-induced dissociation (CID) of the [M + Na]- adduct ions of phospholipids can be isolated and subjected to subsequent gas-phase ozonolysis (OzID) in a linear ion-trap mass spectrometer.
Abstract: The complete structural elucidation of complex lipids, including glycerophospholipids, using only mass spectrometry represents a major challenge to contemporary analytical technologies. Here, we demonstrate that product ions arising from the collision-induced dissociation (CID) of the [M + Na]+ adduct ions of phospholipids can be isolated and subjected to subsequent gas-phase ozonolysis – known as ozone-induced dissociation (OzID) – in a linear ion-trap mass spectrometer. The resulting CID/OzID experiment yields abundant product ions that are characteristic of the acyl substitution on the glycerol backbone (i.e., sn-position). This approach is shown to differentiate sn-positional isomers, such as the regioisomeric phosphatidylcholine pair of PC 16:0/18:1 and PC 18:1/16:0. Importantly, CID/OzID provides a sensitive diagnostic for the existence of an isomeric mixture in a given sample. This is of very high value for the analysis of tissue extracts since CID/OzID analyses can reveal changes in the relative abundance of isomeric constituents even within different tissues from the same animal. Finally, we demonstrate the ability to assign carbon–carbon double bond positions to individual acyl chains at specific backbone positions by adding subsequent CID and/or OzID steps to the workflow and that this can be achieved in a single step using a hybrid triple quadrupole-linear ion trap mass spectrometer. This unique approach represents the most complete and specific structural analysis of lipids by mass spectrometry demonstrated to date and is a significant step towards comprehensive top-down lipidomics.

Journal ArticleDOI
17 Feb 2014-Analyst
TL;DR: A method for the long-term dry storage of ELISA reagents, including horseradish peroxidase conjugated antibody label and its colorimetric substrate diaminobenzidine, which demonstrates the potential of enzyme-based signal amplification for enhanced sensitivity in POC devices for low resource settings.
Abstract: Lateral flow devices are commonly used for many point-of-care (POC) applications in low-resource settings. However, they lack the sensitivity needed for many analytes relevant in the diagnosis of diseases. One approach to achieve higher sensitivity is signal amplification, which is commonly used in laboratory assays, but uses reagents that require refrigeration and inherently requires multiple assay steps not normally compatible with POC settings. Enzyme-based signal amplification, such as the one used in ELISA, could greatly improve the limit of detection if it were translated to a format compatible with POC requirements. A signal-amplified POC device not only requires the reagents to be stored in a stable form, but also requires automation of the multiple sequential steps of signal amplification protocols. Here, we describe a method for the long-term dry storage of ELISA reagents: horseradish peroxidase (HRP) conjugated antibody label and its colorimetric substrate diaminobenzidine (DAB). The HRP conjugate retained ∼80% enzymatic activity after dry storage at 45 °C for over 5 months. The DAB substrate was also stable at 45 °C and exhibited no detectable loss of activity over 3 months. These reagents were incorporated into a two-dimensional paper network (2DPN) device that automated the steps of ELISA for the detection of a malarial biomarker. These results demonstrate the potential of enzyme-based signal amplification for enhanced sensitivity in POC devices for low resource settings.

Journal ArticleDOI
26 Aug 2014-Analyst
TL;DR: Compared with some promising variable selection methods such as competitive adaptive reweighted sampling (CARS), Monte Carlo uninformative variable elimination (MCUVE) and iteratively retaining informative variables (IRIV), VISSA showed better prediction ability for the calibration of NIR data.
Abstract: In this study, a new optimization algorithm called the Variable Iterative Space Shrinkage Approach (VISSA) that is based on the idea of model population analysis (MPA) is proposed for variable selection. Unlike most of the existing optimization methods for variable selection, VISSA statistically evaluates the performance of variable space in each step of optimization. Weighted binary matrix sampling (WBMS) is proposed to generate sub-models that span the variable subspace. Two rules are highlighted during the optimization procedure. First, the variable space shrinks in each step. Second, the new variable space outperforms the previous one. The second rule, which is rarely satisfied in most of the existing methods, is the core of the VISSA strategy. Compared with some promising variable selection methods such as competitive adaptive reweighted sampling (CARS), Monte Carlo uninformative variable elimination (MCUVE) and iteratively retaining informative variables (IRIV), VISSA showed better prediction ability for the calibration of NIR data. In addition, VISSA is user-friendly; only a few insensitive parameters are needed, and the program terminates automatically without any additional conditions. The Matlab codes for implementing VISSA are freely available on the website: https://sourceforge.net/projects/multivariateanalysis/files/VISSA/.

Journal ArticleDOI
14 Jul 2014-Analyst
TL;DR: It is demonstrated how a protein chemical image of the amide I band of a 2 × 2.4 cm(2) breast tissue microarray containing over 200 cores can be measured in 9 min, indicating that applications requiring chemical images from a few key wavelengths would be ideally served by laser-based microscopes.
Abstract: Chemical imaging in the field of vibrational spectroscopy is developing into a promising tool to complement digital histopathology. Applications include screening of biopsy tissue via automated recognition of tissue/cell type and disease state based on the chemical information from the spectrum. For integration into clinical practice, data acquisition needs to be speeded up to implement a rack based system where specimens are rapidly imaged to compete with current visible scanners where 100's of slides can be scanned overnight. Current Fourier transform infrared (FTIR) imaging with focal plane array (FPA) detectors are currently the state-of-the-art instrumentation for infrared absorption chemical imaging, however recent development in broadly tunable lasers in the mid-IR range is considered the most promising potential candidate for next generation microscopes. In this paper we test a prototype quantum cascade laser (QCL) based spectral imaging microscope with a focus on discrete frequency chemical imaging. We demonstrate how a protein chemical image of the amide I band (1655 cm−1) of a 2 × 2.4 cm2 breast tissue microarray (TMA) containing over 200 cores can be measured in 9 min. This result indicates that applications requiring chemical images from a few key wavelengths would be ideally served by laser-based microscopes.