The 50 most-cited articles in The Analyst, 2016

Carbon quantum dots (CQDs) are attracting tremendous interest owing to their low toxicity, water dispersibility, biocompatibility, optical properties and wide applicability. Herein, CQDs with an average diameter of (4.0 ± 0.2) nm and high crystallinity were produced simply from the electrochemical oxidation of a graphite electrode in alkaline alcohols. The as-formed CQDs dispersion was colourless but the dispersion gradually changed to bright yellow when stored in ambient conditions. Based on UV-Vis absorption, fluorescence spectroscopy, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR) and high-resolution transmission electron microscopy (HRTEM), this colour change appeared to be due to oxygenation of surface species over time. Furthermore, the CQDs were used in specific and sensitive detection of ferric ion (Fe(3+)) with broad linear ranges of 10-200 μM with a low limit of detection of 1.8 μM (S/N = 3). The application of the CQDs for Fe(3+) detection in tap water was demonstrated and the possible mechanism was also discussed. Finally, based on…

Metal nanoclusters (NCs) are emerging as a new class of functional nanomaterials in the area of biological sensing, labelling, imaging and therapy due to their unique physical and chemical properties, such as ultrasmall size, HOMO-LUMO transition, strong luminescence together with good photostability and biocompatibility. A recent surge of interest in this field is the surface functionalization of these metal NCs through which one can tailor their physicochemical properties, such as stability in solution, and strong luminescence, as well as their biodistribution and toxicity in biological systems, which in turn can empower these functionalized NCs with desirable targeting, imaging, and therapeutic ability for biomedical applications. In this review, we first introduce the functionalization strategies for the metal NCs developed in the past few years, followed by highlighting some biomedical applications of these functionalized metal NCs. We then discuss the difference of in vitro and in vivo fate as well as toxicity between various functionalized metal NCs. Finally, we present a…

The field of paper-based microfluidics has experienced rapid growth over the past decade. Microfluidic paper-based analytical devices (μPADs), originally developed for point-of-care medical diagnostics in resource-limited settings, are now being applied in new areas, such as environmental analyses. Low-cost paper sensors show great promise for on-site environmental analysis; the theme of ongoing research complements existing instrumental techniques by providing high spatial and temporal resolution for environmental monitoring. This review highlights recent applications of μPADs for environmental analysis along with technical advances that may enable μPADs to be more widely implemented in field testing.

Exosomes are secreted nanovesicles shed by almost all kinds of cells. Recently, increased interest has been focused on these extracellular vesicles as natural carriers transporting biological contents for intercellular communication. However, current isolation techniques, such as ultracentrifugation, are not convenient and often require specialized equipment. Herein, we describe a polyethylene glycol (PEG)-based approach, which could permit facile, low-cost and effective isolation of exosomes from cell culture supernatant. High-resolution electron microscopes clearly visualized the size and morphology of isolated exosome aggregates, implying the mechanism of PEG-based precipitation. Combined with tandem mass spectrometry analysis, 6299 protein groups encoded by 5120 genes were successfully characterized from HeLa cell culture supernatant, including numerous exosome proteins which could overlap 97% of the Top 100 exosome marker proteins recorded in the ExoCarta database, as well as a series of low-abundance cytokines and biomarkers. Furthermore, we found a higher ratio of…

One of the most exciting developments in Raman spectroscopy in the last decade has been its application to cells and tissues for diagnostic and pharmaceutical applications, and in particular its use in the analysis of cellular dynamics. Raman spectroscopy is rapidly advancing as a cell imaging method that overcomes many of the limitations of current techniques and is earning its place as a routine tool in cell biology. In this review we focus on important developments in Raman spectroscopy that have evolved into the exciting technique of live-cell Raman microscopy and highlight some of the most recent and significant applications to cell biology.

Advances in liquid chromatography-mass spectrometry (LC-MS) instruments and analytical strategies have brought about great progress in targeted metabolomics analysis. This methodology is now capable of performing precise targeted measurement of dozens or hundreds of metabolites in complex biological samples. Classic targeted quantification assay using the multiple reaction monitoring (MRM) mode has been the foundation of high-quality metabolite quantitation. However, utilization of this strategy in biological studies has been limited by its relatively low metabolite coverage and throughput capacity. A number of methods for large-scale targeted metabolomics assay which have been developed overcome these limitations. These strategies have enabled extended metabolite coverage which is defined as targeting of large numbers of metabolites, while maintaining reliable quantification performance. These recently developed techniques thus bridge the gap between traditional targeted metabolite quantification and untargeted metabolomics profiling, and have proven to be powerful tools for…

Understanding how biological molecules are generated, metabolized and eliminated in living systems is important for interpreting processes such as immune response and disease pathology. While genomic and proteomic studies have provided vast amounts of information over the last several decades, interest in lipidomics has also grown due to improved analytical technologies revealing altered lipid metabolism in type 2 diabetes, cancer, and lipid storage disease. Mass spectrometry (MS) measurements are currently the dominant approach for characterizing the lipidome by providing detailed information on the spatial and temporal composition of lipids. However, interpreting lipids' biological roles is challenging due to the existence of numerous structural and stereoisomers (i.e. distinct acyl chain and double-bond positions), which are often unresolvable using present approaches. Here we show that combining liquid chromatography (LC) and structurally-based ion mobility spectrometry (IMS) measurement with MS analyses distinguishes lipid isomers and allows insight into biological and disease…

Nucleic acid aptamers are promising alternatives to antibodies in analytics. They are generally obtained through an iterative SELEX protocol that enriches a population of synthetic oligonucleotides to a subset that can recognize the chosen target molecule specifically and avidly. A wide range of targets is recognized by aptamers. Once identified and optimized for performance, aptamers can be reproducibly synthesized and offer other key features, like small size, low cost, sensitivity, specificity, rapid response, stability, and reusability. This makes them excellent options for sensory units in a variety of analytical platforms including those with electrochemical, optical, and mass sensitive transduction detection. Many novel sensing strategies have been developed by rational design to take advantage of the tendency of aptamers to undergo conformational changes upon target/analyte binding and employing the principles of base complementarity that can drive the nucleic acid structure. Despite their many advantages over antibodies, surprisingly few aptamers have yet been integrated…

This paper describes the modification of microfluidic paper-based analytical devices (μPADs) with chitosan to improve the analytical performance of colorimetric measurements associated with enzymatic bioassays. Chitosan is a natural biopolymer extensively used to modify biosensing surfaces due to its capability of providing a suitable microenvironment for the direct electron transfer between an enzyme and a reactive surface. This hypothesis was investigated using glucose and uric acid (UA) colorimetric assays as model systems. The best colorimetric sensitivity for glucose and UA was achieved using a chromogenic solution composed of 4-aminoantipyrine and sodium 3,5-dichloro-2-hydroxy-benzenesulfonate (4-AAP/DHBS), which provided a linear response for a concentration range between 0.1 and 1.0 mM. Glucose and UA were successfully determined in artificial serum samples with accuracies between 87 and 114%. The limits of detection (LODs) found for glucose and UA assays were 23 and 37 μM, respectively. The enhanced analytical performance of chitosan-modified μPADs allowed the colorimetric…

A dynamic development of methodologies of analytical flow injection measurements during four decades since their invention has reinforced the solid position of flow analysis in the arsenal of techniques and instrumentation of contemporary chemical analysis. With the number of published scientific papers exceeding 20,000, and advanced instrumentation available for environmental, food, and pharmaceutical analysis, flow analysis is well established as an extremely vital field of modern flow chemistry, which is developed simultaneously with methods of chemical synthesis carried out under flow conditions. This review work is based on almost 300 original papers published mostly in the last decade, with special emphasis put on presenting novel achievements from the most recent 2-3 years in order to indicate current development trends of this methodology. Besides the evolution of the design of whole measuring systems, and including especially new applications of various detections methods, several aspects of implications of progress in nanotechnology, and miniaturization of measuring…

Intracellular pH values are some of the most important factors that govern biological processes and the acid-base homeostasis in cells, body fluids and organs sustains the normal operations of the body. Subcellular organelles including the acidic lysosomes and the alkalescent mitochondria undergo various processes such as intracellular digestion, ATP production and apoptosis. Due to their precise imaging capabilities, fluorescent probes have attracted great attention for the illustration of pH modulated processes. Furthermore, based on the unique acidic extracellular environment of acidic lysosomes, fluorescent probes can specifically be activated in cancer cells or tumors. In this review, recently reported lysosome and mitochondria specific pH imaging probes as well as pH-activatable cancer cell-targetable probes have been discussed.

Janus particles, named after the two-faced Roman god Janus, have different surface makeups, structures or compartments on two sides. This review highlights recent advances in employing Janus particles as novel analytical tools for live cell imaging and biosensing. Unlike conventional particles used in analytical science, two-faced Janus particles provide asymmetry and directionality, and can combine different or even incompatible properties within a single particle. The broken symmetry enables imaging and quantification of rotational dynamics, revealing information beyond what traditional measurements offer. The spatial segregation of molecules on the surface of a single particle also allows analytical functions that would otherwise interfere with each other to be decoupled, opening up opportunities for novel multimodal analytical methods. We summarize here the development of Janus particles, a few general methods for their fabrication and, more importantly, the emerging and novel applications of Janus particles as multi-functional imaging probes and sensors.

Due to the biological and industrial importance of hypochlorous acid, the development of optical probes for HOCl has been an active research area. Hypochlorous acid and hypochlorite can oxidize electron-rich analytes with accompanying changes in molecular sensor spectroscopic profiles. Probes for such processes may monitor HOCl levels in the environment or in an organism and via bio-labeling or bioimaging techniques. This review summarizes recent developments in the area of chromogenic and fluorogenic chemosensors for HOCl.

The measurement of sulfide, especially hydrogen sulfide, has held the attention of the analytical community due to its unique physiological and pathophysiological roles in biological systems. Electrochemical detection offers a rapid, highly sensitive, affordable, simple, and real-time technique to measure hydrogen sulfide concentration, which has been a well-documented and reliable method. This review details up-to-date research on the electrochemical detection of hydrogen sulfide (ion selective electrodes, polarographic hydrogen sulfide sensors, etc.) in biological samples for potential therapeutic use.

Confocal Raman microscopy has been used to measure depth-dependent profiles of human SC in vivo in the high wavenumber (HWN) region. In order to keep the linearity of HWN region boundaries and to not remove an informative signal from Raman spectra, a new baseline subtraction procedure has been introduced. After baseline subtraction, the HWN spectrum was deconvoluted using 10 Gaussian functions with individual chemical meanings. The results show that the hydrogen bound water molecule types contributed differently to the water diffusion process in the SC. The most concentrated double donor-double acceptor (DDAA) and single donor-single acceptor (DA) water molecule types in the SC represent more than 90% of the SC's water and mostly contribute to the water flux in the skin. Single donor-double acceptor (DAA) and weakly-bound water molecule types represent less than 10% of the SC's water content. The most tightly hydrogen bound water molecule type, DAA, reaches its maximum concentration near the skin surface and does not take part in the water diffusion process via the SC. The results…

The detection of nitroaromatic explosives is of great importance owing to their strong explosive power and harmfulness in terms of the environment, homeland security and public safety. Herein, rare earth-doped carbon dots with multifunctional features were firstly prepared by simply keeping the mixture of terbium(iii) nitrate pentahydrate and citric acid at 190 °C for 30 min. The as-prepared terbium doped carbon dots (Tb-CDs), through a rapid and simple direct carbonization route, have a size of about 3 nm, and exhibit excitation wavelength dependent emission of blue fluorescence, are stable, and can be applied for the selective and colorimetric detection of 2,4,6-trinitrophenol (TNP) in the range of 500 nM-100 μM with a limit of detection of 200 nM based on the inner filtering effect (IFE) of the excitation and emission bands of Tb-CDs by TNP and the electron transfer (ET) from Tb-CDs to TNP, giving a precise and highly reproducible result for detecting complex water samples.

cells per mL. Therefore, our CEE three-step method offered the significant advantages of short assay time, simple operating procedure, and higher sensitivity than previously reported methods of SERS-based bacteria detection.

We report on a smartphone spectrometer for colorimetric biosensing applications. The spectrometer relies on a sample cell with an integrated grating substrate, and the smartphone's built-in light-emitting diode flash and camera. The feasibility of the smartphone spectrometer is demonstrated for detection of glucose and human cardiac troponin I, the latter in conjunction with peptide-functionalized gold nanoparticles.

Adulteration of high quality food products with sub-standard and cheaper grades is a world-wide problem taxing the global economy. Currently, many traditional tests suffer from poor specificity, highly complex outputs and a lack of high-throughput processing. Metabolomics has been successfully used as an accurate discriminatory technique in a number of applications including microbiology, cancer research and environmental studies and certain types of food fraud. In this study, we have developed metabolomics as a technique to assess the adulteration of meat as an improvement on current methods. Different grades of beef mince and pork mince, purchased from a national retail outlet were combined in a number of percentage ratios and analysed using GC-MS and UHPLC-MS. These techniques were chosen because GC-MS enables investigations of metabolites involved in primary metabolism whilst UHPLC-MS using reversed phase chromatography provides information on lipophilic species. With the application of chemometrics and statistical analyses, a panel of differential metabolites were found for…

nanobelts with different oxygen vacancy concentrations calculated based on the model matches very well with experimental results. As an extension, some potential metal oxide semiconductor SERS-active substrates were predicted based on the model. Our results clearly demonstrate that, through defect engineering, the metal oxide semiconductors can be made SERS-active substrates with high stability and high biocompatibility.

Food safety is a global health objective, and foodborne diseases represent a major crisis in health. Techniques that are simple and suitable for fast screening to detect and identify pathogenic factors in the food chain are vital to ensure food safety. At present, a variety of analytical methods have been reported for the detection of pathogenic agents. Whereas the sensitivity of detection and quantification are still important challenges, we expect major advances from new assay formats and synthetic bio-recognition elements, such as aptamers. Owing to the specific folding capability of aptamers in the presence of an analyte, aptasensors have substantially and successfully been exploited for the detection of a wide range of small and large molecules (e.g., toxins, antibiotics, heavy metals, bacteria, viruses) at very low concentrations. Here, we review the use of aptasensors for the development of highly sensitive and affordable detection tools for food analysis.

Rapid detection of nitroaromatic explosives is of utmost importance from the perspectives of civilian safety, the natural environment and human health. Herein, we design a simple yet powerful optical sensing motif based on an electrodeposited luminescent MOF thin film of {[Eu2(TDC)3(CH3OH)2]·CH3OH} (Eu-TDC, where TDC is thiophene-2,5-dicarboxylate). The parameters of the electrodeposition process were investigated and optimized. The phase and elemental composition of the Eu-TDC film were confirmed using powder X-ray diffraction (PXRD) and energy dispersive spectrometry (EDS), and its morphology and structure were examined using scanning electron microscopy (SEM). This thin film shows extremely good luminescence properties with characteristics of Eu(3+) that are sensitized by the electron-rich ligands, which allows for high efficiency in electron-deficient nitroaromatic explosive sensing. Selective and sensitive responses to a series of nitroaromatic explosives were exhibited, especially for nitrophenols (TNP/DNP/4-NP, KSV > 10(4) M(-1)). It is also an efficient luminescent sensor…

wavenumber region. Subsequently, this enabled us to attribute the main influences in the regression coefficients plots. This study demonstrated that the performance of NIR spectroscopy with benchtop and miniaturized devices as a fast and non-invasive technique is able to replace time- and resource-consuming analytical tools. Referring to the developed application of the RA content quantification this work is especially interesting for the continuous growing phytopharmaceutical industry and its quality control. The results reveal the importance of monitoring the performances of available NIR-spectrometers in every analytical area.

In this minireview, we discuss advancements in ion concentration polarization (ICP)-based preconcentration, separation, desalination, and dielectrophoresis that have been made over the past three years. ICP as a means of controlling the distribution of the ions and electric field in a microfluidic device has rapidly expanded its areas of application. Recent advancements have focused on the development of ion-permselective materials with tunable dimensions and surface chemistry, adaptation to paper microfluidics, higher-throughput device geometries, and coupling ICP with other separation (isotachophoresis and dielectrophoresis) and fluidic (valve and droplet microfluidic) strategies. These studies have made great strides toward solving real-world problems such as low-cost and rapid analysis, accessible desalination technology, and single-cell research tools.

The analysis of thin films is of central importance for functional materials, including the very large and active field of nanomaterials. Quantitative elemental depth profiling is basic to analysis, and many techniques exist, but all have limitations and quantitation is always an issue. We here review recent significant advances in ion beam analysis (IBA) which now merit it a standard place in the analyst's toolbox. Rutherford backscattering spectrometry (RBS) has been in use for half a century to obtain elemental depth profiles non-destructively from the first fraction of a micron from the surface of materials: more generally, "IBA" refers to the cluster of methods including elastic scattering (RBS; elastic recoil detection, ERD; and non-Rutherford elastic backscattering, EBS), nuclear reaction analysis (NRA: including particle-induced gamma-ray emission, PIGE), and also particle-induced X-ray emission (PIXE). We have at last demonstrated what was long promised, that RBS can be used as a primary reference technique for the best traceable accuracy available for non-destructive…

Colloidal inorganic nanoparticles are being used in an increasingly large number of applications ranging from biological imaging to television displays. In all cases, nanoparticle surface chemistry can significantly impact particle physical properties, processing, and performance. The first step in leveraging this tunability is to develop analytical approaches to describe surface chemical features. Some of the most basic descriptors of particle surface chemistry include the quantity, identity, and arrangement of ligands appended to the particle core. Here, we review approaches to quantify molecular ligand densities on nanoparticle surfaces and consider fundamental barriers to the accuracy of this analysis including parameters such as dispersity in colloidal nanoparticle samples, particle-ligand interactions, and currently available analytical techniques. Techniques reviewed include widely studied methods such as optical, atomic, vibrational, and nuclear magnetic resonance spectroscopies as well as emerging or niche approaches including electrospray-differential mobility analysis,…

Infrared spectroscopy and spectroscopic imaging, are robust, label free and inherently non-destructive methods with a high chemical specificity and sensitivity that are frequently employed in forensic science research and practices. This review aims to discuss the applications and recent developments of these methodologies in this field. Furthermore, the use of recently emerged Fourier transform infrared (FT-IR) spectroscopic imaging in transmission, external reflection and Attenuated Total Reflection (ATR) modes are summarised with relevance and potential for forensic science applications. This spectroscopic imaging approach provides the opportunity to obtain the chemical composition of fingermarks and information about possible contaminants deposited at a crime scene. Research that demonstrates the great potential of these techniques for analysis of fingerprint residues, explosive materials and counterfeit drugs will be reviewed. The implications of this research for the examination of different materials are considered, along with an outlook of possible future research avenues…

Depending on the situation, metal ions may either play beneficial roles or be harmful to human health and ecosystems. Sensitive and accurate detection of metal ions is thus a critical issue in the field of analytical sciences and great efforts have been devoted to the development of various metal ion sensors. Photoelectrochemical (PEC) detection is an emerging technique for the bio/chemical detection of metal ions, and features a fast response, low cost and high sensitivity. Using representative examples, this review will first introduce the fundamentals and summarize recent progress in the PEC detection of metal ions. In addition, interesting strategies for the design of particular PEC metal ion sensors are discussed. Challenges and opportunities in this field are also presented.

Ion mobility mass spectrometry experiments enable the characterization of mass, assembly, and shape of biological molecules and assemblies. Here, a new radio-frequency confining drift cell is characterized and used to measure the mobilities of peptide, protein, and protein complex ions. The new drift cell replaced the traveling-wave ion mobility cell in a Waters Synapt G2 HDMS. Methods for operating the drift cell and determining collision cross section values using this experimental set up are presented within the context of the original instrument control software. Collision cross sections for 349 cations and anions are reported, 155 of which are for ions that have not been characterized previously using ion mobility. The values for the remaining ions are similar to those determined using a previous radio-frequency confining drift cell and drift tubes without radial confinement. Using this device under 2 Torr of helium gas and an optimized drift voltage, denatured and native-like ions exhibited average apparent resolving powers of 14.2 and 16.5, respectively. For ions with high…

Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is an emerging technique for microbial identification, characterization, and typing. The single colony method can be used for obtaining a protein fingerprint or profile unique to each microorganism. This technique has been mainly used in the clinical field, but it also has significant potential in the environmental field. The applications of MALDI-TOF MS in environmental microbiology are discussed in this review. An overview on the use of MALDI-TOF MS for environmental proteomics and metabolomics is given as well as its use for bacterial strain typing and bioremediation research. A more detailed review on the use of this technique for the identification, differentiation, and categorization of environmental microorganisms is given. Some of the parameters that can influence the results and reproducibility of MALDI-TOF MS are also discussed.

It is widely appreciated that widespread antibiotic resistance has significantly reduced the utility of today's antibiotics. Many antibiotics now fail to cure infectious diseases, although they are classified as effective bactericidal agents based on antibiotic susceptibility tests. Here, via kinetic growth assays, we evaluated the effects of 12 commonly used antibiotics on the lag phase of a range of pure environmental isolates and of sludge bacterial communities with a high diversity. We show that an extended lag phase offers bacteria survival advantages and promotes regrowth upon the removal of antibiotics. By utilizing both lag phase extension and IC50, the killing efficiency of an antibiotic on a strain or a community can be easily revealed. Interestingly, for several strains of relevance to endemic nosocomial infections (e.g. Acinetobacter sp. and Pseudomonas aeruginosa) and the diverse sludge communities, tetracycline and quinolone antibiotics are most likely to be resisted via extended lag phase. This discovery is significant from a clinical point view since underestimation…

Cortisol has long been recognized as the "stress biomarker" in evaluating stress related disorders. Plasma, urine or saliva are the current source for cortisol analysis. The sampling of these biofluids is either invasive or has reliability problems that could lead to inaccurate results. Sweat has drawn increasing attention as a promising source for non-invasive stress analysis. A sensitive HPLC-MS/MS method was developed for the quantitation of cortisol ((11β)-11,17,21-trihydroxypregn-4-ene-3,20-dione) in human eccrine sweat. At least one unknown isomer that has previously not been reported and could potentially interfere with quantification was separated from cortisol with mixed mode RP HPLC. Detection of cortisol was carried out using atmospheric pressure chemical ionization (APCI) and selected reaction monitoring (SRM) in positive ion mode, using cortisol-9,11,12,12-D4 as internal standard. LOD and LOQ were estimated to be 0.04 ng ml(-1) and 0.1 ng ml(-1), respectively. Linear range of 0.10-25.00 ng ml(-1) was obtained. Intraday precision (2.5%-9.7%) and accuracy (0.5%-2.1%),…

In the last decade Raman-spectroscopy has become an invaluable tool for biomedical diagnostics. However, a manual rating of the subtle spectral differences between normal and abnormal disease states is not possible or practical. Thus it is necessary to combine Raman-spectroscopy with chemometrics in order to build statistical models predicting the disease states directly without manual intervention. Within chemometrical analysis a number of corrections have to be applied to receive robust models. Baseline correction is an important step of the pre-processing, which should remove spectral contributions of fluorescence effects and improve the performance and robustness of statistical models. However, it is demanding, time-consuming, and depends on expert knowledge to select an optimal baseline correction method and its parameters every time working with a new dataset. To circumvent this issue we proposed a genetic algorithm based method to automatically optimize the baseline correction. The investigation was carried out in three main steps. Firstly, a numerical quantitative marker was…

We report the first use of a paper-based device coated with nanoceria as a simple, low-cost and rapid detection platform for the analysis of organophosphate (OP) pesticides using an enzyme inhibition assay with acetylcholinesterase (AChE) and choline oxidase (ChOX). In the presence of acetylcholine, AChE and ChOX catalyze the formation of H2O2, which is detected colorimetrically by a nanoceria-coated device resulting in the formation of a yellow color. After incubation with OP pesticides, the AChE activity was inhibited, producing less H2O2, and a reduction in the yellow intensity. The assay is able to analyze OP pesticides without the use of sophisticated instruments and gives detection limits of 18 ng mL(-1) and 5.3 ng mL(-1) for methyl-paraoxon and chlorpyrifos-oxon, respectively. The developed method was successfully applied to detect methyl-paraoxon in spiked vegetables (cabbage) and a dried seafood product (dried green mussel), obtaining ∼95% recovery values for both sample types. The spiked samples were also analyzed using LC-MS/MS as a comparison to the developed method and…

The immunoglobulin degrading enzyme from Streptococcus pyogenes, IdeS, was discovered as a mechanism by which pathogenic bacteria circumvent antibody mediated immune defense. IdeS was found to rapidly and specifically cleave IgG into F(ab')2 and Fc/2 fragments. The enzymatic specificity has led to a range of recent developments in the analytical strategies for characterization of monoclonal therapeutic antibodies and related products such as antibody-drug conjugates, bispecific antibodies, antibody mixtures and Fc-fusion proteins. In this review article we describe the discovery and properties of IdeS, discuss the current challenges in characterizing antibody therapeutics and review the methodologies using IdeS to improve the characterization of therapeutic antibodies. The review is focused on critical quality attributes of the final antibody product as studied by IdeS fragmentation such as Fab- and Fc-glycosylation, oxidation, glycation, C-terminal lysine and others. In summary, this review presents a wide range of IdeS-based applications for improved characterization of…

Small molecule neurotransmitters are essential for the function of the nervous system, and neurotransmitter imbalances are often connected to neurological disorders. The ability to quantify such imbalances is important to provide insights into the biochemical mechanisms underlying the disorder. This proof-of-principle study presents online quantification of small molecule neurotransmitters, specifically acetylcholine, γ-aminobutyric acid (GABA) and glutamate, in rat brain tissue sections using nanospray desorption electrospray ionization (nano-DESI) mass spectrometry imaging. By incorporating deuterated internal standards in the nano-DESI solvent we show identification, accurate mapping, and quantification of these small neurotransmitters in rat brain tissue without introducing any additional sample preparation steps. We find that GABA is about twice as abundant in the medial septum-diagonal band complex (MSDB) as in the cortex, while glutamate is about twice as abundant in the cortex as compared to the MSDB. The study shows that nano-DESI is well suited for imaging of small…

Graphene aerogel (GA)-supported metal-organic framework (MOF) particles with a three-dimensional (3D) architecture were fabricated for the first time via a facile template-free "sol-cryo" method. The prepared MOFs@graphene hybrid aerogels exhibit a 3D interconnected macroporous framework of graphene sheets with uniform dispersion of MOF particles. We also report the first attempt at using the hybrid aerogels as adsorbents for the solid-phase extraction (SPE) of non-steroidal anti-inflammatory drugs (NSAIDs) and the selective enrichment of proteins. The macroporous skeletons of GA provide both low backpressure and rapid mass transfer in SPE application, thus overcoming the obstacle of high backpressure caused by directly packing submicron or micron sized MOF particles into SPE cartridges. Excellent performances including satisfactory recoveries, high sensitivity and good reproducibility were achieved in the extraction of five NSAIDs. The hybrid aerogels also showed an interesting ability for selective enrichment of ribonuclease A (RNase A) and simultaneous exclusion of cytochrome C…

Gold nanoparticles (Au NPs) have become one of the most popular materials for sensing of analytes of interest in the last decade, mainly because of their ease in preparation and conjugation, stability, biocompatibility, and size-dependent optical properties. We have witnessed many sensitive and selective Au NP based optical systems for the quantitation of metal ions, anions, proteins, and DNA, based on analyte induced changes in their absorption, fluorescence, and scattering. In this tutorial review, we briefly discuss wet chemical approaches for the preparation of Au NPs. Sensing mechanisms and strategies of Au NP based optical systems are provided to show basic concepts in designing sensitive and selective sensing systems. Strategies for signal amplification applied in Au NP based systems are emphasized for the analysis of trace amounts of analytes in real samples. Many excellent Au NP based optical sensing systems are discussed to highlight their practicality for the analysis of complicated biological and environmental samples. The tutorial review ends with the discussion of the…

Key to native ion mobility/mass spectrometry is to prevent the structural denaturation of biological molecules in the gas phase. Here, we systematically assess structural changes induced in the protein ubiquitin during a trapped ion mobility spectrometry (TIMS) experiment. Our analysis shows that the extent of structural denaturation induced in ubiquitin ions is largely proportional to the amount of translational kinetic energy an ion gains from the applied electric field between two collisions with buffer gas particles. We then minimize the efficiency of the structural denaturation of ubiquitin ions in the gas phase during a TIMS experiment. The resulting "soft" TIMS spectra of ubiquitin are found largely identical to those observed on "soft" elevated-pressure ion mobility drift tubes and the corresponding calibrated cross sections are consistent with structures reported from NMR experiments for the native and A-state of ubiquitin. Thus, our analysis reveals that TIMS is useful for native ion mobility/mass spectrometry analysis.

Mesoporous silica films consisting of highly ordered and vertically aligned nanochannels (abbreviated as VMSFs) have received considerable attention because of their high surface area, long-range order, thermal and mechanical stability, controllable pore size and more importantly good molecular accessibility for rapid mass transport. These characteristics are ideal for electroanalytical chemistry and separation science. In this review, we firstly present briefly the strategies for the synthesis of VMSFs on the electrode surface, mainly the electrochemically assisted self-assembly and Stöber-solution growth approaches, as well as the surface modification of channel walls with diverse terminal groups for various functionalities. In the next section, recent progress on the applications of VMSFs in electroanalytical chemistry and sensing is summarized, in terms of the spatial confinement and permselective effects (size, charge and lipophilicity selectivity of the mesochannels). We then present the preparation and application of perforated free-standing VMSFs for fast and precise…

Recently, the nanoarchitectonics concept has been proposed to fabricate functional materials on the basis of concerted harmonization actions to control materials organization. In this review, we introduce recent several sensor designs from viewpoints of nanoarchitectonics that are classified into three categories: (i) inside nanoarchitectonics, (ii) outside nanoarchitectonics, and (iii) hierarchic nanoarchitectonics. In addition, various nanocarbons, such as carbon nanotubes, graphene derivatives, fullerene assemblies and nanoporous carbons, are selected as standard materials. In the first section, design and fabrication of mesoporous or nanoporous materials for enhanced sensing are especially exemplified as inside nanoarchitectonics. In the next section, recent examples of layer-by-layer nanoarchitectures for sensor fabrications are explained for outside nanoarchitectonics. Finally, their combined strategies for hierarchic carbon nanoarchitectonics are introduced as advanced sensor materials designs.

Surface-enhanced Raman spectrocopy (SERS) offers ultrasensitive vibrational fingerprinting at the nanoscale. Its non-destructive nature affords an ideal tool for interrogation of the intracellular environment, detecting the localisation of biomolecules, delivery and monitoring of therapeutics and for characterisation of complex cellular processes at the molecular level. Innovations in nanotechnology have produced a wide selection of novel, purpose-built plasmonic nanostructures capable of high SERS enhancement for intracellular probing while microfluidic technologies are being utilised to reproducibly synthesise nanoparticle (NP) probes at large scale and in high throughput. Sophisticated multivariate analysis techniques unlock the wealth of previously unattainable biomolecular information contained within large and multidimensional SERS datasets. Thus, with suitable combination of experimental techniques and analytics, SERS boasts enormous potential for cell based assays and to expand our understanding of the intracellular environment. In this review we trace the pathway to…

In this review, various inorganic materials were summarized for the analysis of small molecules by laser desorption/ionization mass spectrometry (LDI-MS). Due to its tremendous advantages, such as simplicity, high speed, high throughput, small analyte volumes and tolerance towards salts, LDI-MS has been widely used in various analytes. During the ionization process, a suitable agent is required to assist the ionization, such as an appropriate matrix for matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). However, it is normally difficult to analyze small molecules with the MALDI technique because conventional organic matrices may produce matrix-related peaks in the low molecular-weight region, which limits the detection of small molecules (m/z < 700 Da). Therefore, more and more inorganic materials, including carbon-based materials, silicon-based materials and metal-based materials, have been developed to assist the ionization of small molecules. These inorganic materials can transfer energy and improve the ionization efficiency of analytes. In addition,…

Lateral flow assays (LFAs) have been extensively explored in nucleic acid testing (NAT) for medical diagnostics, food safety analysis and environmental monitoring. However, the amount of target nucleic acid in a raw sample is usually too low to be directly detected by LFAs, necessitating the process of amplification. Even though cost-effective paper-based amplification techniques have been introduced, they have always been separately performed from LFAs, hence increasing the risk of reagent loss and cross-contaminations. To date, integrating paper-based nucleic acid amplification into colorimetric LFA in a simple, portable and cost-effective manner has not been introduced. Herein, we developed an integrated LFA with the aid of a specially designed handheld battery-powered system for effective amplification and detection of targets in resource-poor settings. Interestingly, using the integrated paper-based loop-mediated isothermal amplification (LAMP)-LFA, we successfully performed highly sensitive and specific target detection, achieving a detection limit of as low as 3 × 10(3)…

This paper presents a portable quantitative method for the on-site determination of uric acid in urine using surface-enhanced Raman spectroscopy (SERS) and gold nanoparticle-coated paper as a substrate. A procedure was developed for the rapid preparation of cost-effective SERS substrates that enabled the adequate control of a homogeneous active area and the use of small quantities of gold nanoparticles per substrate. The standard addition method and multivariate curve resolution-alternating least squares (MCR-ALS) were applied to compensate for the matrix effect and to address overlapping bands between uric acid and interference SERS spectra. The proposed methodology demonstrated better performance than conventional univariate methods (in terms of linearity, accuracy and precision), a wide linear range (0-3.5 mmol L(-1)) and an adequate limit of detection (0.11 mmol L(-1)). For the first time, a portable SERS method coupled with chemometrics was developed for the routine analysis of uric acid at clinically relevant concentrations with minimal sample preparation and easy extension…

Electrospray ion mobility-mass spectrometry (IM-MS) data show that for some small molecules, two (or even more) ions with identical sum formula and mass, but distinct drift times are observed. In spite of showing their own unique and characteristic fragmentation spectra in MS/MS, no configurational or constitutional isomers are found to be present in solution. Instead the observation and separation of such ions appears to be inherent to their gas-phase behaviour during ion mobility experiments. The origin of multiple drift times is thought to be the result of protonation site isomers ('protomers'). Although some important properties of protomers have been highlighted by other studies, correlating the experimental collision cross-sections (CCSs) with calculated values has proven to be a major difficulty. As a model, this study uses the pharmaceutical compound melphalan and a number of related molecules with alternative (gas-phase) protonation sites. Our study combines density functional theory (DFT) calculations with modified MobCal methods (e.g. nitrogen-based Trajectory Method…

Highly sensitive detection of mercury( ii ) ions by using a fluorescence spectrometer using MoS 2 nanosheets and DNA labeled carbon dots is presented.

3D printing, an upcoming technology, has vast potential to transform conventional fabrication processes due to the numerous improvements it can offer to the current methods. To date, the employment of 3D printing technology has been examined for applications in the fields of engineering, manufacturing and biological sciences. In this study, we examined the potential of adopting 3D printing technology for a novel application, electrochemical DNA biosensing. Metal 3D printing was utilized to construct helical-shaped stainless steel electrodes which functioned as a transducing platform for the detection of DNA hybridization. The ability of electroactive methylene blue to intercalate into the double helix structure of double-stranded DNA was then exploited to monitor the DNA hybridization process, with its inherent reduction peak serving as an analytical signal. The designed biosensing approach was found to demonstrate superior selectivity against a non-complementary DNA target, with a detection range of 1-1000 nM.

The presence of pathogenic bacteria in foods has always been a great threat to the wellbeing of people and the revenue of food manufacturers. Therefore, the demand for advanced detection methods that can sensitively and rapidly detect these pathogens has been of great importance. This study reports an electrochemical biosensor for rapid detection of E. coli O157:H7 with the integration of bifunctional glucose oxidase (GOx)-polydopamine (PDA) based polymeric nanocomposites (PMNCs) and Prussian blue (PB) modified screen-printed interdigitated microelectrodes (SP-IDMEs). The core-shell magnetic beads (MBs)-GOx@PDA PMNCs were first synthesized by the self-polymerization of dopamine (DA). Gold nanoparticles (AuNPs) were dispersed on the surface of PMNCs through biochemical synthesis to achieve further highly efficient adsorption of antibodies (ABs) and GOx. The final product ABs/GOxext/AuNPs/MBs-GOx@PDA PMNCs served as the carrier to separate target bacteria from food matrices as well as the amplifier for electrochemical measurement. The unbound PMNCs were separated by a filtration step…

The intercellular lipid structure of the stratum corneum (SC) plays a key role in skin barrier function. A depth profile of the intercellular lipid conformation and the lipid lateral packing order were measured in vivo in the human SC using confocal Raman microscopy. The depth profiles of the 2880 cm(-1)/2850 cm(-1) peak ratio intensity, which represent the C-H stretching and lateral packing order of lipids, and the 1080 cm(-1)/(1130 cm(-1) + 1060 cm(-1)) peak ratio, which represents the C-C skeleton vibration and trans-gauche conformation order of lipids, were investigated. The influence of keratin on the lipid peaks at 2850 cm(-1) and 2880 cm(-1) was excluded by the developed mathematical algorithm. The results show that the trans-conformation and lateral packing order of the intercellular lipids reach their maximum value in the SC at 20-40% of its depth and then decrease towards the stratum granulosum. These results show that at a depth of 20-40% (normally corresponding to a depth of 4-8 μm) the SC exhibits the most ordered lipids and therefore the highest skin barrier function.…