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A novel Fe3O4@coPPy-PTH nanocomposite-based sorbent was prepared via in situ oxidative polymerization using Fe3O4 nanoparticles with spherical and flower-like morphologies of thiophene and pyrrole as the feedstocks. The synthesized nanocomposite displayed sensitive extraction and determination of metal ions Co(II), Cr(III), and Ni(II) without a chelating agent, followed by microsample injection system-flame atomic absorption spectrometry. Advanced spectroscopic and imaging techniques including scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy were used to characterize the composition and morphology of the Fe3O4@coPPy-PTH nanocomposite. SEM observations showed that the size of the Fe3O4 nanoparticles changed from 30 nm to 120 nm in diameter after copolymer (PPy-PTH) coating. The Fe3O4@coPPy-PTH nanocomposite has good dispersion properties and stability in strong acid solutions. For effective extraction of the studied analytes, the influence of sample pH, volume of sample solution and eluent, amount of adsorbent, and interference of coexisting metal ions were optimized. Under the optimum conditions, preconcentration factors were obtained as 25 for all analytes. The calibration curves were linear in the range of 0.0-10.0 µg L-1 with coefficients of determination (R2) greater than 0.9957 for all three analytes. Limits of detection (S/N = 3) were calculated in the range of 0.17-0.23 µg L-1. Precision values, expressed as relative standard deviations, were lower than 3.0%, and relative recoveries were obtained in the range of 88.6%-103.6%. The proposed method (Fe3O4@coPPy-PTH/MSPE/MIS-FAAS) was successfully applied to extract and determine the studied metal ions in beer, wine, and nonalcoholic beverage samples.

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The overarching challenge of this research is setting up a procedure to select the most appropriate fraction from complex, heterogeneous materials such as historic mortars in case of radiocarbon dating. At present, in the international community, there is not a unique and fully accepted way of mortar sample preparation to systematically obtain accurate results. With this contribution, we propose a strategy for selecting suitable mortar samples for radiocarbon dating of anthropogenic calcite in binder or lump. A four-step procedure is proposed: (I) good sampling strategies along with architectural and historical surveys; (II) mineralogical, petrographic, and chemical characterization of mortars to evaluate the feasibility of sample dating; (III) a non-destructive multi-analytical characterization of binder-rich portions to avoid geogenic calcite contamination; (IV) carbonate micro-sample preparation and accelerator mass spectrometer (AMS) measurements. The most innovative feature of the overall procedure relies on the fact that, in case of positive validation in step III, exactly the same material is treated and measured in step IV. The paper aims to apply this procedure to the ancient mortar of the Florentine historical building (Trebbio Castle), selecting micro-samples suitable for dating in natural hydraulic mortars. The discussion of the mortar dating results with the historical-archaeological hypotheses provided significant insights into the construction history of the building.

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OBJECTIVE: A simple, highly specific, accurate and fast method by smartphone-based digital imaging was developed for estimating lidocaine hydrochloride in pharmaceutical formulations. MATERIAL AND METHODS: To obtain the images, a Galaxy A03 Core smartphone and an image acquisition device developed in the laboratory were used to control the incident factors in reproducibility of the measurements. The processing of the images was carried out with the Color Grab application. Finally, the absorbance values were calculated using the RGB intensity values of blank, standard, and sample solutions. The proposed method was compared with spectroscopic and chromatographic methods. RESULTS: The reaction between copper and lidocaine hydrochloride was characterized, showing better results in an equimolar ratio and maintaining the pH of the solution above 11.5. The use of the device for the capture of digital images allowed to control those sensitive parameters for reproducibility so that the analytical measurements showed adequate precision and accuracy. Validation of the main parameters of the method showed compliance with acceptance criteria. The application of the method for the analysis of injectable samples achieved reliable results, which were statistically similar to other reference instrumental methods. CONCLUSION: The proposed method presented figures of merit in relation to linearity, precision, selectivity, accuracy, and robustness; it was carried out by designing and manufacturing a device for capturing digital images on a smartphone, which were analyzed to obtain RGB intensity values. These data are finally used to calculate absorbance values of solutions. All these elements provide this work with innovative characteristics in the field of analysis for control of pharmaceutical formulations.

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Introduction: Determination of folate insufficiency is of considerable interest given its importance in fetal development and red blood cell formation; however, access to blood tests may be limited due to the requirement for phlebotomy as well as controlled temperature shipping of blood specimens to laboratories for testing due to the inherent instability of folate and its vitamers. Methods: An LC-MS/MS test was developed and validated for the measurement of 5-methyltetrahydrofolate (5MTHF) in dried plasma specimens collected from fingerstick blood using a laminar flow blood separation device, as well as liquid venous plasma for comparison. Two pre-analytical factors investigated influencing the measurement of 5MTHF in dried plasma were hemolysis of the fingerstick blood during collection and storage/shipment of the dried plasma. Results: Although observed infrequently, hemolysis >10 % resulted in elevated 5MTHF measurements, but hemolysis >1 % resulted in elevated chloride measurements, which were necessary to normalize 5MTHF measurements for variation in volume of dried plasma specimens. Stability of 5MTHF was improved in dried plasma relative to liquid plasma at ambient temperatures, but not sufficiently to allow for uncontrolled temperature shipping despite controlling for humidity and light exposure. Shipping studies emulating ISTA procedure 7D were conducted with a reusable cold packaging solution. The packaging failed to stabilize 5MTHF in dried plasma specimens during a 2-day summer shipping evaluation, but did provide sufficient temperature control to stabilize 5MTHF during the overnight shipping evaluation. Conclusion: Our studies provide boundary conditions with respect to hemolysis, storage, and shipping for successful analysis of 5MTHF from dried plasma specimens.

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Tyrosine phosphorylation is one of the important protein phosphorylations in eukaryotes responsible for a variety of biological processes including cell signaling transduction, cell migration, and apoptosis. In the study of phosphoproteomics, due to the low stoichiometry of tyrosine phosphorylation (pTyr) proteins and sometimes limited initial sample, traditional phosphoproteomics enrichment technology is inefficient for the enrichment of pTyr peptides. Here, we review the substantial progress in tyrosine phosphoproteomics by preparation of limited amount sample and the newly introduced SH2 superbinder.

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Quantitative cellular in vitro nanoparticle uptake measurements are possible with a large number of different techniques, however, all have their respective restrictions. Here, we demonstrate the application of synchrotron-based X-ray fluorescence imaging (XFI) on prostate tumor cells, which have internalized differently functionalized gold nanoparticles. Total nanoparticle uptake on the order of a few hundred picograms could be conveniently observed with microsamples consisting of only a few hundreds of cells. A comparison with mass spectroscopy quantification is provided, experimental results are both supported and sensitivity limits of this XFI approach extrapolated by Monte-Carlo simulations, yielding a minimum detectable nanoparticle mass of just 5 pg. This study demonstrates the high sensitivity level of XFI, allowing non-destructive uptake measurements with very small microsamples within just seconds of irradiation time.

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One of the most cited limitations of biochemical detection is its poor sensitivity, owing to the relatively high complexity of micro-samples. Moreover, some samples cannot be easily self-replicated and their abundance cannot be increased through traditional technologies. Therefore, the preconcentration of low-abundance samples is a key requirement for microfluidic biological analysis. In recent years, the ion-concentration polarization phenomenon has aroused widespread interest in the application of microfluidic technology. In addition, paper-based materials are readily available, easy to modify, and exhibit good hydrophilicity. The study of the ion-concentration polarization preconcentration of micro-samples in paper-based microfluidic chips is of considerable significance. In this review, we discuss the development and applications of ion-concentration polarization paper-based preconcentrator in the past 5 years, with emphasis on key progresses in chip fabrication and performance optimization under different conditions. The current needs and development prospects in this field have also been discussed.

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Tyrosine phosphorylation is one of the important protein phosphorylations in eukaryotes responsible for a variety of biological processes including cell signaling transduction, cell migration, and apoptosis. In the study of phosphoproteomics, due to the low stoichiometry of tyrosine phosphorylation (pTyr) proteins and sometimes limited initial sample, traditional phosphoproteomics enrichment technology is inefficient for the enrichment of pTyr peptides. Here, we review the substantial progress in tyrosine phosphoproteomics by preparation of limited amount sample and the newly introduced SH2 superbinder.

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A targeted, bottom-up proteomic assay was developed for the qualitative detection of apolipoprotein L1 (ApoL1) protein variants (G0, G1, and G2) in blood plasma for identification of high and low renal risk genotypes. Following trypsin digestion of liquid or dry plasma, surrogate peptides specific to each ApoL1 variant were detected by liquid chromatography-tandem mass spectrometry along with two surrogate peptides common among all variants which served as internal (positive) controls to verify correct sample processing. Using isotopically labeled peptide internal standards, the presence or absence of each surrogate peptide was determined using multiple objective metrics including: 1) retention time confirmation relative to its internal standard, 2) comparison of the internal standard-normalized response relative to pre-established thresholds for confident detection, and 3) ion ratio monitoring. Based on the pattern of variant-specific surrogate peptides detected, the genotype was accurately inferred. The final, optimized method was fully validated for liquid plasma specimens, as well as dry plasma specimens collected on a laminar flow blood separation device. For both specimen types, the latter which can be self-collected for remote or in-home sampling, the assay was shown to be reproducible, interference-free with the exception of gross hemolysis, and accurate relative to Sanger sequencing (100% agreement). This targeted, qualitative bottom-up proteomic assay for detection of ApoL1 variants in blood provides a high-throughput, cost-effective alternative to molecular methods and has potential implications to improve organ allocation by facilitating screening kidney donors for high-risk ApoL1 genotypes, but could be applicable for genotyping other clinically relevant blood proteins variants.

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Art forgeries have existed since antiquity, but with the recent rapidly expanding commercialization of art, the approach to art authentication has demanded increasingly sophisticated detection schemes. So far, the most conclusive criterion in the field of counterfeit detection is the scientific proof of material anachronisms. The establishment of the earliest possible date of realization of a painting, called the terminus post quem, is based on the comparison of materials present in an artwork with information on their earliest date of discovery or production. This approach provides relative age information only and thus may fail in proving a forgery. Radiocarbon (14C) dating is an attractive alternative, as it delivers absolute ages with a definite time frame for the materials used. The method, however, is invasive and in its early days required sampling tens of grams of material. With the advent of accelerator mass spectrometry (AMS) and further development of gas ion sources (GIS), a reduction of sample size down to microgram amounts of carbon became possible, opening the possibility to date individual paint layers in artworks. Here we discuss two microsamples taken from an artwork carrying the date of 1866: a canvas fiber and a paint chip (<200 µg), each delivering a different radiocarbon response. This discrepancy uncovers the specific strategy of the forger: Dating of the organic binder delivers clear evidence of a post-1950 creation on reused canvas. This microscale 14C analysis technique is a powerful method to reveal technically complex forgery cases with hard facts at a minimal sampling impact.

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Piperacillin-tazobactam is a beta-lactam/beta-lactamase combination antibiotic used in patients with moderate to severe infection. Dosing of piperacillin-tazobactam requires an understanding of this patient group to maximise the effectiveness of this antibiotic and limit a further emergence of resistant pathogens. This is the first method that measures piperacillin and tazobactam simultaneously, across this range of clinically-relevant biological matrices. The calibration line was linear across the concentration range of 0.5-500µg/mL for piperacillin and 0.625-62.5µg/mL for tazobactam. All validation testing for matrix effects, precision and accuracy, specificity and stability were within 15%. A calibration equivalence study was performed to investigate the suitability of applying calibration curves prepared in an alternative matrix, with a mean bias of -10.8% identified for the application of a calibration line prepared for tazobactam in plasma only. Bias for all other calibration lines prepared in alternate matrices was within the 5% acceptance criteria. The method was successfully applied to a pharmacokinetic study of a critically ill patient receiving renal replacement therapy, with the results included.

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Plastic deformation of micron-scale crystalline materials differs considerably from bulk samples as it is characterized by stochastic strain bursts. To obtain a detailed picture of the intermittent deformation phenomena, numerous micron-sized specimens must be fabricated and tested. An improved focused ion beam fabrication method is proposed to prepare non-tapered micropillars with excellent control over their shape. Moreover, the fabrication time is less compared with other methods. The in situ compression device developed in our laboratory allows high-accuracy sample positioning and force/displacement measurements with high data sampling rates. The collective avalanche-like motion of the dislocations is observed as stress decreases on the stress-strain curves. An acoustic emission (AE) technique was employed for the first time to study the deformation behavior of micropillars. The AE technique provides important additional in situ information about the underlying processes during plastic deformation and is especially sensitive to the collective avalanche-like motion of the dislocations observed as the stress decreases on the deformation curves.

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Improved understanding of dental enamel development will benefit not only dentistry but also biomedicine more generally. Rat and mouse models of enamel development are relatively well characterized and experimentally powerful. However, the diminutive size of murine teeth makes them difficult to study using standard proteomics approaches. Here, we describe gel-based proteomic methods that enable parallel quantification, identification, and functional characterization of proteins from developing rat and mouse teeth. These refined methods are applicable to other scarce samples including human enamel defects.

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A rapid, sensitive, selective and validated reverse phase high-performance liquid chromatography (RP-HPLC) method for the estimation of paclitaxel in micro-sample of rat plasma and in culture of cancer cells was performed in this study. The mobile phase consisted of an optimized mixture of methanol:water: trifluroacetic acid (80: 20: 0.1, v/v/v). Column elution at a flow rate of 1 mL/minute with UV detection at 225 nm at room temperature was used. The RP-HPLC method was successfully applied for the determination of paclitaxel in plasma samples and in culture of cancer cells with nano-quantity of estimation. The validation studies were performed in accordance with the International Conference on Harmonization (ICH) guidelines. The intra- and inter-day precision showed that the coefficients of variation ranged from 1.07% to 4.27% at different levels of concentrations. To the best of our knowledge, this study also reported for the first time the optimization of different solvents for effective extraction of paclitaxel wherein tert.-butyl methyl ether (TBME): diethyl ether (DEE) in 50: 50 v/v composition was found most efficient with extraction efficiency ranging between 77.99% and 91.74% and between 76.14 and 93.66% in the plasma and cell culture, respectively. This proposed method was successfully applied to study the pharmacokinetics of paclitaxel and the influence of verapamil and all-trans retinoic acid (atRA) on paclitaxel pharmacokinetics in rat models. This proposed method might emerge as a valuable aid in the laboratory monitoring of paclitaxel in a variety of in vitro as well as in vivo scenarios.