Tuning the properties of peptide imprinted nanoparticles for protein immunoprecipitation using magnetic streptavidin beads.

Maximizing the binding properties of thermoresponsive molecularly imprinted nanoparticles (MIN) was aimed to explore their feasibility as antibody substitutes in protein immunoprecipitation (IPP) with magnetic streptavidin beads (MSB). Thermoresponsive MIN targeting the cannabinoid CB1 receptor were produced by epitope imprinting through solid-phase synthesis. It was intended to determine how different variables influenced physicochemical features, binding behaviour and immunoprecipitation of the target recombinant glutathione S-transferase tagged fusion protein (GST-CTer). Such variables included the cross-linking degree of MIN, and variables like pH, temperature or the use of Tween-20 for binding and IPP experiments. The cross-linker (CL) amount influenced the coil-to-globule transition of thermoresponsive MIN, making the lower critical solution temperature (LCST) decrease from 37.2 °C using 5% of CL, to 29.0 °C using 25%, also suggesting higher plasticity on the former. Temperature influence on size was corroborated by dynamic light scattering, observing size reductions from 250-450 nm (RT) to 70-100 nm (> LCST) for MIN produced with 5-15% of CL. However, binding behaviour did not clearly  improve for more than 10% CL. Further experiments revealed that temperature and pH control were critical for efficient binding and release, selecting 40 °C and pH 5 as appropriate. Following binding experiments, the GST-CTer-MIN complex was successfully immunoprecipitated using MSB, achieving an IPP efficiency of 11.48% over the initial input protein concentration, which was calculated after SDS-PAGE separation and Western blot analysis. The methodology may be exploited for selective protein extraction and quantification from complex tissue homogenates.

Revealing novel biomarkers for diagnosing chronic kidney disease in pediatric patients.

Pediatric chronic kidney disease (CKD) is a clinical condition characterized by progressive renal function deterioration. CKD diagnosis is based on glomerular filtration rate, but its reliability is limited, especially at the early stages. New potential biomarkers (citrulline (CIT), symmetric dimethylarginine (SDMA), S-adenosylmethionine (SAM), n-butyrylcarnitine (nC4), cis-4-decenoylcarnitine, sphingosine-1-phosphate and bilirubin) in addition to creatinine (CNN) have been proposed for early diagnosis. To verify the clinical value of these biomarkers we performed a comprehensive targeted metabolomics study on a representative cohort of CKD and healthy pediatric patients. Sixty-seven children with CKD and forty-five healthy children have been enrolled in the study. Targeted metabolomics based on liquid chromatography-triple quadrupole mass spectrometry has been used for serum and plasma samples analysis. Univariate data analysis showed statistically significant differences (p < 0.05) in the concentration of CNN, CIT, SDMA, and nC4 among healthy and CKD pediatric patients. The predictive ability of the proposed biomarkers was also confirmed through specificity and sensitivity expressed in Receiver Operating Characteristic curves (AUC = 0.909). In the group of early CKD pediatric patients, AUC of 0.831 was obtained, improving the diagnostic reliability of CNN alone. Moreover, the models built on combined CIT, nC4, SDMA, and CNN allowed to distinguish CKD patients from healthy control regardless of blood matrix type (serum or plasma). Our data demonstrate potential biomarkers in the diagnosis of early CKD stages.

Biostimulants as an Alternative to Improve the Wine Quality from Vitis vinifera (cv. Tempranillo) in La Rioja.

The application of biostimulants appears to be an environmentally friendly, innovative, and sustainable agronomical tool to mitigate the negative effects induced by adverse climatology in traditional grape-growing regions such as La Rioja (Spain). However, their mechanism of action in grapevines is still unclear. We evaluated how commercial substances (two from Ascophyllum nodosum extraction and one amino acids-based biostimulant) and the non-proteinogenic amino acid ß-aminobutyric acid (BABA) affect the quality and quantity of musts and grapes in Vitis vinifera L. cv. Tempranillo from a semi-arid region of La Rioja during two seasons. We hypothesized an enhancement in organic metabolites in berries and leaves in response to these treatments, changing the organoleptic characteristics of the final products. The treatments altered the primary metabolites such as carbohydrates, organic acids (AcOrg), and free amino acids, first in the leaves as the effect of the foliar application and second in grapes and musts. As the main result, the biostimulant efficiency depended on the climatology and vineyard location to improve the final yield. Whereas biostimulant application enhanced the yield in 2018 (less dry year), it did not help production in 2019 (dry year). BABA was the most efficient biostimulant, enhancing plant production. Regarding yield quality, the biostimulant application improved the musts mainly by enhancing the fumaric acid content and by reducing carbohydrates, except in BABA-treated plants, where they were accumulated. These results corroborate biostimulants as an exciting approach in wine production, especially for improving wine quality.

Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis.

The production of artificial anti-CB1 antibodies in nanoparticle format is described using the solid-phase imprinting approach. Instead of whole protein imprinting, a linear C-terminus sequence of the receptor comprising 15 amino acids (458-KVTMSVSTDTSAEAL-472) has been used as template, in accordance with the epitope imprinting approach. This sequence is located intracellularly, and it is involved in coupling to Gi/o proteins, being responsible for CB1 receptor desensitisation and internalisation. Developed molecularly imprinted materials were found to be in the nanometre scale, with a particle size of 126.4 ± 10.5 nm at pH 3 (25 ºC) and spherical shape. It was also observed that the size was sensible to temperature changes being reduced to 106.3 ± 15.2 nm at 35 °C. Lower critical solution temperature of this polymer was found to be ≈ 33.4 °C. The affinity and selectivity of the artificial antibody were assessed through dot blot and Western blot experiments. For the latter, recombinant fusion proteins GST-CB1414-472 and GST-CB1414-442 were produced to work respectively as target and negative control proteins. The control protein did not carry the target epitope for being devoid of last 30 amino acids at the C-terminus. The results demonstrated that the anti-CB1 material recognised selectively the target protein, thanks to the presence of the 15-amino acid sequence selected as epitope, which revealed that binding occurred at the C-terminus of the receptor itself. The methodology presented may pave the way for the development of novel imprinted nanomaterials for other proteins included in the superfamily of the G-protein-coupled receptors (GPCR).

Interplay between 1-aminocyclopropane-1-carboxylic acid, γ-aminobutyrate and D-glucose in the regulation of high nitrate-induced root growth inhibition in maize.

Nitrogen is one of the main factors that affect plant growth and development. However, high nitrogen concentrations can inhibit both shoot and root growth, even though the processes involved in this inhibition are still unknown. The aim of this work was to identify the metabolic alterations that induce the inhibition of root growth caused by high nitrate supply, when the whole plant growth is also reduced. High nitrate altered nitrogen and carbon metabolism, reducing the content of sugars and inducing the accumulation of Ca2+ and amino acids, such as glutamate, alanine and γ-aminobutyrate (GABA), that could act to replenish the succinate pool in the tricarboxylic acid cycle and maintain its activity. Other metabolic alterations found were the accumulation of the polyamines spermidine and spermine, and the reduction of jasmonic acid (JA) and the ethylene precursor aminocyclopropane-1-carboxylic acid (ACC). These results indicate that the growth root inhibition by high NO3- is a complex metabolic response that involves GABA as a key link between C and N metabolism which, together with plant growth regulators such as auxins, cytokinins, abscisic acid, JA, and the ethylene precursor ACC, is able to regulate the metabolic response of root grown under high nitrate concentrations.

Particle Analysis for the Detection of Gunshot Residue (GSR) in Nasal Samples Using Scanning Laser Ablation and Inductively Coupled Plasma-Mass Spectrometry (SLA-ICPMS).

Currently, aluminum stub with carbon adhesive devices are used to collect inorganic gunshot residues (GSR) from the hands of a shooter. In an ideal shooting case, the gunshot particles do not persist for more than 2 h in the hands of the shooter, provided that the hands have not been washed. However, for forensic analysis and inference, the extended persistence of GSR would be desirable. This study investigates a novel GSR sampling and detection protocol. Sampling was performed in the nostrils using swab devices impregnated in ethylenediaminetetraacetic acid (EDTA). The GSRs persisted for longer periods in nasal mucus than on the hands, and particles were detected 6 h after shooting occurred. The analytical determination was conducted by scanning laser ablation-inductively coupled plasma-mass spectrometry (SLA-ICPMS) which enable the identification of the number of particles and their elemental composition. Seventeen isotope signals corresponding to 13 C, 205 Tl and 15 analytes that are usually associated with the composition of GSR residues were monitored: 27 Al, 29 Si, 31 P, 33 S, 35 Cl, 39 K, 44 Ca, 57 Fe, 60 Ni, 63 Cu, 66 Zn, 118 Sn, 121 Sb, 137 Ba, and 208 Pb. The SLA technique enabled the reduction of the swab analysis time to 40 min. The effectiveness of this methodology was evaluated with two types of firearms: a pistol and a shotgun. The results indicated that the methodology proposed for the analysis of the nasal GSR was effective and that it can improve or complement the forensic analyses and inferences presented in a court.

Fungicide distribution in vitiviniculture ecosystems according to different application strategies to reduce environmental impact.

Systematic fungicides treatments in vine-growing European ecosystems have been conducted for decades. The goal of this study was to determine the mobility and persistence of 20 fungicides used in two viticultural zones in Atlantic and Mediterranean climates, from the moment of their application until their distribution throughout different compartments of the ecosystem: soil, water, grapes, musts and wines. This study also sought to obtain valuable information to reduce the usage of these products without affecting the health of the vines. For this purpose, different phytosanitary treatments were applied, using dosing criteria based on data provided by meteorological stations, degree-day accumulation, phenological state, and growers' criteria. The observed differences between studied geographical areas were not significant with regard to chemical accumulation in the soil and water; however, they were significantly different regarding to grapes, musts, and wines.

Characterization of ancient lipids in prehistoric organic residues: Chemical evidence of livestock-pens in rock-shelters since early neolithic to bronze age.

The characterization of ancient lipids from prehistoric sediments (fumiers) located in a rock-selter has been possible after the optimization of an analytical method based on the microwave-assisted extraction and solid-phase extraction clean-up step and a final derivatization step followed by gas chromatography with mass spectrometry. Eight sterols and two bile acids were detected just in the partially burned and unburned layers of the fumiers (animal organic residues deriving from manure/dung). The relationship between some of these compounds can be used to distinguish the biogenic origin of the samples, concluding that these strata (from Early Neolithic to Late Chalcolithic/Early Bronze Age) can be classified as ruminant residues. Three main periods of activity are observed over a period of 2000 years: one from 3990 ± 40 before present (4530-4410 calibrated before present) to 4100 ± 40 before present (4820-4750/4730-4510/4470-4450 calibrated before present), the second from 4470 ± 40 before present (5300-4970 calibrated before present) to 5490 ± 30 before present (6310-6275/6230-6220 calibrated before present) and the third from 5880 ± 30 before present (6775-6765/6750-6645 calibrated before present) to 6010 ± 30 before present (6940-6780/6765-6755 calibrated before present). Chemical data obtained are in concordance with the previous results obtained in the area.

Molecularly imprinted nanoparticles grafted to porous silica as chiral selectors in liquid chromatography.

The work presented here explores the grafting of molecularly imprinted nanoparticles (MIN) on silica beads for the development of new chiral stationary phases (CSP). Both solid-phase imprinting and precipitation polymerisation were tested for MIN synthesis though the latter approach was the only one that provided efficient chiral selectors. MIN particles were prepared by iniferter polymerisation initiated by UV radiation, using itaconic acid as functional monomer and ethylene dimethacrylate as cross-linker. This resulted in particles with an average size of 249.0±4.0nm which were covalently immobilised onto chromatographic silica beads. The resultant CSP based on the composite silica beads-MIN was capable of resolving the racemate of the antidepressant drug citalopram and also separating its major metabolites by liquid chromatography, with better efficiency and peak symmetry than other MIP based CSP. The methodology presented here allowed for the quantification of the pharmacologically active enantiomer (+)-(S)-citalopram (SCIT) and its main metabolites (+)-(S)-desmethylcitalopram (SDCIT) and (+)-(S)-didesmethylcitalopram (SDDCIT) in urine, registering mean recoveries that ranged from 91.5 to 103.7% with RSD values that were below 10% in all tested concentration levels (0.1, 0.75 and 5µgmL-1), which confirmed method suitability for the intended application.

Determination of mercury(ii) in water at sub-nanomolar levels by laser ablation-ICPMS analysis of screen printed electrodes used as a portable voltammetric preconcentration system.

Environmental pollution by mercury in ambient water samples is a recognized problem worldwide. Sample preservation and transport to the laboratory lead to uncertain analytical results. This study outlines the development of a procedure for on-site electrodeposition of mercury from water samples on a screen-printed gold electrode (SPGE) using portable voltammetric techniques. Once in the laboratory, Hg is analyzed by laser ablation inductively coupled plasma-mass spectrometry (LA-ICPMS) in order to ensure that the required sensitivity and precision levels for environmental sample analysis are reached. A new ablation chamber was intentionally designed for the analysis of SPGE's gold electrode. This cell has a small internal volume of 15 cm3 and the SPGE device perfectly fits inside. This design assures signal stability, avoids elemental fractionation and reduces wash-out time to a few seconds, reducing the analysis time considerably. The proposed method is capable of measuring dissolved mercury at the ng L-1 level (quantification limit 200 ng L-1) with good precision (RSD < 7.6%). The proposed method was tested with the NCS ZC 76303 (NIM-GBW08603) Mercury in water Certified Reference Material.

Liquid chromatography-quadrupole time of flight tandem mass spectrometry-based targeted metabolomic study for varietal discrimination of grapes according to plant sterols content.

Grapevine and derived products are rich in a wide range of compounds and its quality mainly depends on its metabolites, as a result of viticulture practices. Plant sterols, also called phytosterols (PS), are secondary metabolites regarded as bioactive substance present in grape berries and other plant-based food. The present study deals with a metabolomic approach focusing on phytosterols family in six varieties of Rioja grapes (Cabernet Sauvignon, Tempranillo, Graciano, Garnacha, White Garnacha and Viura), in order to find significant differences among them. Liquid chromatography- mass spectrometry with a quadrupole-time of flight mass analyzer (LC-QTOF) was used to find as many metabolites as possible in the different grape berry fractions, and using statistics to help finding significant clustering of the metabolic profile of pulp, peel and seeds in relation to the variety. The best chromatographic and detection conditions were achieved by gas phase ionization via atmospheric pressure chemical ionization (APCI) in positive mode. Furthermore, analysis with electrospray (ESI) is also needed for phytosterol derivatives confirmation. Putative compounds of interest in the analyzed samples were found by an automated compound extraction algorithm (Molecular Feature Extraction, MFE) and an initial differential expression from the data was created with the aid of commercial software. Once the data were collected, the results were filtered, aligned and normalized, and evaluating applying one-way analysis of variance (ANOVA) with a 95% significance level. For sample class prediction, partial least square-discriminant analysis (PLS-DA) is used as a supervised pattern recognition method and excellent separation among the grape varieties is shown. An overall accuracy of 93.3% (pulp samples), 100.0% (peel) or 96.7% (seeds) in discriminating between grape varieties was achieved when comparing the different fractions. In general, 7 PS derivatives were identified with ID scores higher than 84%.

A novel strategy for Cr(III) and Cr(VI) analysis in dietary supplements by speciated isotope dilution mass spectrometry.

In recent years, Cr speciation in dietary supplements has become decisive in the evaluation of their health risks. Despite being an beneficial micronutrient, Cr(III) can be toxic at living organisms at high concentrations, while Cr(VI) is known to be highly toxic and carcinogenic. The main objective of this work was to optimize an analytical methodology for the extraction and accurate quantification of Cr(III) and Cr(VI) in dietary supplements. The extraction of Cr species was carried out with 50mM EDTA solution on a hotplate under optimized conditions. Special attention was paid to bidirectional species transformations. No noticeable oxidation of Cr(III) into Cr(VI) was observed and the reduction to Cr(III) only occurred at very high Cr(VI) concentrations. Cr(III) as Cr(EDTA)(-) complex was chromatographically separated from Cr(VI), retained as CrO4(2-), on an anion exchange column coupled to inductively coupled plasma mass spectrometry (LC-ICP-MS). The limit of quantification (0.08µgg(-1)) was below the limit established for Cr enriched yeasts by the European Union. Eleven dietary supplements were analyzed and Cr(III) and Cr(VI) quantification was carried out by external calibration monitoring (52)Cr isotope and by speciated isotope dilution mass spectrometry (SIDMS) adding (50)Cr(III) and (53)Cr(VI) spikes. Total Cr was also quantified by ICP-MS and mass balance between total Cr and the sum of Cr(III) and Cr(VI) was achieved. In eight of the eleven tested supplements Cr(III) calculated amounts were higher than those indicated by the manufacturer, but only one of them exceeded the 250µgday(-1) recommended by World Health Organization (WHO). In contrast, it is worth noting that Cr(VI) amounts beyond the recommendations of the European Union for Cr enriched yeasts were found in five supplements. These results revealed that more accurate and rigorous quality assurance protocols should be applied to the testing of the final products, including the analysis of both Cr(III) and Cr(VI).

Water compatible stir-bar devices imprinted with underivatised glyphosate for selective sample clean-up.

This paper reports the development of stir bars with a new MIP based coating, for the selective sorptive extraction of the herbicide glyphosate (GLYP). Molecular imprinting of the polymer has directly been carried out employing underivatised GLYP as the template molecule. Due to the poor solubility of the target compound in organic solvents, the MIP methodology has been optimised for rebinding in aqueous media, being the synthesis and the rebinding steps carried out in water:methanol mixtures and pure aqueous media. The coating has been developed by radical polymerisation initiated by UV energy, using N-allylthiourea and 2-dimethyl aminoethyl methacrylate as functional monomers and ethylene glycol dimethacrylate as the cross-linker. Mechanical stability of the coating has been improved using 1,3-divinyltetramethyldisiloxane in the polymerisation mixture. Under the optimised conditions, the MIP has demonstrated excellent selectivity for the target compound in the presence of structural analogues, including its major metabolites. The applicability of the proposed method to real matrices has also been assessed using river water and soil samples. Registered mean recoveries ranged from 90.6 to 97.3% and RSD values were below 5% in all cases, what confirmed the suitability of the described methodology for the selective extraction and quantification of GLYP.

LC-QTOF-MS-based targeted metabolomics of arginine-creatine metabolic pathway-related compounds in plasma: application to identify potential biomarkers in pediatric chronic kidney disease.

Chronic kidney disease (CKD) is a major epidemiologic problem which causes several disturbances in adults and in pediatrics. Despite being a worldwide public health problem, information available for CKD in the pediatric population is scarce. For that reason, an ion-pair reversed-phase liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) method has been developed and validated in order to analyze 16 amino acids, amino acid derivatives, and analogous compounds related to the arginine-creatine metabolic pathway that are suspicious of being increased or decreased in plasma from patients with CKD. The analytical method involved the addition of dithiothreitol, a reducing agent which reduces disulfide and thus giving total aminothiol concentration, as well as a simple precipitation of plasma proteins. Moreover, despite amino acids being usually derivatized to improve their retention time and to enhance their signal, for this method, an ion-pairing reagent was used, thus avoiding the need for derivatization. Subsequently, analysis of plasma from pediatric patients suffering from CKD and control pediatrics was carried out. As a result, glycine, citrulline, creatinine, asymmetric dimethylarginine (ADMA), and symmetric dimethylarginine (SDMA) were significantly increased in patients with CKD, regardless of their creatinine level, whereas in addition to these compounds dimethylglycine was also increased when CKD patients had plasma creatinine concentrations above 12 µg mL(-1), thus all are suggested as potential biomarkers for renal impairment.

Iniferter-mediated grafting of molecularly imprinted polymers on porous silica beads for the enantiomeric resolution of drugs.

A surface-imprinted chiral stationary phase for the enantiomeric resolution of the antidepressant drug, citalopram, is presented in this work. N, N'-diethylaminodithiocarbamoylpropyl(trimethoxy)silane has been used as silane iniferter for the surface functionalization of the solid silica support. A molecularly imprinted polymer thin film, in the nm scale, was then grafted on the silanized silica using itaconic acid as the functional monomer and ethylene glycol dimethacrylate as the cross-linker in the presence of the template S-citalopram. The total monomer amount was calculated to obtain the desired thickness. Non-imprinted stationary phases were prepared similarly in the absence of S-citalopram. Characterization of the materials was carried out by scanning electron microscopy, thermogravimetric analysis, elemental analysis and Fourier transform infrared spectroscopy. Stationary phases have been applied to the chromatographic separation of the target. Conditions for best chromatographic resolution of the enantiomers were optimized, and it was found that a mobile phase consisting of a mixture of formate buffer (40 mM, pH 3) and acetonitrile (30:70 v/v) at 40 °C provided best results. Binding behaviour of the developed material was finally assessed by batch rebinding experiments. The obtained binding isotherm was fitted to different binding models being the Freundlich-Langmuir model, the one that best fitted the experimental data. The developed material has shown high selectivity for the target enantiomer, and the stationary phase could be undoubtedly exploited for chiral separation of the drug.

Molecularly imprinted polymers as a tool for the study of the 4-ethylphenol metabolic pathway in red wines.

A molecularly imprinted polymer (MIP) based methodology is described here for the determination of compounds that belong to the 4-ethylphenol (4EP) metabolic pathway in red wines. To this end, two MIP materials have been developed: a 4EP MIP as a class-selective material to extract phenols that belong to the 4EP metabolic pathway and a coumaric acid (CA) imprinted polymer as a MIP-based stationary phase capable of selectively separating these phenols on HPLC analysis, obtaining clean chromatograms. 4-vinyl pyridine and ethylene glycol dimethacrylate were respectively used as functional monomer and cross-linker for both MIPs. Once polymer compositions were optimised, the 4EP MIP was packed into SPE cartridges for wine sample clean-up and CA MIP was packed into HPLC columns to chromatographically separate the compounds present in the eluates obtained after SPE extraction. The accuracy of the proposed method was tested spiking wine samples with known concentrations of target compounds and subsequently, analytes were quantified by the standard addition method. Registered mean recoveries ranged from 95.2 to 109.2% and RSD values were below 10% in most cases. The described methodology was found to be suitable for the selective extraction and quantification of the compounds that belong to the 4EP metabolic pathway in red wines with minimal matrix effects and could be undoubtedly exploited to monitor 4EP and its precursors in wines.

Nuclear diacylglycerol lipase-α in rat brain cortical neurons: evidence of 2-arachidonoylglycerol production in concert with phospholipase C-ß activity.

In this report, we describe the localization of diacylglycerol lipase-α (DAGLα) in nuclei from adult cortical neurons, as assessed by double-immunofluorescence staining of rat brain cortical sections and purified intact nuclei and by western blot analysis of subnuclear fractions. Double-labeling assays using the anti-DAGLα antibody and NeuN combined with Hoechst staining showed that only nuclei of neuronal origin were DAGLα positive. At high resolution, DAGLα-signal displayed a punctate pattern in nuclear subdomains poor in Hoechst's chromatin and lamin B1 staining. In contrast, SC-35- and NeuN-signals (markers of the nuclear speckles) showed a high overlap with DAGLα within specific subdomains of the nuclear matrix. Among the members of the phospholipase C-ß (PLCß) family, PLCß1, PLCß2, and PLCß4 exhibited the same distribution with respect to chromatin, lamin B1, SC-35, and NeuN as that described for DAGLα. Furthermore, by quantifying the basal levels of 2-arachidonoylglycerol (2-AG) by liquid chromatography and mass spectrometry (LC-MS), and by characterizing the pharmacology of its accumulation, we describe the presence of a mechanism for 2-AG production, and its PLCß/DAGLα-dependent biosynthesis in isolated nuclei. These results extend our knowledge about subcellular distribution of neuronal DAGLα, providing biochemical grounds to hypothesize a role for 2-AG locally produced within the neuronal nucleus.

Characterization of organic gunshot residues in lead-free ammunition using a new sample collection device for liquid chromatography-quadrupole time-of-flight mass spectrometry.

The identification of characteristic organic gunshot residues (OGSR) provides conclusive evidence in the elucidation of elemental profiles when lead-free ammunition is fired. OGSR also prevents false negatives. Toward this aim, a quick and efficient method based on liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF) was developed to detect and identify 18 gunpowder additives in gunshot residues (GSR). The unequivocal identification of target analytes was assured by using MS/MS mode. Swabs were compared with home-modified tape lift supports covered with a PTFE layer to determine the better sampling technique. The modified tape lift provided better extraction recoveries and enabled the analysis of inorganic and organic GSR simultaneously. The developed method was applied to the analysis of GSR from four different lead-free ammunitions. Diphenylamine and its nitrated degradation products and centralites were identified in all samples, providing strong evidence of GSR.

A novel method for the identification of inorganic and organic gunshot residue particles of lead-free ammunitions from the hands of shooters using scanning laser ablation-ICPMS and Raman micro-spectroscopy.

A method based on scanning laser ablation and inductively coupled plasma-mass spectrometry (SLA-ICPMS) and Raman micro-spectroscopy for the detection and identification of compounds consistent with gunshot residue particles (GSR) has been developed. The method has been applied to the characterization of particles resulting from the discharge of firearms using lead-free ammunition. Modified tape lifts were used to collect the inorganic and organic residues from skin surfaces in a single sample. Using SLA-ICPMS, aggregates related to the composition of the ammunition, such as Cu-Zn-Sn, Zr-Sr, Cu-Zn, Al-Ti, or Al-Sr-Zr were detected, but this composition is only consistent with GSR from lead-free ammunitions. Additional evidence was provided by micro-Raman spectroscopy, which identified the characteristic organic groups of the particles as centralite, diphenylamine or their nitrated derivatives, which are indicative of GSR.

Direct potentiometric quantification of histamine using solid-phase imprinted nanoparticles as recognition elements.

A new potentiometric sensor based on molecularly imprinted nanoparticles produced via the solid-phase imprinting method was developed. For histamine quantification, the nanoparticles were incorporated within a membrane, which was then used to fabricate an ion-selective electrode. The use of nanoparticles with high affinity and specificity allowed for label-free detection/quantification of histamine in real samples with short response times. The sensor could selectively quantify histamine in presence of other biogenic amines in real wine and fish matrices. The limit of detection achieved was 1.12×10(-6)molL(-1), with a linear range between 10(-6) and 10(-2)molL(-1) and a response time below 20s, making the sensor as developed a promising tool for direct quantification of histamine in the food industry.