Detailed investigation of the lipidome remodeling upon normal weight conditions, obesity, or weight loss, as well as the influence of physical activity, can help to understand the mechanisms underlying dyslipidemia in metabolic conditions correlated to the emergence and progression of non-alcoholic fatty liver disease (NAFLD). C57BL/6 male mice were fed a normal diet (ND) or a high-fat diet (HFD) for 20 weeks. Subgroups within the high-fat diet (HFD) group underwent different interventions: some engaged in exercise (HFDex), others were subjected to weight loss (WL) by changing from the HFD to ND, and some underwent a combination of weight loss and exercise (WLex) during the final 8 weeks of the 20-week feeding period. To support our understanding, not only tissue-specific lipid remodeling mechanisms but also the cross-talk between different tissues and their impact on the systemic regulation of lipid metabolism are essential. Exercise and weight loss-induced specific adaptations in the liver and visceral adipose tissue lipidomes of mice were explored by the UPLC–TOF–MS/MS untargeted lipidomics methodology. Lipidomic signatures of ND and HFD-fed mice undergoing weight loss were compared with animals with and without physical exercise. Several lipid classes were identified as contributing factors in the discrimination of the groups by multivariate analysis models, such as glycerolipids, glycerophospholipids, sphingolipids, and fatty acids, with respect to liver samples, whereas triglycerides were the only lipid class identified in visceral adipose tissue. Lipids found to be dysregulated in HFD animals are related to well-established pathways involved in the biosynthesis of PC, PE, and TG metabolism. These show a reversing trend back to basic levels of ND when animals change to a normal diet after 12 weeks, whereas the impact of exercise, though in some cases it slightly enhances the reversing trend, is not clear. Full article

A method was developed for the determination of 26 drugs of abuse from different classes, including illicit drugs in quantitative dried blood spots (qDBSs), with the aim to provide a convenient method for drug testing by using only 10 μL of capillary blood. A satisfactory limit of quantification (LOQ) of 2.5 ng/mL for 9 of the compounds and 5 ng/mL for 17 of the compounds and a limit of detection (LOD) of 0.75 ng/mL for 9 of the compounds and 1.5 ng/mL for 17 of the compounds were achieved for all analytes. Reversed-phase liquid chromatography was applied on a C18 column coupled to MS, providing selective detections with both +ESI and -ESI modes. Extraction from the qDBS was performed using AcN-MeOH, 1:1 (v/v), with recovery ranging from 84.6% to 106%, while no significant effect of the hematocrit was observed. The studied drugs of abuse were found to be stable over five days under three different storage conditions (at ambient temperature 21 °C, at −20 °C, and at 35 °C), thus offering a highly attractive approach for drug screening by minimally invasive sampling for individuals that could find application in forensic toxicology analysis. Full article

Food and fish adulteration is a major public concern worldwide. Apart from economic fraud, health issues are in the forefront mainly due to severe allergies. Sardines are one of the most vulnerable-to-adulteration fish species due to their high nutritional value. Adulteration comprises the substitution of one fish species with similar species of lower nutritional value and lower cost. The detection of adulteration, especially in processed fish products, is very challenging because the morphological characteristics of the tissues change, making identification by the naked eye very difficult. Therefore, new analytical methods and (bio)sensors that provide fast analysis with high specificity, especially between closely related fish species, are in high demand. DNA-based methods are considered as important analytical tools for food adulteration detection. In this context, we report the first DNA sensors for sardine species identification. The sensing principle involves species recognition, via short hybridization of PCR-amplified sequences with specific probes, capture in the test zone of the sensor, and detection by the naked eye using gold nanoparticles as reporters; thus, avoiding the need for expensive instruments. As low as 5% adulteration of Sardina pilchardus with Sardinella aurita was detected with high reproducibility in the processed mixtures simulating canned fish products. Full article

The Bland–Altman approach is one of the most widely used mathematical approaches for method comparison and analytical agreement. This work describes, for the first time, the application of Bland–Altman to study ¹⁴N/¹⁵N and ¹H/²H (D) chromatographic isotope effects of endogenous analytes of the L-arginine/nitric oxide pathway in human plasma, serum and urine samples in GC-MS. The investigated analytes included arginine, asymmetric dimethylarginine, dimethylamine, nitrite, nitrate and creatinine. There was a close correlation between the percentage difference of the retention times of the isotopologs of the Bland–Altman approach and the area under the curve (AUC) values of the receiver operating characteristic (ROC) approach (r = 0.8619, p = 0.0047). The results of the study suggest that the chromatographic isotope effects in GC-MS result from differences in the interaction strengths of H/D isotopes in the derivatives with the hydrophobic stationary phase of the GC column. D atoms attenuate the interaction of the skeleton of the molecules with the lipophilic GC stationary phase. Differences in isotope effects in plasma or serum and urine in GC-MS are suggested to be due to a kind of matrix effect, and this remains to be investigated in forthcoming studies using Bland–Altman and ROC approaches. Full article

In this work, a simple and cost-efficient aptasensor strip is developed for the rapid detection of OTA in food samples. The biosensor is based on the lateral flow assay concept using an OTA-specific aptamer for biorecognition of the target analyte. The strip consists of a sample pad, a conjugate pad, a nitrocellulose membrane (NC) and an absorbent pad. The conjugate pad is loaded with the OTA-specific aptamer conjugated with gold nanoparticles (AuNPs). The test line of the NC membrane is loaded with a specific OTA-aptamer probe and the control line is loaded with a control probe. The assay is based on a competitive format, where the OTA present in the sample combines with the OTA aptamer-AuNP conjugate and prevents the interaction between the specific probe immobilized on the test line and the OTA aptamer-AuNP conjugates; therefore, the color intensity of the test line decreases as the concentration of OTA in the sample increases. Qualitative detection of OTA is performed visually, while quantification is performed by reflectance colorimetry using a commercial scanner and image analysis. All the parameters of the assay are investigated in detail and the analytical features are established. The visual limit of detection (LOD) of the strip is 0.05 ng mL⁻¹, while the LOD for semi-quantitative detection using reflectance colorimetry is 0.02 ng mL⁻¹. The lateral flow strip aptasensor is applied to the detection of OTA in wine, beer, apple juice and milk samples with recoveries in the range from 91 to 114%. The assay exhibits a satisfactory selectivity for OTA with respect to other mycotoxins and lasts 20 min. Therefore, the lateral flow strip aptasensor could be useful for the rapid, low-cost and fit-for-purpose on-site detection of OTA in food samples. Full article

Significant efforts have been spent in the modern era towards implementing environmentally friendly procedures like composting to mitigate the negative effects of intensive agricultural practices. In this context, a novel fertilizer was produced via the hydrolysis of an onion-derived compost, and has been previously comprehensively chemically characterized. In order to characterize its efficacy, the product was applied to tomato plants at five time points to monitor plant health and growth. Control samples were also used at each time point to eliminate confounding parameters due to the plant’s normal growth process. After harvesting, the plant leaves were extracted using aq. MeOH (70:30, v/v) and analyzed via UPLC-QToF-MS, using a C18 column in both ionization modes (±ESI). The data-independent (DIA/bbCID) acquisition mode was employed, and the data were analyzed by MS-DIAL. Statistical analysis, including multivariate and trend analysis for longitudinal monitoring, were employed to highlight the differentiated features among the controls and treated plants as well as the time-point sequence. Metabolites related to plant growth belonging to several chemical classes were identified, proving the efficacy of the fertilizer product. Furthermore, the efficiency of the analytical and statistical workflows utilized was demonstrated. Full article

In this work, a simple, fast, and sensitive voltammetric method for the trace determination of the alkaloid drug colchicine (Colc) using a 3D-printed device is described. The electrochemical method was based on the adsorptive accumulation of the drug at a carbon-black polylactic acid (CB/PLA) working electrode, followed by voltammetric determination of the accumulated species. The plastic sensor was printed in a single step by a low-cost dual extruder 3D-printer and featured three CB/PLA electrodes (serving as working, reference, and counter electrodes) and a holder, printed from a non-conductive PLA filament. The electrochemical parameters that affected the response of the device towards Colc determination, such as accumulation time and potential, solution pH, and other variables, were optimized. Under the selected conditions, the oxidation current of Colc was proportional to the concentration of Colc, and its quantification was conducted in the concentration range of 0.6–2.2 μmol L⁻¹ with a limit of detection of 0.11 μmol L⁻¹ in phosphate buffer (pH 7.0). Both within-device and between-device reproducibility were lower than 9%, revealing satisfactory operational and fabrication reproducibility. Furthermore, the 3D-printed device was employed for the voltammetric determination of Colc in pharmaceutical tablets and in human urine with satisfactory results, justifying its suitability for low-cost routine analysis of Colc. Full article

In this work, the total content of seven metallic elements (Fe, Cu, Zn, Mg, Pb, Ni, and Co) in common edible herbs was determined and related to their bioaccessibility by an in vitro human digestion model. Specifically, the unified bioaccessibility protocol developed by the BioAccessibility Research Group of Europe (BARGE) was used to determine the release of each element during gastric and gastrointestinal digestion. The results show that Fe, Zn, and Mg are released during gastric digestion (34–57% Fe, 28–80% Zn, 79–95% Mg), but their overall bioaccessibility is reduced in the gastrointestinal tract (<30%). On the contrary, Cu is more bioaccessible during gastrointestinal digestion (38–60%). Pb, Ni, and Co exhibited similar bioaccessibility in both gastric and gastrointestinal fluids. Principle component analysis of the data shows that the classification of the nutritional value of herbs differs between the total and the gastrointestinal concentration, suggesting that the total concentration alone is not an adequate indicator for drawing secure conclusions concerning the nutritional benefits of edible plant species. Full article

This study reports a new approach for the determination of copper ions in water samples that exploits the complexation reaction with diethyldithiocarbamate (DDTC) and uses widely available imaging devices (i.e., flatbed scanners or smartphones) as detectors. Specifically, the proposed approach is based on the ability of DDTC to bind to copper ions and form a stable Cu-DDTC complex with a distinctive yellow color detected with the camera of a smartphone in a 96-well plate. The color intensity of the formed complex is linearly proportional to the concentration of copper ions, resulting in its accurate colorimetric determination. The proposed analytical procedure for the determination of Cu²⁺ was easy to perform, rapid, and applicable with inexpensive and commercially available materials and reagents. Many parameters related to such an analytical determination were optimized, and a study of interfering ions present in the water samples was also carried out. Additionally, even low copper levels could be noticed by the naked eye. The assay performed was successfully applied to the determination of Cu²⁺ in river, tap, and bottled water samples with detection limits as low as 1.4 µM, good recoveries (89.0–109.6%), adequate reproducibility (0.6–6.1%), and high selectivity over other ions present in the water samples. Full article

A simple and rapid method for the extraction of D-series resolvins (RvD1, RvD2, RvD3, RvD4, RvD5) released into Leibovitz’s L-15 complete medium by head kidney cells from Atlantic salmon and the further determination of liquid chromatography triple quadrupole mass spectrometry is proposed. A three-level factorial design was proposed to select the optimal concentrations of internal standards that were used in the evaluation of the performance parameters, such as linear range (0.1–50 ng mL⁻¹), limits of detection and quantification (0.05 and 0.1 ng mL⁻¹, respectively), and recovery values ranging from 96.9 to 99.8%. The optimized method was used to determine the stimulated production of resolvins by head kidney cells exposed to docosahexaenoic acid, and the results indicated that it is possible that the production was controlled by circadian responses. Full article

Nitrite (O=N-O⁻, NO₂⁻) and nitrate (O=N(O)-O⁻, NO₃⁻) are ubiquitous in nature. In aerated aqueous solutions, nitrite is considered the major autoxidation product of nitric oxide (^●NO). ^●NO is an environmental gas but is also endogenously produced from the amino acid L-arginine by the catalytic action of ^●NO synthases. It is considered that the autoxidation of ^●NO in aqueous solutions and in O₂-containing gas phase proceeds via different neutral (e.g., O=N-O-N=O) and radical (e.g., ONOO^●) intermediates. In aqueous buffers, endogenous S-nitrosothiols (thionitrites, RSNO) from thiols (RSH) such as L-cysteine (i.e., S-nitroso-L-cysteine, CysSNO) and cysteine-containing peptides such as glutathione (GSH) (i.e., S-nitrosoglutathione, GSNO) may be formed during the autoxidation of ^●NO in the presence of thiols and dioxygen (e.g., GSH + O=N-O-N=O → GSNO + O=N-O⁻ + H⁺; pK_a^HONO, 3.24). The reaction products of thionitrites in aerated aqueous solutions may be different from those of ^●NO. This work describes in vitro GC-MS studies on the reactions of unlabeled (¹⁴NO₂⁻) and labeled nitrite (¹⁵NO₂⁻) and RSNO (RS¹⁵NO, RS¹⁵N¹⁸O) performed in pH-neutral aqueous buffers of phosphate or tris(hydroxyethylamine) prepared in unlabeled (H₂¹⁶O) or labeled H₂O (H₂¹⁸O). Unlabeled and stable-isotope-labeled nitrite and nitrate species were measured by gas chromatography–mass spectrometry (GC-MS) after derivatization with pentafluorobenzyl bromide and negative-ion chemical ionization. The study provides strong indication for the formation of O=N-O-N=O as an intermediate of ^●NO autoxidation in pH-neutral aqueous buffers. In high molar excess, HgCl₂ accelerates and increases RSNO hydrolysis to nitrite, thereby incorporating ¹⁸O from H₂¹⁸O into the SNO group. In aqueous buffers prepared in H₂¹⁸O, synthetic peroxynitrite (ONOO⁻) decomposes to nitrite without ¹⁸O incorporation, indicating water-independent decomposition of peroxynitrite to nitrite. Use of RS¹⁵NO and H₂¹⁸O in combination with GC-MS allows generation of definite results and elucidation of reaction mechanisms of oxidation of ^●NO and hydrolysis of RSNO. Full article

Mixtures of anionic sodium oleate (NaOl) and nonionic ethoxylated or alkoxylated surfactants improve the selective separation of magnesite particles from mineral ores during the process of flotation. Apart from triggering the hydrophobicity of magnesite particles, these surfactant molecules adsorb to the air–liquid interface of flotation bubbles, changing the interfacial properties and thus affecting the flotation efficiency. The structure of adsorbed surfactants layers at the air–liquid interface depends on the adsorption kinetics of each surfactant and the reformation of intermolecular forces upon mixing. Up to now, researchers use surface tension measurements to understand the nature of intermolecular interactions in such binary surfactant mixtures. Aiming to adapt better to the dynamic character of flotation, the present work explores the interfacial rheology of NaOl mixtures with different nonionic surfactants to study the interfacial arrangement and viscoelastic properties of adsorbed surfactants under the application of shear forces. Interfacial shear viscosity results reveal the tendency on nonionic molecules to displace NaOl molecules from the interface. The critical nonionic surfactant concentration needed to complete NaOl displacement at the interface depends on the length of its hydrophilic part and on the geometry of its hydrophobic chain. The above indications are supported by surface tension isotherms. Full article

Sol-gel graphene oxide-coated polyester fabric platforms were synthesized and used for the on-line sequential injection fabric disk sorptive extraction (SI-FDSE) of toxic (i.e., Cd(II), Cu(II) and Pb(II)) metals in different distilled spirit drinks prior to their determination by electrothermal atomic absorption spectrometry (ETAAS). The main parameters that could potentially influence the extraction efficiency of the automatic on-line column preconcentration system were optimized and the SI-FDSE-ETAAS method was validated. Under optimum conditions, enhancement factors of 38, 120 and 85 were achieved for Cd(II), Cu(II) and Pb(II), respectively. Method precision (in terms of relative standard deviation) was lower than 2.9% for all analytes. The limits of detection for Cd(II), Cu(II) and Pb(II) were 1.9, 7.1 and 17.3 ng L⁻¹, respectively. As a proof of concept, the proposed protocol was employed for the monitoring of Cd(II), Cu(II), and Pb(II) in distilled spirit drinks of different types. Full article

The use of e-cigarettes (ECs) has become increasingly popular worldwide, even though scientific results have not established their safety. Diacetyl (DA) and acetylpropionyl (AP), which can be present in ECs, are linked with lung diseases. Ethyl maltol (EM)—the most commonly used flavoring agent—can be present in toxic concentrations. Until now, there is no methodology for the determination of nicotine, propylene glycol (PG), vegetable glycerin (VG), EM, DA, and acetylpropionyl in e-liquids that can be used as a quality control procedure. Herein, gas chromatography coupled with mass spectrometry (GC-MS) was applied for the development of analytical methodologies for these substances. Two GC-MS methodologies were developed and fully validated, fulfilling the standards for the integration in a routine quality control procedure by manufacturers. As proof of applicability, the methodology was applied for the analysis of several e-liquids. Differences were observed between the labeled and the experimental levels of PG, VG, and nicotine. Three samples contained EM at higher concentrations compared to the other samples, while only one contained DA. These validated methodologies can be used for the quality control analysis of EC liquid samples regarding nicotine, PG, and VG amounts, as well as for the measurement of the EM. Full article

The technique of two-dimensional high-performance liquid chromatography has managed to gain the recognition it deserves thanks to the advantages of satisfactory separations it can offer compared to simple one-dimensional. This review presents in detail key features of the technique, modes of operation, and concepts that ensure its optimal application and consequently the best possible separation of even the most complex samples. Publications focusing on the separation of antibiotics and their respective impurities are also presented, providing information concerning the analytical characteristics of the technique related to the arrangement of the instrument and the chromatographic conditions. Full article

Reliable quantification in biological systems of endogenous low- and high-molecular substances, drugs and their metabolites, is of particular importance in diagnosis and therapy, and in basic and clinical research. The analytical characteristics of analytical approaches have many differences, including in core features such as accuracy, precision, specificity, and limits of detection (LOD) and quantitation (LOQ). Several different mathematic approaches were developed and used for the comparison of two analytical methods applied to the same chemical compound in the same biological sample. Generally, comparisons of results obtained by two analytical methods yields different quantitative results. Yet, which mathematical approach gives the most reliable results? Which mathematical approach is best suited to demonstrate agreement between the methods, or the superiority of an analytical method A over analytical method B? The simplest and most frequently used method of comparison is the linear regression analysis of data observed by method A (y) and the data observed by method B (x): y = α + βx. In 1986, Bland and Altman indicated that linear regression analysis, notably the use of the correlation coefficient, is inappropriate for method-comparison. Instead, Bland and Altman have suggested an alternative approach, which is generally known as the Bland–Altman approach. Originally, this method of comparison was applied in medicine, for instance, to measure blood pressure by two devices. The Bland–Altman approach was rapidly adapted in analytical chemistry and in clinical chemistry. To date, the approach suggested by Bland–Altman approach is one of the most widely used mathematical approaches for method-comparison. With about 37,000 citations, the original paper published in the journal The Lancet in 1986 is among the most frequently cited scientific papers in this area to date. Nevertheless, the Bland–Altman approach has not been really set on a quantitative basis. No criteria have been proposed thus far, in which the Bland–Altman approach can form the basis on which analytical agreement or the better analytical method can be demonstrated. In this article, the Bland–Altman approach is re-valuated from a quantitative bioanalytical perspective, and an attempt is made to propose acceptance criteria. For this purpose, different analytical methods were compared with Gold Standard analytical methods based on mass spectrometry (MS) and tandem mass spectrometry (MS/MS), i.e., GC-MS, GC-MS/MS, LC-MS and LC-MS/MS. Other chromatographic and non-chromatographic methods were also considered. The results for several different endogenous substances, including nitrate, anandamide, homoarginine, creatinine and malondialdehyde in human plasma, serum and urine are discussed. In addition to the Bland–Altman approach, linear regression analysis and the Oldham–Eksborg method-comparison approaches were used and compared. Special emphasis was given to the relation of difference and mean in the Bland–Altman approach. Currently available guidelines for method validation were also considered. Acceptance criteria for method agreement were proposed, including the slope and correlation coefficient in linear regression, and the coefficient of variation for the percentage difference in the Bland–Altman and Oldham–Eksborg approaches. Full article