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Oxidative stress is a pathological condition that contributes to the onset of various diseases. In this way, studying oxidative stress could lead to significant discoveries in the field of therapeutic and preventive medicine. Lipid peroxidation is the most significant event in the oxidative stress process and the gold standard biomarkers for endogenous oxidative damage to lipids are isoprostanes (IsoPs). This project aims to develop a reliable analytical method for the liquid-liquid microextraction technique, parallel artificial liquid membrane extraction (PALME) and LC-MS/MS analysis. PALME allowed to obtain a significant enrichment factor and, at the same time, a good sample purification by removing compounds that cause signal suppression, thereby reducing matrix effect. The chromatographic and mass spectrometric conditions have been fine tuned to improve the sensitivity of the method and therefore obtaining very low LOD and LOQ values. The recovery values obtained for the analytes are slightly above 50 %, except for 6-keto Prostaglandin F1A (24 %). Matrix effects were ≤ -10 %, with LODs ranging between 1 and 5 pg mL -1 . The developed method is characterized by high sensitivity and low consumption of organic solvents, according to the principles of Green Analytical Chemistry and enables the determination of basal levels of IsoPs in oral fluid by processing 96 samples simultaneously., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The present study encompasses the development of a fast and reliable analytical method to quantify the main endocannabinoids and some of their conjugated congeners, particularly N-arachidonoyl amino acids, in brain tissue. Samples were homogenized and a micro solid phase extraction (μSPE) procedure was developed for brain homogenate clean-up. Miniaturized SPE was selected as it allowed to work with reduced sample amounts, while maintaining high sensitivity; this last feature was mandatory due to the low concentration of endocannabinoids in biological matrices that makes their determination a challenging analytical task. UHPLC-MS/MS was used for the analysis as it provided a great sensitivity, especially for conjugated forms that were detected by negative ionization. Polarity switching was applied during the run; low limits of quantification were between 0.003 ng g -1 and 0.5 ng g -1 . This method provided also low matrix effect (lower than 30%) and good extraction recoveries in the brain. To the best of our knowledge, this is the first time that μSPE is applied on this matrix for this class of compounds. The method was validated according to international guidelines, and then tested on real cerebellum samples from mice, which were sub-chronically treated with URB597, a well-known inhibitor of the fatty acid amide hydrolase., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Europe is an important market for illegal drugs, and nowadays a lot of new different psychoactive substances (NPS) are widespread. This work reports the development of a method to determine simultaneously different classes of NPS, as synthetic cannabinoids (SC) and their metabolites, cathinones and phenetylamines, directly on whole blood (WB) without anti-coagulants and using miniaturized solid phase extraction (μ-SPE). In order to demonstrate the feasibility of the method 16 different NPS belonging to the mentioned classes were selected for the analysis. Recoveries ranged from 21% to 70% while matrix effect was lower than 15% for all the analytes. LOQ values were 5ngmL -1 for cathinones and phenetylamines, between 0.25 and 1ngmL -1 for SCs and up to 2.5ngmL -1 for SC metabolites. The performance of μ-SPE was compared with different clean-up strategies (i.e. protein precipitation (PPT), liquid liquid extraction, PPT/SPE hybrid) in term of recovery, matrix effect and suitability for multi-class analysis. The developed method was validated according to SWGTOX guidelines. The validation data demonstrated that this approach is potentially very useful as confirmation method for multiclass analysis in WB and post mortem specimens. In fact only 100μL of human WB are used, sample preparation involves few rapid steps and the method is easily implementable for the determination of other NPS., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The objective of the present work is to demonstrate a rational way to prepare selective sorbents able to extract simultaneously several structural analogs. For this purpose the binding specificity of two hexapeptides computationally designed (VYWLVW and YYIGGF) versus four synthetic cannabinoids Naphthalen-1-yl-(1-pentylindol-3-yl)methanone (JWH 018), naphthalen-1-yl-(1-butylindol-3-yl)methanone (JWH 073), (R)-(1-((1-methylpiperidin-2-yl)methyl)-1H-indol-3-yl)(naphthalen-1-yl)methanone (AM 1220) and (R)-(+)-[2,3-Dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-napthalenylmethanone (WIN 55) was computationally studied and then experimentally tested by solid-phase extraction (SPE) clean-up and ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis. The two peptides were chosen using a semi combinatorial virtual technique by generating 4 cycles of peptide libraries (around 2.3×10 4 elements). To select the two peptides, the simulated binding scores between synthetic cannabinoids and peptides was used by maximizing the recognition properties of amino acid motif between the two JWH and the other synthetic cannabinoids. In particular, the peptide YYIGGF, having also affinity for AM 120, was selected as control because it was the only one without tryptophan residues within the best peptides obtained from simulation. Experimentally, the two hexapeptides were tested as SPE sorbent using nanomolar solutions of the four drugs. After optimization of best retentions the binding constants were calculated by loading synthetic cannabinoids solutions at different concentrations. The results indicated a strong interaction between hexapeptide VYWLVW and JWH 018 (15.58±2.03×10 6 M -1 ), 3-fold and 40-fold larger compared to the analog JWH 073 and both AM 1220 and the WIN 55. Similar trend was observed for the hexapeptide YYIGGF but the binding constants were at least three times lower highlighting the key role of the tryptophan. To demonstrate the hexapeptides specific interaction with only synthetic cannabinoids, a cross-reactivity study was carried out using other drugs (cocaine, morphine, phencyclidine and methamphetamine) in the same SPE condition. Finally the practical utility of these peptide modified sorbent materials was further demonstrated by detecting the synthetic cannabinoids in real samples using hair matrix., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The binding specificity of a bio-inspired hexapeptide (QHWWDW) versus cocaine and four other drugs such as 3,4-methylenedioxy-N-methylamphetamine (MDMA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA), phencyclidine and morphine was computationally studied and then experimentally confirmed in solid phase extraction (SPE) followed by liquid chromatography-mass spectrometry (LC/MS) detection. In simulation, the hexapeptide-drug complexes were docked with different scoring functions and considering pH chemical environment. In experimental, the cross reactivity of the selected hexapeptide was tested as SPE sorbent versus cocaine and other four drugs using buffer solutions at pH 4 and 7. Significant differences in specific retention were found between cocaine (97% of recovery) and both morphine (45% of recovery) and phencyclidine (60% of recovery), but less for ecstasies (average recovery 69%). In agreement with docking simulation, the hexapeptide showed the highest recovery with best specificity versus cocaine at pH 7 with an experimentally binding constant of 2.9 × 10(6)M(-1). The bio-inspired sorbent material analytical performances were compared with a commercial reversed phase cartridge confirming the hexapeptide specificity to cocaine and validating simulated data., (Copyright © 2014 Elsevier B.V. All rights reserved.)