César Raposo

Synthetic cathinones have emerged as the predominant New Psychoactive Substances (NPS) seized within the European Union. Substituted methylmethcathinones (MMCs), chloromethcathinones (CMCs), and the emerging bromomethcathinones (BMCs) currently account for over 70% of total cathinone seizures. While Nuclear Magnetic Resonance (NMR) is the gold standard for determining ring substitution, its application is often hindered by the presence of cutting agents in forensic samples, which necessitate laborious purification steps. This study introduces a novel, dual-analytical framework for the rapid profiling of 2-MMC, 3-MMC, 4-MMC, 2-CMC, 3-CMC, 4-CMC, 4-BMC and N-ethylpentedrone (NEP) using authentic seized samples provided by the Spanish National Police. We report a high-throughput, eco-friendly GC-MS method using ethanol as a sustainable solvent, achieving baseline resolution of all isomers within a 6-minute run time, even in the presence of common adulterants such as caffeine. Furthermore, we describe, for the first time, a systematic FTIR spectroscopic analysis of in-plane C-H bending vibrations (1000–950 cm−1) as a robust diagnostic tool for discriminating between ortho, meta, and para positional isomers. This integrated approach —combining 'green' chromatography for rapid screening with definitive FTIR isomeric identification— offers forensic laboratories a reliable, rapid and efficient analytical tool to fight the evolving challenges of the illicit drug market.

In this study, sixteen secondary metabolites, including two chromones, four dibenzofurans, three lipids, three depsides, two aromatic compounds, a quinone, and a terpene, were detected in the methanol:acetone (1:1 v/v) extract of the lichen Hypotrachyna enderythraea (Zahlbr.) Hale, using High-Performance Liquid Chromatography coupled to Orbitrap Electrospray Ionization tandem Mass Spectrometry (HPLC-Orbitrap ESI tandem MS/MS). These metabolites were characterized by analysis of their exact molecular masses and corresponding fragmentation patterns. The retention times of the identified metabolites were compared with those of standard compounds, confirming the presence of naturally occurring bioactive compounds. Density Functional Theory (DFT) calculations were employed to investigate preferential deprotonation sites in representative polyprotic metabolites. All these findings may contribute to expanding the spectrum of compounds identified within the genus Hypotrachyna and to evaluating their potential biological activities.

The phytochemical investigation and evaluation of the antioxidant activity of the leaves, bark, and roots of Cochlospermum angolense Welw ex Oliv—a valued plant that is widely used in traditional Angolan medicine—hold significant importance. Compounds were extracted from the aforementioned plant using acetone and ethanol and identified by HPLC-ESI-MSn. Both extracts demonstrated notable abilities to scavenge 2,2-diphenyl-1-picrylhydrazyl, nitric oxide, and superoxide radicals, as well as to inhibit lipid peroxidation. A HPLC analysis revealed a diverse array of bioactive compounds, including flavonoids, phenols, alkaloids, quinones, and terpenes, which help neutralize free radicals and protect cells against oxidative stress, thereby contributing to the prevention of various diseases. Moreover, the acetone and ethanol extracts proved to be excellent sources of antioxidants. For the first time, the present study identified new compounds never reported in this species, such as (+)-abscisic acid, angustine B, pinobanksin, dihydrogenistein, (−)-8-prenylnaringenin, isoquercetin, samandarine, dihydromyricetin, and eupatoriocromene, in the leaves, bark, and roots, marking a significant advance in the chemical characterization of C. angolense. These findings enhance our understanding of the bioactive phytochemicals and antioxidant properties of C. angolense and open new avenues for future therapeutic and pharmacological research, further supporting its traditional use in Angolan medicine.

Water mint (Mentha aquatica) is used in many formulations worldwide as a functional food and natural remedy to treat gastrointestinal disorders, lung diseases, and certain mental disorders such as epilepsy and depression. This study assessed the bioactivity of its infusion extract (INF) and hydroethanolic extract (HE) to highlight its health benefits. These extracts were analyzed for their chemical composition by HPLC-DAD-ESI-MSn, their antioxidant and antidiabetic properties, and their capacities to protect human erythrocytes against induced hemoglobin oxidation and lipid peroxidation. The effect on normal human dermal fibroblast (NHDF) cells and on the N27 rat dopaminergic neuron cell line was also assessed. The chromatographic analysis identified 57 compounds belonging to hydroxycinnamic acids, flavanones, flavone, and isoflavonoids. In respect to the biological potential, the Mentha aquatica extracts revealed a notable capacity for 2,2-diphenyl-1-picrylhydrazyl, nitric oxide, and superoxide radicals, as well as for the inhibition of α-glucosidase action and the protection of human erythrocytes against oxidative damage. Quantification revealed noteworthy phenolic content in both extracts. Additionally, the extracts demonstrated less cytotoxic effects regarding the NHDF and N27 cell lines. Overall, Mentha aquatica presents promising antioxidant activity and a spectrum of potential biological activities, underscoring its significance as a novel antioxidant candidate for applications in animal nutrition, human medicine, and natural product research in the pharmaceutical and nutraceutical industries.

In this work, we carried out studies of the chemical composition of hexane, chloroform and ethanol extracts from two samples of the lichen Parmotrema hypoleucinum collected in Algeria. Each sample of the lichen P. hypoleucinum was collected on two different supports: Olea europaea and Quercus coccifera. Hexane extracts were prepared, in Soxhlet; each hexane extract was fractionated by its solubility in methanol; the products soluble in methanol were separated (cold): 1-Hexane, 2-Hexane; and the products insoluble in methanol (cold): 1-Cires, 2-Cires. A diazomethane esterified sample of 1-Hexane, 2-Hexane, 1-Cires and 2-Cires was analyzed by GC-MS, and the components were identified as methyl esters. In the 1-Hexane and 2-Hexane fractions, the methyl esters of the predominant fatty acids in the lichen were identified: palmitic acid, linoleic acid, oleic acid and stearic acid; a hydrocarbon was also identified: 13-methyl-17-norkaur-15-ene and several derivatives of orsellinic acid. In the 1-Cires and 2-Cires fractions, the previous fatty acids were no longer observed, and only the derivatives of orsellinic acid were found. The analysis of the 1-Hexane, 2-Hexane fractions by HPLC-MS/MS allows us to identify different chemical components, and the most characteristic products of the lichen were identified, such as Atranol, Chloroatranol, Atranorin and Chloroatranorin. In the fractions of 1-Cires and 2-Cires, the HPLC-MS/MS analysis reveals that they are very similar in their chemical components; the characteristic products of this lichen in this fraction are Atranorin and Chloroatranorin. In the extracts of chloroform, 1-Chloroform and 2-Chloroform, the analysis carried out by HPLC-MS/MS shows small differences in their chemical composition at the level of secondary products; among the products to be highlighted for this work, we have chloroatranorin, the stictic acid, norstictic acid and other derivatives. In the analysis of the most polar extracts carried out in ethanol: 1-Ethanol and 2-Ethanol, HPLC-MS/MS analysis shows very similar chemical compositions in these two extracts with small differences. In these extracts, the following acids were identified as characteristic compounds of this lichen: constictic acid, stictic acid, substictic acid and methylstictic acid. In the HPLC–MS/MS analysis of all these extracts, alectoronic acid was not found.

The present study provides new data concerning the chemical characterisation of Physcia mediterranea Nimis, a rare Mediterranean species belonging to the family Physciaceae. The phytochemical screening was carried out using GC-MS, HPLC-ESI-MS-MS, and NMR techniques. Hot extraction of n-hexane was carried out, followed by separation of the part insoluble in methanol: wax (WA-hex), from the part soluble in methanol (ME-hex). GC-MS analysis of the ME-hex part revealed the presence of methylbenzoic acids such as sparassol and atraric acid and a diterpene with a kaurene skeleton which has never been detected before in lichen species. Out of all the compounds identified by HPLC-ESI-MS-MS, sixteen compounds are common between WA-hex and ME-hex. Most are aliphatic fatty acids, phenolic compounds and depsides. The wax part is characterised by the presence of atranorin, a depside of high biological value. Proton 1H and carbon 13C NMR have confirmed its identification. Atranol, chloroatranol (depsides compound), Ffukinanolide (sesquiterpene lactones), leprolomin (diphenyl ether), muronic acid (triterpenes), and ursolic acid (triterpenes) have also been identified in ME-hex. The results suggested that Physcia mediterranea Nimis is a valuable source of bioactive compounds that could be useful for several applications as functional foods, cosmetics, and pharmaceuticals.

English. Foeniculum sanguineum Triano & A. Pujadas, sp. nov., from the south western Mediterranean Region (Spain & Morocco) is described. Its characterization and a comparative study with the related species Foeniculum vulgare Mill., has been carried out through morphological, cytological, chemical and molecular analysis. F. sanguineum is distinguished primarily for its red petals, pink pollen in fresh, and red stylopod. It is a diploid species (2n= 22). A high proportion of limonene and piperitenone oxide (absent in F. vulgare) has been found in the essential oil composition of the dry fruits of F. sanguineum and a high amount (about 50 %) of α-phellandrene in its roots and stems. Phylogenetic analyses were performed using the internal transcribed spacer sequences of nuclear ribosomal DNA (ITS) and the chloroplast rbcL gene sequences. ITS analysis supports the existence of the new species, while revealing sequence divergence both at the intraspecific and at the interspecific levels. A Single-nucleotide-polymorphism (SNP) sequence divergence found in the slow evolving chloroplast gene provided additional support for the novel species characterization, for which the name Foeniculum sanguineum is proposed.Español. Se describe Foeniculum sanguineum Triano & A. Pujadas, sp. nov., del suroeste de la Región Mediterránea (España y Marruecos). Para su caracterización se ha realizado el análisis morfológico, citológico, fitoquímico y molecular. Se ha llevado a cabo el studio comparativo con Foeniculum vulgare Mill. La nueva especie F. sanguineum se distingue principalmente por sus pétalos rojos, polen rosado en fresco y por su estilopodio rojo. Es una especie diploide (2n= 22). Se ha encontrado una alta proporción de óxido de limoneno y piperitenona (ausente en F. vulgare) en la composición de aceite esencial de los frutos secos de F. sanguineum y una elevada cantidad (aproximadamente 50%) de α-felandreno en sus raíces y tallos. El análisis filogenético se realizó utilizando las secuencias del espaciador transcrito interno de ADN ribosomal nuclear (ITS) y las secuencias del gen cloroplástico rbcL. El análisis ITS apoya la existencia de la nueva especie, al tiempo que revela la divergencia de secuencias tanto a nivel intraespecífico como a nivel interespecífico. La divergencia de secuencia encontrada en el gen cloroplástico, aunque reducida a un nucleótido, proporcionó apoyo adicional para la caracterización de la nueva especie, para la que se propone el nombre de Foeniculum sanguineum.

Taste, quality and perceived health benefits result in a high demand for both green and mature pork products derived from the Iberian bred swine. These characteristics are directly related to diet and breeding style (free ranging in a particular Mediterranean ecosystem: the Dehesa). Given that current demand is not matched by the available resources, animal feed is increasingly used. GC-C-IRMS analysis of Palmitic, Stearic, Oleic and Linoleic Acid methyl esters allows for differentiating between pork raised in the traditional way and that fattened on animal feed. Although a value of δ13CC18:1 = –25.9‰ is a good discriminator, exploratory analysis of principal canonical components is required to properly assign any unknown sample.

Urine is a suitable biological fluid to look for markers of physiological and pathological processes, including renal and nonrenal diseases. In addition, it is an optimal body sample for diagnosis, because it is easily obtained without invasive procedures and can be sampled in large quantities at almost any time. Rats are frequently used as a model to study human diseases, and rat urine has been analyzed to search for disease biomarkers. The normal human urinary proteome has been studied extensively, but the normal rat urinary proteome has not been studied in such depth. In light of this, we were prompted to analyze the normal rat urinary proteome using three complementary proteomics platforms: SDS‐PAGE separation, followed by LC‐ESI‐MS/MS; 2DE, followed by MALDI‐TOF‐TOF and 2D‐liquid chromatography‐chromatofocusing, followed by LC‐ESI‐Q‐TOF. A total of 366 unique proteins were identified, of which only 5.2% of unique proteins were identified jointly by the three proteomics platforms used. This suggests that simultaneous proteomics techniques provide complementary and nonredundant information. Our analysis affords the most extensive rat urinary protein database currently available and this may be useful in the study of renal physiology and in the search for biomarkers related to renal and nonrenal diseases.

Ras small GTPases function as transducers of extracellular signals regulating cell survival, growth and differentiation. There are three major ras isoforms: H‐, N‐ and K‐Ras. To improve the understanding of H‐ and N‐Ras protein signalling networks, we compared total proteome changes in mouse embryonic fibroblasts knock out for H‐ras and/or N‐ras, using proteomics tools combining 2DE, semi‐quantitative image analysis, in‐gel trypsin digestion and mass spectrometry. There are four up‐regulated proteins due to the loss of expression of H‐Ras (including cyclin‐dependent kinase inhibitor 2A) and eight down‐regulated (including stress‐70 protein, dihydropyrimidinase‐related‐protein 3, heat shock cognate 71 kDa protein, tropomyosin beta chain, Rho GDP‐dissociation inhibitor 1) and six up‐regulated proteins (e.g. leukocyte elastase inhibitor A, L‐lactate dehydrogenase B chain, c‐Myc‐responsive protein Rcl, interleukin‐1 receptor antagonist protein) due to the loss of expression of both N‐ and H‐Ras. Most of these proteins are related to Ras signalling in one way or another. Changes in expression of some of these proteins were further confirmed by Western blot. This proteomic comparative analysis from loss of function of H‐ and N‐Ras knockout fibroblasts yields interpretable data to elucidate the differential protein expression, and contributes to evaluate the possibilities for physiological and therapeutic targets.