Centro de Química Estrutural

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Publications

Synthesis of emerging cathinones and validation of a SPE GC–MS method for their simultaneous quantification in blood

Publication . Júlio, Sara; Ferro, Raquel A.; Santos, Susana; Alexandre, Andrea; Caldeira, Maria João; Franco, João; Barroso, Mário; Gaspar, Helena

Over the past 15 years, synthetic cathinones have emerged as an important class of new psychoactive substances (NPS) worldwide. The proliferation of these psychostimulants and their sought-after effects among recreational drug users pose a serious threat to public health and enormous challenges to forensic laboratories. For forensic institutions, it is essential to be one step ahead of covert laboratories, foreseeing the structural changes possible to introduce in the core skeleton of cathinones while maintaining their stimulating activity. In this manner, it is feasible to equip themselves with standards of possible new cathinones and validated analytical methods for their qualitative and quantitative detection. Therefore, the aim of the work herein described was to synthesize emerging cathinones based on the evolving patterns in the illicit drug market, and to develop an analytical method for their accurate determination in forensic situations. Five so far unreported cathinones [4′-methyl-N-dimethylbuphedrone (4-MDMB), 4′-methyl-N-ethylbuphedrone (4-MNEB), 4′-methyl-N-dimethylpentedrone (4-MDMP), 4′-methyl-N-dimethylhexedrone (4-MDMH), and 4′-methyl-N-diethylbuphedrone (4-MDEB)] and a sixth one, 4′-methyl-N-ethylpentedrone, already reported to EMCDDA and also known as 4-MEAP, were synthesized and fully characterized by nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS). An analytical method for the simultaneous quantification of these cathinones in blood, using solid phase extraction (SPE) combined with gas chromatography-mass spectrometry (GC–MS) was developed and validated. The results prove that this methodology is selective, linear, precise, and accurate. For all target cathinones, the extraction efficiency was higher than 73%, linearity was observed in the range of 10 (lower limit of quantification, LLOQ) to 800 ng/mL, with coefficients of determination higher than 0.99, and the limits of detection (LODs) were 5 ng/mL for all target cathinones. The stability of these cathinones in blood matrices is dependent on the storage conditions; 4-MNEB is the most stable compound and 4-MDMH is the least stable compound. The low limits obtained allow the detection of the compounds in situations where they are involved, even if present at low concentrations.

2023Journal article

Restricted access

Mesoporous silica nanoparticles with manganese and lanthanide salts: synthesis, characterization and cytotoxicity studies

Publication . Forte, Andreia; Gago, Sandra; Carrott, Manuela Ribeiro; Carrott, Peter; Alves, Celso; Teodoro, Fernando; Pedrosa, Rui; Marrucho, Isabel M.; Branco, Luis C

Several organic salts based on the combination of two different choline derivative cations and MnCl3−, GdCl4− and TbCl4− as anions were immobilized in mesoporous silica nanoparticles (MSNs) by a two-step synthetic method. Firstly, MSNs were functionalized with choline derivative cations with chloride anions and then the metals were incorporated by the reaction of the chloride with the respective metal chloride salts. These nanomaterials were fully characterized by different characterization techniques such as 1H-NMR, FT-IR, elemental analysis, TEM, TGA, N2 adsorption, XRD and DLS. These characterization data were important to confirm the successful functionalization of the nanomaterials and to access their textural properties and colloidal stability. The final materials were also characterized by ICP-MS that indicated the metal contents. The cytotoxicity profile was evaluated in four different cell lines (3T3, 293T, HepG2 and Caco-2), which shows some relevant differences between the metal organic salts and their immobilized analogues.

2021Journal article

Restricted access

Valorisation of Sargassum muticum through the extraction of phenolic compounds using eutectic solvents and intensification techniques

Publication . Jesus, Bárbara C.; Sáenz de Miera, Blanca; Santiago, Rubén; Martins, Alice; Pedrosa, Rui; Gonzalez-Miquel, Maria; Marrucho, Isabel M.

Seaweeds are naturally abundant and spread all over the globe. They have several biologically active secondary metabolites of great interest. In this work, Sargassum muticum was the algae employed as biomass and the aim was to extract phenolic compounds (PCs) using eutectic solvents (ESs). Several betaine-based, proline-based, and choline-based ESs were tested for the extraction of PCs. All extracts were evaluated according to the total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity (DPPH). Afterwards, the extracts were characterized using HPLC in terms of 9 target PCs (gallic acid, 3,4-dihydroxybenzoic acid, caffeic acid, syringic acid, p-coumaric acid, ferulic acid, salicylic acid, catechin and quercetin). Proline combined with propylene glycol (Pro[thin space (1/6-em)]:[thin space (1/6-em)]PPG) exhibited a higher yield according to HPLC results, followed by proline[thin space (1/6-em)]:[thin space (1/6-em)]1,2-butanediol (Pro[thin space (1/6-em)]:[thin space (1/6-em)]1,2-But) and choline[thin space (1/6-em)]:[thin space (1/6-em)]citric acid (ChCl[thin space (1/6-em)]:[thin space (1/6-em)]CA). Pro[thin space (1/6-em)]:[thin space (1/6-em)]PPG also presented high selectivity towards salicylic acid, while ChCl[thin space (1/6-em)]:[thin space (1/6-em)]CA towards gallic acid. Optimization studies of water content and temperature were performed for the three best ESs, the optimum conditions being 30% (v/v) water and 60 °C extraction temperature. Ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) were two intensification methods evaluated to enhance the extraction process, proving their ability to reduce the extraction time when compared with the conventional solid–liquid extraction (SLE) process. In particular, Pro[thin space (1/6-em)]:[thin space (1/6-em)]PPG-based MAE provided a significantly higher extraction yield in comparison with conventional extraction and with the other extraction solvents. In summary, the combination of ESs with intensification techniques was shown to be a valuable valorization strategy of a marine macroalgae waste, in particular Sargassum muticum.

2023Journal article

Open access

Adverse outcome pathways induced by 3,4‑dimethylmethcathinone and 4‑methylmethcathinone in differentiated human SH‑SY5Y neuronal cells

Publication . Soares, Jorge; Costa, Vera Marisa; Gaspar, Helena; Santos, Susana; Bastos, Maria de Lourdes

Cathinones (β-keto amphetamines), widely abused in recreational settings, have been shown similar or even worse toxicological profile than classical amphetamines. In the present study, the cytotoxicity of two β-keto amphetamines [3,4-dimethylmethcathinone (3,4-DMMC) and 4-methylmethcathinone (4-MMC)], was evaluated in differentiated dopaminergic SH-SY5Y cells in comparison to methamphetamine (METH). MTT reduction and NR uptake assays revealed that both cathinones and METH induced cytotoxicity in a concentration- and time-dependent manner. Pre-treatment with trolox (antioxidant) partially prevented the cytotoxicity induced by all tested drugs, while N-acetyl-l-cysteine (NAC; antioxidant and glutathione precursor) and GBR 12909 dopamine transporter inhibitor) partially prevented the cytotoxicity induced by cathinones, as evaluated by the MTT reduction assay. Unlike METH, cathinones induced oxidative stress evidenced by the increase on intracellular levels of reactive oxygen species (ROS), and also by the decrease of intracellular glutathione levels. Trolox prevented, partially but significantly, the ROS generation elicited by cathinones, while NAC inhibited it completely. All tested drugs induced mitochondrial dysfunction, since they led to mitochondrial membrane depolarization and to intracellular ATP depletion. Activation of caspase-3, indicative of apoptosis, was seen both for cathinones and METH, and confirmed by annexin V and propidium iodide positive staining. Autophagy was also activated by all drugs tested. Pre-incubation with bafilomycin A1, an inhibitor of the vacuolar H+- ATPase, only protected against the cytotoxicity induced by METH, which indicates dissimilar toxicological pathways for the tested drugs. In conclusion, the mitochondrial impairment and oxidative stress observed for the tested cathinones may be key factors for their neurotoxicity, but different outcome pathways seem to be involved in the adverse effects, when compared to METH.

2020Journal article

Restricted access

Enhancement of the antioxidant and antimicrobial activities of porphyran through chemical modification with tyrosine derivatives

Publication . Adão, Pedro; Reboleira, João; Teles, Marco; Santos, Beatriz; Ribeiro, Nádia; Teixeira, Carlos M.; Guedes, Mafalda; Pessoa, João Costa; Bernardino, Susana M.

The chemical modification of porphyran hydrocolloid is attempted, with the objective of enhancing its antioxidant and antimicrobial activities. Sulfated galactan porphyran is obtained from commercial samples of the red algae Porphyra dioica using Soxhlet extraction with water at 100ºC and precipitation with isopropyl alcohol. The extracted porphyran is then treated with modified L-tyrosines in aqueous medium in the presence of NaOH, at ca. 70ºC. The modified tyrosines L1 and L2 are prepared through a Mannich reaction with either thymol or 2,4-di-tert-butylphenol, respectively. While the reaction with 2,4-di-tert-butylphenol yields the expected tyrosine derivative, a mixture of products is obtained with thymol. The resulting polysaccharides are structurally characterized and the respective antioxidant and antimicrobial activities are determined. Porphyran treated with the N-(2-hydroxy-3,5-di-tert-butyl-benzyl)-L-tyrosine derivative, POR-L2, presents a noticeable superior radical scavenging and antioxidant activity compared to native porphyran, POR. Furthermore, it exhibited some antimicrobial activity against S. aureus. The surface morphology of films prepared by casting with native and modified porphyrans is studied by SEM/EDS. Both POR and POR-L2 present potential applicability in the production of films and washable coatings for food packaging with improved protecting characteristics.

2021Journal article

Open access