Lacosamide intake during pregnancy increases the incidence of foetal malformations and symptoms associated with schizophrenia in the ofspring of mice

The use of frst and second generation antiepileptic drugs during pregnancy doubles the risk of major congenital malformations and other teratogenic defects. Lacosamide (LCM) is a third-generation antiepileptic drug that interacts with collapsing response mediator protein 2, a protein that has been associated with neurodevelopmental diseases like schizophrenia. The aim of this study was to test the potential teratogenic efects of LCM on developing embryos and its efects on behavioural/histological alterations in adult mice. We administered LCM to pregnant mice, assessing its presence, and that of related compounds, in the mothers’ serum and in embryonic tissues using liquid chromatography coupled to quadrupole/time of fight mass spectrometry detection. Embryo morphology was evaluated, and immunohistochemistry was performed on adult ofspring. Behavioural studies were carried out during the frst two postnatal weeks and on adult mice. We found a high incidence of embryonic lethality and malformations in mice exposed to LCM during embryonic development. Neonatal mice born to dams treated with LCM during gestation displayed clear psychomotor delay and behavioural and morphological alterations in the prefrontal cortex, hippocampus and amygdala that were associated with behaviours associated with schizophrenia spectrum disorders in adulthood. We conclude that LCM and its metabolites may have teratogenic efects on the developing embryos, refected in embryonic lethality and malformations, as well as behavioural and histological alterations in adult mice that resemble those presented by patients with schizophreniaWe are very thankful to Mark Sefton and Miguel Ruiz-Veguilla for revising the manuscript and valuable comments on the manuscript. We also gratefully acknowledge Servicio de Microanalisis de la Universidad de Sevilla (CITIUS) for the support ofered with UPLC-QTOF/MS. Tis work was supported by the Andalusian Regional Ministry of Economy, Science and Innovation (P11-cts-7634); the Andalusian Regional Ministry of Health (PI-0438-2010) and the Instituto de Salúd Carlos III (PS09/00050, CP08/00111, CPII14/00033) cofnanced by European Development Regional Fund “A way to achieve Europe” ERDF to PYG; Fundación Ramón Areces, DGICYT BFU2011-27207, and the Junta de Andalucía CTS-2257 to AMC; Funding FEDER Project UNSE10–1E-429 to RFT and MVN; Francisco José de Caldas fellowship by COLCIENCIAS to V.V-LDepartamento de Biología Ambiental y Salud Públic

A microfluidic liquid phase microextraction method for drugs and parabens monitoring in human urine

Frequent consumption of pharmaceuticals and personal care products (PPCPs) have emerged as a current problem that highlights the pressing need for new multi-residue analytical methods that allow their simultaneous determination to assess their overall effect on human health. In this regard and for the first time, a versatile microfluidic based- liquid phase microextraction (LPME) method was developed for simultaneous monitoring of ten compounds from six different classes: amoxicillin, sulfadiazine, sulfamerazine, tiamphenicol, ethyl 4-hydroxybenzoate, flumequine, propyl 4-hydroxybenzoate, 5-hydroxydiclofenac, 3-hydroxydiclofenac and diclofenac. The microfluidic device was combined with a HPLC-UV system for the separation and determination of the model analytes in the sample. Optimal conditions were reached using 2-nitrophenyl octyl ether as supported liquid membrane, pH 3.5 as donor phase, pH 11.5 as acceptor phase, 0.5 µL min−1 as donor flow rate and 1 µL min−1 as acceptor flow rate. Under optimal method conditions, the extraction efficiency was between 85 and 100% for most compounds after 10 min extraction, and it was successfully applied in non-diluted human urine, with recoveries between 70 and 100% for all analytes except for sulfamerazine (52% recovery). In addition, the extraction of metabolites (3-hydroxydiclofenac and 5-hydroxydiclofenac) was also demonstrated in microfluidic systems with recoveries between 71 and 100% in human urine. The proposed method allowed consecutive extraction and only requires 5 µL of organic solvent and less than 15 µL of sample volume

Uptake Study in Juncus sp. and Salicornia Europaea of Six Pharmaceuticals by Liquid Chromatography Quadrupole Time-of-flight Mass Spectrometry

In this work, eight plants of Juncus sp. and ten of Salicornia europaea were used for an uptake assay of pharmaceuticals (flumequine, cirpofloxacin, enrofloxacin, carbamazepine, diclofenac and ibuprofen) by irrigation at three concentration levels: 10 ng mL−1 (low level); 700 ng mL−1 (medium level) and 10 μg mL−1 (high level). Two plants irrigated with pharmaceutical-free water were set up as controls. For each level, two plants were watered every day with 50 mL (Juncus sp.) and every two days with 20 mL (Salicornia europaea) of aqueous solutions containing all the analytes at the described concentrations. Plants irrigated at 10 μg mL−1 were significantly the most affected, whereas the rest of the plants remained, in general, largely displayed no apparent physiological effects throughout the 30 days (Juncus sp.) and 21 days (Salicornia europaea) assays. Leaves and stems were cut every seven days and roots were collected at the end of the assay. The samples were lyophilized, submitted to a microwave assisted extraction using 5 mL of acetonitrile:water mixture (1:1, v/v) and they were analyzed (in triplicate) in a liquid chromatography-quadrupole time of flight mass spectrometry instrument. Most of the analytes were quantified in many of the samples corresponding to the three exposure levels with the highest concentrations obtained at high exposure levels. Ibuprofen was not detected in any sample and enrofloxacin, ciprofloxacin and diclofenac were not detected in the samples from Salicornia europaea.Ministerio de Economía y Competitividad CTM2015-67902-C-1-PMinisterio de Ciencia e Innovación PGC2018-096608-B-C2

Use of Polymer Inclusion Membranes (PIMs) as Support for Electromembrane Extraction of Non-steroidal Anti-inflammatory Drugs and Highly Polar Acidic Drugs

The use of polymer inclusion membranes (PIMs) as support of 1-octanol liquid membrane in electromembrane extraction (EME) procedure is proposed. Synthesis of PIMs were optimized to a composition of 29% (w/w) of cellulose triacetate as base polymer and 71% (w/w) of Aliquat®336 as cationic carrier. Flat PIMs of 25 µm thickness and 6 mm diameter were used. EME protocol was implemented for the simultaneous extraction of four non-steroidal anti-inflammatory drugs (NSAIDs) (salicylic acid, ketoprofen, naproxen and ibuprofen) and four highly polar acidic drugs (anthranilic acid, nicotinic acid, amoxicillin and hippuric acid). Posterior HPLC separation of the extracted analytes was developed with diode array detection. Recoveries in the 81–34% range were obtained. EME procedure was applied to human urine samples.Ministerio de Educación y Ciencia TM2015-67902-C-1-PPremio Mensual Publicación Científica Destacada de la US. Facultad de Químic

Liquid Chromatography Quadrupole Time-of-flight Mass Spectrometry Determination of Six Pharmaceuticals in Vegetal Biota. Uptake Study in Lavandula Dentata

A procedure based on microwave assisted extraction for the determination of 6 pharmaceuticals in samples of Lavandula dentata, Salicornia ramosissima and Juncus sp. by liquid chromatography-quadrupole time of flight mass spectrometry (LC-QTOF/MS) was optimized and validated. Best results were obtained using microwave assisted extraction of 1.0 g of homogeneous lyophilized samples and 5 mL of a mixture ACN:H2O (1:1 v/v) as extracting solvent. Analytical recoveries ranged from 60 to 107% with relative standard deviation (RSD) lower than 15%. Limits of quantitation (LOQ) for the 6 pharmaceuticals flumequine (FLM), carbamazepine (CBZ), ciprofloxacin (CPR), enrofloxacin (ENR), diclofenac (DCL), and ibuprofen (IBU) were in the range 20.8–125 ng g− 1. The method was satisfactory applied for an uptake study in Lavandula dentata samples finding quantifying concentrations of FLM and CBZ in roots, leaf and stem.Ministerio de Educación, Cultura y Deporte CTM2015-67902-C-1-PEuropean Commission. Fondo Europeo de Desarrollo Regional UNSE10-1E-42

Effect of Aliquat®336 on supported liquid membrane on electromembrane extraction of non-steroidal anti-inflammatory drugs

Up to now, most electromembrane extraction methods describe the use of pure organic solvents or mixtures as supported liquid membrane. However, the need to incorporate carriers in the supported liquid membrane to achieve the extraction of high polar compounds, seems to indicate that the presence of certain additives in the organic solvent may improve the extraction yield. For this reason, some studies have tried to enhance electrokinetic migration in different ways, modifying either the supported liquid membrane or even the donor solution. In this work, it has been studied and optimized an electromembrane extraction of five widely used non-steroidal anti-inflammatory drugs: salicylic acid, ketoprofen, naproxen, diclofenac and ibuprofen. The thickness and porosity of the support, the supported liquid membrane composition, the donor and acceptor phase pH, the voltage, the extraction time and the electrode configuration were optimized. supported liquid membrane was modified by adding different amounts of Aliquat®336, a cationic carrier commonly used in electromembrane extraction procedure for anionic compounds. The results compared with those obtained in the same extraction conditions using the pure organic solvent as supported liquid membrane, showed better extraction recoveries. The highest recoveries were achieved using a pH 5 donor phase and an acceptor phase at pH 12. The recoveries were within the range of 39 and 53% after 12 min extraction, using a voltage of 80 V, a stirring speed of 400 rpm and 1-nonanol modified with Aliquat®336 2.5% (w/v) as support liquid membrane. Detection and quantitation limits were within 0.02–1.0 ng mL−1 and 0.05–3.0 ng mL−1, respectively. The selected analytes were extracted by electromembrane extraction using a home-made device designed with a flat configuration. The analyses were carried out by high performance liquid chromatography with diode array and fluorescence detection and finally, applied to the analysis of human urine samples.Ministerio de Educación y Ciencia (MEC). España CTM2015-67902-C-1-

Urine and saliva biomonitoring by HF-LPME-LC/MS to assess dinitrophenols exposure

In this work, the determination of 2,4-, 2,5- and 2,6-dinitrophenols and the identification of some of their metabolites in human urine and saliva is proposed. A three phase hollow fiber based liquid phase microextraction prior to ultra-high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry allowed low detection and quantitation limits of the target analytes, as well as the investigation and tentatively identification of some metabolites by accurate mass full-spectrum measurements. The chromatographic separation was accomplished on an Acquity BEH C18 column (50 mm × 2.1 mm i.d., 1.7 μm particle size) at 25 ºC using water and acetonitrile (with 0.1 % (v/v) formic acid) 20:80 v/v as mobile phase, at a flow rate of 0.5 mL/min in isocratic elution mode for 5 min. Hollow fiber liquid phase microextraction was achieved at donor phase pH 2, acceptor phase pH 13 and dihexylether as supported liquid membrane. Under the optimal conditions, detection limits for 2,4-, 2,5- and 2,6-dinitrophenol, respectively, were 0.18 μg·L-1, 0.38 μg·L-1 and 0.14 μg·L-1 in urine samples and 0.32 μg·L-1, 0.67 μg·L-1 and 0.24 μg·L-1 in saliva samples. The proposed methodology was applied on urine and saliva samples from laboratory staff likely to be or not occupationally exposed to dinitrophenols, finding quantitative levels of 2,4- and 2,6-dinitrophenol and identifying some metabolites previously reported in literature.Ministerio de Economía y Competitividad (MINECO). España CTM2015-67902-C-1-PMinisterio de Ciencia e Innovación (MICIN). España GC2018-096608-B-C22European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER) GC2018-096608-B-C2

Búsqueda de alimentos ricos en provitamina D (Ergosterol)

En este trabajo se presentan los resultados obtenidos para el contenido en provitamina D2 (ergosterol) en diferentes muestras de origen vegetal. Muestras de lechuga, dátiles, uvas, arroz y champiñones fueron sometidas a un tratamiento de extracción con hexano y el contenido en ergosterol medido mediante análisis por cromatografía líquida de alta resolución con detección ultravioleta de diodos en fila, obteniéndose los mayores contenidos en ergosterol en las muestras de champiñón y lechuga

Targeted and untargeted metabolomic analysis of Procambarus clarkii exposed to a “chemical cocktail” of heavy metals and diclofenac

La contaminación del agua plantea un problema importante, pero se dispone de información limitada sobre los efectos conjuntos de xenobióticos de diferentes grupos químicos para evaluar la respuesta biológica real. Procambarus clarkii (P. clarkii) ha demostrado ser un buen bioindicador para evaluar la calidad de los ecosistemas acuáticos. En este trabajo, se ha estudiado la bioacumulación de cadmio (Cd), arsénico (As) y diclofenaco (DCF) en diferentes tejidos durante 21 días después de la exposición a un “cóctel químico” de As, Cd y DCF, y hasta 28 días considerando un período de purificación. Además, se llevó a cabo un análisis metabolómico dirigido y no dirigido para profundizar en las alteraciones metabólicas provocadas así como en la metabolización del DCF.Water pollution poses an important problem, but limited information is available about the joined effects of xenobiotics of different chemical groups to evaluate the real biological response. Procambarus clarkii (P. clarkii) has been demonstrated to be a good bioindicator for assessing the quality of aquatic ecosystems. In this work, we studied the bioaccumulation of cadmium (Cd), arsenic (As) and diclofenac (DCF) in different tissues of P. clarkii during 21 days after the exposure to a “chemical cocktail” of As, Cd and DCF, and until 28 days considering a depuration period. In addition, a combined untargeted and targeted metabolomic analysis was carried out to delve the metabolic impairments caused as well as the metabolization of DCF. Our results indicate that As and Cd were mainly accumulated in the hepatopancreas followed by gills and finally abdominal muscle. As and Cd show a general trend to increase the concentration throughout the exposure experience, while a decrease in the concentration of these elements is observed after 7 days of the depuration process. This is also the case in the abdominal muscle for Cd, but not for As and DCF, which increased the concentration in this tissue in the depuration phase. The hepatopancreas showed the greatest number of metabolic pathways affected. Thus, we observed a crucial bioaccumulation of xenobiotics and impairments of metabolites in different tissues. This is the first study combining the exposure to metals and pharmaceutically active compounds in P. clarkii by untargeted metabolomics including the biotransformation of DCF.This work was supported by the coordinated projects PGC 2018- 096608-B-C21 and PGC 2018-096608-B-C22 from the Spanish Ministry of Science and innovation (MCIN). (Generacion ´ del Conocimiento. MCIN/AEI/10.13039/501100011033/FEDER “Una manera de hacer Europa”). Authors are grateful to FEDER (European Community) for financial support, Grant UNHU13-1E-1611. Rodríguez-Moro, G. thanks to Plan Andaluz de Investigacion, ´ Desarrollo e Innovacion ´ (PAIDI 2020) and Fondo Social Europeo for a postdoctoral grant (DOC_01115). Au thors are grateful to Servicio General de Investigacion ´ de Microanalisis, ´ CENTRO DE INVESTIGACION ´ TECNOLOG´IA E INNOVACION ´ (CITIUS). Universidad de Sevilla, for the use of some of the chromatographic equipment. Funding for open access charge: Universidad de Huelva / CBUA

Desarrollo de líneas experimentales para su aplicación en los Trabajos de Fin de Grado en Química

Desde el punto de vista académico, los Trabajos Fin de Grado en Ciencias Químicas se presentan como proyectos experimentales relacionados con problemas reales, orientados a progresar en conocimientos y fortalecer competencias adquiridas, convirtiéndose en claves del currículo para el mundo profesional. Se abordan una serie de proyectos de laboratorio como una aproximación más realista al aprendizaje integral. La metodología docente propuesta favorece la búsqueda de información, pensamiento crítico, superación continua y éxito de los estudiantes, similar a la que se realiza en el mundo real, estimulando su interés y motivación durante el proceso de aprendizaje.Des del punt de vista acadèmic, els Treballs Fi de Grau en Ciències Químiques es presenten com a projectes experimentals relacionats amb problemes reals, orientats a progressar en coneixements i enfortir competències adquirides, convertint-se en claus del currículum per al món professional. S'aborden una sèrie de projectes integrals de laboratori com una aproximació més realista a l'aprenentatge integral. La metodologia docent proposta afavoreix la busca d'informació, pensament crític, superació contínua i èxit dels estudiants, semblant a la què es realitza en el món real, estimulant el seu interès i motivació durant el procés d'aprenentatge.From the academic point of view, the bachelor thesis in Chemistry is presented as a project based on a real problem. The goal is to progress in strengthening knowledge and skills already acquired. This is a key element for the students curricula related to their integration into the professional world. A comprehensive series of laboratory projects have been developed as a more realistic approach to the concept of integral learning. Teaching methodology promotes information searching, critical thinking, continuous improvement and success. The procedure is performed in conditions close to the real world, stimulating interest and motivation during the learning process