¿Son fiables los productos que encontramos en los supermercados?

Duración (en horas): De 21 a 30 horas. Destinatario: EstudianteEn este recurso didáctico se presenta la metodología en base a proyectos utilizada en el proceso de enseñanza-aprendizaje de la asignatura optativa "Cromatografía y Técnicas Afines" que se imparte en el segundo ciclo de la Licenciatura de Química. El proyecto, que lleva como título "¿Son fiables los productos que encontramos en los supermercados?", permite al alumno aprender los distintos métodos analíticos para el analisis de contaminantes en los productos alimenticios, pero a su vez comprender todos aquellos aspectos teórico-prácticos de la cromatografía y las técnicas afines. En este sentido, cada vez es mayor la información y las noticias que nos llegan sobre la contaminación de los productos alimenticios. Podemos encontrar un amplio número de ejemplos en la última década como la contaminación de pollos con dioxinas en Bélgica y Alemania, la contaminación de leche para niños con melanina en China, etc. Estos casos suscitan una gran preocupación en la sociedad sobre la calidad de los productos alimenticios así como el interés de plantear reglamentación sobre los alimentos que garanticen el control de la calidad de los mismos. Este proyecto, por lo tanto, irá encaminado al desarrollo de determinados métodos analíticos que pueden ser aplicados en el departamento de la unidad de análisis de alimentos que garanticen su calidad y el bienestar del consumidor, como son, por ejemplo: a) Objetivos marcados por el responsable del laboratorio de cromatografía de gases: análisis de bifenilos policlorados en peces, la elección de un inyector adecuado para el análisis de furano en café, y el análisis de melanina en leche en polvo. b) Con lo que respecta al laboratorio de Cromatografía de Líquidos los objetivos marcados son: determinación de clenbuterol, salbutamol y cimateol en carne y la detección de determinados problemas analíticos detectados en distintos métodos analíticos y su resolución

From target analysis to suspect and non-target screening of endocrine-disrupting compounds in human urine

[EN] In the present work, a target analysis method for simultaneously determining 24 diverse endocrine-disrupting compounds (EDCs) in urine (benzophenones, bisphenols, parabens, phthalates and antibacterials) was developed. The target analysis approach (including enzymatic hydrolysis, clean-up by solid-phase extraction and analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS)) was optimized, validated and applied to volunteers' samples, in which 67% of the target EDCs were quantified. For instance, benzophenone-3 (0.2-13 ng g(-1)), bisphenol A (7.7-13.7 ng g(-1)), methyl 3,5-dihydroxybenzoate (8-254 ng g(-1)), mono butyl phthalate (2-17 ng g(-1)) and triclosan (0.3-9 ng g(-1)) were found at the highest concentrations, but the presence of other analogues was detected as well. The developed target method was further extended to suspect and non-target screening (SNTS) by means of LC coupled to high-resolution MS/MS. First, well-defined workflows for SNTS were validated by applying the previously developed method to an extended list of compounds (83), and then, to the same real urine samples. From a list of approximately 4000 suspects, 33 were annotated at levels from 1 to 3, with food additives/ingredients and personal care products being the most abundant ones. In the non-target approach, the search was limited to molecules containing S, Cl and/or Br atoms, annotating 4 pharmaceuticals. The results from this study showed that the combination of the lower limits of detection of MS/MS and the identification power of high-resolution MS/MS is still compulsory for a more accurate definition of human exposome in urine samples.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work has been financially supported by the Ministry of Science and Innovation of the Spanish Government through project PID2020-117686RB-C31, and by the Education Department of the Basque Government as a consolidated group of the Basque Research System (IT1213-19)

Dilute-and-shoot coupled to mixed mode liquid chromatography-tandem mass spectrometry for the analysis of persistent and mobile organic compounds in human urine

In this work, a comprehensive method for the simultaneous determination of 33 diverse persistent and mobile organic compounds (PMOCs) in human urine was developed by dilute-and-shoot (DS) followed by mixed-mode liquid chromatography coupled with tandem mass spectrometry (MMLC-MS/MS). In the sample preparation step, DS was chosen since it allowed the quantification of all targets in comparison to lyophilization. For the chromatographic separation, Acclaim Trinity P1 and P2 trimodal columns provided greater capacity for retaining PMOCs than reverse phase and hydrophilic interaction liquid chromatography. Therefore, DS was validated at 5 and 50 ng/mL in urine with both mixed mode columns at pH = 3 and 7. Regarding figures of merit, linear calibration curves (r2 > 0.999) built between instrumental quantification limits (mostly below 5 ng/mL) and 500 ng/mL were achieved. Despite only 60% of the targets were recovered at 5 ng/mL because of the dilution, all PMOCs were quantified at 50 ng/mL. Using surrogate correction, apparent recoveries in the 70–130% range were obtained for 91% of the targets. To analyse human urine samples, the Acclaim Trinity P1 column at pH = 3 and 7 was selected as a consensus between analytical coverage (i.e. 94% of the targets) and chromatographic runs. In a pooled urine sample, industrial chemicals (acrylamide and bisphenol S), biocides and their metabolites (2-methyl-4-isothiazolin-3-one, dimethyl phosphate, 6-chloropyridine-3-carboxylic acid, and ammonium glufosinate) and an artificial sweetener (aspartame) were determined at ng/mL levels. The outcomes of this work showed that humans are also exposed to PMOCs due to their persistence and mobility, and therefore, further human risk assessment is needed.Authors gratefully acknowledge financial support from the State Research Agency of the Ministry of Science and Innovation (Government of Spain) through project PID2020–117686RB-C31 and the Basque Government as a consolidated group of the Basque Research System (IT-1446–22). M. Musatadi also acknowledges the Basque Government for his predoctoral grant

Integrated biological response to environmentally-relevant concentration of amitriptyline in Sparus aurata

[EN]Amitriptyline (AMI) is a commonly tricyclic antidepressant to treat depression, anxiety, and other conditions. Like many other pharmaceuticals, AMI and its by-products are incompletely removed during wastewater treatment and therefore they are released to rivers, estuaries and coastal waters. The presence of this kind of compounds in the water environment may involve a negative impact on non-target aquatic organisms at relatively low concentrations. However, the knowledge of AMI effects on aquatic organisms in the environment is scarce. Thus, the objective of this work is to determine the effects of environmentally-relevant concentrations of AMI on biological responses at biochemical and cellular levels in marine teleost. Gilt-head seabream (S. aurata) were exposed to AMI for 7 days at 0.2 mu g/L in an open flow system and a battery of biomarkers were investigated: acetylcholinesterase, catalase, superoxide dismutase, glutathione S-transferase, cytochrome C oxidase, P450 CYP1A1 ethoxyresorufin (O) dealkylation, and lysosomal biomarkers. Biomarkers were integrated as IBR/n (biological response index). Overall, it can be concluded that AMI exposure at environmentally-relevant concentration induces significant biological responses to stress in marine teleost, especially in lysosomal biomarkers. However, further research is needed about the effects of AMI and other pharmaceuticals on biomarkers in nontargeted species, to raise the knowledge about the toxicity of this type of emerging pollutants.The authors are indebted to Ainhoa Camille for helping us with her invaluable technical support. This work was financially supported by the Ministry of Economy and Competitiveness through the project CTM2017-84763-C3-1-R and the Basque Government (IT-742-13 & IT1213-19). H. Ziarrusta is grateful to the Spanish Ministry predoctoral fellowship and L. Mijangos to the University of the Basque Country for her postdoctoral fellowship

Lurzoruetako kutsagarri organikoen erauzketaren auzia

Egun lurzoruetako kutsagarri organikoen determinazio analitikoan arazo gehiendakarren etapa erauzketa da. Etapa honi dagozkion auziak bi dira: erauzketa-prozesuaren etekinaren kalkulua, hots, nola jakin zenbatekoa den erreskuratutako kutsagarrien etekina lurzoruan dagoen kontzentrazioa ezezaguna izanik, eta erauzketa-teknika berrien egokiera, batez ere, teknika horiek dakartzaten prozedura erosoagoak eta ziurragoak baitira eta aurreko auziaren argibide izan baitaitezke. Kontuan izan behar da ohiko erauzketa-teknikek erabiltzen dituztela disolbatzailearen bolumen altuak, erauzketa-denbora luzeak eta prozedura gogaikarriak. Honenbestez, azken hamarkadetan konposatu organikoen erauzketarako metodo berrien garapena eta optimizazioa nabarmenki ugaritu da

Multi-Target Analysis and Suspect Screening of Xenobiotics in Milk by UHPLC-HRMS/MS

The development of suspect or non-target screening methods to detect xenobiotics in biological fluids is essential to properly understand the exposome and assess its adverse health effects on humans. In order to fulfil that aim, the biomonitorization of human fluids is compulsory. However, these methods are not yet extensively developed, especially for polar organic xenobiotics in biofluids such as milk, as most works are only focused on certain analytes of interest. In this work, a multi-target analysis method to determine 245 diverse xenobiotics in milk by means of Ultra High Performance Liquid Chromatography (UHPLC)-qOrbitrap was developed. Under optimal conditions, liquid milk samples were extracted with acetonitrile in the presence of anhydrous Na2SO4 and NaCl, and the extracts were cleaned-up by protein precipitation at low temperature and Captiva Non-Drip (ND)—Lipids filters. The optimized method was validated at two concentration-levels (10 ng/g and 40 ng/g) obtaining satisfactory figures of merit for more than 200 compounds. The validated multi-target method was applied to several milk samples, including commercial and breast milk, provided by 4 healthy volunteers. Moreover, the method was extended to perform suspect analysis of more than 17,000 xenobiotics. All in all, several diverse xenobiotics were detected, highlighting food additives (benzothiazole) or phytoestrogens (genistein and genistin) in commercial milk samples, and stimulants (caffeine), plasticizers (phthalates), UV filters (benzophenone), or pharmaceuticals (orlistat) in breast milk samples.This research was funded by the Agencia Estatal de Investigación (AEI) of Spain, the European Regional Development Fund through projects CTM2017-84763-C3-1-R and CTM2017-90890-REDT (AEI/FEDER, EU) and the Basque Government through the financial support as consolidated group of the Basque Research System (IT1213-19). B.G. acknowledges a Juan de la Cierva-Formación fellowship by the Spanish Ministry for the Economy, Industry and Competitiveness (MINECO)

Exploratory optimisation of a LC-HRMS based analytical method for untargeted metabolomic screening of Cannabis Sativa L. through Data Mining

Background Recent increase in public acceptance of cannabis as a natural medical alternative for certain neurological pathologies has led to its approval in different regions of the world. However, due to its previous illegal background, little research has been conducted around its biochemical insights. Therefore, in the current framework, metabolomics may be a suitable approach for deepening the knowledge around this plant species. Nevertheless, experimental methods in metabolomics must be carefully handled, as slight modifications can lead to metabolomic coverage loss. Hence, the main objective of this work was to optimise an analytical method for appropriate untargeted metabolomic screening of cannabis. Results We present an empirically optimised experimental procedure through which the broadest metabolomic coverage was obtained, in which extraction solvents for metabolite isolation, chromatographic columns for LC-qOrbitrap analysis and plant-representative biological tissues were compared. By exploratory means, it was determined that the solvent combination composed of CHCl3:H2O:CH3OH (2:1:1, v/v) provided the highest number of features from diverse chemical classes, as it was a two-phase extractant. In addition, a reverse phase 2.6 μm C18 100 Å (150 × 3 mm) chromatographic column was determined as the appropriate choice for adequate separation and further detection of the diverse metabolite classes. Apart from that, overall chromatographic peak quality provided by each column was observed and the need for batch correction methods through quality control (QC) samples was confirmed. At last, leaf and flower tissues resulted to provide complementary metabolic information of the plant, to the detriment of stem tissue, which resulted to be negligible. Significance It was concluded that the optimised experimental procedure could significantly ease the path for future research works related to cannabis metabolomics by LC-HRMS means, as the work was based on previous plant metabolomics literature. Furthermore, it is crucial to highlight that an optimal analytical method can vary depending on the main objective of the research, as changes in the experimental factors can lead to different outcomes, regardless of whether the results are better or worse.This work was financially supported by the Education Department of the Basque Country as a consolidated group of the Basque Research System (IT1213-19) and by Sovereign Fields S.L., in the framework of the project Metabolomic study of Cannabis Sativa L. cultivations and determination of contaminants in medical cannabis plants

Comparison of conventional and dispersive solid phase extraction clean-up approaches for the simultaneous analysis of tetracyclines and sulfonamides in a variety of fresh vegetables

he extensive use of antibiotics in agriculture has led to the occurrence of residual drugs in different vegetables frequently consumed by humans. This could pose a potential threat to human health, not only because of the possible effects after ingestion but also because the transmission of antibiotic-resistant genes could occur. In this work, two accurate sample preparation procedures were developed and validated for the simultaneous analysis of sulfonamides (SAs) and tetracyclines (TCs) in four of the most widely consumed vegetables (lettuce, onion, tomato, and carrot) in Europe. The evaluated protocols were based on QuECHERS for extraction and subsequent clean-up by SPE (solid phase extraction) or dispersive SPE. Parameters affecting both extraction and clean-up were carefully evaluated and selected for accuracy of results and minimal matrix effect. Overall, apparent recoveries were above 70% for most of the target analytes with both analytical procedures, and adequate precision (RSD<30%) was obtained for all the matrices. The procedural limits of quantification (LOQPRO) values for SPE clean-up remained below 4.4 μg kg−1 for TCs in all vegetables except for chlortetracycline (CTC) in lettuce (11.3 μg kg−1) and 3.0 μg kg−1 for SAs, with the exception of sulfadiazine (SDZ) in onion (3.9 μg kg−1) and sulfathiazole (STZ) in carrot (5.0 μg kg−1). Lower LOQPRO values (0.1–3.7 μg kg−1) were obtained, in general, when dSPE clean-up was employed. Both methods were applied to twenty-five market vegetable samples from ecological and conventional agriculture and only sulfamethazine (SMZ) and sulfapyridine (SPD) were detected in lettuce at 1.2 μg kg−1 and 0.5 μg kg−1, respectively.Authors acknowledge financial support from the Elkartek project entitled “Emergencia y diseminación de resistencia a los antibióticos: vínculos entre salud humana, ganadería, alimentación y medioambiente (elkartek 20/88)”, the projects “Evaluación del riesgo de aparición y diseminación de resistencias a antibióticos en productos vegetales frescos y suelos de cultivo de la comunidad autónoma del País Vasco (PA21/05 and PA22/05)” inside the “Research projects targeted to agriculture 2020 program” of the Basque Government (Basque Country, Spain); and the Basque Government through the financial support as consolidated group of the Basque Research System (IT1446-22). I. Vergara-Luis and B. Gonzalez-Gaya are grateful to the University of the Basque Country (UPV/EHU) for their pre-doctoral and post-doctoral fellowships. I. Baciero thanks to the Basque Government for her pre-doctoral fellowship

Characterization of the contamination fingerprint of wastewater treatment plant effluents in the Henares River Basin (central Spain) based on target and suspect screening analysis

The interest in contaminants of emerging concern (CECs) has increased lately due to their continued emission and potential ecotoxicological hazards. Wastewater treatment plants (WWTPs) are generally not capable of eliminating them and are considered the main pathway for CECs to the aquatic environment. The number of CECs in WWTPs effluents is often so large that complementary approaches to the conventional target analysis need to be implemented. Within this context, multitarget quantitative analysis (162 compounds) and a suspect screening (>40,000 suspects) approaches were applied to characterize the CEC fingerprint in effluents of five WWTPs in the Henares River basin (central Spain) during two sampling campaigns (summer and autumn). The results indicated that 76% of the compounds quantified corresponded to pharmaceuticals, 21% to pesticides and 3% to industrial chemicals. Apart from the 82 compounds quantified, suspect screening increased the list to 297 annotated compounds. Significant differences in the CEC fingerprint were observed between summer and autumn campaigns and between the WWTPs, being those serving the city of Alcala de Henares the ones with the largest number of compounds and concentrations. Finally, a risk prioritization approach was applied based on risk quotients (RQs) for algae, invertebrates, and fish. Azithromycin, diuron, chlortoluron, clarithromycin, sertraline and sulfamethoxazole were identified as having the largest risks to algae. As for invertebrates, the compounds having the largest RQs were carbendazim, fenoxycarb and eprosartan, and for fish acetaminophen, DEET, carbendazim, caffeine, fluconazole, and azithromycin. The two WWTPs showing higher calculated Risk Indexes had tertiary treatments, which points towards the need of increasing the removal efficiency in urban WWTPs. Furthermore, considering the complex mixtures emitted into the environment and the low dilution capacity of Mediterranean rivers, we recommend the development of detailed monitoring plans and stricter regulations to control the chemical burden created to freshwater ecosystems.Authors acknowledge financial support from the Agencia Estatal de Investigación (AEI) of Spain and the European Regional Development Fund through project CTM2017-84763-C3-1-R project and the Basque Government through the financial support as consolidated group of the Basque Research System (IT1213-19). NLH is grateful to the Spanish Ministry of Economy, Industry and Competitivity for her predoctoral scholarship FPI 2018. BGG acknowledge an EHU/UPV postdoctoral fellowship. AR is supported by the Talented Researcher Support Programme - Plan GenT (CIDEGENT/2020/043) of the Generalitat Valenciana. Finally, the authors acknowledge support from the AEI and the Ministry of Science, Innovation and Universities (MICIU) to support the Thematic Network of Excellence (NET4SEA) on emerging contaminants in marine settings (CTM2017-90890-REDT, MICIU/AEI/ FEDER, EU)