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Pòster presentat al congrés "Understanding Human Diseases Through Metabolomics: Interactions Among the Genome, Proteome, Gut Microbiome and Nutrition", Metabolomics and Human Health, Gordon Research Conference (February 3 - 8, 2019 Ventura, CA, United States) Diet is one of the most important lifestyle factors associated with health status. Currently, one of the main limitations of nutritional epidemiology and nutritional genomics is the difficulty in the measurements of dietary intake. In observational studies carried out in a large number of participants, the most commonly applied tools for estimating dietary intake are based on self-reporting, including food frequency questionnaires (FFQ) for the assessment of regular consumption (usually 1-year), or 24-h recalls for 1-day assessment. However, such instruments for data collection may contain several recall bias and other systematic or random errors that may have a great effect in the subsequent associations found. Although in recent years, it has been an improvement in increasing the validity and precision of food questionnaires due to the use of the new information technologies (Figure 1), these instruments are still biased and additional information based on objective biomarkers of food intake is needed.

Trabajo presentado en la XV Reunión Científica de la Sociedad Española de Cromatografía y Técnicas Afines (SECyTA 2015) junto con la VII Reunión Nacional de la Sociedad Española de Espectrometría de Masas (SEEM 2015), celebrada en Castellón de la Plana (España), del 27 al 30 de octubre de 2015, The high power of new analytical techniques hold their expansion to new application fields as health research, nutrition or food safety, extremely interesting for human welfare. This widening in analytical chemistry has been supported with new non-targeted approaches as ¿omic¿ techniques. The relevance of these techniques encourages opening new perspectives in other subjects as the nutrition of farmed animals. The assessment of nutrients and energy sources that animals need to grow ensures a correct life quality for both animals and consumers. In order to ensure this quality, metabolomics are being currently explored. On the last years, NMR based metabolomics have been typically used because of its versatility, robustness, universality and elucidation power. On the other hand, their high price and maintenance cost together with a low sensitivity have raised the search for alternative methods like LC-HRMS. This hyphenated technique is considered ¿ideal¿ for this kind of research thanks to its full spectrum acquisition with high sensitivity (around ppb) from small sample quantities (microliters) of animal bio-fluids (blood, urine¿). In our case, juveniles of gilthead sea bream, one of the most important fish species in European aquaculture, were fed with four different diets. In these diets, marine derived ingredients were partially substituted by alternative plant materials in order to have more sustainable diets for fish farming. On the other hand, the potential introduction of mineral additives might also induce extraordinary nutritional-dependent effects on animals. Serum samples were treated with acetonitrile followed by centrifugation. An aliquot was directly injected in HILIC chromatography and the rest was dried and reconstituted for RP chromatography with H2O:MeOH (90:10). UHPLC-HRMS data was extracted, aligned and normalized with XCMS package and finally analyzed by multivariate analysis. Some potential biomarkers from the different experiences highlighted different metabolic pathways relevant to the diet intake and composition.

Treball Final de Màster Universitari en Tècniques Cromatogràfiques Aplicades (Pla de 2013). Codi: SIY009. Curs: 2015/2016 The aim of this work was to determine if the multimodal analgesia by intravenous infusion of the mixture of drugs composed by dipyrone, tramadol and metoclopramide is an effective way of analgesia administration to postoperative patients. Depending on the pain level suffered by the patient after a surgical intervention, the department of Anaesthesiology, Resuscitation and Pain therapy of the General University Hospital of Castellon must evaluate two procedures of drugs administration in post-operatory analgesia. The first one is selected for patients with acute pain, and consist on a mixture of 6 g of dipyrone, 300 mg of tramadol and 30 mg of metoclopramide in 500 mL of physiological saline and infused intravenously during 24 hours. The other one, 2 g of dipyrone, 100 mg of tramadol and 10 mg of metoclopramide is dissolved in 100 mL of saline and infused for 20 minutes every 8 hours to patients with moderate pain (the template for the analgesic treatment order can be found in Figure S 1 of supplementary information). This work was divided in three sections: The first step was to develop a Liquid Chromatography coupled to tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of dipyrone, tramadol and metoclopramide in physiological saline with the purpose of performing a stability study during 24 hours of the mixture prior to intravenous administration. In this stability study, High Performance Liquid Chromatography coupled to triple quadrupole mass spectrometry (HPLC-QqQ) was used, butsupplementary information about possible transformation products was obtained from liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) and HPLC-diode-array detector (HPLCDAD). The next step was to develop and validate an analytical method by HPLC-QqQ for quantification of tramadol, metoclopramide, dipyrone and their metabolites in blood plasma of patients to make a pharmacokinetic study and compare the therapeutic effect of the two procedures of analgesia administrated to post-operatory patients. The last study goal is to find out non-reported metoclopramide metabolites to better understanding the human metabolism of that drug, since it was founded less information about metabolism of metoclopramide compared to dipyrone or tramadol. To this end, blood plasma samples prior and after post-operatory analgesia administration where analysed by LC-HRMS and compared.

Trabajo presentado en el 10º Congreso de la Asociación Ibérica de Endocrinología Comparada (AIEC), celebrado en Castellón (España), del 23 al 25 de septiembre de 2015, Up-stream regulators of the endocrine cascade are largely altered by malnutrition due to feed deprivation or unbalanced diet composition. This includes many changes in circulating catabolic and growth-promoting factors, also related to plasma metabolite levels. This complex trade-off has been addressed in both fish and higher vertebrates, although modern omic approaches are required to identify and validate the most robust biomarkers with either diagnostic and/or prognostic value. For this goal, a non-targeted metabolomics approach is developed. Nuclear magnetic resonance has been often used for routine metabolomic profiling in a vast array of species due to its high elucidation power and universality. However, liquid chromatography-high resolution mass spectrometry (LC-HRMS) has emerged as a reliable alternative technique with greater sensitivity and selectivity. This methodological advantage has been exploited to assess the serum metabolome of two-year old gilthead sea bream (Sparus aurata) in continuously fed fish and 10-days fasted fish. Feed deprivation resulted in a significant reduction of condition factor (2.91 vs 2.63), hepatosomatic index (1.32 vs 0.90) and mesenteric fat index (1.90 vs 1.64). Deproteinized serum samples were analysed by LC-HRMS for individual fish (n=10) and data was peak picked, aligned and normalized with XCMS. Partial Least Squares¿Discriminant Analysis (PLS-DA) clearly separated the two experimental groups, explaining the first component more than 90% of total variance. Orthogonal PLS-DA highlighted more than 850 differentiation ions (P>0.80), which primarily yielded 46 compounds that were classified into different physiological pathways (phospholipid metabolism, lysophosphatidic acid metabolism, fatty acid oxidation, bile salt metabolism, methionine-cysteine metabolism, amino acid catabolism, biotin metabolism). A targeted refined search in combination with transcriptional validation procedures were conducted to go further into fish biomarker research.

Olive oil is a fundamental component of the Mediterranean area diet and it is widely used as a condiment, cooking medium, emulsion component and in the storage of vegetable and animal food [1]. Olive oil cultivation originated in Asia Minor and has spread to Greece, Italy, Spain and North Africa. Recently it was also extended in other countries with temperate climate like France. In contrast to other vegetables oils, extra virgin olive oil (EVOO), produced by modern milling technologies, can be directly consumed without any further manipulation. Once a year, the International Olive Council (IOC) updates statistics on production, imports, exports and consumption of olive oil. The statistics of 2013 shows that EU is the responsible for 71.9% of world production of olive oil and Spain produces the 59.1% of Europe’s olive oil. Many areas where high-quality olive oils are produced are located in Spain. The characteristics of these oils are determined by a series of factors: the nature of the soil, the climate variety of the plant, cultivation and oil extraction techniques [2]. The Food Quality Certification are products that are protected by a European Union legislation ensuring compliance of higher requirements than those required for other products. Protected Designations of Origin (PDO), Protected Geographical Indications (PGI) and others are official EU classifications, and a system used in Spain for the recognition of distinct quality [3]. Only some works have been conducted on the identification of markers for the origin of oils by analytical methods [4–6]. Metabolomics (or more precisely metabolic fingerprinting) is a feasible approach for the discovering of those markers. In the Universidad Jaime I (UJI) some metabolomics projects have been carried out in the field of Food Authenticity. Very recently an article was published in Food Chemistry where some biomarkers were identified for correct authentication of Spanish (Valencian) oranges. In this project, 91 Extra Virgin Olive Oil (EVOO) samples from different regions of Spain with Protected Designation Origin (PDO) certificate have been analyzed using multivariate analysis from the data obtained by UHPLC-QTOF MS with the objective to discover useful biomarkers for origin differentiation. QTOF MS has been used under MSE mode, i.e. simultaneous acquisition at low (LE) and high collision energy (HE), which provides useful information on the molecular mass (commonly at LE) and on the main fragments ions (commonly at HE). The non-lipidic (unsaponifiable) and lipidic (saponifiable) portion of the oil (see in Figure 1 the olive oil composition) were analyzed separately because of the difference in polarity, that means, two separation methods were performed in UHPLC under reversed phase conditions but using different mobile phases. Furthermore, the samples were analyzed in both positive and negative electrospray ionization mode in the QTOF MS. This work only deals on the analysis of lipidic portion of the EVOO, composed mainly by triglycerides (a triglyceride is an ester derived from glycerol and three fatty acids). A freeware R-package has been used to extract and integrate peaks from the Total Ion Chromatogram (TIC), and to al techniques have been employed to classify samples according to their origin significant biomarkers have been highlighted in order different Spanish Regions.