Combining data acquisition modes in liquid-chromatography–tandem mass spectrometry for comprehensive determination of acylcarnitines in human serum

Acylcarnitines (ACs) are metabolites involved in fatty acid β-oxidation and organic acid metabolism. Metabolic disorders associated to these two processes can be evaluated by determining the complete profile of ACs. In this research, we present an overall strategy for identification, confirmation, and quantitative determination of acylcarnitines in human serum. By this strategy we identified the presence of 47 ACs from C2 to C24 with detection of the unsaturation degree by application of a data-independent acquisition (DIA) liquid chromatography–tandem mass spectrometry (LC–MS/MS) method. Complementary, quantitative determination of ACs is based on a high-throughput and fully automated method consisting of solid-phase extraction on-line coupled to LC–MS/MS in data-dependent acquisition (DDA) to improve analytical features avoiding the errors associated to sample processing. Quantitation limits were at pg mL–1 level, the intra-day and between-day variability were below 15–20%, respectively; and the accuracy, expressed as bias, was always within ± 25%. The proposed method was tested with 40 human volunteers to determine the relative concentration of ACs in serum and identify predominant forms. Significant differences were detected by comparing the ACs profile of obese versus non-obese individuals

Strategies for proteomic analysis of blood glycated proteins

Comunicaciones a congreso

Quantitative analysis of protein glycation in clinical samples

Comunicaciones a congreso

Comparison of drying techniques for extraction of bioactive compounds from olive-tree materials

Olive tree vegetal materials are considered a powerful source for the isolation of bioactive compounds—mainly phenols and triterpenic acids. However, the high humidity content of them reduces their preservation and extractability to a liquid solvent. Accordingly, a drying step is crucial to homogenize the material and to obtain an efficient extraction. We studied the influence of the drying process on the extraction efficiency of bioactive compounds from olive vegetal material. For this purpose, we evaluated the effects of four drying processes on the solid–liquid extraction of bioactive compounds from two by-products, olive leaves and pomace, and olive fruits harvested from two cultivars, Alfafara and Koroneiki. Infrared-assisted drying (IAD) was the most suited approach to obtain extracts enriched in oleuropein from leaves (28.5 and 22.2% dry weight in Alfafara and Koroneiki, respectively). In the case of pomace, lyophilization and microwave-assisted drying led to extracts concentrated in oleacein and oleuropein aglycone, whereas IAD and oven-drying led to extracts with enhanced contents of hydroxytyrosol glucoside and hydroxytyrosol, respectively. The drying process considerably affected the chemical composition of extracts obtained from fruits. Changes in the composition of the extracts were explained essentially by the drying process conditions using auxiliary energies, temperature, and time, which promoted chemical alterations and increased the extractability of the compounds. Therefore, the drying protocol should be selected depending on the phenolic content and initial raw material

Glycated platelets proteome

Comunicaciones a congreso

Distribution of Main Bioactive Compounds from Saffron Species as a Function of Infusion Temperature and Time in an Oil/Water System

Most research on saffron has focused on its composition and beneficial effects, while the culinary perspective to enhance its gastronomic potential remains unexplored. This study aims to define the transfer of the main compounds responsible for color, flavor, and aromatic properties, evaluating three critical variables: temperature (60 °C, 80 °C and 100 °C), infusion time (ranging from 10 to 30 min), and the composition of the medium (water, oil, and water/oil). Samples were analyzed using the LC-QTOF MS/MS and ISO 3632-1:2011 methods. The major compounds were crocins, including trans-crocin and picrocrocin. Among the flavonoids, kaempferol 3-O-sophoroside stands out. Regarding extraction conditions, crocins, glycoside flavonoids, and picrocrocin were enhanced in water, the former in 100% water and at low temperatures, while picrocrocin proved to be the most stable compound with extraction favored at high temperatures. The variable with the greatest incidence of picrocrocin isolation seemed to be the concentration of water since water/oil compositions reported higher concentrations. Safranal and kaempferol were enriched in the oil phase and at lower temperatures. This study provides a chemical interpretation for the appropriate gastronomic use of saffron according to its versatility. Finally, the determination of safranal using the ISO method did not correlate with that obtained using chromatography

Perfil fenólico de aceites de oliva vírgenes obtenidos de selecciones avanzadas en un programa de mejora

The evaluation of the phenolic composition in advanced selections in breeding programs constitutes the first approach for selecting genotypes with improved olive oil quality. In this work, the influence of genotype and ripening index on the phenolic profile of olive oils from advanced selections in comparison to their genitors was studied. Fruit samples were collected in genotypes from crosses between ‘Arbequina’ × ‘Picual’, ‘Picual’ × ‘Arbequina’ and ‘Frantoio’ × ‘Picual’ at five dates from 1st October to 26th November 2009. Characterization of the phenolic profile was performed by liquid–liquid extraction with 60:40 (v/v) methanol–water and subsequent chromatographic analysis with absorption and fluorescence detection in a sequential configuration. A dual effect of genotype and fruit ripening on the phenolic profile has been observed with more pronounced genetic influence in both total (34.73% and 20.45%, respectively) and individual phenols (16.99% to 49.25% and 1.58% to 23.77%, respectively). A higher degree of variability between genotypes at early ripening stages was also observed (p<0.05). The obtained results also allow identification of selections with high content of total and individual phenols. These results suggest a strategy based on early harvesting of fruits (at the first three ripening indexes) for better comparison and selection of genotypes in further crosses in olive breeding programs aiming at improving the quality of virgin olive oil.La evaluación de la composición fenolica en selecciones avanzadas en programas de mejora constituye el primer paso para la selección de genotipos cuyos aceites son de mejor calidad. Se ha estudiado la influencia del genotipo y del índice de madurez en el perfil fenólico del aceite de oliva de selecciones avanzadas en comparación con sus genitores. Se recogieron muestras de genotipos obtenidos de cruzamientos entre 'Arbequina' × 'Picual', 'Picual' × 'Arbequina' y 'Frantoio' × 'Picual' en cinco fechas entre el 1 de octubre y el 26 noviembre de 2009. Se realizó la caracterización del perfil fenólico por extracción líquido–líquido con metanol–agua al 60:40 (v/v) seguido de un análisis cromatográfico con detección por absorción y fluorescencia en una configuración secuencial. Los resultados obtenidos mostraron un mayor grado de variabilidad entre genotipos en las primeras etapas de maduración de los frutos (p<0.05), así como el efecto de la madurez de los frutos y del genotipo en el perfil fenólico, con una influencia genética más pronunciada tanto para fenoles totales (34.73% and 20.45%, respectivamente) como individuales (entre 16.99% y 49.25% y entre 1.58% y 23.77%, respectivamente). Los resultados obtenidos han permitido la identificación de selecciones con alto contenido en fenoles totales e individuales. Ambos resultados sugieren una estrategia basada en la evaluación de los fenoles totales e individuales en las primeras etapas de la maduración para comparar y seleccionar genotipos en programas de mejora por cruzamiento encaminados a mejorar la calidad del aceite de oliva virgen

Changes in human sweat metabolome conditioned by severity of obstructive sleep apnea and intermittent hypoxemia

Obstructive sleep apnea (OSA) is a sleep disorder that has been associated with the incidence of other pathologies. Diagnosis is mainly based on the apnea–hypopnea index (AHI) obviating other repercussions such as intermittent hypoxemia, which has been found to be associated to cardiovascular complications. Blood-based samples and urine have been the most utilised biofluids in metabolomics studies related to OSA, while sweat could be an alternative due to its non-invasive and accessible sampling, its reduced complexity, and comparability with other biofluids. Therefore, this research aimed to evaluate metabolic overnight changes in sweat collected from patients with OSA classified according to the AHI and oxygen desaturation index (ODI), looking for potential cardiovascular repercussions. Pre- and post-sleeping sweat samples from all individuals (n = 61) were analysed by gas chromatography coupled to high-resolution mass spectrometry after appropriate sample preparation to detect as many metabolites as possible. Permanent significant alterations in the sweat were reported for pyruvate, serine, lactose, and hydroxybutyrate. The most relevant overnight metabolic alterations in sweat were reported for lactose, succinate, urea, and oxoproline, which presented significantly different effects on factors such as the AHI and ODI for OSA severity classification. Overall metabolic alterations mainly affected energy production-related processes, nitrogen metabolism, and oxidative stress. In conclusion, this research demonstrated the applicability of sweat for evaluation of OSA diagnosis and severity supported by the detected metabolic changes during sleep

Metabolic patterns in the lipoxygenase pathway associated to fruitiness attributes of extra virgin olive oil

Fruitiness is one of the most appreciated organoleptic features of extra virgin olive oils (EVOOs), which are categorized as a function of the fruitiness intensity and typology. In this research, we determined the main C5 and C6 volatiles formed through the lipoxygenase pathway (LOX) in 200 EVOOs produced in two agronomic seasons. Multivariate analysis was applied to find discrimination patterns in the concentrations of volatiles in EVOOs, which were tasted and classified in terms of fruitiness by the Panel Test. Multivariate ROC analysis of metabolic ratios governing the formation of C5 and C6 volatiles allowed identifying the main enzymatic steps contributing to explain chemical differences in the volatile composition of EVOOs. Fruitiness intensity was specially explained by the conversion of 3-hexenal to 2-hexenol and 3-hexenol, whereas the type of fruitiness was conditioned by the three branches of the LOX pathway that lead to the formation of C5 and C6 volatiles