Desarrollo y validación de un método HPLC para la determinación simultánea de paracetamol, ácido acetilsalicílico y cafeína en productos farmacéuticos

Los métodos usados en los análisis farmacéuticos deben ser validados para demostrar que son lo suficientemente exactos, específicos, sensibles y precisos para cumplir con los requisitos reglamentarios de la industria farmacéutica, garantizando así la seguridad y la calidad de los productos. La validación es una forma sistemática de evaluar que el procedimiento analítico es adecuado para el propósito previsto, que produce resultados confiables y precisos y cumple con los requisitos de los organismos reguladores. El ácido acetilsalicílico (AAS) y el paracetamol (PCM) se utilizan como agentes analgésicos y antipiréticos y aparecen generalmente asociados con la cafeína (CFN). Estos compuestos pueden determinarse simultáneamente en medicamentos y en otras formulaciones utilizando la técnica RP-HPLC-UV-Vis. Las mediciones se realizaron con un cromatógrafo de líquidos Agilent Technologies serie 1200 equipado con una columna RP C18 y un detector UV-Vis de longitud de onda variable. Antes de la validación del método, las condiciones cromatográficas se optimizaron utilizando un diseño experimental de Taguchi para identificar los factores más significativos y sus niveles. Se seleccionaron seis factores a tres niveles (composición de metanol, acetonitrilo y tampón de fosfato 0,01 M en la fase móvil, pH, velocidad de flujo y longitud de onda) y un factor a dos niveles (temperatura de columna) para elaborar un diseño L18(2137) de matriz ortogonal. La sacarosa se utilizó como factor de ruido para simular el efecto del excipiente con el fin de aumentar la robustez del método analítico. El análisis de varianza de las áreas de los picos para los tres compuestos indicó que el factor que más influyó fue la longitud de onda, sin embargo la composición y el pH de la fase móvil fueron los factores que más influyeron en la resolución de los picos. La sacarosa no afectó ni a las áreas de los picos ni a los tiempos de retención. Las condiciones de RP-HPLC optimizadas y posteriormente utilizadas en la realización de experimentos fueron: temperatura, 25 ºC; composición de la fase móvil MeOH: Acetonitrilo: Tampón fosfato, 30: 5: 25; pH, 2,7; velocidad de flujo, 1 mL·min-1 y longitud de onda, 230, 246 y 273 nm para AAS, PCM y CFN, respectivamente. Los parámetros de validación estimados han sido: exactitud, precisión, selectividad, robustez, límite de detección y límite de cuantificación, linealidad e intervalo lineal. La veracidad se evaluó para AAS y PCM mediante la participación en una prueba de aptitud para estudiantes de Química Analítica. Todos los parámetros de validación cumplieron los objetivos previamente especificados. La incertidumbre de medida se evaluó a partir de los resultados de la validación. La incertidumbre obtenida superó ligeramente el objetivo propuesto. El método RP-HPLC optimizado es simple, rápido, sensible y preciso y, por lo tanto, adecuado para el análisis rutinario del ácido acetilsalicílico, paracetamol y cafeína en analgésicos. Este se aplicó a la determinación de estos analitos en formulaciones comerciales vendidas en España.Grado en Químic

Development and validation of a gas chromatography-mass spectrometry method for determining acaricides in bee pollen

Producción CientíficaPesticides can be found in beehives for several reasons, including contamination from surrounding crops or for their use by beekeepers, which poses a risk to bee ecosystems and consumers. Therefore, efficient and sensitive methods are needed for determining pesticide residues in bee products. In this study, a new analytical method has been developed and validated to determine seven acaricides (atrazine, chlorpyrifos, chlorfenvinphos, α-endosulfan, bromopropylate, coumaphos, and τ-fluvalinate) in bee pollen using gas chromatography coupled to mass spectrometry. After an optimization study, the best sample treatment was obtained when using a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) method employing an ethyl acetate and cyclohexane as the extractant mixture, and a mixture of salts for the clean-up step. A chromatographic analysis (<21 min) was performed in an Agilent DB-5MS column, and it was operated under programmed temperature conditions. The method was fully validated in terms of selectivity, limits of detection (0.2–3.1 µg kg−1) and quantification (0.6–9.7 µg kg−1), linearity, matrix effect (<20% in all cases), trueness (recoveries between 80% and 108%), and precision. Finally, the proposed method was applied to analyze commercial bee pollen samples, and some of the target pesticides (chlorfenvinphos, α-endosulfan, coumaphos, and τ-fluvalinate) were detected.Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) y Fondo Europeo de Desarrollo Regional (FEDER) - (grant RTA2017-00004-C02-0

Development and validation of an analytical methodology based on solvent extraction and gas chromatography for determining pesticides in royal jelly and propolis

Producción CientíficaWe propose a new analytical methodology to determine seven pesticides (atrazine, chlorpyrifos, chlorfenvinphos, α-endosulfan, bromopropylate, coumaphos, and τ-fluvalinate) in royal jelly and propolis products using gas chromatography-mass spectrometry. Sample treatment, with minor modifications for propolis, consisted of a solvent extraction with a hexane and isopropanol mixture, and a further clean-up step. Meanwhile, chromatographic analysis (<25 min) was performed in a DB-5MS column under programmed temperature conditions. In all cases we validated the method in terms of selectivity, limits of detection (0.1–2.8 μg kg−1) and quantification (0.3–9.2 μg kg−1), linearity, matrix effect (<±20 %), trueness (recoveries between 93 % and 118 %), and precision (relative standard deviation < 11 %). All royal jelly liquid dietary supplements were positive for chlorfenvinphos and, in the case of one of them, for α-endosulfan; chlorfenvinphos was determined in some fresh royal jelly samples, and no pesticide residues were detected in the propolis samples analysedPlan Nacional de Investigación e Innovación Científica y Técnica, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria-INIA-FEDER (grant number RTA2017-00004-C02-02

Evaluation of the potential migration of acaricides from stamped beeswax to honey simulating beehive conditions: a pilot study

Producción CientíficaThere is an issue concerning the migration of pesticides from stamped beeswax to other bee products, mainly, honey. This implies that honey and other beehive-based products could be contaminated, which would represent a potential risk to consumers and food safety. To determine the degree to which acaricides can be transferred from the stamped beeswax to the honey, a pilot experiment was designed consisting of placing beeswax that contained acaricides (endogenous content, τ-fluvalinate, chlorfenvinphos, and coumaphos; spiked samples, coumaphos) in contact with multifloral honey in an incubator. Temperature and shaking conditions were settled simulating beehive conditions. The isolation of the analytes from beeswax involved the use of a QuEChERS (quick, easy, cheap, effective, rugged & safe) method, and honey was analyzed through a solvent extraction procedure. Acaricides were determined in the resulting extracts by gas chromatography with mass spectrometry detection. Coumaphos was the only acaricide transferred into honey in beeswaxes with endogenous acaricide content; meanwhile, results of the experiments with spiked beeswax samples showed that the initial concentration present in the contaminated beeswax significantly influences the transfer to honey, as it was observed that in the spiked beeswax sheets with high levels of coumaphos, the migration from beeswax to honey arose.Plan Nacional de Investigación e Innovación Científica y Técnica 2013–2016, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria-INIA–FEDER (RTA2017-00004-C02-02

Comprehensive overview of the analytical methods for determining pyrrolizidine alkaloids and their derived oxides in foods

Producción CientíficaPyrrolizidine alkaloids and their derived oxides are toxins naturally produced by plants when they are exposed to stress factors. In particular, the unsaturated pyrrolizidine alkaloids exhibiting a double bond between the 1,2-positions have caused great interest in recent years since they have pneumotoxic, genotoxic and carcinogenic effects upon ingestion. In this review, focus on their chemical structure justifying their toxicity is provided as well as insight on the recently adopted EU regulatory framework. In addition, to reveal current trends and highlight the research effort in the field, an all-time bibliometric analysis was performed for the first time. It was found that tea, infusions, honey, spices, and cereals are the most common foodstuffs in which pyrrolizidine alkaloids have been detected. In terms of pre-analytical steps, the application of efficient sample preparation is necessary to detect pyrrolizidine alkaloids in complex food matrices. Solvent extraction followed by a clean-up stage, e.g., solid-phase extraction was the most applied option. Focusing on the analytical methods, liquid chromatography combined with various mass spectrometry detectors has been the golden standard in the field. Gas chromatography methods were also applied, but the need for analyte derivatisation has hindered their application. All in all, this review provides an overview on the analysis of pyrrolizidine alkaloids and their derived oxides, comprehensively discussing the up-to-date advances and highlighting the analytical challenges to be faced.METROFOOD-CZ research infrastructure project (MEYS Grant No: LM2018100

XXXIX REUNIÓN BIENAL DE LA SOCIEDAD ESPAÑOLA DE QUÍMICA

Certificado de presentación de comunicación, y copia de la comunicación oral-flash presentada al congreso

Determination of acaricides in honeys from different botanical origins by gas chromatography-mass spectrometry

Producción CientíficaAn analytical method has been proposed and validated to determine seven acaricides (atrazine, chlorpyrifos, chlorfenvinphos, α-endosulfan, bromopropylate, coumaphos, and τ-fluvalinate) in honeys from different botanical origins (multifloral, heather and rosemary) by means of gas chromatography-mass spectrometry. An efficient and simple sample treatment was proposed that involved a solvent extraction with an ethyl acetate and cyclohexane (50:50, v/v) mixture. Chromatographic analysis (<25 min) was performed in a DB-5MS column under programmed temperature conditions. The method was validated in terms of selectivity, limits of detection (0.2–2.0 μg kg-1) and quantification (0.5–7.6 μg kg-1), linearity (limit of quantification-700 (heather) or 800 (multifloral and rosemary) μg kg-1), matrix effect (<20 % in most cases), trueness (recoveries between 81 % and 108 %), and precision (relative standard deviation < 15 %). Finally, of the seven acaricides investigated in several honey samples only τ-fluvalinate residues (<limit of quantification - 23 μg kg-1) were found.Este trabajo forma parte del proyecto de investigación financiado por el Ministerio de Economía y Competitividad e INIA-FEDER (RTA2017-00004-C02-02)

VII Jornada de iNNovación docente Uva

Trabajo poster presentad

VII Jornada de innovación Docente (UVa)

Trabajo presentado en las Jornadas de iNNovación Docente 2023 de la Uv