Development of an HPLC-MS method for the determination of natural pteridines in tomato samples

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Development of an HPLC-MS method for the determination of natural pteridines in tomato samples

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Title: Development of an HPLC-MS method for the determination of natural pteridines in tomato samples
Author: Martín Tornero, Elísabet; González Gómez, David; Durán Merás, Isabel; Espinosa Mansilla, Anunciación
Abstract: En las plantas, las pteridinas reducidas son intermedios de la biosíntesis de folato, y la presencia de estos analitos en los procesos de biofortificación se considera crucial. Se optimizó un método simple de cromatografía líquida-espectrometría de masas (LC-ESI-MS) para la determinación de pteridinas naturales en muestras de tomate. Se ha empleado una etapa de extracción en fase sólida (SPE) usando cartuchos ISOLUTE ENV para limpiar las muestras. Once pteridinas han sido analizadas pero solo cuatro de ellas han sido detectadas y cuantificadas en tomates. La estabilidad de las pteridinas e hidropteridinas en extractos de tomate ha sido estudiada. Se evaluaron los parámetros de validación y se obtuvo una buena linealidad ( R 2 > 0,99 en todos los casos) y precisión (los valores de la desviación estándar relativa interdía fueron inferiores al 10%).Las cantidades (como μg por g de muestra fresca) encontradas de cada pteridina fueron 0,019, 0,44, 0,043 y 0,087 para neopterina, 7,8-dihidroneopterina, 6-hidroximetilpterina y ácido pterin-6-carboxílico, respectivamenteIn plants, reduced pteridines are folate biosynthesis intermediates, and the presence of these analytes in biofortification processes is considered crucial. A simple liquid chromatography-mass spectrometry (LC-ESI-MS) method has been optimized for the determination of natural pteridines in tomato samples. A solid phase extraction (SPE) step using ISOLUTE ENV cartridges has been employed for cleaning up the samples. Eleven pteridines have been assayed but only four of them have been detected and quantified in tomatoes. The stability of the pteridines and hydropteridines in tomato extracts has been studied. Validation parameters have been evaluated and good linearity (R2 > 0.99 in all cases) and precision (interday relative standard deviation values were lower than 10%) were obtained. The amounts (as μg per g of fresh sample) found of each pteridine were 0.019, 0.44, 0.043 and 0.087 for neopterin, 7,8-dihydroneopterin, 6-hydroxymethylpterin and pterin-6-carboxylic acid, respectively.
URI: http://hdl.handle.net/10662/7571
Date: 2016


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