Determination of Resveratrol and Piceid Isomers in Bee Pollen by Liquid Chromatography Coupled to Electrospray Ionizatio
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Determination of Resveratrol and Piceid Isomers in Bee Pollen by Liquid Chromatography Coupled to Electrospray Ionization-Mass Spectrometry Ana M. Ares & María E. Soto & María J. Nozal & José L. Bernal & Mariano Higes & José Bernal
Received: 2 October 2014 / Accepted: 7 November 2014 # Springer Science+Business Media New York 2014
Abstract A new method has been developed to determine resveratrol and piceid isomers in bee pollen using liquid chromatography (LC) coupled to electrospray ionization-mass spectrometry (ESI-MS). An efficient extraction procedure has also been proposed (average analyte recoveries were between 89 and 96 %); this involved a cleaning step with hexane, a solid–liquid extraction using a mixture of ethanol and water (80:20, v/v), and a concentration step in a rotary evaporator. The separation of all the compounds was achieved using a C18 column (150×4.6 mm) and a mobile phase composed of 1 % (v/v) formic acid in water and acetonitrile at a flow rate of 0.8 mL/min. The method was fully validated in terms of selectivity, limits of detection (LOD) and quantification (LOQ), linearity, precision, and accuracy. The LOD and LOQ values ranged from 10 to 25 and 35 to 80 μg/kg, respectively. Finally, the proposed method was applied to analyze bee pollen samples from different origins.
Keywords Bee pollen . LC-ESI-MS . Piceid isomers . Resveratrol isomers . Solid–liquid extraction
A. M. Ares : M. E. Soto : M. J. Nozal : J. L. Bernal : J. Bernal (*) I.U. CINQUIMA, Analytical Chemistry Group, University of Valladolid, 47011 Valladolid, Spain e-mail: [email protected] URL: http://tesea.uva.es M. E. Soto Faculty of Sciences and Technology, National Autonomous University of Nicaragua, León, Nicaragua M. Higes Centro Apícola Regional, Bee Pathology Laboratory, Consejería de Agricultura, JCCM, Marchamalo 19180 Guadalajara, Spain
Introduction Resveratrol (3,4′,5-trihydroxystilbene) is the most representative member of the stilbenes, which are polyphenolic phytoalexins produced by plants as defense substances against biotic or abiotic stress (Cacho et al. 2013). In nature, it can be found in two isomeric forms (cis and trans; see Fig. 1), which may be transformed into the corresponding cis or trans piceids (resveratrol-3-O-β-D-glucopyranoside), respectively (Liu et al. 2013). Resveratrol and its related compounds have all been positively linked to health benefits, including anticancer, anti-inflammatory, antioxidant, antimicrobial, and antiaging properties (Cacho et al. 2013; Fernández-Mar et al. 2012; Leiro et al. 2010; Liu et al. 2013; Shukitt-Hale et al. 2006; Weng et al. 2010). It must be specified that the biological activity of trans-resveratrol has been more widely researched than the other isomers (cis-resveratrol, transpiceid, and cis-piceid), but it appears that all of them may also have health benefits (Liu et al. 2013). What is more, the supposed benefits of these natural compounds have been investigated not only in humans, but also in animals such as honeybees. Some studies, for example, have
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