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Pressurized liquid extraction as a sample preparation method for the analysis of isoflavones in pulses

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Abstract

In this work, we describe a rapid and simple analytical method that exploits pressurized liquid extraction (PLE) and liquid chromatography with diode array detection for the determination of isoflavones in samples of Spanish pulses. Confirmation of the analytes present was performed using ion-trap mass spectrometry. To optimize the PLE extraction, variables such as the dispersing agent, type of solvent and sample amount, and the experimental parameters, such as temperature and the number of extraction cycles, were studied. Separation was carried out using a reverse-phase C18 with polar endcapping as the stationary phase and acetonitrile/water with 0.2 % of formic acid, under a gradient regime, as the mobile phase. Optimal extraction of formononetin and biochanin-A from chickpeas with PLE was achieved using Hydromatrix as a dispersant agent, methanol/water (50:50), a temperature of 90 °C, and three cycles. The same optimal conditions—except methanol/water (75:25)—for solvent extraction were obtained for the extraction of daidzin, genistin, and formononetin from lentils. Recoveries ranged from 97 to 110 %, and standard deviations lower than 20 % were obtained. The contents obtained for daidzin in lentils using the proposed method were not significantly different from those obtained using another official method of analysis.

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Acknowledgments

The authors acknowledge the financial support of the Ministerio de Ciencia e Innovación, Spain (Project CTQ 2011-24075), and L.P.M. is also grateful to Junta de Castilla-León for an awarded doctoral fellowship.

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Authors and Affiliations

  1. Departamento de Química Analítica, Nutrición y Bromatología, Facultad de Ciencias Químicas, Plaza de los Caídos s/n, 37008, Salamanca, Spain

    Ma. Milagros Delgado-Zamarreño, Lara Pérez-Martín, Myriam Bustamante-Rangel & Rita Carabias-Martínez

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  1. Ma. Milagros Delgado-Zamarreño
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  2. Lara Pérez-Martín
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  3. Myriam Bustamante-Rangel
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  4. Rita Carabias-Martínez
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Correspondence to Ma. Milagros Delgado-Zamarreño.

Additional information

Published in the special paper collection Progress on Environmental and Bioanalysis in Spain with guest editors Alfredo Sanz-Medel and Elena Domínguez.

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Delgado-Zamarreño, M.M., Pérez-Martín, L., Bustamante-Rangel, M. et al. Pressurized liquid extraction as a sample preparation method for the analysis of isoflavones in pulses. Anal Bioanal Chem 404, 361–366 (2012). https://doi.org/10.1007/s00216-012-5912-z

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  • DOI: https://doi.org/10.1007/s00216-012-5912-z

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