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Comparison of different types of stationary phases for the analysis of soy isoflavones by HPLC

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Abstract

Nowadays, there are new technologies in high-performance liquid chromatography columns available enabling faster and more efficient separations. In this work, we compared three different types of columns for the analysis of main soy isoflavones. The evaluated columns were a conventional reverse phase particle column, a fused-core particle column, and a monolithic column. The comparison was in terms of chromatographic parameters such as resolution, asymmetry, number of theoretical plates, variability of retention time, and peak width. The lower column pressure was provided by the monolithic column, although lower chromatographic performance was achieved. Conventional and fused-core particle columns presented similar pressure. Results also indicate that direct transfer between particle and monolithic columns is not possible requiring adjustment of conditions and a different method optimization strategy. The best chromatographic performance and separation speed were observed for the fused-core particle column. Also, the effect of sample solvent on the separation and peak shape was evaluated and indicated that monolithic column is the most affected especially when using higher concentrations of acetonitrile or ethanol. Sample solvent that showed the lowest effect on the chromatographic performance of the columns was methanol. Overall evaluation of methanol and acetonitrile as mobile phase for the separation of isoflavones indicated higher chromatographic performance of acetonitrile, although methanol may be an attractive alternative. Using acetonitrile as mobile phase resulted in faster, higher resolution, narrower, and more symmetric peaks than methanol with all columns. It also generated the lower column pressure and flatter pressure profile due to mobile phase changes, and therefore, it presents a higher potential to be explored for the development of faster separation methods.

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Acknowledgments

The authors acknowledge funding from the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) projects AT 07-003 and RTA2009-00049-00-00. The authors also would like thank the invaluable work of Rosario Vicente in the experimental part of this work.

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Authors and Affiliations

  1. Centro para la Calidad de los Alimentos, Instituto Nacional de Investigación Y Tecnología Agraria y Alimentaria (INIA), c/José Tudela s/n, 42004, Soria, Spain

    N. Manchón, M. D’Arrigo, A. García-Lafuente, E. Guillamón, A. Villares, J. A. Martínez, A. Ramos & M. A. Rostagno

  2. Dpto. Fisiología y Nutrición, Edificio de Investigación, Universidad de Navarra, C/Irunlarrea, 1, 31008, Pamplona, Spain

    J. A. Martínez

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  1. N. Manchón
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  2. M. D’Arrigo
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  3. A. García-Lafuente
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  4. E. Guillamón
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  5. A. Villares
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  6. J. A. Martínez
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  7. A. Ramos
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  8. M. A. Rostagno
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Correspondence to M. A. Rostagno.

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Published in the special issue Advances in Analytical Separations with Guest Editors Yolanda Pico and Joan O. Grimalt.

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Manchón, N., D’Arrigo, M., García-Lafuente, A. et al. Comparison of different types of stationary phases for the analysis of soy isoflavones by HPLC. Anal Bioanal Chem 400, 1251–1261 (2011). https://doi.org/10.1007/s00216-011-4673-4

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  • DOI: https://doi.org/10.1007/s00216-011-4673-4

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