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Monitoring the large-scale production of the antihypertensive peptides RYLGY and AYFYPEL by HPLC-MS

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Abstract

This work reports the quantitative analysis of two novel antihypertensive peptides αs1-CN f(90-94), with sequence RYLGY, and αs1-CN f(143-149), with sequence AYFYPEL, by high-performance liquid chromatography–mass spectrometry in food-grade hydrolysates of milk proteins. The method was validated and showed sufficient specificity, reproducibility, linearity and recovery. Linear calibrations of the molecular ions m/z 671.2 and 902.3 were selected for the determination of the peptides RYLGY and AYFYPEL, respectively, and showed good statistical results (R 2 ≥ 0.995 and with no significant lack-of-fit). The simplicity of RP-HPLC-MS method allowed the automated quantification of both antihypertensive peptides without any sample pretreatment. The application of this method permitted the evaluation of some hydrolysis variables, i.e., substrate, temperature, hydrolysis time or enzyme/substrate ratio, on the formation of antihypertensive peptides. The quantitative analysis of RYLGY and AYFYPEL showed that ultrafiltration was not effective to improve the content in active peptides, containing the hydrolysates and their respective permeates similar peptide amounts.

HPLC-MS allows the specific quantitative determination of the antihypertensive peptides RYLGY and AYFYPEL in milk casein hydrolysates.

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Acknowledgments

This work has received financial supported from projects AGL2007-65035, AGL2008-01713, P2009/AGR-1469, Consolider Ingenio 2010 FUN-C-Food CSD2007-00063, and CENIT DEV-MET-FUN. M. M. Contreras was the recipient of a fellowship of Danone Institute.

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Authors and Affiliations

  1. Instituto de Fermentaciones Industriales (CSIC), Juan de la Cierva 3, 28006, Madrid, Spain

    María del Mar Contreras, Pedro Jesús Martín-Álvarez, Lourdes Amigo, Mercedes Ramos & Isidra Recio

  2. Innaves S.A. Polígono industrial A Granxa, Paralela 3, parcela 142, 36475, O Porriño, Pontevedra, Spain

    Beatriz Gómez-Sala

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  1. María del Mar Contreras
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  2. Beatriz Gómez-Sala
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  3. Pedro Jesús Martín-Álvarez
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  4. Lourdes Amigo
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  5. Mercedes Ramos
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  6. Isidra Recio
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Correspondence to Isidra Recio.

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del Mar Contreras, M., Gómez-Sala, B., Martín-Álvarez, P.J. et al. Monitoring the large-scale production of the antihypertensive peptides RYLGY and AYFYPEL by HPLC-MS. Anal Bioanal Chem 397, 2825–2832 (2010). https://doi.org/10.1007/s00216-010-3647-2

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  • DOI: https://doi.org/10.1007/s00216-010-3647-2

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