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Determination of degradation degree of ovalbumin in hen’s egg enzymatic hydrolysates using liquid chromatography–tandem mass spectrometry

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Abstract

Enzymatic hydrolysis of hen egg confers beneficial functionalities, but it is essential to control the progress of hydrolysis to enhance the quality and reduce allergy risk. However, a reliable method for directly controlling the progress of egg protein hydrolysis is unavailable. Focusing on ovalbumin, a major protein and allergen in egg white, we demonstrated the applicability of liquid chromatography–tandem mass spectrometry (LC–MS/MS) to assess the degradation of ovalbumin in its enzymatic hydrolysates. Using immobilized Alcalase or Everlase, the degradation degree (DD) of ovalbumin was determined using LC–MS/MS; the result (8.8–95.3%) was highly correlated with the DD (8.9–92.3%) obtained using size-exclusion chromatography (SEC-HPLC) (r > 0.96, p < 0.001). In egg white hydrolysates with a DD of 16.1–99.5%, ovalbumin recovery when spiked with 1250 ng/mL was 81.1–114.8% with excellent dilution linearity (r2 > 0.99). These results indicate that LC–MS/MS is a reliable method for increasing the quality of enzymatically hydrolyzed egg products for food, nutraceutical, pharmaceutical, and cosmetic industries.

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Acknowledgements

The authors thank Mr. Kotaro Uchida, Dr. Keiji Kajimura, and Dr. Kazuhiko Akutsu for their sincere encouragement.

Funding

This work was supported by the Japan Society for the Promotion of Science KAKENHI [Grant number 18K13044].

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Authors and Affiliations

  1. Division of Hygienic Chemistry, Osaka Institute of Public Health, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-0025, Japan

    Kyohei Kiyota, Masato Yoshimitsu & Hiroshi Matsui

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  1. Kyohei Kiyota
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  2. Masato Yoshimitsu
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  3. Hiroshi Matsui
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Contributions

KK: Conceptualization, Methodology, Validation, Formal analysis and investigation, Resources, Writing—original draft preparation, Funding acquisition. MY: Conceptualization, Resources, Writing—review and editing, Supervision. HM: Visualization, Writing—review and editing.

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Correspondence to Kyohei Kiyota.

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Kiyota, K., Yoshimitsu, M. & Matsui, H. Determination of degradation degree of ovalbumin in hen’s egg enzymatic hydrolysates using liquid chromatography–tandem mass spectrometry. Food Measure 17, 46–53 (2023). https://doi.org/10.1007/s11694-022-01591-z

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  • DOI: https://doi.org/10.1007/s11694-022-01591-z

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