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Estimation of Degree of Hydrolysis of Protein Hydrolysates by Size Exclusion Chromatography

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Abstract

Estimating the degree of hydrolysis (DH) of commercial protein hydrolysates from their molecular weight (MW) median values generated by high performance size exclusion chromatography with UV detection (HPSEC/UV) was evaluated. DH is defined as the proportion (%) of cleaved peptide bonds in a protein hydrolysate. Several methods for DH determination are available, and protein hydrolysates are often tested both for DH by one of these methods and for MW profile by an HPSEC/UV method. We describe a means of estimating the DH of a protein hydrolysate from its MW profile, thereby eliminating the need for the performance of a second, separate test. In this study, a plot of average DH (%) vs. MW median (Da) was constructed from pancreatin digestion time data for seven protein substrates, determined by an HPSEC/UV system with a peptide gel filtration column. A general formula obtained from the plot enables DH estimation for a protein hydrolysate from its MW median value. By using this approach, the DH values for a total of thirteen commercial protein hydrolysates derived from whey, casein, and soy were estimated. Method accuracy was assessed by comparing the MW median-estimated DH to the DH determined by TNBS colorimetry, which generally agreed well, with R2 > 0.95. This study demonstrated that a reliable estimate of protein hydrolysate DH may be obtained from its MW median so that the performance of a separate DH test may not be necessary.

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Authors and Affiliations

  1. Abbott Nutrition Research and Development, 20 Biopolis Way, #09-01/02 Centros Building, Singapore, 138668, Singapore

    Ding Yi & Qi Lin

  2. Abbott Laboratories, Abbott Nutrition Division, 3300 Stelzer Road, Columbus, OH, 43219, USA

    Paul W. Johns

Authors
  1. Ding Yi
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  2. Qi Lin
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  3. Paul W. Johns
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Contributions

Ding Yi: conceptualization; data curation; methodology; writing, original draft preparation; writing, review and editing. Qi Lin: conceptualization; data curation; methodology; writing, original draft preparation; writing, review and editing. Paul Johns: conceptualization; data curation; formal analysis; validation; methodology; writing, original draft preparation; writing, review and editing.

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Correspondence to Paul W. Johns.

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Ding Yi declares that he has no conflict of interest. Qi Lin declares that she has no conflict of interest. Paul Johns declares that he has no conflict of interest.

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Yi, D., Lin, Q. & Johns, P.W. Estimation of Degree of Hydrolysis of Protein Hydrolysates by Size Exclusion Chromatography. Food Anal. Methods 14, 805–813 (2021). https://doi.org/10.1007/s12161-020-01936-8

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  • DOI: https://doi.org/10.1007/s12161-020-01936-8

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