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A collaborative study to validate in vitro assays for protein digestibility assessment using pH-drop and pH-stat methods.

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Title: A collaborative study to validate in vitro assays for protein digestibility assessment using pH-drop and pH-stat methods.
Authors: Goldberg, Erin M.1 (AUTHOR), Sá, Amanda G.A.1 (AUTHOR), Franczyk, Adam J.1 (AUTHOR) umfranc3@myumanitoba.ca, Krul, Elaine S.2 (AUTHOR), Lyle, Barbara3 (AUTHOR), Jing, Mingyan1 (AUTHOR), Liu, Fiona4 (AUTHOR), Wu, Xin4 (AUTHOR), Bandara, Nandika1 (AUTHOR), Brisson, Guillaume5 (AUTHOR), Campayo, Vicenta Garcia6 (AUTHOR), Chen, Lingyun7 (AUTHOR), Hooper, Sharon8 (AUTHOR), L'Hocine, Lamia9 (AUTHOR), Nickerson, Mike10 (AUTHOR), Nosworthy, Matthew11 (AUTHOR), House, James D.1 (AUTHOR)
Source: Journal of Food Composition & Analysis. Mar2026, Vol. 151, pN.PAG-N.PAG. 1p.
Subjects: In vitro studies, Acidification, Proteolysis, Food composition, Cooperative research
Abstract: The diversification of protein sources used in food formulation has increased the need to assess protein quality beyond traditional food forms. Protein digestibility is a key component of the Protein Digestibility Corrected Amino Acid Score (PDCAAS), the regulatory metric derived from rodent bioassays. Ethical concerns and the high cost of animal testing limit the ability of food formulators to screen protein ingredients and evaluate processing effects on digestibility. Findings from an international collaborative study are presented to position two in vitro methods, the pH-drop and pH-stat assays, as accredited approaches for determining protein digestibility. Nine laboratories participated in the study and analyzed 12 protein ingredients. Relative standard deviations for repeatability ranged from 0.8 to 2.1 % and 0.5–4.8 %, while reproducibility ranged from 1.2 to 3.6 % and 1.1–4.9 % for the pH-drop and pH-stat methods, respectively. Comparison between the two assays demonstrated strong correlation and moderate agreement, with the pH-stat assay yielding slightly lower digestibility values. In vitro protein digestibility coefficients aligned well with literature reported true faecal protein digestibility values for comparable, non-identical protein ingredients. Both methods received official AOCS Uniform Methods Committee approval, offering simple, affordable, reliable, and ethical tools to support informed decisions on protein digestibility during food formulation. [Display omitted] ● 9 laboratories successfully evaluated protein digestibility of 12 diverse proteins. ● Both pH assays showed strong repeatability and reproducibility across laboratories. ● pH-stat assay yielded slightly lower digestibility values than the pH-drop method. ● Validated in vitro assays offer ethical, affordable tools for protein screening. ● American Oil Chemists' Society approved pH-drop and pH-stat methods in 2025. [ABSTRACT FROM AUTHOR]
Copyright of Journal of Food Composition & Analysis is the property of Academic Press Inc. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
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DbLabel: Engineering Source
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PubTypeId: academicJournal
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  Data: A collaborative study to validate in vitro assays for protein digestibility assessment using pH-drop and pH-stat methods.
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  Data: <searchLink fieldCode="AR" term="%22Goldberg%2C+Erin+M%2E%22">Goldberg, Erin M.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Sá%2C+Amanda+G%2EA%2E%22">Sá, Amanda G.A.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Franczyk%2C+Adam+J%2E%22">Franczyk, Adam J.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<i> umfranc3@myumanitoba.ca</i><br /><searchLink fieldCode="AR" term="%22Krul%2C+Elaine+S%2E%22">Krul, Elaine S.</searchLink><relatesTo>2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Lyle%2C+Barbara%22">Lyle, Barbara</searchLink><relatesTo>3</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Jing%2C+Mingyan%22">Jing, Mingyan</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Liu%2C+Fiona%22">Liu, Fiona</searchLink><relatesTo>4</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Wu%2C+Xin%22">Wu, Xin</searchLink><relatesTo>4</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Bandara%2C+Nandika%22">Bandara, Nandika</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Brisson%2C+Guillaume%22">Brisson, Guillaume</searchLink><relatesTo>5</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Campayo%2C+Vicenta+Garcia%22">Campayo, Vicenta Garcia</searchLink><relatesTo>6</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Chen%2C+Lingyun%22">Chen, Lingyun</searchLink><relatesTo>7</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Hooper%2C+Sharon%22">Hooper, Sharon</searchLink><relatesTo>8</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22L'Hocine%2C+Lamia%22">L'Hocine, Lamia</searchLink><relatesTo>9</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Nickerson%2C+Mike%22">Nickerson, Mike</searchLink><relatesTo>10</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Nosworthy%2C+Matthew%22">Nosworthy, Matthew</searchLink><relatesTo>11</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22House%2C+James+D%2E%22">House, James D.</searchLink><relatesTo>1</relatesTo> (AUTHOR)
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  Data: <searchLink fieldCode="JN" term="%22Journal+of+Food+Composition+%26+Analysis%22">Journal of Food Composition & Analysis</searchLink>. Mar2026, Vol. 151, pN.PAG-N.PAG. 1p.
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  Data: <searchLink fieldCode="DE" term="%22In+vitro+studies%22">In vitro studies</searchLink><br /><searchLink fieldCode="DE" term="%22Acidification%22">Acidification</searchLink><br /><searchLink fieldCode="DE" term="%22Proteolysis%22">Proteolysis</searchLink><br /><searchLink fieldCode="DE" term="%22Food+composition%22">Food composition</searchLink><br /><searchLink fieldCode="DE" term="%22Cooperative+research%22">Cooperative research</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: The diversification of protein sources used in food formulation has increased the need to assess protein quality beyond traditional food forms. Protein digestibility is a key component of the Protein Digestibility Corrected Amino Acid Score (PDCAAS), the regulatory metric derived from rodent bioassays. Ethical concerns and the high cost of animal testing limit the ability of food formulators to screen protein ingredients and evaluate processing effects on digestibility. Findings from an international collaborative study are presented to position two in vitro methods, the pH-drop and pH-stat assays, as accredited approaches for determining protein digestibility. Nine laboratories participated in the study and analyzed 12 protein ingredients. Relative standard deviations for repeatability ranged from 0.8 to 2.1 % and 0.5–4.8 %, while reproducibility ranged from 1.2 to 3.6 % and 1.1–4.9 % for the pH-drop and pH-stat methods, respectively. Comparison between the two assays demonstrated strong correlation and moderate agreement, with the pH-stat assay yielding slightly lower digestibility values. In vitro protein digestibility coefficients aligned well with literature reported true faecal protein digestibility values for comparable, non-identical protein ingredients. Both methods received official AOCS Uniform Methods Committee approval, offering simple, affordable, reliable, and ethical tools to support informed decisions on protein digestibility during food formulation. [Display omitted] ● 9 laboratories successfully evaluated protein digestibility of 12 diverse proteins. ● Both pH assays showed strong repeatability and reproducibility across laboratories. ● pH-stat assay yielded slightly lower digestibility values than the pH-drop method. ● Validated in vitro assays offer ethical, affordable tools for protein screening. ● American Oil Chemists' Society approved pH-drop and pH-stat methods in 2025. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
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  Data: <i>Copyright of Journal of Food Composition & Analysis is the property of Academic Press Inc. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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RecordInfo BibRecord:
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      – Type: doi
        Value: 10.1016/j.jfca.2026.108918
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      – Code: eng
        Text: English
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        PageCount: 1
        StartPage: N.PAG
    Subjects:
      – SubjectFull: In vitro studies
        Type: general
      – SubjectFull: Acidification
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      – SubjectFull: Proteolysis
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      – SubjectFull: Food composition
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      – SubjectFull: Cooperative research
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              Text: Mar2026
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