{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T15:15:31Z","timestamp":1787238931403,"version":"build-2736575974"},"reference-count":25,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2024,12,19]],"date-time":"2024-12-19T00:00:00Z","timestamp":1734566400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32201254"],"award-info":[{"award-number":["32201254"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["U24A20419"],"award-info":[{"award-number":["U24A20419"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["ift.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Food Science"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:sec>\n                    <jats:title>Abstract<\/jats:title>\n                    <jats:p>\n                      Phytohemagglutinin (PHA), a natural tetramer comprising PHA\u2010E and PHA\u2010L subunits that preferentially bind to red and white blood cells, respectively, constitutes a significant antinutritional and allergenic factor in common bean seeds. The accurate measurement of PHA content is a prerequisite for ensuring food safety inspections and facilitating genetic improvements in common bean cultivars with reduced PHA levels. Currently, mainstream methods for PHA quantification involve hemagglutination assays and immunodetection, but these methods often require fresh animal blood and lack specificity and accuracy. In this study, we present a novel liquid chromatography\u2010mass spectrometry\/MS\u2010based method for PHA quantification, leveraging the advantages of isotope dilution mass spectrometry (IDMS). Two signature peptides each for PHA\u2010E and PHA\u2010L, along with a common signature peptide, were identified and employed for quantification, allowing differentiation between PHA\u2010E and PHA\u2010L subunits. The incorporation of amino acid analysis isotope IDMS enabled accurate determination of the synthetic signature peptides' purity during measurement, enhancing metrological accuracy. In addition, the trichloroacetic acid\u2013acetone protocol was established as the optimized method for total protein extraction from dry bean seeds. Quantitative analysis of PHA\u2010E and PHA\u2010L subunits in six common bean varieties using the developed method demonstrated excellent linearity (\n                      <jats:italic>R<\/jats:italic>\n                      <jats:sup>2<\/jats:sup>\n                      \u00a0&gt;\u00a00.999), sensitivity (limit of detection and quantitation as low as 2.32\u00a0ng\u00b7mg\n                      <jats:sup>\u2212<\/jats:sup>\n                      \u00b9 and 7.73\u00a0ng\u00b7mg\n                      <jats:sup>\u2212<\/jats:sup>\n                      \u00b9, respectively), recovery (94.18%\u2013104.47%), and precision (relative standard deviation\u00a0&lt;\u00a03.45%). Collectively, we offer a precise and highly reproducible method that may be used as a standard for the determination of PHA content in common beans and other agricultural products containing PHA.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/1750-3841.17590","type":"journal-article","created":{"date-parts":[[2024,12,19]],"date-time":"2024-12-19T22:49:10Z","timestamp":1734648550000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["An accurate IDMS\u2010based method for absolute quantification of phytohemagglutinin, a major antinutritional component in common bean"],"prefix":"10.1111","volume":"90","author":[{"given":"Lan","family":"Li","sequence":"first","affiliation":[{"name":"Key Laboratory of Specialty Agri\u2010Product Quality and Hazard Controlling Technology of Zhejiang, College of Life Sciences China Jiliang University  Hangzhou P.R. China"},{"name":"Technology Innovation Center of Mass Spectrometry for State Market Regulation, Center for Advanced Measurement Science National Institute of Metrology  Beijing P.R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhanying","family":"Chu","sequence":"additional","affiliation":[{"name":"Technology Innovation Center of Mass Spectrometry for State Market Regulation, Center for Advanced Measurement Science National Institute of Metrology  Beijing P.R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kang","family":"Ning","sequence":"additional","affiliation":[{"name":"Key Laboratory of Specialty Agri\u2010Product Quality and Hazard Controlling Technology of Zhejiang, College of Life Sciences China Jiliang University  Hangzhou P.R. 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