{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T13:49:56Z","timestamp":1792590596791,"version":"4.5.0"},"reference-count":33,"publisher":"American Chemical Society (ACS)","issue":"41","license":[{"start":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T00:00:00Z","timestamp":1791590400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T00:00:00Z","timestamp":1791590400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T00:00:00Z","timestamp":1791590400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"name":"National Basic Research Program of China (973 Program)","award":["2023YFF0613400"],"award-info":[{"award-number":["2023YFF0613400"]}]},{"name":"Capital's Funds for Health Improvement and Research","award":["CFH2024-1G-3011"],"award-info":[{"award-number":["CFH2024-1G-3011"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,21]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Accurate quantification of cow\u2019s milk allergens in processed foods by UPLC-MS\/MS is challenged by enzymatic digestion variability and complex matrix effects. \u03b2-Casein, a major milk allergen, requires reliable quantification. Herein, we developed a novel method for quantifying bovine \u03b2-casein using a recombinant, isotope-labeled, full-length protein (\u03b2-casein-I) as an internal standard. With a high isotopic labeling rate and sequence homology to the native analyte, \u03b2-casein-I corrects errors from digestion, sample preparation, and MS analysis. The validated method showed excellent linearity from 0.1 to 50 ng\/mL. In three processed food matrices, mean recoveries were 89\u2013116% with RSD &amp;lt; 13% at three spiking levels, and LODs were 0.2\u20130.5 \u03bcg\/g. Specificity was ensured by simultaneous monitoring of two signature peptides, minimizing false positives from cross-reactivity with goat milk. \u03b2-Casein was detected in all 11 commercial samples analyzed. This study provides an accurate, reliable approach for milk allergen quantification, significantly advancing traditional peptide-based methods.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c09713","type":"journal-article","created":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T15:02:00Z","timestamp":1791644520000},"page":"32677-32686","source":"Crossref","is-referenced-by-count":0,"title":["Accurate \u03b2-Casein\nAllergen Quantification\nin Processed Foods via an Isotope-Labeled Full-Length Protein Internal\nStandard"],"prefix":"10.1021","volume":"74","author":[{"given":"Yige","family":"Wu","sequence":"first","affiliation":[{"name":"Zhejiang Provincial Center for Disease Control and Prevention , , ,","place":["Hangzhou, China, 310051"]},{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qibin","family":"Liang","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]},{"name":"Capital Medical University , , ,","place":["Beijing, China, 101300"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Chang","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]},{"name":"Capital Medical University , , ,","place":["Beijing, China, 100069"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yizhu","family":"Chen","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]},{"name":"Capital Medical University , , ,","place":["Beijing, China, 100069"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuncai","family":"Liang","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]},{"name":"Sanya Institute of China Agricultural University , , ,","place":["Sanya, China, 572025"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenkai","family":"Cao","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]},{"name":"Capital Medical University , , ,","place":["Beijing, China, 100069"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuan","family":"Wu","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]},{"name":"Capital Medical University , , ,","place":["Beijing, China, 100069"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sai","family":"Fan","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]},{"name":"Capital Medical University , , ,","place":["Beijing, China, 100069"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5631-6889","authenticated-orcid":true,"given":"Bing","family":"Shao","sequence":"additional","affiliation":[{"name":"Xihua University , , ,","place":["Chengdu, China, 610039"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6480-5866","authenticated-orcid":true,"given":"Hui","family":"Li","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control , , ,","place":["Beijing, China, 100013"]},{"name":"Capital Medical University , , ,","place":["Beijing, China, 100069"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,10]]},"reference":[{"issue":"4","key":"2026102109073210400_cit1","doi-asserted-by":"publisher","first-page":"401","DOI":"10.1016\/j.anai.2023.02.002","article-title":"Food Allergy and Travel","volume":"130","author":"Yuan","year":"2023","journal-title":"Ann.\nAllergy, Asthma Immunol."},{"issue":"7","key":"2026102109073210400_cit2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/nu11071492","article-title":"Molecular Approaches\nfor Diagnosis, Therapy and Prevention\nof Cow\u2032s Milk Allergy","volume":"11","author":"Linhart","year":"2019","journal-title":"Nutrients"},{"key":"2026102109073210400_cit3","doi-asserted-by":"publisher","DOI":"10.3389\/fped.2020.00440","article-title":"Dietary Prevention\nof Atopic March in Pediatric Subjects With Cow\u2019s Milk Allergy","volume":"8","author":"Carucci","year":"2020","journal-title":"Front. Pediatr."},{"issue":"23","key":"2026102109073210400_cit4","doi-asserted-by":"publisher","DOI":"10.1002\/mnfr.202100536","article-title":"A Systematic Review of Metabolic Alterations Underlying\nIgE-Mediated Food Allergy in Children","volume":"65","author":"De Paepe","year":"2021","journal-title":"Mol.\nNutr. Food Res."},{"key":"2026102109073210400_cit5","unstructured":"EU.\n          . Regulation (EU)\nNo 1169\/2011-Food\nInformation to Consumers. 2011."},{"key":"2026102109073210400_cit6","unstructured":"FDA.\n          . Food Allergen Labeling and Consumer\nProtection Act of 2004. 2004."},{"key":"2026102109073210400_cit7","unstructured":"NHC of PRC.\n          . National\nFood Safety Standard - General Rules for Pre-Packaged Food Labeling\nGB7718\u201320252025https:\/\/sppt.cfsa.net.cn:8086\/db?type=2&guid=63F005D7-BA6F-428C-8098-826735B6C13E."},{"issue":"1","key":"2026102109073210400_cit8","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1016\/j.jaci.2017.11.003","article-title":"Food Allergy: A\nReview and Update on Epidemiology,\nPathogenesis, Diagnosis, Prevention, and Management","volume":"141","author":"Sicherer","year":"2018","journal-title":"J. Allergy Clin. Immunol."},{"issue":"235","key":"2026102109073210400_cit9","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2024.139462","article-title":"Lipid Matrix-Specific\nPretreatment\nMethod for Enhancing the Extractability and Allergenicity Maintenance\nof Bovine Milk Allergens in ELISA Detection","volume":"452","author":"Hu","year":"2024","journal-title":"Food Chem."},{"issue":"19","key":"2026102109073210400_cit10","doi-asserted-by":"publisher","first-page":"7495","DOI":"10.1021\/acs.jafc.3c00392","article-title":"Impact\nof Physicochemical Modifications in Casein Promoted by UV-C on the\nPeptide Profile of Gastric Digestion and the Transepithelial Transport\nof Peptides","volume":"71","author":"Morais","year":"2023","journal-title":"J. Agric. Food Chem."},{"issue":"22","key":"2026102109073210400_cit11","doi-asserted-by":"publisher","DOI":"10.3390\/foods11223648","article-title":"Acid and Rennet Coagulation Properties of A2Milk","volume":"11","author":"Juan","year":"2022","journal-title":"Foods (Basel, Switzerland)"},{"key":"2026102109073210400_cit12","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2024.138510","article-title":"Recent Advances\nin the Bovine \u03b2-Casein Gene Mutants on Functional Characteristics\nand Nutritional Health of Dairy Products: Status, Challenges, and\nProspects","volume":"443","author":"Sun","year":"2024","journal-title":"Food Chem."},{"key":"2026102109073210400_cit13","doi-asserted-by":"publisher","DOI":"10.1016\/j.jff.2025.107144","article-title":"A1 \u03b2-Casein and\nA2 \u03b2-Casein: Effects on the Gastrointestinal Motility, Digestion,\nand Absorption","volume":"136","author":"Li","year":"2026","journal-title":"J. Funct. Foods"},{"key":"2026102109073210400_cit14","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/j.aca.2018.11.048","article-title":"Specific and\nSensitive ELISA for Measurement of IgE-Binding Variations of Milk\nAllergen \u03b2-Lactoglobulin in Processed Foods","volume":"1052","author":"Orcajo","year":"2019","journal-title":"Anal. Chim. Acta"},{"issue":"9","key":"2026102109073210400_cit15","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2016.06.006","article-title":"Simultaneous Quantification\nof \u03b1-Lactalbumin and \u03b2-Casein in Human Milk Using Ultra-Performance\nLiquid Chromatography with Tandem Mass Spectrometry Based on Their\nSignature Peptides and Winged Isotope Internal Standards","volume":"1864","author":"Chen","year":"2016","journal-title":"Biochim. Biophys. Acta, Proteins Proteomics"},{"key":"2026102109073210400_cit16","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2015.12.058","article-title":"Development\nand Inter-Laboratory Transfer of a Decaplex Polymerase Chain Reaction\nAssay Combined with Capillary Electrophoresis for the Simultaneous\nDetection of Ten Food Allergens","volume":"199","author":"Cheng","year":"2016","journal-title":"Food Chem."},{"key":"2026102109073210400_cit17","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2022.132592","article-title":"Targeted\nProteomics for Rapid and Robust Peanut Allergen Quantification","volume":"383","author":"Nelis","year":"2022","journal-title":"Food Chem."},{"issue":"8","key":"2026102109073210400_cit18","doi-asserted-by":"publisher","first-page":"2964","DOI":"10.1021\/acs.jproteome.9b00844","article-title":"Parallel\nReaction Monitoring Mass\nSpectrometry Method for Detection of Both Casein and Whey Milk Allergens\nfrom a Baked Food Matrix","volume":"19","author":"Ramachandran","year":"2020","journal-title":"J. Proteome Res."},{"issue":"38","key":"2026102109073210400_cit19","doi-asserted-by":"publisher","first-page":"9889","DOI":"10.1021\/acs.jafc.8b03657","article-title":"Protein or\nNo Protein? Opportunities for DNA-Based Detection of Allergenic Foods","volume":"66","author":"Holzhauser","year":"2018","journal-title":"J. Agric. Food Chem."},{"issue":"3","key":"2026102109073210400_cit20","doi-asserted-by":"publisher","first-page":"728","DOI":"10.1016\/j.fshw.2022.09.006","article-title":"Food Science\nand Human Wellness Determination of Egg and Milk Allergen\nin Food Products by Liquid Chromatography-Tandem Mass Spectrometry\nBased on Signature Peptides and Isotope-Labeled Internal Standard","volume":"12","author":"Fan","year":"2023","journal-title":"Food Sci. Hum. Wellness"},{"key":"2026102109073210400_cit21","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1016\/j.foodchem.2018.08.095","article-title":"Development of a Strategy\nfor the Quantification of\nFood Allergens in Several Food Products by Mass Spectrometry in a\nRoutine Laboratory","volume":"274","author":"Planque","year":"2019","journal-title":"Food Chem."},{"issue":"250","key":"2026102109073210400_cit22","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2022.134479","article-title":"Effect of Species,\nMuscle Location, Food Processing\nand Refrigerated Storage on the Fish Allergens, Tropomyosin and Parvalbumin","volume":"402","author":"Tsai","year":"2023","journal-title":"Food Chem."},{"issue":"30","key":"2026102109073210400_cit23","doi-asserted-by":"publisher","first-page":"18974","DOI":"10.1021\/acs.jafc.5c03044","article-title":"Development and Validation of a QconCAT-Based LC-MS\/MS\nMethod for\nthe Quantification of 12 Food Allergens in Foods","volume":"73","author":"Shencheng","year":"2025","journal-title":"J. Agric. Food Chem."},{"issue":"8","key":"2026102109073210400_cit24","doi-asserted-by":"publisher","first-page":"4429","DOI":"10.1021\/acs.analchem.5b00331","article-title":"Quantitative\nPerformance of Internal\nStandard Platforms for Absolute Protein Quantification Using Multiple\nReaction Monitoring-Mass Spectrometry","volume":"87","author":"Scott","year":"2015","journal-title":"Anal.\nChem."},{"key":"2026102109073210400_cit25","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2023.138170","article-title":"A SILAC-Based Accurate\nQuantification of Shrimp Allergen Tropomyosin in Complex Food Matrices\nUsing UPLC-MS\/MS","volume":"439","author":"Wu","year":"2024","journal-title":"Food Chem."},{"issue":"6","key":"2026102109073210400_cit26","doi-asserted-by":"publisher","first-page":"2650","DOI":"10.1038\/nprot.2006.427","article-title":"A Practical Recipe\nfor Stable Isotope Labeling by Amino Acids in\nCell Culture (SILAC)","volume":"1","author":"Ong","year":"2006","journal-title":"Nat. Protoc."},{"key":"2026102109073210400_cit27","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2024.105109","article-title":"Optimal\nConditions for Carrying out Trypsin Digestions on Complex Proteomes:\nFrom Bulk Samples to Single Cells","volume":"297","author":"Mansuri","year":"2024","journal-title":"J. Proteomics"},{"key":"2026102109073210400_cit28","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijms.2019.116186","article-title":"International Journal of Mass Spectrometry\nCharacterization of Collision-Induced Dissociation of Deprotonated\nPeptides of 4 e 16 Amino Acids Using High-Resolution Mass Spectrometry","volume":"445","author":"Zuo","year":"2019","journal-title":"Int. J. Mass Spectrom."},{"issue":"4","key":"2026102109073210400_cit29","doi-asserted-by":"publisher","first-page":"1122","DOI":"10.5740\/jaoacint.SMPR2016.002","article-title":"AOAC SMPR 2016.002 Standard Method Performance Requirements\nfor Detection and Quantitation of Selected Food Allergens","volume":"99","author":"Paez","year":"2016","journal-title":"J. AOAC Int."},{"issue":"2","key":"2026102109073210400_cit30","doi-asserted-by":"publisher","first-page":"570","DOI":"10.5740\/jaoacint.19-0112","article-title":"Detection and Quantitation of Selected\nFood Allergens by Liquid Chromatography with Tandem Mass Spectrometry:\nFirst Action 2017.17","volume":"103","author":"New","year":"2020","journal-title":"J. AOAC Int."},{"issue":"12","key":"2026102109073210400_cit31","doi-asserted-by":"publisher","first-page":"5278","DOI":"10.1002\/jsfa.11177","article-title":"Quantification of Crustacean Tropomyosin in Foods Using\nHigh-Performance Liquid Chromatography\u2013Tandem Mass Spectrometry\nMethod","volume":"101","author":"Wang","year":"2021","journal-title":"J. Sci. Food Agric."},{"key":"2026102109073210400_cit32","doi-asserted-by":"publisher","DOI":"10.1016\/j.lwt.2021.112367","article-title":"Development and Validation of Mass\nSpectrometry-Based Method for\nDetecting Shrimp Allergen Tropomyosin","volume":"152","author":"Ho","year":"2021","journal-title":"Lwt"},{"issue":"11","key":"2026102109073210400_cit33","doi-asserted-by":"publisher","DOI":"10.3390\/foods13111637","article-title":"A Dual and\nRapid RPA-CRISPR\/Cas12a Method for Simultaneous Detection of Cattle\nand Soybean-Derived Adulteration in Goat Milk Powder","volume":"13","author":"Wen","year":"2024","journal-title":"Foods"}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c09713","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/41\/32677\/68811055\/acs.jafc.6c09713.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/41\/32677\/68811055\/acs.jafc.6c09713.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T13:30:27Z","timestamp":1792589427000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/41\/32677\/5448508\/Accurate-Casein-Allergen-Quantification-in"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,10]]},"references-count":33,"journal-issue":{"issue":"41","published-online":{"date-parts":[[2026,10,10]]},"published-print":{"date-parts":[[2026,10,21]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c09713","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,10]]}}}