{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T18:16:39Z","timestamp":1787508999292,"version":"build-2736575974"},"reference-count":38,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2019,5,18]],"date-time":"2019-05-18T00:00:00Z","timestamp":1558137600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology, Taiwan","doi-asserted-by":"publisher","award":["MOST 102-2313-B-002-041-MY3"],"award-info":[{"award-number":["MOST 102-2313-B-002-041-MY3"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology, Taiwan","doi-asserted-by":"publisher","award":["MOST 107-2314-B-002-184-MY3"],"award-info":[{"award-number":["MOST 107-2314-B-002-184-MY3"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Stable-isotope dimethyl labeling is a highly reactive and cost-effective derivatization procedure that could be utilized in proteomics analysis. In this study, a liquid chromatography\u2013 tandem mass spectrometry in multiple reaction monitoring mode (LC-MS-MRM) platform for the quantification of kiwi allergens was first developed using this strategy. Three signature peptides for target allergens Act d 1, Act d 5, and Act d 11 were determined and were derivatized with normal and deuterated formaldehyde as external calibrants and internal standards, respectively. The results showed that sample preparation with the phenol method provided comprehensive protein populations. Recoveries at four different levels ranging from 72.5\u2013109.3% were achieved for the H-labeled signature peptides of Act d 1 (SPA1-H) and Act d 5 (SPA5-H) with precision ranging from 1.86\u20139.92%. The limit of quantification (LOQ) was set at 8 pg mL\u22121 for SPA1-H and at 8 ng mL\u22121 for SPA5-H. The developed procedure was utilized to analyze seven kinds of hand-made kiwi foods containing 0.0175\u20130.0515 mg g\u22121 of Act d 1 and 0.0252\u20130.0556 mg g\u22121 of Act d 5. This study extended the applicability of stable-isotope dimethyl labeling to the economical and precise determination of food allergens and peptides.<\/jats:p>","DOI":"10.3390\/molecules24101920","type":"journal-article","created":{"date-parts":[[2019,5,20]],"date-time":"2019-05-20T11:05:07Z","timestamp":1558350307000},"page":"1920","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Feasibility of Utilizing Stable-Isotope Dimethyl Labeling in Liquid Chromatography\u2013Tandem Mass Spectrometry-Based Determination for Food Allergens\u2014Case of Kiwifruit"],"prefix":"10.3390","volume":"24","author":[{"given":"Yi-Chen","family":"Shih","sequence":"first","affiliation":[{"name":"Department of Horticulture and Landscape Architecture, National Taiwan University, Taipei 10673, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jhih-Ting","family":"Hsiao","sequence":"additional","affiliation":[{"name":"Department of Horticulture and Landscape Architecture, National Taiwan University, Taipei 10673, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fuu","family":"Sheu","sequence":"additional","affiliation":[{"name":"Department of Horticulture and Landscape Architecture, National Taiwan University, Taipei 10673, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,5,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1007\/s00216-007-1353-5","article-title":"Food allergen detection methods and the challenge to protect food-allergic consumers","volume":"389","year":"2007","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/02652030310001620423","article-title":"Methods for allergen analysis in food: A review","volume":"21","author":"Poms","year":"2004","journal-title":"Food Addit. Contam."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1016\/j.trac.2009.02.013","article-title":"Mass spectrometry-based proteomics methods for analysis of food allergens","volume":"28","author":"Monaci","year":"2009","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/j.ymeth.2004.08.018","article-title":"The absolute quantification strategy: A general procedure for the quantification of proteins and post-translational modifications","volume":"35","author":"Kirkpatrick","year":"2005","journal-title":"Methods"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3499","DOI":"10.1021\/pr500247r","article-title":"Next Generation of Food Allergen Quantification Using Mass Spectrometric Systems","volume":"13","author":"Koeberl","year":"2014","journal-title":"J. Proteome"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1038\/nchembio736","article-title":"Mass spectrometry\u2013based proteomics turns quantitative","volume":"1","author":"Ong","year":"2005","journal-title":"Nat. Methods"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Leung, H.C., Man, T.K., and Flores, R. (2012). Integrative Proteomics, InTech.","DOI":"10.5772\/2473"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"6843","DOI":"10.1021\/ac0348625","article-title":"Stable-Isotope Dimethyl Labeling for Quantitative Proteomics","volume":"75","author":"Hsu","year":"2003","journal-title":"Anal. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"20150364","DOI":"10.1098\/rsta.2015.0364","article-title":"Stable isotope dimethyl labelling for quantitative proteomics and beyond","volume":"374","author":"Hsu","year":"2016","journal-title":"Philos. Trans. Soc. A: Math. Phys. Eng. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4359","DOI":"10.1016\/S0021-9258(17)30016-9","article-title":"Labeling of proteins by reductive methylation using sodium cyanoborohydride","volume":"254","author":"Jentoft","year":"1979","journal-title":"J. Boil. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7198","DOI":"10.1021\/acs.analchem.6b01388","article-title":"Intact Protein Quantitation Using Pseudo-isobaric Dimethyl Labeling","volume":"88","author":"Fang","year":"2016","journal-title":"Anal. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1002\/pmic.201100308","article-title":"Quantification of protein isoforms in mesenchymal stem cells by reductive dimethylation of lysines in intact proteins","volume":"12","author":"She","year":"2012","journal-title":"Proteomics"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4164","DOI":"10.1021\/pr500630a","article-title":"Comparing SILAC- and stable isotope dimethyl-labeling approaches for quantitative proteomics","volume":"13","author":"Lau","year":"2014","journal-title":"J. Proteome"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2056","DOI":"10.1021\/pr401202d","article-title":"Quantification of the Brain Proteome in Alzheimer\u2019s Disease Using Multiplexed Mass Spectrometry","volume":"13","author":"Musunuri","year":"2014","journal-title":"J. Proteome"},{"key":"ref_15","unstructured":"(2019, May 17). WHO\/IUIS Allergen Nomenclature Home Page. Available online: http:\/\/www.webcitation.org\/78RHwatZR."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1016\/S0091-6749(98)70360-4","article-title":"Identification of actinidin as the major allergen of kiwi fruit","volume":"101","author":"Pastorello","year":"1998","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1046\/j.1398-9995.2003.00421.x","article-title":"Thaumatin-like proteins\u2014A new family of pollen and fruit allergens","volume":"59","author":"Breiteneder","year":"2004","journal-title":"Allergy"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1220","DOI":"10.1111\/j.1365-2222.2007.02927.x","article-title":"Immunoglobulin E recognition patterns to purified Kiwifruit (Actinidinia deliciosa) allergens in patients sensitized to Kiwi with different clinical symptoms","volume":"38","author":"Palacin","year":"2008","journal-title":"Clin. Exp. Allergy"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1169","DOI":"10.1016\/j.jaci.2004.07.016","article-title":"IgE sensitization profiles toward green and gold kiwifruits differ among patients allergic to kiwifruit from 3 European countries","volume":"114","author":"Bublin","year":"2004","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1007\/s10930-005-7638-7","article-title":"Kiwellin, a Novel Protein from Kiwi Fruit. Purification, Biochemical Characterization and Identification as an Allergen","volume":"24","author":"Tamburrini","year":"2005","journal-title":"Protein J."},{"key":"ref_21","first-page":"230","article-title":"Characterization of Bet v 1-related allergens from kiwifruit relevant for patients with combined kiwifruit and birch pollen allergy","volume":"52","author":"Oberhuber","year":"2008","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1016\/j.jaci.2009.10.017","article-title":"Component-resolved diagnosis of kiwifruit allergy with purified natural and recombinant kiwifruit allergens","volume":"125","author":"Bublin","year":"2010","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Bernardi, M.L., Giangrieco, I., Camardella, L., Ferrara, R., Palazzo, P., Panico, M.R., Crescenzo, R., Carratore, V., Zennaro, D., and Liso, M. (2011). Allergenic lipid transfer proteins from plant-derived foods do not immunologically and clinically behave homogeneously: The kiwifruit LTP as a model. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0027856"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1111\/j.1398-9995.2011.02555.x","article-title":"Kiwifruit Act d 11 is the first member of the ripening-related protein family identified as an allergen","volume":"66","author":"Bernardi","year":"2011","journal-title":"Allergy"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1765","DOI":"10.1016\/j.jaci.2014.01.035","article-title":"Act d 12 and Act d 13: Two novel, masked, relevant allergens in kiwifruit seeds","volume":"133","author":"Sirvent","year":"2014","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1002\/mnfr.200700286","article-title":"Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA","volume":"52","author":"Spasic","year":"2008","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1016\/j.jchromb.2007.07.015","article-title":"Quantification of Art v 1 and Act c 1 being major allergens of mugwort pollen and kiwi fruit extracts in mass-units by ion-exchange HPLC-UV method","volume":"857","author":"Blanusa","year":"2007","journal-title":"J. Chromatogr. B"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1016\/j.foodcont.2010.11.022","article-title":"Screening and confirmatory methods for the determination of melamine in cow\u2019s milk and milk-based powdered infant formula: Validation and proficiency-tests of ELISA, HPLC-UV, GC-MS and LC-MS\/MS","volume":"22","author":"Lutter","year":"2011","journal-title":"Food Control"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1039\/c0ay00319k","article-title":"Mass spectrometry analysis of soybean seed proteins: Optimization of gel-free quantitative workflow","volume":"2","author":"Lee","year":"2010","journal-title":"Anal. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2369","DOI":"10.1002\/elps.200305500","article-title":"Protein extraction for two-dimensional electrophoresis from olive leaf, a plant tissue containing high levels of interfering compounds","volume":"24","author":"Wang","year":"2003","journal-title":"Electrophoresis"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1038\/msb.2008.61","article-title":"Selected reaction monitoring for quantitative proteomics: A tutorial","volume":"4","author":"Lange","year":"2008","journal-title":"Mol. Syst. Boil."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.aca.2010.09.040","article-title":"Absolute quantification method and validation of airborne snow crab allergen tropomyosin using tandem mass spectrometry","volume":"681","author":"Rahman","year":"2010","journal-title":"Anal. Chim. Acta"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1007\/s11095-008-9532-4","article-title":"Quantitative Atlas of Membrane Transporter Proteins: Development and Application of a Highly Sensitive Simultaneous LC\/MS\/MS Method Combined with Novel In-silico Peptide Selection Criteria","volume":"25","author":"Kamiie","year":"2008","journal-title":"Pharm. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1007\/s00216-012-6012-9","article-title":"Isobaric tagging approaches in quantitative proteomics: The ups and downs","volume":"404","author":"Christoforou","year":"2012","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Chen, T., Zhang, L., Shang, H., Liu, S., Peng, J., Gong, W., Shi, Y., Zhang, S., Li, J., and Gong, J. (2016). iTRAQ-Based Quantitative Proteomic Analysis of Cotton Roots and Leaves Reveals Pathways Associated with Salt Stress. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0148487"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.jprot.2014.12.017","article-title":"Proteomic characterization and comparison of mammalian milk fat globule proteomes by iTRAQ analysis","volume":"116","author":"Yang","year":"2015","journal-title":"J. Proteom."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2717","DOI":"10.1021\/ac9914869","article-title":"Relating Electrospray Ionization Response to Nonpolar Character of Small Peptides","volume":"72","author":"Cech","year":"2000","journal-title":"Anal. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1021\/jf304289f","article-title":"Diversity and Relative Levels of Actinidin, Kiwellin, and Thaumatin-Like Allergens in 15 Varieties of Kiwifruit (Actinidia)","volume":"61","author":"Maddumage","year":"2013","journal-title":"J. Agric. Food Chem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/24\/10\/1920\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:53:23Z","timestamp":1760187203000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/24\/10\/1920"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,5,18]]},"references-count":38,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2019,5]]}},"alternative-id":["molecules24101920"],"URL":"https:\/\/doi.org\/10.3390\/molecules24101920","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,5,18]]}}}