{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T15:51:34Z","timestamp":1761061894105,"version":"build-2065373602"},"reference-count":38,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2023,11,20]],"date-time":"2023-11-20T00:00:00Z","timestamp":1700438400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Drug Testing and Analysis"],"published-print":{"date-parts":[[2024,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Plasma proteins have been a valuable source of biomarkers for clinical uses and for monitoring of the illicit use of prohibited substances or practices in equine sports. We have previously reported the first use of label\u2010free proteomics in profiling equine plasma proteome. This study aimed to refine the method by systematically evaluating various plasma fractionation methods and the use of narrower precursor mass ranges in data\u2010independent acquisition (DIA) mass spectrometry (MS). Tandem fractionations of equine plasma with octanoic acid precipitation followed by solid\u2010phase extraction (SPE) with C4 cartridges provided the largest increase in the number of new proteins identified. The use of two narrow precursor mass ranges of <jats:italic>m\/z<\/jats:italic> 400\u2013600 and 600\u2013800 in DIA not only identified most proteins detectable by using a single mass range of <jats:italic>m\/z<\/jats:italic> 350\u20131500 but also identified ~27% more proteins. The improved method was applied to analyse the plasma proteome of \u2018postrace\u2019 samples which, unlike other samples, had been collected from racehorses soon after racing. Multivariate data analysis has identified upregulation of 14 proteins and downregulation of six proteins in postrace plasma compared with the non\u2010postrace plasma samples. Literature review of these proteins has provided evidence of exercise\u2010induced haemolysis and changes in antioxidant enzyme activities, kinin system, insulin signalling and energy metabolism after strenuous exercise. The improved method has enabled a deeper profiling of the equine plasma proteome and identified the proteins associated with normal physiological changes after racing which are potential confounding factors in the development of a biomarker approach for doping control.<\/jats:p>","DOI":"10.1002\/dta.3606","type":"journal-article","created":{"date-parts":[[2023,11,21]],"date-time":"2023-11-21T19:55:01Z","timestamp":1700596501000},"page":"841-854","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["An enhanced label\u2010free proteomics approach for deep\u2010diving into equine plasma proteome, including the discovery of protein biomarkers for strenuous exercise"],"prefix":"10.1002","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2471-1790","authenticated-orcid":false,"given":"Hiu Wing","family":"Cheung","sequence":"first","affiliation":[{"name":"Racing Laboratory The Hong Kong Jockey Club, Sha Tin Racecourse  Sha Tin Hong Kong China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7136-4712","authenticated-orcid":false,"given":"Kin\u2010Sing","family":"Wong","sequence":"additional","affiliation":[{"name":"Racing Laboratory The Hong Kong Jockey Club, Sha Tin Racecourse  Sha Tin Hong Kong China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ning Sum","family":"To","sequence":"additional","affiliation":[{"name":"Racing Laboratory The Hong Kong Jockey Club, Sha Tin Racecourse  Sha Tin Hong Kong China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3779-2649","authenticated-orcid":false,"given":"Terence S. M.","family":"Wan","sequence":"additional","affiliation":[{"name":"Racing Laboratory The Hong Kong Jockey Club, Sha Tin Racecourse  Sha Tin Hong Kong China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emmie N. M.","family":"Ho","sequence":"additional","affiliation":[{"name":"Racing Laboratory The Hong Kong Jockey Club, Sha Tin Racecourse  Sha Tin Hong Kong China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,11,20]]},"reference":[{"key":"e_1_2_10_2_1","unstructured":"International Federation of Horseracing Authorities.International agreement on breeding racing and wagering and appendixes. Article 6https:\/\/ifhaonline.org\/Default.asp?section=IABRW&area=2#article6. Accessed 4 Nov 2022."},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tvjl.2014.01.006"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1159\/000460722"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2011.162271"},{"key":"e_1_2_10_6_1","unstructured":"World Anti\u2010Doping Agency.Athlete Biological Passport operating guidelines version 8.0.2021;https:\/\/www.wada-ama.org\/sites\/default\/files\/resources\/files\/guidelines_abp_v8_final.pdf. Accessed 4 Nov 2022."},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.forsciint.2007.04.001"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2013.208538"},{"key":"e_1_2_10_9_1","unstructured":"World Anti\u2010Doping Agency.Laboratory guidelines\u2014human growth hormone (hGH) biomarkers test version 3.0.2021;https:\/\/www.wada-ama.org\/sites\/default\/files\/resources\/files\/wada_guidelines_hgh_biomarkers_test_v3_jan_2021_eng.pdf. Accessed 4 Nov 2022."},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/evj.13183"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.200800737"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2012.04.014"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1294\/jes.26.141"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jproteome.9b00724"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.2988"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200406189"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.15252\/msb.20167357"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010017\u201009968\u20107"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2018.04.045"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.4172\/2155\u20109872.1000198"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1039\/B713712E"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.24539"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1294\/jes.29.53"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.3390\/life11090982"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0001078"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1021\/pr050300l"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1152\/japplphysiol.00631.2001"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.cdd.4400905"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2016.00279"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1155\/2011\/540458"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2018.01006"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.3390\/biom5020356"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1155\/2016\/3708905"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1590\/1414\u2010431x20176400"},{"issue":"3","key":"e_1_2_10_35_1","first-page":"124","article-title":"The kinin system\u2010bradykinin: biological effects and clinical implications. Multiple role of the kinin system\u2010bradykinin","volume":"11","author":"Golias C","year":"2007","journal-title":"Hippokratia"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014\u20105793(77)80847\u20108"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1055\/s\u20102007\u20101010879"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2020.00727"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.49.30729"}],"container-title":["Drug Testing and Analysis"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/dta.3606","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T18:08:30Z","timestamp":1760378910000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/dta.3606"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,20]]},"references-count":38,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2024,8]]}},"alternative-id":["10.1002\/dta.3606"],"URL":"https:\/\/doi.org\/10.1002\/dta.3606","archive":["Portico"],"relation":{},"ISSN":["1942-7603","1942-7611"],"issn-type":[{"type":"print","value":"1942-7603"},{"type":"electronic","value":"1942-7611"}],"subject":[],"published":{"date-parts":[[2023,11,20]]},"assertion":[{"value":"2023-02-10","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-10-30","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-11-20","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}