{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T07:39:59Z","timestamp":1792568399853,"version":"4.5.0"},"reference-count":25,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T00:00:00Z","timestamp":1792454400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T00:00:00Z","timestamp":1792454400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Drug Testing and Analysis"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Following the patent expiration of the originator darbepoetin alfa, three biosimilar products have been marketed in Japan; however, their detectability by current anti\u2010doping methods has not yet been verified. The present study assessed whether these biosimilars can be detected in human urine, serum, plasma, and dried blood spots (DBS) using the established initial testing procedure and confirmation analysis for erythropoietin\u2010receptor agonists. The originator product and three biosimilars (5\u2009\u03bcg, i.v.) were administered to healthy male volunteers. Samples collected up to 72\u2009h postdose were subjected to immunopurification followed by sodium\n                    <jats:italic>N<\/jats:italic>\n                    \u2010lauroyl sarcosinate polyacrylamide\u2010gel electrophoresis (SAR\u2010PAGE) as the initial testing procedure. Confirmation was performed by SAR\u2010PAGE with double blotting for serum and plasma. DBS and urine samples were confirmed by SAR\u2010PAGE using an alternative antibody, and urine samples were further examined by liquid chromatography\u2013tandem mass spectrometry (LC\u2013MS\/MS). SAR\u2010PAGE analysis of the pharmaceutical preparations revealed product\u2010specific electrophoretic profiles: two biosimilars migrated slightly above one originator product as diffused smears, whereas the third produced a sharp band comparable to that of another originator product. Similar profiles were observed in postdose samples from all biological matrices, and all biosimilars remained detectable in urine, serum, plasma, and DBS for at least 72\u2009h. Although some biosimilars showed distinct SAR\u2010PAGE banding patterns, product attribution remained limited. LC\u2013MS\/MS confirmed administered darbepoetin alfa in the 24\u2009h urine samples but did not detect analyte in lower concentration samples, indicating that its performance was concentration\u2010dependent. Accordingly, SAR\u2010PAGE should incorporate reference standards beyond the originator product, particularly those with different band morphologies.\n                  <\/jats:p>","DOI":"10.1002\/dta.70175","type":"journal-article","created":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T06:54:11Z","timestamp":1792565651000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Doping Control Analysis of Darbepoetin Alfa Biosimilars in Human Urine, Serum, Plasma, and Dried Blood Spots"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-5665-4644","authenticated-orcid":false,"given":"Kentaro","family":"Akiyama","sequence":"first","affiliation":[{"name":"Anti\u2010Doping Laboratory, LSI Medience Corporation  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hikaru","family":"Sakurai","sequence":"additional","affiliation":[{"name":"Anti\u2010Doping Laboratory, LSI Medience Corporation  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mitsuhiko","family":"Sato","sequence":"additional","affiliation":[{"name":"Anti\u2010Doping Laboratory, LSI Medience Corporation  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-1224-3309","authenticated-orcid":false,"given":"Atsushi","family":"Momobayashi","sequence":"additional","affiliation":[{"name":"Anti\u2010Doping Laboratory, LSI Medience Corporation  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3614-4121","authenticated-orcid":false,"given":"Masato","family":"Okano","sequence":"additional","affiliation":[{"name":"Anti\u2010Doping Laboratory, LSI Medience Corporation  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,20]]},"reference":[{"issue":"3","key":"e_1_2_8_2_1","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1182\/blood.V77.3.419.419","article-title":"Erythropoietin","volume":"77","author":"Krantz S. B.","year":"1991","journal-title":"Blood"},{"issue":"25","key":"e_1_2_8_3_1","doi-asserted-by":"crossref","first-page":"12059","DOI":"10.1016\/S0021-9258(18)45317-3","article-title":"Carbohydrate Structure of Erythropoietin Expressed in Chinese Hamster Ovary Cells by a Human Erythropoietin cDNA","volume":"262","author":"Sasaki H.","year":"1987","journal-title":"Journal of Biological Chemistry"},{"key":"e_1_2_8_4_1","unstructured":"U.S. Food and Drug Administration \u201cEpogen Prescribing Information \u201d accessed June 17 2026 https:\/\/www.accessdata.fda.gov\/drugsatfda_docs\/label\/2024\/103234s5378lbl.pdf."},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1054\/bjoc.2001.1746"},{"key":"e_1_2_8_6_1","unstructured":"Pharmaceuticals and Medical Devices Agency \u201cNESP Injection Plastic Syringe \u201d accessed July 8 2026 https:\/\/www.info.pmda.go.jp\/go\/interview\/1\/230124_3999425S5028_1_013_1F.pdf."},{"key":"e_1_2_8_7_1","unstructured":"European Medicines Agency \u201cBinocrit EPAR Product Information \u201d accessed June 17 2026 https:\/\/www.ema.europa.eu\/en\/documents\/product\u2010information\/binocrit\u2010epar\u2010product\u2010information_en.pdf."},{"key":"e_1_2_8_8_1","unstructured":"Pharmaceuticals and Medical Devices Agency \u201cEpoetin Alfa BS Injection Syringe \u201cJCR\u201d \u201d accessed June 17 2026 https:\/\/www.pmda.go.jp\/PmdaSearch\/iyakuDetail\/ResultDataSetPDF\/530210_3999427A1026_1_12."},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.2165\/11593730\u2010000000000\u201000000"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.7b00351"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.369"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.1941"},{"key":"e_1_2_8_13_1","unstructured":"Pharmaceuticals and Medical Devices Agency \u201cDarbepoetin Alfa BS Injection Syringe \u201cMYL\u201d \u201d accessed June 17 2026 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