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In this study, we assessed the impact of different BCTs on exercise\u2010induced cfDNA dynamics. Eleven participants [25 (SD 2.3) years of age] performed three different treadmill exercise protocols, including an all\u2010out test and combinations of constant and interval load. Blood samples were collected before, 5\u00a0min and 30\u00a0min post\u2010exercise using EDTA, lithium\u2013heparin (LH) and serum BCTs. Concentrations of cfDNA were quantified using quantitative PCR. The cfDNA increased significantly across all protocols and BCTs. A significant effect of BCT on cfDNA concentrations (<jats:italic>P<\/jats:italic>\u00a0=\u00a00.034) was found, with serum showing higher concentrations than EDTA and LH. Although absolute differences from pre\u2010 to post\u2010exercise were comparable across BCTs (<jats:italic>P<\/jats:italic>\u00a0=\u00a00.476), fold changes differed significantly (<jats:italic>P<\/jats:italic>\u00a0=\u00a00.012), with the highest observed in EDTA and the lowest in serum. Bland\u2013Altman analyses demonstrated better agreement between EDTA and LH compared with serum. Significant correlations of cfDNA with energy expenditure and peak oxygen uptake were found. These correlations were stronger in EDTA and LH than in serum. Our findings highlight the crucial influence of BCT choice on cfDNA measurements in exercise settings. Given that EDTA and LH reflected exercise load better, they could be preferred for exercise physiology research. This work underscores the need to account for the choice of BCT to improve data comparability across studies. Additionally, these findings might have broader implications for clinical settings where cfDNA is used as a biomarker.<\/jats:p>","DOI":"10.1113\/ep092284","type":"journal-article","created":{"date-parts":[[2025,3,4]],"date-time":"2025-03-04T08:55:48Z","timestamp":1741078548000},"page":"1087-1098","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Pre\u2010analytical pitfalls: How blood collection tubes influence exercise\u2010induced cell\u2010free DNA concentrations"],"prefix":"10.1113","volume":"110","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-7178-4994","authenticated-orcid":false,"given":"Kira","family":"Enders","sequence":"first","affiliation":[{"name":"Department of Sports Medicine, Disease Prevention and Rehabilitation Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5090-2286","authenticated-orcid":false,"given":"Barlo","family":"Hillen","sequence":"additional","affiliation":[{"name":"Department of Sports Medicine, Disease Prevention and Rehabilitation Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nils","family":"Haller","sequence":"additional","affiliation":[{"name":"Department of Sports Medicine, Disease Prevention and Rehabilitation Johannes Gutenberg University Mainz  Mainz Germany"},{"name":"Department of Sport and Exercise Science University of Salzburg  Salzburg Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandra","family":"Brahmer","sequence":"additional","affiliation":[{"name":"Department of Sports Medicine, Disease Prevention and Rehabilitation Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vincent","family":"Weber","sequence":"additional","affiliation":[{"name":"Department of Sports Medicine, Disease Prevention and Rehabilitation Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7996-4034","authenticated-orcid":false,"given":"Perikles","family":"Simon","sequence":"additional","affiliation":[{"name":"Department of Sports Medicine, Disease Prevention and Rehabilitation Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2021-6477","authenticated-orcid":false,"given":"Elmo W. 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