Affiliation:
1. Division of Bio‐Medical Science & Technology, KIST School University of Science and Technology Seoul South Korea
2. Doping Control Center Korea Institute of Science and Technology Seoul South Korea
3. Center for Biomaterials, Biomedical Research Institute Korea Institute of Science and Technology Seoul South Korea
4. Center for Advanced Biomolecular Recognition Korea Institute of Science and Technology Seoul South Korea
Abstract
AbstractThymosin β4 (Tβ4) was reported to exert various beneficial bioactivities such as tissue repair, anti‐inflammation, and reduced scar formation, and it is listed on the prohibited substances in sports by the World Anti‐Doping Agency. However, no metabolism studies of Tβ4 were reported yet. Previously, our lab reported in in vitro experiment that a total of 13 metabolites were found by using multiple enzymes, and six metabolites (Ac‐Tβ31–43, Ac‐Tβ17–43, Ac‐Tβ1–11, Ac‐Tβ1–14, Ac‐Tβ1–15, and Ac‐Tβ1–17) were confirmed by comparing with the synthetic standards. This study was aimed at identifying new metabolites of Tβ4 leucine aminopeptidase (LAP), human kidney microsomes (HKM), cultured huvec cells, and rats after administration of Tβ4 protein to develop biomarkers for detecting doping drugs in sports. A method for detecting and quantifying Ac‐Tβ1–14 was developed and validated using Q‐Exactive orbitrap mass spectrometry. The limit of detection (LOD) and limit of quantification (LOQ) of the Ac‐Tβ1–14 were 0.19 and 0.58 ng/mL, respectively, and showed a good linearity (r2 = 0.9998). As a result, among the six metabolites above, Ac‐Tβ1–14, as a common metabolite, was found in LAP, HKM, huvec cells exposed to Tβ4, and the urine of rats intraperitoneally treated with 20‐mg/kg Tβ4. And the metabolite Ac‐Tβ1–14 was quantitatively determined by 48 h in rats, with the highest concentration occurring between 0 and 6 h. Ac‐Tβ1–14 was not detected in non‐treated control groups, including human blank urine. These results suggest that Ac‐Tβ1–14 in urine is a potential biomarker for screening the parent Tβ4 in doping tests.
Funder
Korea Institute of Science and Technology
Subject
Spectroscopy,Pharmaceutical Science,Environmental Chemistry,Analytical Chemistry
Cited by
2 articles.
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