TY - JOUR
T1 - Comprehensive insights into the formation of metabolites of the ghrelin mimetics capromorelin, macimorelin and tabimorelin as potential markers for doping control purposes
AU - Lange, Tobias
AU - Thomas, Andreas
AU - Görgens, Christian
AU - Bidlingmaier, Martin
AU - Schilbach, Katharina
AU - Fichant, Eric
AU - Delahaut, Philippe
AU - Thevis, Mario
N1 - © 2021 The Authors. Biomedical Chromatography published by John Wiley & Sons Ltd.
PY - 2021/6
Y1 - 2021/6
N2 - Analytical methods to determine the potential misuse of the ghrelin mimetics capromorelin (CP-424,391), macimorelin (macrilen, EP-01572) and tabimorelin (NN703) in sports were developed. Therefore, different extraction strategies, i.e. solid-phase extraction, protein precipitation, as well as a "dilute-and-inject" approach, from urine and EDTA-plasma were assessed and comprehensive in vitro/in vivo experiments were conducted, enabling the identification of reliable target analytes by means of high resolution mass spectrometry. The drugs' biotransformation led to the preliminary identification of 51 metabolites of capromorelin, 12 metabolites of macimorelin and 13 metabolites of tabimorelin. Seven major metabolites detected in rat urine samples collected post-administration of 0.5-1.0 mg of a single oral dose underwent in-depth characterization, facilitating their implementation into future confirmatory test methods. In particular, two macimorelin metabolites exhibiting considerable abundances in post-administration rat urine samples were detected, which might contribute to an improved sensitivity, specificity, and detection window in case of human sports drug testing programs. Further, the intact drugs were implemented into World Anti-Doping Agency-compliant initial testing (limits of detection 0.02-0.60 ng/ml) and confirmation procedures (limits of identification 0.18-0.89 ng/ml) for human urine and blood matrices. The obtained results allow extension of the test spectrum of doping agents in multitarget screening assays for growth hormone-releasing factors from human urine.
AB - Analytical methods to determine the potential misuse of the ghrelin mimetics capromorelin (CP-424,391), macimorelin (macrilen, EP-01572) and tabimorelin (NN703) in sports were developed. Therefore, different extraction strategies, i.e. solid-phase extraction, protein precipitation, as well as a "dilute-and-inject" approach, from urine and EDTA-plasma were assessed and comprehensive in vitro/in vivo experiments were conducted, enabling the identification of reliable target analytes by means of high resolution mass spectrometry. The drugs' biotransformation led to the preliminary identification of 51 metabolites of capromorelin, 12 metabolites of macimorelin and 13 metabolites of tabimorelin. Seven major metabolites detected in rat urine samples collected post-administration of 0.5-1.0 mg of a single oral dose underwent in-depth characterization, facilitating their implementation into future confirmatory test methods. In particular, two macimorelin metabolites exhibiting considerable abundances in post-administration rat urine samples were detected, which might contribute to an improved sensitivity, specificity, and detection window in case of human sports drug testing programs. Further, the intact drugs were implemented into World Anti-Doping Agency-compliant initial testing (limits of detection 0.02-0.60 ng/ml) and confirmation procedures (limits of identification 0.18-0.89 ng/ml) for human urine and blood matrices. The obtained results allow extension of the test spectrum of doping agents in multitarget screening assays for growth hormone-releasing factors from human urine.
KW - Animals
KW - Biomarkers/metabolism
KW - Chromatography, Liquid/methods
KW - Dipeptides/metabolism
KW - Doping in Sports
KW - Female
KW - Ghrelin
KW - Humans
KW - Indoles/metabolism
KW - Limit of Detection
KW - Male
KW - Piperidines/metabolism
KW - Pyrazoles/metabolism
KW - Rats
KW - Reproducibility of Results
KW - Solid Phase Extraction
KW - Tandem Mass Spectrometry/methods
KW - Tryptophan/analogs & derivatives
UR - https://www.mendeley.com/catalogue/0c128e4e-80a8-3d9e-b530-a5ffdd1af564/
U2 - 10.1002/bmc.5075
DO - 10.1002/bmc.5075
M3 - Journal articles
C2 - 33458843
SN - 0269-3879
VL - 35
SP - 1
EP - 18
JO - Biomedical chromatography : BMC
JF - Biomedical chromatography : BMC
IS - 6
M1 - e5075
ER -