Universität zu Köln

Thomas, Andreas, Krombholz, Sophia, Wolf, Carina and Thevis, Mario (2021). Determination of ghrelin and desacyl ghrelin in human plasma and urine by means of LC-MS/MS for doping controls. Drug Test. Anal., 13 (11-12). S. 1862 - 1871. HOBOKEN: WILEY. ISSN 1942-7611

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Abstract

The hunger hormone ghrelin (G) is classified as prohibited substance in professional sport by the World Anti-Doping Agency (WADA), due to its known growth hormone releasing properties. The endogenous bioactive peptide consists of 28 amino acids with a caprylic acid attached to serine at position 3. Within this study, it was aimed to develop methods to determine G and desacyl ghrelin (DAG) in plasma and urine by means of LC-MS/MS. Two strategies were applied with a bottom-up approach for plasma and top-down analyses for urine. Both sample preparation procedures were based on solid-phase extraction for enrichment and sample clean-up. Method validation showed good results for plasma and urine with limits of detection (LODs) for G and DAG between 30 and 50 pg/ml, recoveries between 45-50%, and imprecisions (intra- and inter-day) between 3% and 24%. Plasma analysis was also valid for quantification with accuracies determined with similar to 100% for G and similar to 106% for DAG. The minimum required performance level for doping control laboratories is set to 2 ng/ml in urine, and the herein established method yielded acceptable results even at 5% of this level. As proof-of-concept, plasma levels (G and DAG) of healthy volunteers were determined and ranged between 30 and 100 pg/ml for G and 100-1200 pg/ml for DAG. In contrast to earlier reported studies using ligand binding assays for urinary G and DAG, in this mass spectrometry-based study, no endogenous urinary G and DAG were found, although the LODs should enable this.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Thomas, Andreas UNSPECIFIED UNSPECIFIED UNSPECIFIED Krombholz, Sophia UNSPECIFIED UNSPECIFIED UNSPECIFIED Wolf, Carina UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-572120
DOI:	10.1002/dta.3176
Journal or Publication Title:	Drug Test. Anal.
Volume:	13
Number:	11-12
Page Range:	S. 1862 - 1871
Date:	2021
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1942-7611
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DES-ACYL; QUANTIFICATION; SERUM Multiple languages Biochemical Research Methods; Chemistry, Analytical; Pharmacology & Pharmacy Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/57212