Universität zu Köln

Thomas, Andreas ORCID: 0000-0003-1199-0743, Vogel, Matthias, Piper, Thomas ORCID: 0000-0002-7462-6693, Krug, Oliver, Beuck, Simon, Schanzer, Wilhelm and Thevis, Mario (2013). Quantification of AICAR-ribotide concentrations in red blood cells by means of LC-MS/MS. Anal. Bioanal. Chem., 405 (30). S. 9703 - 9710. HEIDELBERG: SPRINGER HEIDELBERG. ISSN 1618-2650

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Abstract

AICAR (5-amino-4-imidazolecarboxyamide ribonucleoside) arguably provides performance-enhancing properties even in the absence of physical exercise and, therefore, the substance is banned in elite sports since 2009. Due to the natural presence of AICAR in human blood and urine, uncovering the misuse by direct qualitative analysis is not possible. Entering the circulation, the riboside is immediately incorporated into red blood cells (RBCs) and transformed into the corresponding ribotide (5'-monophosphate) form. Within the present study, an analytical method was developed to determine AICAR-ribotide concentrations in RBC concentrates by means of liquid chromatography-tandem mass spectrometry. The method was validated enabling quantitative result interpretation considering the parameters specificity, precision (intra- and interday), linearity, recovery, accuracy (LOD/LOQ), stability and ion suppression. By analysing 99 RBC samples of young athletes, normal physiological levels of AICAR-ribotide were determined (10-500 ng/mL), and individual levels were found to be stable for several days. Employing in vitro incubation experiments with AICAR riboside in fresh whole blood samples, the ribotide concentrations were observed to increase significantly within 30 min from baseline to 1-10 mu g/mL. These levels are considered conserved for the lifetime of the erythrocyte and, thus, the results of the in vitro model strongly support the hypothesis that measuring abnormally high AICAR-ribotide concentrations in RBC of elite athletes has the potential to uncover the misuse of this substance for a long period of time.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Thomas, Andreas UNSPECIFIED orcid.org/0000-0003-1199-0743 UNSPECIFIED Vogel, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Piper, Thomas UNSPECIFIED orcid.org/0000-0002-7462-6693 UNSPECIFIED Krug, Oliver UNSPECIFIED UNSPECIFIED UNSPECIFIED Beuck, Simon UNSPECIFIED UNSPECIFIED UNSPECIFIED Schanzer, Wilhelm UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-471035
DOI:	10.1007/s00216-013-7162-0
Journal or Publication Title:	Anal. Bioanal. Chem.
Volume:	405
Number:	30
Page Range:	S. 9703 - 9710
Date:	2013
Publisher:	SPRINGER HEIDELBERG
Place of Publication:	HEIDELBERG
ISSN:	1618-2650
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language MASS-SPECTROMETRY; RIBOSIDE; DRUGS Multiple languages Biochemical Research Methods; Chemistry, Analytical Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/47103