Paper in Forefront

Analytical and Bioanalytical Chemistry

, Volume 396, Issue 8, pp 2899-2908

Quantification of urinary AICAR concentrations as a matter of doping controls

  • Andreas ThomasAffiliated withInstitute of Biochemistry, Center for Preventive Doping Research, German Sport University Cologne Email author 
  • , Simon BeuckAffiliated withInstitute of Biochemistry, Center for Preventive Doping Research, German Sport University Cologne
  • , Jens Christian EickhoffAffiliated withDepartment of Statistics, Colorado State University
  • , Sven GuddatAffiliated withInstitute of Biochemistry, Center for Preventive Doping Research, German Sport University Cologne
  • , Oliver KrugAffiliated withInstitute of Biochemistry, Center for Preventive Doping Research, German Sport University Cologne
  • , Matthias KamberAffiliated withDepartment of Doping Prevention, Federal Office of Sports
  • , Wilhelm SchänzerAffiliated withInstitute of Biochemistry, Center for Preventive Doping Research, German Sport University Cologne
  • , Mario ThevisAffiliated withInstitute of Biochemistry, Center for Preventive Doping Research, German Sport University Cologne

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Abstract

Influencing the endurance in elite sports is one of the key points in modern sports science. Recently, a new class of prohibited substances reached in the focus of doping control laboratories and their misuse was classified as gene doping. The adenosine monophosphate activated protein kinase activator 5-amino-4-imidazolecarboxyamide ribonucleoside (AICAR) was found to significantly enhance the endurance even in sedentary mice after treatment. Due to endogenous production of AICAR in healthy humans, considerable amounts were present in the circulation and, thus, were excreted into urine. Considering these facts, the present study was initiated to fix reference values of renally cleared AICAR in elite athletes. Therefore a quantitative analytical method by means of isotope-dilution liquid chromatography (analytical column: C6-phenyl) coupled to tandem mass spectrometry, after a sample preparation consisting of a gentle dilution of native urine, was developed. Doping control samples of 499 athletes were analysed, and AICAR concentrations in urine were determined. The mean AICAR value for all samples was 2,186 ng/mL with a standard deviation of 1,655 ng/mL. Concentrations were found to differ depending on gender, type of sport and type of sample collection (in competition/out of competition). The method was fully validated for quantitative purposes considering the parameters linearity, inter- (12%, 7% and 10%) and intraday precision (14%, 9% and 12%) at low, mid and high concentration, robustness, accuracy (approx. 100%), limit of quantification (100 ng/mL), stability and ion suppression effects, employing an in-house synthesised 13C5-labelled AICAR as internal standard.

http://static-content.springer.com/image/art%3A10.1007%2Fs00216-010-3560-8/MediaObjects/216_2010_3560_Figa_HTML.gif

Urinary AICAR concentrations (ng/mL)

Keywords

Gene doping Sports drug testing LC-MS/M Mass spectrometry