Presenter Bio: I finished in 2013 the Faculty of Chemistry and in 2015 I obtained my master’s degree in Biochemistry and Molecular Biology. From 2015 I am a PhD student in Chemistry. My PhD thesis is based on the development of new biosensors (both electrochemical and immunosensors) for the improved detection of prohibited substances in sport. Since 2012, I am part of the WADA accredited laboratory from Bucharest. Starting as a part-time assistant, now I am responsible for the Doping Analysis department and also, as a research scientist, for the detection of anabolic androgenic steroids using gas chromatography coupled with mass spectrometry.
Relevant Financial Disclosures
(within past 24 months)
No relevant financial relationship(s) to disclose.
Abstract
Introduction: Zearalenone or F-2 mycotoxin is a heat-stable, potent estrogenic metabolic product of some Fusarium and Gibberella species, found in cereal crops like maize, oats, wheat, rice and barley. Due to its similarity to naturally-occurring estrogens (17β-estradiol), zearalenone and its main metabolites (α-zearalenol, β-zearalenol, zeranol and taleranol) are considered by World Health Organization and European Commission as endocrine-disrupting chemicals and a possible cause of carcinogenesis.
Objectives: Due to its anabolic effects, zeranol and taleranol are banned in sport. The presence of this metabolites in athlete’s urine must be clearly attributed to illegal use or unintended contamination. The primary objective of this study is to develop and optimize a gas chromatography - mass spectrometry application for the detection of zeranol and/or taleranol in the presence of zearalenone mycotoxin.
Methods: The method was developed for gas chromatography –tandem mass spectrometry (GC-MS/MS) GC Trace 1310 connected to a TSQ Quantum XLS Ultra from Thermo Scientific. The GC was equipped with an HP-Ultra 1 (17m x 200µm and 0.11µm film tickness) from Agilent Technologies (USA). The temperature program was as follows: the initial temperature was 160°C hold time 2 min, increased by 5°C/min to 255°C and then by 30°C/min to 285°C (hold time 5 min) and finally by 60°C/min to a final temperature of 300°C (held 3.75 min). The transfer line temperature was set at 310°C. Helium was used as carrier gas (constant flow rate aprox.1 mL/min).
Results:The method for the detection of zearalenone and its main metabolites α-zearalenol, β-zearalenol, zeranol and taleranol was developed and validated. Both urine samples spiked with the compounds and real excretion urines were analyzed in order to test the validated method for limit of detection and matrix interference. Using this GC-MS/MS method, the analysts can discriminate between a urine sample resulting from illegal use of anabolic agent zeranol or unintended contamination.
Conclusion: Contamination of food with mycotoxins and veterinary products used as growth promoters is a global and dangerous phenomenon that could lead to unfair sanctions in sport for athletes. The detection of metabolic profile using mass spectrometry applications developed towards the differentiation between mycotoxin contamination and anabolic agent illegal administration is the solution in this case for a correct and accurate interpretation of analytical findings of zeranol in urine samples.
= Discovery stage.
= Translation stage.
= Clinically available.
