Volume: 4, 2025
4th International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland:
ENVIRONMENT – PLANT – ANIMAL – PRODUCT
Abstract number: H020
DOI: https://doi.org/10.24326/ICDSUPL4.H020
Published online: 9 April 2025
ICDSUPL, 4, H020 (2025)
Application of the QuEChERS technique for determination of methandienone in biological samples by GC-MS/MS
Krystian Siwek1*, Michał P. Dybowski1
1Department of Chromatography, Institute of Chemical Sciences, Faculty of Chemistry, Maria Curie Sklodowska University in Lublin, Marii Skłodowskiej-Curie 3, 20-031 Lublin, Poland
* Corresponding author: krystian.siwek0511@gmail.com
Abstract
Methandienone is a substance on the World Anti-Doping Agency’s (WADA) List of Prohibited Substances and Methods. It is included in the S1.1 group, anabolic-androgenic steroids (AAS), the drugs that dominate doping cases in sport. Despite their medical applications in treating various conditions and dysfunctions, these substances are also used by healthy individuals to enhance physical performance, thereby improving results in sport. Given the prevalence of AAS utilization for enhancing competitive advantage, there is an imperative for research facilities to enhance existing techniques, such as GC-MS, to optimize the efficacy in the determination of prohibited substances. A promising approach involves the implementation of the QuEChERS technique, a method that has demonstrated its ability to extract these substances from biological material more effectively. GC-MS/MS method for determination of methandienone containing samples was developed and validated. The extraction of analytes from blood plasma was accomplished by utilizing the QuEChERS technique on biological samples. The optimization of the technique was achieved through the consideration of three distinct factors (collision energy (CE), addition of extraction salts and addition of organic solvent). The findings presented in this study suggest that the most suitable conditions for performing the QuEChERS technique on a 900 µl blood plasma sample are as follows: addition of extraction salts: MgSO4 (350 mg) and NaCl (87.5 mg), followed by the addition of 500 µl of organic solvent and after centrifugation, the addition of 17.5 mg of C18 sorbent. The fragmentation transitions of methandienone were selected by analysis in Q3-SCAN (TIC) mode, while optimization of their CE was performed in MRM (multiple reaction monitoring) mode. The analysis of the following transitions is most convenient at the following energies: 161.00>147.00 (CE = 12 eV), 161.00 > 134.00 (CE = 12 eV), 147.00 > 134.00 (CE = 12 eV). As the QuEChERS technique is comprised of two sequential stages, the first involving the extraction of the analytes from the sample matrix and the second involving their purification, it unquestionably demonstrates its applicability in the determination of anabolic-androgenic steroids, such as methandienone, in a matrix as complex as blood plasma.
Keywords: anabolic-androgenic steroids, Methandienone, anti-doping, QuEChERS, GC-MS/MS
How to cite
K. Siwek, M.P. Dybowski, 2025. Application of the QuEChERS technique for determination of methandienone in biological samples by GC-MS/MS. In: 4th International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland: Environment – Plant – Animal – Product. https://doi.org/10.24326/ICDSUPL4.H020