High sensitive analysis of steroids in doping control using gas chromatography/time-of-flight mass-spectrometry

Area of Science:

• Analytical Chemistry
• Biochemistry
• Mass Spectrometry

Background:

• Steroid analysis is crucial in various fields, including clinical diagnostics and doping control.
• Existing methods may lack the sensitivity and throughput required for comprehensive steroid profiling.
• The development of advanced analytical techniques is essential for accurate and efficient steroid detection.

Purpose of the Study:

• To develop a highly sensitive method for steroid analysis using gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS).
• To improve the detection limits and the number of identified steroid-like compounds in complex biological matrices.
• To optimize sample preparation and injection techniques for enhanced analyte recovery and sensitivity.

Main Methods:

• Development of a novel GC/TOF-MS method involving silyl derivatization of steroids.
• Utilized tert-butylmethylether for reagent exchange and offline large volume injection with sorption concentration.
• Employed solvent-free concentrate transfer of analytes into the gas chromatograph injector.
• Utilized a low-resolution TOF-MS instrument with a fast spectral acquisition rate for analysis.

Main Results:

• Achieved detection limits of 0.5-2 pg/µl for steroid derivatives with a 100 µl sample solution volume.
• The GC/TOF-MS method, utilizing the TruTOF model and ChromaTOF software, enabled full mass spectra extraction and resolution of coeluted peaks.
• Analysis of a urine extract using the proposed method (10 µl aliquot) identified twice as many steroid-like compounds compared to a standard 1 µl injection.

Conclusions:

• The developed high-sensitivity GC/TOF-MS method offers superior performance for steroid analysis.
• This technique significantly enhances the capability to detect and identify a broader range of steroid-like compounds in biological samples.
• The optimized sample preparation and injection strategy contribute to improved sensitivity and comprehensive profiling in steroid research.