A modified LC-MS/MS method to simultaneously quantify glycerol and mannitol concentrations in human urine for doping...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Forensic Science

Background:

• Glycerol and mannitol are prohibited masking agents by the World Anti-Doping Agency (WADA).
• Accurate and rapid detection of these substances in urine is crucial for anti-doping efforts.
• Existing detection methods may lack the speed or comprehensive analysis required.

Purpose of the Study:

• To develop and validate an improved, rapid analytical strategy for simultaneous determination of urinary glycerol and mannitol.
• To achieve baseline separation of mannitol and other potential hexitols for confirmation.
• To establish a sensitive and reliable method for anti-doping testing.

Main Methods:

• Liquid chromatography/tandem mass spectrometry (LC-ESI-MS/MS) was employed for analysis.
• Glycerol and mannitol were derivatized using benzoyl chloride.
• A rapid method with limited sample preparation was developed, achieving analysis within 7 minutes.

Main Results:

• The method demonstrated limits of detection (LOD) below 50 ng/mL and limits of quantitation (LOQ) below 150 ng/mL.
• Assay linearity was established from 0.15 to 1000 μg/mL for both glycerol and mannitol.
• High accuracy and precision were achieved, with coefficients of variation better than 13% for glycerol and 15% for mannitol.

Conclusions:

• The validated LC-MS/MS method provides a fast and accurate approach for detecting urinary glycerol and mannitol.
• The method allows for simultaneous detection and confirmation of mannitol and related hexitols.
• This strategy significantly enhances the efficiency of anti-doping testing procedures for prohibited masking agents.