High resolution accurate mass screening of prohibited substances in equine plasma using liquid chromatography–Orbitrap mass spectrometry.

This research article discusses the development of a high resolution accurate mass screening (HRAMS) process used with liquid chromatography and Orbitrap mass spectrometry to detect prohibited substances in the plasma of horses participating in sports. The method is designed to screen over 320 substances in a single test, aiming to strengthen doping control.

Methodology

• Researchers use liquid chromatography-high resolution mass spectrometry (LC-HRMS) for plasma sample analysis, in a bid to identify over 320 prohibited substances in one analytical run.
• The plasma samples were initially diluted and further processed by solid-phase extraction.
• LC-HRMS analysis is used in full-scan positive electrospray ionization mode, employing a mass resolution of 60,000.
• The researchers used benzyldimethylphenylammonium as an internal lock mass.
• Identifying drug targets involved verifying the retention time and accurate mass, with a mass tolerance window of ±3 ppm (parts per million).

Results

• The LC-HRMS method is capable of detecting over 320 targeted substances in a run lasting 13 minutes.
• The paper includes validation data such as sensitivity, specificity, extraction recovery, and precision.
• Since the method uses full-scan mass spectrometry, there is no restriction on the number of drug targets that can be integrated.
• The generated HRAMS data allows for retrospective reprocessing to search for untargeted drugs during the initial analysis.

Significance

• This research leads to a leap forward in doping control, especially in the arena of equine sports, allowing for faster and wider coverage in the detection of drugs in blood samples.
• The technology’s sensitivity and breadth of coverage also facilitate reanalysis of data, helping to detect substances that might have been missed or not targeted in the first place.
• The high sensitivity, mass accuracy, mass resolution, and mass stability associated with high-resolution mass spectrometry make this method extremely effective and accurate.
• This method may be beneficial in human sports as well, where doping cases are regularly reported, enhancing the overall standard of fair play in sports.