Development and validation of a hydrophilic interaction liquid chromatography with tandem mass spectrometry method for the simultaneous detection and quantification of etilefrine and oxilofrine in equine blood plasma and urine.

This research developed and validated a method using hydrophilic interaction liquid chromatography and tandem mass spectrometry to successfully detect and measure levels of etilefrine and oxilofrine(equine medication) in horse blood plasma and urine.

Research Methodology

• The researchers developed a method using hydrophilic interaction liquid chromatography paired with tandem mass spectrometry to detect and measure etilefrine and oxilofrine in horse blood and urine.
• The compatibility and precision of this method were tested and validated to ensure it can accurately measure these substances.

Sensitivity and Specificity

• The method proved to be both highly sensitive and specific, meaning it can accurately identify these substances even in relatively small amounts.
• In horse plasma, the smallest amounts that could be detected were 0.03 and 0.1 ng/mL, respectively.
• For horse urine, the method could detect even slightly larger amounts of 0.3 and 1 ng/mL, respectively.

Doping Control Analysis

• This new method could be used in doping control analysis for horses.
• The researchers confirmed this by using their method to analyse samples taken after a horse was intravenously administered with 50 mg of etilefrine.
• Etilefrine could be detected in horse urine up to 120 hours post administration and up to 48 hours in plasma.

Recommended Enzyme Hydrolysis

• The research noted that etilefrine is highly conjugated only in horse urine, not in its plasma.
• Because of this, they recommend using enzyme hydrolysis before sample preparation to effectively detect and measure the levels of etilefrine and oxilofrine in horse urine.