Detection of myo-inositol trispyrophosphate in equine urine and plasma by hydrophillic interaction chromatography-tandem mass spectrometry.

This research article presents a newly developed method for detecting the drug myo-inositol trispyrophosphate (ITPP) in racehorse urine and blood samples, a chemical suspected to be used for doping in the horse-racing industry. The detection process includes isolating ITPP and analysing it via hydrophilic interaction chromatography-tandem mass spectrometry.

Overview of the Drug and Its Effects

• The drug in question, myo-inositol trispyrophosphate (ITPP), has the capability to increase oxygen levels in hypoxic tissues. This could, in theory, enhance performance in racehorses by enabling them to exert more effort without tiring.
• Prior studies have confirmed that the drug improves maximum exercise capacity in normal mice as well as those suffering from severe heart failure, suggesting a similar impact could be expected in horses.
• Given these properties of ITPP, researchers identified the need to develop a detection method for this drug to control its potential misuse in horse racing.

Challenges in Detecting ITPP

• Because of its highly hydrophilic nature, detecting ITPP poses a significant challenge. Hydrophilic substances tend to dissolve in water, meaning that ITPP can easily disperse in watery biological samples, such as urine.
• Additionally, the complexity of equine biological matrices—meaning the various substances found in horse urine or blood—further complicates the process of identifying and isolating the presence of ITPP.

Development of a New Detection Method

• The researchers developed a process for identifying and confirming the presence of ITPP in equine biology. They first isolate ITPP from biological samples using solid-phase extraction. In this process, a solid material is used to absorb the ITPP from the liquid sample.
• This ITPP-containing extract is then analysed using hydrophilic interaction chromatography-tandem mass spectrometry. This highly accurate method separates, identifies, and quantifies the compound, allowing researchers to confirm the presence of ITPP.
• The developed method showed good precision, fast turnaround time, and negligible matrix interferences, making it a reliable tool for detecting ITPP.

Significance of the Development

• This is a significant development as it stands as the first-ever method for the detection and unequivocal confirmation of ITPP levels, regardless of how low, in any biological fluid.
• Such a detection method is crucial for maintaining fair and ethical practices in the horse-racing industry, as ITPP is a strong candidate for being used as a performance-enhancing drug.