Screening analysis of doping agents in horse urine and plasma with dilute and shoot using liquid chromatography high resolution mass spectrometry.

The research primarily focuses on developing a fast, efficient, and cost-effective method for identifying banned substances in horse urine and plasma by utilizing a technique called “dilute and shoot” screening in conjunction with high-performance liquid chromatography and high-resolution mass spectrometry.

Objective of the Study

• The main goal of the study was to develop an accurate, quick, and cost-efficient method to identify substances in biological samples, especially those that are susceptible to chemical degradation, can be analyzed without hydrolysis, or have short-term stability.
• Such a method is of paramount importance for laboratories where many samples are analyzed simultaneously with a need for easy sample preparation, short analysis times, and low analysis costs.

Methodology

• The researchers developed and validated a “dilute and shoot” screening procedure that employs high-performance liquid chromatography coupled with high-resolution mass spectrometry. This method was used to analyze a variety of pharmacological compounds present in horse urine and plasma.
• Chromatographic separation was performed using a C18 column, with two mobile phases: 5 mM ammonium bicarbonate at pH 9 (phase A), and methanol (phase B). The flow rate was maintained at 0.3 mL min.

Validation and Detection Capability

• The method was verified according to the specifications laid out by the multi-screening requirements of 2002/657/EC.
• The detection ability of the developed method was found to range from less than or equal to 1 to 200 ng mL for prohibited substances. In a single 13 minute run, the method was able to scan over 200 banned substances across various categories.

Practical Applicability

• The developed method was applied to actual positive case samples to underscore its practical applicability.
• The screening method was also implemented in doping analysis, further demonstrating its practical usage in real-world scenarios.
• According to the researchers, this developed method stands as a rare technique that can be used to analyze both urine and plasma samples, offering rapid, comprehensive, high-throughput analysis of banned substances in horse plasma and urine in a cost-effective manner.

In conclusion, the research offers a promising and efficient method for doping analysis, especially in the field of horse racing, where quick and accurate detection of prohibited substances is essential.