Simultaneous separation and confirmation of amphetamine and related drugs in equine plasma by non-aqueous capillary-electrophoresis-tandem mass spectrometry.

This research outlines a method for the simultaneous separation and identification of 12 amphetamine and related compounds in horse plasma via a technique called non-aqueous capillary electrophoresis-mass spectrometry (NACE-MS).

Methodology

• The team of researchers harnessed the power of a non-aqueous capillary electrophoresis-mass spectrometry (NACE-MS) technique as an innovative method for the simultaneous separation and identification of 12 amphetamine and related compounds in equine (horse) plasma.
• The substances being measured (analytes) were recovered from the plasma through a process called liquid-liquid extraction, which used methyl tertiary butyl ether (MTBE).
• A bare fused-silica capillary, a small, delicate tube, was employed for the separation of these analytes.
• The detection of the analytes was enabled by a full-scan data acquisition procedure, which involved the addition of a sheath liquid to the capillary effluent.

Results and Parameters

• The limit of detection (LOD) for the target analytes was identified as 10-200 ng/mL and the limit of confirmation (LOC) was pegged at 50-1000 ng/mL in the equine plasma.
• The mass spectrometry (MS) parameters, together with the capillary electrophoresis (CE), were fine-tuned to facilitate the full CE separation and MS detection of the analytes.
• The buffer used for separation comprised of 25mM ammonium formate in acetonitrile/methanol (20:80, v/v) plus 1M formic acid, and sheath liquid contained isopropanol, water, and formic acid (50:50:0.5, v/v/v).

Application and Conclusion

• The researchers harnessed a hydrodynamic injection and managed to separate the samples at 25 kV, and the analytes were subsequently separated electrokinetically and identified/confirmed through mass spectrometry.
• This research has practical implications in that it pioneers a method that is not only simple, fast, and reproducible, but is also inexpensive.
• The method was successfully applied to post-race equine plasma and research samples, demonstrating its potential for screening for amphetamine and related drugs in horses, a crucial aspect of horse racing and veterinary sciences.