Simultaneous detection of xenon and krypton in equine plasma by gas chromatography‐tandem mass spectrometry for doping control

Abstract

Xenon can activate the hypoxia-inducible factors (HIFs). As such, it has been allegedly used in human sports for increasing erythropoiesis. Krypton, another noble gas with reported narcosis effect, can also be expected to be a potential and less expensive erythropoiesis stimulating agent. This has raised concern about the misuse of noble gases as doping agents in equine sports. The aim of the present study is to establish a method for the simultaneous detection of xenon and krypton in equine plasma for the purpose of doping control. Xenon- or krypton-fortified equine plasma samples were prepared according to reported protocols. The target noble gases were simultaneously detected by gas chromatography-triple quadrupole mass spectrometry using headspace injection. Three xenon isotopes at m/z 129, 131, and 132, and four krypton isotopes at m/z 82, 83, 84, and 86 were targeted in selected reaction monitoring mode (with the precursor ions and product ions at identical mass settings), allowing unambiguous identification of the target analytes. Limits of detection for xenon and krypton were about 19 pmol/mL and 98 pmol/mL, respectively. Precision for both analytes was less than 15%. The method has good specificity as background analyte signals were not observed in negative equine plasma samples (n = 73). Loss of analytes under different storage temperatures has also been evaluated. Copyright © 2016 John Wiley & Sons, Ltd.

Gas chromatography/mass spectrometry measurement of xenon in gas‐loaded liposomes for neuroprotective applications

“…Almost simultaneously, Kwok et al reported a quantitative GC/MS headspace analytical method for assay of xenon and krypton in equine plasma for the same purpose . They used a PLOT molecular sieve column for GC separation of gases, as we did.…”

“…Almost simultaneously, Kwok et al reported a quantitative GC/MS headspace analytical method for assay of xenon and krypton in equine plasma for the same purpose. [35] They used a PLOT molecular sieve column for GC separation of gases, as we did. They employed the method of Goto et al [27] to saturate plasma with xenon and krypton, using the published solubility values to assign gas concentrations to standard plasma dilutions.…”

Challenges in detecting substances for equine anti‐doping

“…LODs for xenon and krypton were 19 pmol/mL and 98 pmol/mL, respectively. [50] Designer drugs in equine doping (Designer) anabolic steroids Designer steroids are considered chemical structures that originally appeared as imitations of endogenous steroids and later as minor modifications of previously marketed products, with the core intention to be undetectable from the current targeted mass spectrometric procedures. To evade detection, the designer steroids have been synthesized to be marketed freely on the Internet, as their structures do not fall within the scope of known steroid structures/biological activities and thus remain uncontrolled by the legal authorities.…”

Intelligence‐based anti‐doping from an equine biological passport

“…Beyond the steroid and GH axes, the experience of racing laboratories over recent years with potential integrity issues surrounding manipulation of the hypoxia inducible factor‐1α (HIF‐1α) axis may suggest that biomarkers capable of identifying such augmentation will be investigated for inclusion in an EBP. Further to the potential misuse of myo ‐inositol trispyrophosphate and cobalt as HIF‐1α stabilizers or noble gases as HIF‐1α activators, a plethora of small‐molecule drug candidates are being investigated to alter HIF‐1α that ‘target’ testing will unlikely keep pace with . Therefore, indirect measures of HIF‐1α manipulation will be vital to any proactive anti‐doping strategy.…”

“…Beyond the steroid and GH axes, the experience of racing laboratories over recent years with potential integrity issues surrounding manipulation of the hypoxia inducible factor-1α (HIF-1α) axis may suggest that biomarkers capable of identifying such augmentation will be investigated for inclusion in an EBP. Further to the potential misuse of myo-inositol trispyrophosphate [69] and cobalt [39,40] as HIF-1α stabilizers or noble gases [70] as HIF-1α…”