Optimization of solid-phase extraction for the liquid chromatography-mass spectrometry analysis of harpagoside, 8-para-coumaroyl harpagide, and harpagide in equine plasma and urine.

Cyril Colas (1, 2) , Patrice Garcia (2) , Marie-Agnès Popot (2) , Yves Bonnaire (2) , Stéphane Bouchonnet (1)

1 DCMR - Laboratoire des mécanismes réactionnels
2 LCH - Laboratoire des Courses Hippiques

Solid-phase extraction cartridges among those usually used for screening in horse doping analyses are tested to optimize the extraction of harpagoside (HS), harpagide (HG), and 8-para-coumaroyl harpagide (8PCHG) from plasma and urine. Extracts are analyzed by liquid chromatography coupled with multi-step tandem mass spectrometry. The extraction process retained for plasma applies BondElut PPL cartridges and provides extraction recoveries between 91% and 93%, with RSD values between 8 and 13% at 0.5 ng/mL. Two different procedures are needed to extract analytes from urine. HS and 8PCHG are extracted using AbsElut Nexus cartridges, with recoveries of 85% and 77%, respectively (RSD between 7% and 19%). The extraction of HG involves the use of two cartridges: BondElut PPL and BondElut C18 HF, with recovery of 75% and RSD between 14% and 19%. The applicability of the extraction methods is determined on authentic equine plasma and urine samples after harpagophytum or harpagoside administration.

Domaines: Chimie analytique

Cyril Colas, Patrice Garcia, Marie-Agnès Popot, Yves Bonnaire, Stéphane Bouchonnet. Optimization of solid-phase extraction for the liquid chromatography-mass spectrometry analysis of harpagoside, 8-para-coumaroyl harpagide, and harpagide in equine plasma and urine.. Journal of Chromatographic Science, 2008, 46 (2), pp.174-183. ⟨hal-00525486⟩