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.
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.
Colas
Cyril
C
LCH-Laboratoire des Courses Hippiques, 15 rue de Paradis, 91370 Verrières le Buisson, France.
Garcia
Patrice
P
Popot
Marie-Agnès
MA
Bonnaire
Yves
Y
Bouchonnet
Stéphane
S
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D006027:Glycosides / Q000032:analysis* / Q000097:blood / Q000652:urine; D006736:Horses / Q000097:blood* / Q000652:urine*; D006863:Hydrogen-Ion Concentration; D057889:Iridoid Glycosides; D011714:Pyrans / Q000032:analysis* / Q000097:blood / Q000652:urine; D052616:Solid Phase Extraction / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
D006027:Glycosides; D057889:Iridoid Glycosides; D011714:Pyrans; C033249:harpagoside; C033250:harpagide