Simultaneous determination of hydrocortisone, dexamethasone, indomethacin, phenylbutazone and oxyphenbutazone in equine serum by high-performance liquid chromatography.
Ethyl acetate extracts of equine serum, containing 0-5 microg/ml of hydrocortisone (HYD), dexamethasone (DEX), oxyphenbutazone (OPB), indomethacin (IND), phenylbutazone (PB) and probenecid as internal standard, were evaporated with nitrogen, resuspended in methanol and analyzed by HPLC, using a C-18 column equilibrated with 51:49 acetonitrile-water, 0.1% trifluoroacetic acid, at 1 ml/min. The eluate was monitored at 254 nm. The selectivity (inter-assay C.V.<4%), sensitivity (limits of quantitation of 0.25 microg/ml for HYD, DEX and IND, 0.5 microg/ml for PB and 1 microg/ml for OPB, despite the occurrence of significant degradation of OPB and PB during the analysis) and precision (intra-assay and inter-assay C.V.'s of about 3-6 and 9-15%, respectively) of the method appeared appropriate for anti-doping control of racehorses.
Grippa
E
E
Department of Pharmacology of Natural Substances and General Physiology, University of Rome La Sapienza, Italy.
Santini
L
L
Castellano
G
G
Gatto
M T
MT
Leone
M G
MG
Saso
L
L
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000097:blood; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003907:Dexamethasone / Q000097:blood*; D006736:Horses / Q000097:blood*; D006854:Hydrocortisone / Q000097:blood*; D006863:Hydrogen-Ion Concentration; D007213:Indomethacin / Q000097:blood*; D010113:Oxyphenbutazone / Q000097:blood; D010653:Phenylbutazone / Q000097:blood*; D012680:Sensitivity and Specificity
D000893:Anti-Inflammatory Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D003907:Dexamethasone; D010653:Phenylbutazone; D010113:Oxyphenbutazone; D006854:Hydrocortisone; D007213:Indomethacin