Published: Journal of Analytical Toxicology, Volume 21, Number 5, September 1997, pp. 393–396.
The Use of ELISA Tests and Immunoaffinity Chromatography Combined with Reversed-Phase High-Performance Liquid Chromatography for Dexamethasone Detection in Equine Urine
L.M. Ribeiro Neto, H.S. Spinosa, and M.C. Salvadori
Dexamethasone is a corticosteroid drug widely used in racehorses because of its anti-inflammatory effect. It is, therefore, frequently detected in antidoping tests. A method for the antidoping control of dexamethasone in equine urine using screening by ELISA and confirmation by immunoaffinity chromatography combined with reversed-phase high-performance liquid chromatography–diode array detection (HPLC–DAD) is described. The ELISA test is frequently used in antidoping tests for its sensitivity, relative speed, and low cost. The test showed linearity in the range of 4–500 ng/mL of urine, and the intra-assay and interassay imprecision were 9.4 and 9.7%, respectively. The confirmation method showed a limit of detection of 4 ng/mL for dexamethasone. The intra-assay and interassay imprecisions were 10.3 and 14.4%, respectively. The HPLC–DAD showed a limit of detection of 5 ng and linearity in the range of 25–500 ng of dexamethasone. The absolute method recovery was 56.4%. The proposed method detected dexamethasone up to 52 h after administration and proved to be adequate for the antidoping control.
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