Immunoaffinity chromatography combined on-line with high-performance liquid chromatography-mass spectrometry for the determination of corticosteroids.
On-line coupled immunoaffinity chromatography-reversed-phase high-performance liquid chromatography (IAC-HPLC) with detection by quadrupole ion trap mass spectrometry using a particle beam interface has been developed for the determination of the steroids, dexamethasone and flumethasone. HEMA (polyhydroxyethylmethacrylate) was evaluated as a support material for the anti-dexamethasone antibodies used in IAC. Antibody cross-reactivity and non-specific binding have been investigated for the HEMA bound anti-dexamethasone IAC column. The on-line IAC-HPLC-MS determination of dexamethasone and flumethasone in post-administration equine urine samples showed precisions (R.S.D.) of 8.0 and 7.1%, respectively, with limits of detection in the range 3-4 ng/ml.
Creaser
C S
CS
Department of Chemistry and Physics, Nottingham Trent University, UK.
Feely
S J
SJ
Houghton
E
E
Seymour
M
M
D000305:Adrenal Cortex Hormones / Q000032:analysis* / Q000276:immunology; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000032:analysis / Q000276:immunology; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D003907:Dexamethasone / Q000032:analysis / Q000276:immunology; D005443:Flumethasone / Q000032:analysis / Q000276:immunology; D007120:Immunochemistry; D013058:Mass Spectrometry; D011817:Rabbits / Q000276:immunology; D013056:Spectrophotometry, Ultraviolet
D000305:Adrenal Cortex Hormones; D000893:Anti-Inflammatory Agents; D003907:Dexamethasone; D005443:Flumethasone