A sensitive radioimmunoassay, combined with solid-phase extraction, for the sub-nanogram per ml determination of ondansetron in human plasma.
The development of a radioimmunoassay, incorporating solid-phase sample extraction, suitable for the subnanogram per ml determination of ondansetron base in human plasma is described. The antiserum was raised in Soay sheep following primary and booster immunizations with an immunogen prepared by conjugating 9-(carboxypropyl)-ondansetron to bovine thyroglobulin. The radioligand consisted of ondansetron specifically tritium-labelled on the N-methyl group of the indole moiety. The solid-phase extraction method, using a cyanopropyl sorbent, was introduced to remove cross-reacting metabolites and to enhance assay sensitivity. The calibration range is 0.05-2.40 ng ml-1 using a 1 ml sample of human plasma; inter- and intra-assay bias and precision are < +/- 13% and < 10% over this concentration range, respectively. The assay drift, measured as the difference in concentration values for quality control samples assayed immediately before and after the sequence of test plasma samples, is < +/- 10% for run sizes of up to 54 samples.
Division of Drug Metabolism, Glaxo Group Research, Ware, Herts, UK.
S A
SA
Wring
R M
RM
Rooney
C P
CP
Goddard
I
I
Waterhouse
W N
WN
Jenner
D000818:Animals; D000918:Antibody Specificity; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D006801:Humans; D007106:Immune Sera; D017294:Ondansetron / Q000031:analogs & derivatives / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification; D011863:Radioimmunoassay; D011869:Radioligand Assay; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D013056:Spectrophotometry, Ultraviolet
D007106:Immune Sera; D017294:Ondansetron