High-performance liquid chromatographic assay for sematilide in plasma using solid-phase extraction microcolumn technology.
A simple and sensitive high-performance liquid chromatographic assay for quantification of sematilide in rabbit plasma was developed. After extraction of samples via solid-phase extraction on C8 microcolumns, baseline resolution was achieved on a reversed-phase 5 microns Inertsil ODS-2 column using isocratic conditions with mobile phase consisting of water-glacial acetic acid-acetonitrile-methanol-triethylamine (93.5:4.0:1.5:0.5:0.5) and UV detection at 254 nm. The assay did not require evaporation or reconstitution steps. The injection interval was 8 minutes. The inter-day coefficient of variation for replicate analysis of spiked samples was less than 7.6% and the accuracy was more than 97% over the standard curve range (0.128 to 3.191 microM) using 0.5 ml of plasma. The assay has been successfully applied to pharmacokinetic studies in rabbits.
Biopharmaceutics and Pharmacodynamics Division, Department of Health, Ottawa, Ont., Canada.
S
S
Laganière
L
L
Goernert
G
G
Beatch
D000818:Animals; D000889:Anti-Arrhythmia Agents / Q000097:blood* / Q000493:pharmacokinetics; D001681:Biological Assay; D002851:Chromatography, High Pressure Liquid; D006207:Half-Life; D007275:Injections, Intravenous; D011342:Procainamide / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D011817:Rabbits; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet
D000889:Anti-Arrhythmia Agents; D012997:Solvents; C062818:sematilide; D011342:Procainamide