Selective and sensitive high-performance liquid chromatographic assay for the metabolites of nomifensine in human plasma.
A selective high-performance liquid chromatographic method for the determination of the three metabolites of nomifensine in human plasma is described. All metabolites and the internal standard, mexiletine, are extracted with diethyl ether and then back-extracted into an acidic aqueous phase. After subsequent extraction into diethyl ether the metabolites are analysed by high-performance liquid chromatography. A reversed-phase C18 column is used with a mobile phase of dioxane-methanol-potassium phosphate buffer (pH 2.25). The sensitivity of the method is 0.007 micromol/l for all metabolites. Extraction efficiencies are 84.6%, 75.8%, and 78.2% for 4'-hydroxynomifensine, 4'-hydroxy-3'-methoxynomifensine and 3'-hydroxy-4'-methoxynomifensine, respectively. The reproducibility of the method is good, the coefficients of variation (%) varying between 2.1% and 9.9% in the concentration range 0.05-1.00 micromol/l. The procedure was applied to human plasma samples from a volunteer who had received a single oral dose of nomifensine. The method is accurate and sensitive for pharmacokinetic studies on the metabolites of nomifensine.
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007546:Isoquinolines / Q000097:blood*; D007700:Kinetics; D009627:Nomifensine / Q000031:analogs & derivatives / Q000097:blood*; D012997:Solvents
D007546:Isoquinolines; D012997:Solvents; D009627:Nomifensine; C046575:4'-hydroxynomifensine; C046577:3'-hydroxy-4'-methoxynomifensine; C046576:4'-hydroxy-3'-methoxynomifensine