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Abstract
A rapid, simple and reproducible method for sample pretreatment and analysis of disopyramide and its mono-N-dealkylated metabolite in serum is described. Samples for analysis were prepared by using Sep-Pak silica cartridge treated with 1 N sodium hydroxide followed by elution in 8 ml ethyl acetate. The eluate was back-extracted with acidic solution and an aliquot of the solution was injected directly onto the column.
Chromatographic separation using p-chlorodisopyramide as an internal standard was achieved on a radially compressed CN column, with a mobile phase of 0.01 M dibutylamine phosphate(pH 3.0)-acetonitrile (75:25, v/v), and at a flow rate of 2.0 ml/min. The, peaks were monitored by W at 210 nm.
Replicate analysis of serum controls for drug and its metabolite resulted in a within-run coefficient of variation of ≦ 2.1 and ≦ 4.4% and a day-to-day coefficient of variation of ≦ 2.5 and ≦ 4.8%, respectively.
Comparison with an EMIT assay gave a correlation coefficient of 0.950.