Determination of misonidazole and desmethylmisonidazole in plasma by high-performance liquid chromatography with reductive electrochemical detection.

A reversed-phase high-performance liquid chromatographic method with reductive mode electrochemical detection was developed for the determination of misonidazole and desmethylmisonidazole in plasma. A thin-layer amperometric detector with glassy carbon working electrode was used to detect the nitroimidazoles at a potential of -0.60 V. The calibration curves were linear. The within-day and day-to-day coefficients of variation were below 3% for plasma misonidazole concentrations of 6-60 mg/l and 1-15 mg/l for desmethylmisonidazole. Electrochemical detection limits were between 2 and 4 pg, which is about 10-20 times lower than that obtained by detection at 323 nm. Limits of quantitation of the nitroimidazoles in plasma were in the order of 1-2 micrograms/l. Under the described conditions other nitroimidazoles and nitro compounds can also be detected with ultimate sensitivity by reductive mode electrochemical detection.