Add time:08/14/2019         Source:sciencedirect.com

A sensitive high-performance liquid chromatographic (HPLC) method for the analysis of metyrapone [2-methyl-1,2-di-(3-pyridyl)-1-propanone], its reduced metabolite metyrapol (cas 17159-42-1) and metyrapone mono-N-oxide metabolites in biological fluids is reported. These components were extracted into dichloromethane (2 × 5 ml) from alkalinised microsomal incubates, urine and blood (final pH about 12.5), or from cytosolic incubates at pH 7.4 (final aqueous volume 2–4 ml). Recoveries were in the range 70–100% under these conditions. The intact drug and metabolites were separated by reversed-phase HPLC with ultraviolet detection at 261 nm. All calibration curves were linear (correlation coefficient >0.997). For the analysis of hepatic microsomal or cytosolic incubates, the coefficient of variation was less than 10% for samples over the range 2.5–250 nmol/ml N-oxides and 10–250 nmol/ml metyrapol. Measurement of metyrapone and metyrapol in rat blood (0.25-ml sample volume) was linear in the ranges 4.4–265 and 26–263 nmol/ml, respectively, the lower concentration being the limit of detection. The coefficient of variation was less than 20% for samples over the ranges tested for both these compounds. The N-oxide metabolites were not detectable in blood using this assay, their concentrations being below the limit of detection.

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