Determination of Chlormethiazole in Blood by High Performance Liquid Chromatography

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Abstract
Chlormethiazole (CMZ) is a derivative of the thiazole part of thiamine, and has sedative hypnotic and anticonvulsant effects. A simple and sensitive method is described for the determination of chlormethiazole in blood. Separation and quantitation were performed by reversed-phase high performance liquid chromatography utilizing UV detection at 254 nm. The internal standard, 2-amino-4-methylthiazole, was used as an aid to quantitation. The blood sample was deproteinized with phosphotungstic acid reagent and the clear supernatant obtained was directly analyzed in the chromatographic system. Limit of detection was about 0.25 .mu.g/ml blood. Recovery of chlormethiazole from blood was 73 .+-. 2%. The mean between-run coefficient of variation was 4.9% over the range of 0.25-32.0 .mu.g/ml, whereas the within-run coefficient of variation at 0.25 .mu.g/ml was 6.5%. The scale and sensitivity of detection were suitable for use in pharmacokinetic studies in the rat.