Determination of temazepam and temazepam glucuronide by reversed-phase high-performance liquid chromatography |
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Authors: | G W Kunsman J E Manno M A Przekop B R Manno K A Llorens |
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Institution: | G. W. Kunsman, J. E. Manno*, M. A. Przekop, B. R. Manno,K. A. Llorens |
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Abstract: | A rapid and sensitive method for extracting temazepam from human serum and urine is presented. Free temazepam is extracted from plasma and urine samples using n-butyl chloride with nitrazepam as the internal standard. Temazepam glucuronide is analyzed as free temazepam after incubating extracts with β-glucuronidase. Separation is achieved using a C8 reversed-phase column with a methanol—water—phosphate buffer mobile phase. An ultraviolet detector operated at 230 nm is used and a linear response is observed from 20 ng/ml to 10 μg/ml. The limit of detection is 15.5 ng/ml and the limit of quantitation is 46.5 ng/ml. Coefficients of variation are less than 10% for concentrations greater than 50 ng/ml. Application of the methodology is demonstrated in a pharmacokinetic study using eight healthy male subjects. |
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