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  • Determination of nitrazepam and its main metabolites in urine by thin-layer chromatography and direct densitometry.

Determination of nitrazepam and its main metabolites in urine by thin-layer chromatography and direct densitometry.

Journal of chromatography (1985-04-12)
T Inoue, T Niwaguchi
ABSTRACT

A method for the direct quantitative densitometry of nitrazepam and its main metabolites (7-aminonitrazepam, 7-acetamidonitrazepam and 2-amino-5-nitrobenzophenone) in urine was developed. The unchanged drug and its metabolites were extracted with benzene-dichloromethane (4:1), subjected to thin-layer chromatography, and determined by direct ultraviolet densitometry. Recovery experiments showed that the method was quantitative. The limit of detection was 5 ng/ml for 2-amino-5-nitrobenzophenone and 10 ng/ml for other compounds. The method was applied to the determination of nitrazepam and its metabolites excreted in human urine after administration of 10 mg of the drug.

MATERIALS
Product Number
Brand
Product Description

Supelco
7-Aminonitrazepam solution, 1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant®
Sigma-Aldrich
2-Amino-5-nitrobenzophenone, ≥98%