Separation of clonazepam and five metabolites by reverse phase HPLC and quantitation from rat liver microsomal incubations

Rory P. Remmel; Gary W. Elmer

doi:10.1080/01483918308076070

Separation of clonazepam and five metabolites by reverse phase HPLC and quantitation from rat liver microsomal incubations

Rory P. Remmel, Gary W. Elmer

Medicinal Chemistry

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Abstract

Clonazepam, its nitro-reduced metabolites, and the 3-hydroxyl-ated derivatives, were separated by reverse-phase hplc using a four solvent optimization procudure. The separation was used to quantitate the metabolism of³H-clonazepam in rat liver microsomes under aerobic or anaerobic conditions by collection of HPLC eluents and liquid scintillation counting. The predominant metabolite in aerobic incubations was 3-hydroxyclonazepam. The primary anaerobic metabolite was 7-aminoclonazepam. The identity of the microsomal metabolites was confirmed by chemical ionization mass spectrometry. This separation could be applicable for future metabolic studies of clonazepam.

Original language	English (US)
Pages (from-to)	585-598
Number of pages	14
Journal	Journal of Liquid Chromatography
Volume	6
Issue number	3
DOIs	https://doi.org/10.1080/01483918308076070
State	Published - Mar 1 1983

Bibliographical note

Funding Information:
We acknowledge support of this work by the NIGMS National Research Service Award GM-07750 to R. P. Remmel. We also wish to thank Larry Oliver for his able technical assistance.

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abstract = "Clonazepam, its nitro-reduced metabolites, and the 3-hydroxyl-ated derivatives, were separated by reverse-phase hplc using a four solvent optimization procudure. The separation was used to quantitate the metabolism of3H-clonazepam in rat liver microsomes under aerobic or anaerobic conditions by collection of HPLC eluents and liquid scintillation counting. The predominant metabolite in aerobic incubations was 3-hydroxyclonazepam. The primary anaerobic metabolite was 7-aminoclonazepam. The identity of the microsomal metabolites was confirmed by chemical ionization mass spectrometry. This separation could be applicable for future metabolic studies of clonazepam.",

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AU - Elmer, Gary W.

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N2 - Clonazepam, its nitro-reduced metabolites, and the 3-hydroxyl-ated derivatives, were separated by reverse-phase hplc using a four solvent optimization procudure. The separation was used to quantitate the metabolism of3H-clonazepam in rat liver microsomes under aerobic or anaerobic conditions by collection of HPLC eluents and liquid scintillation counting. The predominant metabolite in aerobic incubations was 3-hydroxyclonazepam. The primary anaerobic metabolite was 7-aminoclonazepam. The identity of the microsomal metabolites was confirmed by chemical ionization mass spectrometry. This separation could be applicable for future metabolic studies of clonazepam.

AB - Clonazepam, its nitro-reduced metabolites, and the 3-hydroxyl-ated derivatives, were separated by reverse-phase hplc using a four solvent optimization procudure. The separation was used to quantitate the metabolism of3H-clonazepam in rat liver microsomes under aerobic or anaerobic conditions by collection of HPLC eluents and liquid scintillation counting. The predominant metabolite in aerobic incubations was 3-hydroxyclonazepam. The primary anaerobic metabolite was 7-aminoclonazepam. The identity of the microsomal metabolites was confirmed by chemical ionization mass spectrometry. This separation could be applicable for future metabolic studies of clonazepam.

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Separation of clonazepam and five metabolites by reverse phase HPLC and quantitation from rat liver microsomal incubations

Abstract

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