Determination of isoniazid methanesulphonate and its metabolites in rabbit blood by high-performance liquid chromatography

Kuang Yang Hsu, Yih Ho

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9 Citations (Scopus)

Abstract

A rapid and sensitive high-performance liquid chromatographic method is described for simultaneous determination of isoniazid methanesulphonate (IHMS) and its metabolites, such as isoniazid (INH) and acetylisoniazid (AcINH) in rabbit blood. According to stability studies, IHMS was most stable at pH 3-5. After acidifying the blood to pH 5.0, a suitable amount of acetonitrile was added to the supernatant for extraction and niacinamide served as an internal standard. After evaporation, the residue was reconstituted with phosphate buffer and aliquots of this solution were separated on a reversed-phase phenyl column by a mobile phase consisting of 0.25 mM tetrabutylammonium phosphate as a paired-ion reagent. UV detection was performed at 280 nm. Under these conditions, the between-run coefficients of variation of IHMS, INH and AcINH from 1 to 25 μm/ml were 4.7 ± 2.5, 5.4 ± 1.0 and 5.1 ± 3.1%, respectively. Hence this sensitive, reproducible and accurate method was suitable for pharmacokinetic studies of IHMS.

Original languageEnglish
Pages (from-to)305-312
Number of pages8
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume493
Issue numberC
DOIs
Publication statusPublished - 1989

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High performance liquid chromatography
Metabolites
Blood
Phosphates
High Pressure Liquid Chromatography
Rabbits
Pharmacokinetics
Niacinamide
Isoniazid
Buffers
Evaporation
Ions
Liquids
methaniazide
acetonitrile
acetylisoniazid
factor XIIa inhibitor
tetrabutylammonium

ASJC Scopus subject areas

  • Chemistry(all)
  • Clinical Biochemistry
  • Molecular Medicine

Cite this

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title = "Determination of isoniazid methanesulphonate and its metabolites in rabbit blood by high-performance liquid chromatography",
abstract = "A rapid and sensitive high-performance liquid chromatographic method is described for simultaneous determination of isoniazid methanesulphonate (IHMS) and its metabolites, such as isoniazid (INH) and acetylisoniazid (AcINH) in rabbit blood. According to stability studies, IHMS was most stable at pH 3-5. After acidifying the blood to pH 5.0, a suitable amount of acetonitrile was added to the supernatant for extraction and niacinamide served as an internal standard. After evaporation, the residue was reconstituted with phosphate buffer and aliquots of this solution were separated on a reversed-phase phenyl column by a mobile phase consisting of 0.25 mM tetrabutylammonium phosphate as a paired-ion reagent. UV detection was performed at 280 nm. Under these conditions, the between-run coefficients of variation of IHMS, INH and AcINH from 1 to 25 μm/ml were 4.7 ± 2.5, 5.4 ± 1.0 and 5.1 ± 3.1{\%}, respectively. Hence this sensitive, reproducible and accurate method was suitable for pharmacokinetic studies of IHMS.",
author = "Hsu, {Kuang Yang} and Yih Ho",
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T1 - Determination of isoniazid methanesulphonate and its metabolites in rabbit blood by high-performance liquid chromatography

AU - Hsu, Kuang Yang

AU - Ho, Yih

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N2 - A rapid and sensitive high-performance liquid chromatographic method is described for simultaneous determination of isoniazid methanesulphonate (IHMS) and its metabolites, such as isoniazid (INH) and acetylisoniazid (AcINH) in rabbit blood. According to stability studies, IHMS was most stable at pH 3-5. After acidifying the blood to pH 5.0, a suitable amount of acetonitrile was added to the supernatant for extraction and niacinamide served as an internal standard. After evaporation, the residue was reconstituted with phosphate buffer and aliquots of this solution were separated on a reversed-phase phenyl column by a mobile phase consisting of 0.25 mM tetrabutylammonium phosphate as a paired-ion reagent. UV detection was performed at 280 nm. Under these conditions, the between-run coefficients of variation of IHMS, INH and AcINH from 1 to 25 μm/ml were 4.7 ± 2.5, 5.4 ± 1.0 and 5.1 ± 3.1%, respectively. Hence this sensitive, reproducible and accurate method was suitable for pharmacokinetic studies of IHMS.

AB - A rapid and sensitive high-performance liquid chromatographic method is described for simultaneous determination of isoniazid methanesulphonate (IHMS) and its metabolites, such as isoniazid (INH) and acetylisoniazid (AcINH) in rabbit blood. According to stability studies, IHMS was most stable at pH 3-5. After acidifying the blood to pH 5.0, a suitable amount of acetonitrile was added to the supernatant for extraction and niacinamide served as an internal standard. After evaporation, the residue was reconstituted with phosphate buffer and aliquots of this solution were separated on a reversed-phase phenyl column by a mobile phase consisting of 0.25 mM tetrabutylammonium phosphate as a paired-ion reagent. UV detection was performed at 280 nm. Under these conditions, the between-run coefficients of variation of IHMS, INH and AcINH from 1 to 25 μm/ml were 4.7 ± 2.5, 5.4 ± 1.0 and 5.1 ± 3.1%, respectively. Hence this sensitive, reproducible and accurate method was suitable for pharmacokinetic studies of IHMS.

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