Simultaneous analysis of dextromethorphan and its three metabolites in human plasma using an improved HPLC method with fluorometric detection

Shyr Yi Lin, Chien Ho Chen, Hsiu O. Ho, Hsueh Hui Chen, Ming Thau Sheu

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27 Citations (Scopus)

Abstract

A simple and improved HPLC method with fluorometric detection for simultaneous determination of dextromethorphan (DM) and its three metabolites (dextrorphan (DX), 3-methoxymorphinan (MM), 3-hydroxymorphinan (HM)) in human plasma was developed and validated. The method involved a simple and efficient extraction protocol using an n-heptane/ethyl acetate (1:1) solvent mixture that achieved recoveries of 70-90% with an insignificant interference from the plasma matrix. The analysis was performed on a phenyl column with isocratic elution, a mobile phase composed of 20% methanol, 30% acetonitrile, and 50% KH2PO4 buffer (10 mM, with adding 0.02% of TEA; adjusted with phosphoric acid to pH 3.5), and a run time of only 15 min. Linear calibration curves were constructed in the concentration range of 1-200 nM for DM and its three metabolites. The lower limit of quantitation (LLOQ) in human plasma was 1 nM for each compound. The coefficient of variation and RSE% of the intraday and interday analyses for DM and its three metabolites all complied with USFDA requirements. This analytical method was preliminarily applied to determine the polymorphic functions of CYP2D6 and CYP3A4 in the metabolic pathway of DM to DX and then to HM.

Original languageEnglish
Pages (from-to)141-146
Number of pages6
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume859
Issue number1
DOIs
Publication statusPublished - Nov 1 2007

Fingerprint

Dextromethorphan
Plasma (human)
Metabolites
Dextrorphan
High Pressure Liquid Chromatography
Cytochrome P-450 CYP3A
Cytochrome P-450 CYP2D6
United States Food and Drug Administration
Metabolic Networks and Pathways
Calibration
Methanol
Buffers
Plasmas
Recovery
norlevorphanol

Keywords

  • Dextromethorphan
  • Dextrorphan
  • Hydroxymorphinan
  • Methoxymorphinan
  • Phenyl column

ASJC Scopus subject areas

  • Biochemistry

Cite this

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title = "Simultaneous analysis of dextromethorphan and its three metabolites in human plasma using an improved HPLC method with fluorometric detection",
abstract = "A simple and improved HPLC method with fluorometric detection for simultaneous determination of dextromethorphan (DM) and its three metabolites (dextrorphan (DX), 3-methoxymorphinan (MM), 3-hydroxymorphinan (HM)) in human plasma was developed and validated. The method involved a simple and efficient extraction protocol using an n-heptane/ethyl acetate (1:1) solvent mixture that achieved recoveries of 70-90{\%} with an insignificant interference from the plasma matrix. The analysis was performed on a phenyl column with isocratic elution, a mobile phase composed of 20{\%} methanol, 30{\%} acetonitrile, and 50{\%} KH2PO4 buffer (10 mM, with adding 0.02{\%} of TEA; adjusted with phosphoric acid to pH 3.5), and a run time of only 15 min. Linear calibration curves were constructed in the concentration range of 1-200 nM for DM and its three metabolites. The lower limit of quantitation (LLOQ) in human plasma was 1 nM for each compound. The coefficient of variation and RSE{\%} of the intraday and interday analyses for DM and its three metabolites all complied with USFDA requirements. This analytical method was preliminarily applied to determine the polymorphic functions of CYP2D6 and CYP3A4 in the metabolic pathway of DM to DX and then to HM.",
keywords = "Dextromethorphan, Dextrorphan, Hydroxymorphinan, Methoxymorphinan, Phenyl column",
author = "Lin, {Shyr Yi} and Chen, {Chien Ho} and Ho, {Hsiu O.} and Chen, {Hsueh Hui} and Sheu, {Ming Thau}",
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T1 - Simultaneous analysis of dextromethorphan and its three metabolites in human plasma using an improved HPLC method with fluorometric detection

AU - Lin, Shyr Yi

AU - Chen, Chien Ho

AU - Ho, Hsiu O.

AU - Chen, Hsueh Hui

AU - Sheu, Ming Thau

PY - 2007/11/1

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N2 - A simple and improved HPLC method with fluorometric detection for simultaneous determination of dextromethorphan (DM) and its three metabolites (dextrorphan (DX), 3-methoxymorphinan (MM), 3-hydroxymorphinan (HM)) in human plasma was developed and validated. The method involved a simple and efficient extraction protocol using an n-heptane/ethyl acetate (1:1) solvent mixture that achieved recoveries of 70-90% with an insignificant interference from the plasma matrix. The analysis was performed on a phenyl column with isocratic elution, a mobile phase composed of 20% methanol, 30% acetonitrile, and 50% KH2PO4 buffer (10 mM, with adding 0.02% of TEA; adjusted with phosphoric acid to pH 3.5), and a run time of only 15 min. Linear calibration curves were constructed in the concentration range of 1-200 nM for DM and its three metabolites. The lower limit of quantitation (LLOQ) in human plasma was 1 nM for each compound. The coefficient of variation and RSE% of the intraday and interday analyses for DM and its three metabolites all complied with USFDA requirements. This analytical method was preliminarily applied to determine the polymorphic functions of CYP2D6 and CYP3A4 in the metabolic pathway of DM to DX and then to HM.

AB - A simple and improved HPLC method with fluorometric detection for simultaneous determination of dextromethorphan (DM) and its three metabolites (dextrorphan (DX), 3-methoxymorphinan (MM), 3-hydroxymorphinan (HM)) in human plasma was developed and validated. The method involved a simple and efficient extraction protocol using an n-heptane/ethyl acetate (1:1) solvent mixture that achieved recoveries of 70-90% with an insignificant interference from the plasma matrix. The analysis was performed on a phenyl column with isocratic elution, a mobile phase composed of 20% methanol, 30% acetonitrile, and 50% KH2PO4 buffer (10 mM, with adding 0.02% of TEA; adjusted with phosphoric acid to pH 3.5), and a run time of only 15 min. Linear calibration curves were constructed in the concentration range of 1-200 nM for DM and its three metabolites. The lower limit of quantitation (LLOQ) in human plasma was 1 nM for each compound. The coefficient of variation and RSE% of the intraday and interday analyses for DM and its three metabolites all complied with USFDA requirements. This analytical method was preliminarily applied to determine the polymorphic functions of CYP2D6 and CYP3A4 in the metabolic pathway of DM to DX and then to HM.

KW - Dextromethorphan

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