Quantitative Determination of Dextromethorphan and Three Metabolites in Urine by Reverse-Phase High-Performance Liquid Chromatography

• Published in: Journal of pharmaceutical sciences 1984-01, Vol.73 (1), p.24-29
• Main Authors: Park, Young Han, Peter Kullberg, M., Hinsvark, Orville N.
• Format: Article
• Language: English
• Subjects: Analysis; and metabolites; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Cough suppressants-dextromethorphan; Cough suppressants—dextromethorphan, dextrorphan, and metabolites, quantitation in human urine by high-performance liquid chromatography; Dextromethorphan - metabolism; Dextromethorphan - urine; Dextromethorphan-quantitation with metabolites by high-performance liquid chromatography; Dextromethorphan—quantitation with metabolites by high-performance liquid chromatography, human urine; dextrorphan; Dextrorphan - urine; Dextrorphan-quantitation in human urine after administration of dextromethorphan; Dextrorphan—quantitation in human urine after administration of dextromethorphan, high-performance liquid chromatography; General pharmacology; high-performance liquid chromatography; human urine; Humans; Hydrogen-Ion Concentration; Levorphanol - analogs & derivatives; Medical sciences; Pharmacology. Drug treatments; quantitation in human urine by high-performance liquid chromatography
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1002/jps.2600730107

□ A high-performance liquid chromatographic (HPLC) method for the quantitation of dextromethorphan (I) and its three metabolites, dextrorphan (II), 3-hydroxy-9α,13α,14α -morphinan (III), and 3-methoxy-9α,13α,14α:-morphinan (IV), in urine was developed. For the analysis of nonconjugated compounds, urine samples at pH11-11.5, containing 3-methoxy-17-methyl-10-oxo-9α,13α,14α -morphinan as an internal standard, were applied to an extraction column, and the compounds were eluted with 10% n-butyl alcohol-hexane. The organic eluant was extracted with 0.1M HC1, and an aliquot of the acidic extract was analyzed by HPLC utilizing a 5-μm phenyl column (25×0.46-cm i.d.) with a mobile phase of 10mM potassium phosphate-acetonitrile (45:55, pH4.0); the column effluent was monitored by UV detection at 280nm. Free and conjugated metabolites in the enzyme-treated urine were analyzed by selective extraction of I and IV with hexane from urine samples at pH>12 and extraction of II and III with 10% n-butyl alcohol-hexane from urine samples at pH11-11.5. The minimum quantifiable levels of I-IV ranged from 0.017 to 0.09μg of base/mL and from 0.11 to 0.21μg of base/mL in nonhydrolyzed and hydrolyzed urine, respectively.