RP-HPLC in determination of dextromethorphan and dextrophan in human urine :Phenotype analysis of CYP2D6
10.3724/SP.J.1008.2010.00310
- Author:
Hong ZHANG
1
Author Information
1. Department of Clinical Pharmacology
- Publication Type:Journal Article
- Keywords:
CYP2D6;
Cytochrome P-450;
Dextromethorphans;
Dextrorphan;
High pressure liquid chromatography
- From:
Academic Journal of Second Military Medical University
2010;31(3):310-312
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a RP-HPLC method for determination of the concentrations of dextromethorphan and its metabolites dextrorphan in human urine. Methods Phenacetine was used as internal standard, and the urine sample was hydrolyzed by enzyme, alkalified and extracted with hexane-butanol(9 : 1). The separation was carried out on Diamonsil™ C18 column (250 mm×4. 6 mm, 5 μm) with mobile phase of acetonitrile-1 % triethylamine buffer solution (pH adjusted to 2. 2 with H3PO4). Gradient elution was done for 0-15 min(20%-35% A). The flow rate was 1. 0 ml/min. The detection wavelength was set at 280 nm and the column temperature was 40°C. Results The linear ranges of dextromethorphan and dextrorphan were 0.05-2.0 μg/ml (r = 0. 999 9, n = 5) and 0. 5-20. 0 μg/ml (r= 0. 999 9, n = 5), respectively, and their lowest detecting concentrations were 0. 04 μg/ml and 0. 4 μg/ml, respectively. The intra-day and inter-day precision were both less than 10%. The low, middle and high extraction recoveries were between 94%-108%. Conclusion Our method is accurate and sensitive, and is suitable for the CYP2D6 phenotype analysis and pharmacokinetic studies of dextromethorphan and its metabolites in human.
