Determination of debrisoquine and 4-hydroxydebrisoquine by high-performance liquid chromatography: application to the evaluation of CYP2D6 genotype and debrisoquine metabolic ratio relationship
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Pedro Dorado
Abstract
The drug-metabolizing cytochrome P450 (CYP) enzyme CYP2D6 is involved in the metabolism of several clinically important drugs. So far more than 50 different CYP2D6 allelic variants have been described, and thus there is an increased need for routine high-performance liquid chromatography (HPLC) methods for the evaluation of the functional implication of CYP2D6 polymorphism. Debrisoquine is metabolized to 4-hydroxydebrisoquine by CYP2D6, and therefore it has been used widely to determine the hydroxylation capacity of the enzyme. The aim of the present study was to develop a simple, accurate HPLC method with ultraviolet detection for the measurement of debrisoquine and 4-hydroxydebrisoquine in urine for evaluation of the relationship between CYP2D6 enzyme activity and genotypes. For the HPLC determination, a C18 extraction column was used with a flow rate of 0.8mL/min and detection at 210nm. The compounds were eluted from the column in less than 10min. Coefficients of variation at all concentrations were less than 4% for both compounds. The debrisoquine/4-hydroxydebrisoquine ratio (debrisoquine metabolic ratio) was determined in a panel of 16 Caucasian healthy volunteers with zero (poor metabolizers), one, two or more than two (ultrarapid metabolizers) CYP2D6 active genes. Significant correlation (p<0.05) between the number of CYP2D6 active genes and the hydroxylation capacity of the enzyme was found. The present HPLC method was simple, fast and accurate, and thus will be useful for the evaluation of CYP2D6 hydroxylation capacity in pharmacogenetic studies.
References
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©2005 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- Effect of stress doses of hydrocortisone on S-100B vs. interleukin-8 and polymorphonuclear elastase levels in human septic shock
- Conjugated linoleic acid suppresses the secretion of atherogenic lipoproteins from human HepG2 liver cells
- Determination of debrisoquine and 4-hydroxydebrisoquine by high-performance liquid chromatography: application to the evaluation of CYP2D6 genotype and debrisoquine metabolic ratio relationship
- Assessment of iodine intake in mildly iodine-deficient pregnant women by a new automated kinetic urinary iodine determination method
- Butyrylcholinesterase activity and metabolic syndrome in obese patients
- The non-thyroidal illness syndrome after coronary artery bypass grafting: a 6-month follow-up study
- Advanced oxidation protein products (AOPP) for monitoring oxidative stress in critically ill patients: a simple, fast and inexpensive automated technique
- Quality control in urinary stone analysis: results of 44 ring trials (1980–2001)
- External Quality Assessment in The Netherlands: time to introduce commutable survey specimens. Lessons from the Dutch “Calibration 2000” project
- Evaluation of a high-sensitivity turbidimetric immunoassay for serum C-reactive protein: application to the study of longitudinal changes throughout normal pregnancy
- A quantitative appraisal of interference by icodextrin metabolites in point-of-care glucose analyses
- Preanalytical variability in laboratory testing: influence of the blood drawing technique
- Soluble transferrin receptor and transferrin receptor-ferritin index in iron deficiency anemia and anemia in rheumatoid arthritis
- Elevated plasma cysteinylglycine levels caused by cilastatin-associated antibiotic treatment
- The number of consultant clinical chemists in the 15-nation European Union
- Biological variation of vascular endothelial growth factor
- Terminology, categories and representation of examinations in laboratory medicine
- Reply to W.G. Wood. Questionable results – who directs the EQAS organisers? Clin Chem Lab Med 2004;42:1073
- The predictive power of serum κ/λ ratios for discrimination between monoclonal gammopathy of undetermined significance and multiple myeloma
- 4th Conference on Hyperhomocysteinemia: Saarbrücken, Germany, April, 14–16, 2005, Saarland University
