Determination of debrisoquine and 4-hydroxydebrisoquine by high-performance liquid chromatography

Application to the evaluation of CYP2D6 genotype and debrisoquine metabolic ratio relationship

Pedro Dorado, R. Berecz, Macarena C. Cáceres, Idilio González, Jesús Cobaleda, Adrián Llerana

Research output: Contribution to journalArticle

15 Citations (Scopus)

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 0.8 mL/min and detection at 210 nm. The compounds were eluted from the column in less than 10 min. 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

Original languageEnglish
Pages (from-to)275-279
Number of pages5
JournalClinical Chemistry and Laboratory Medicine
Volume43
Issue number3
DOIs
Publication statusPublished - 2005

Fingerprint

Debrisoquin
Cytochrome P-450 CYP2D6
High performance liquid chromatography
Genotype
High Pressure Liquid Chromatography
Cytochrome P-450 Enzyme System
Hydroxylation
Enzyme activity
Enzymes
Polymorphism
Metabolism
Pharmaceutical Preparations
4-hydroxydebrisoquin
Healthy Volunteers
Genes
Flow rate
Urine

Keywords

  • CYP2D6
  • Debrisoquine
  • HPLC
  • Metabolic ratio

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Determination of debrisoquine and 4-hydroxydebrisoquine by high-performance liquid chromatography : Application to the evaluation of CYP2D6 genotype and debrisoquine metabolic ratio relationship. / Dorado, Pedro; Berecz, R.; Cáceres, Macarena C.; González, Idilio; Cobaleda, Jesús; Llerana, Adrián.

In: Clinical Chemistry and Laboratory Medicine, Vol. 43, No. 3, 2005, p. 275-279.

Research output: Contribution to journalArticle

@article{a7514972d7894dd0a95110eec3804a1f,
title = "Determination of debrisoquine and 4-hydroxydebrisoquine by high-performance liquid chromatography: Application to the evaluation of CYP2D6 genotype and debrisoquine metabolic ratio relationship",
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 0.8 mL/min and detection at 210 nm. The compounds were eluted from the column in less than 10 min. 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",
keywords = "CYP2D6, Debrisoquine, HPLC, Metabolic ratio",
author = "Pedro Dorado and R. Berecz and C{\'a}ceres, {Macarena C.} and Idilio Gonz{\'a}lez and Jes{\'u}s Cobaleda and Adri{\'a}n Llerana",
year = "2005",
doi = "10.1515/CCLM.2005.046",
language = "English",
volume = "43",
pages = "275--279",
journal = "Clinical Chemistry and Laboratory Medicine",
issn = "1434-6621",
publisher = "Walter de Gruyter GmbH & Co. KG",
number = "3",

}

TY - JOUR

T1 - Determination of debrisoquine and 4-hydroxydebrisoquine by high-performance liquid chromatography

T2 - Application to the evaluation of CYP2D6 genotype and debrisoquine metabolic ratio relationship

AU - Dorado, Pedro

AU - Berecz, R.

AU - Cáceres, Macarena C.

AU - González, Idilio

AU - Cobaleda, Jesús

AU - Llerana, Adrián

PY - 2005

Y1 - 2005

N2 - 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 0.8 mL/min and detection at 210 nm. The compounds were eluted from the column in less than 10 min. 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

AB - 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 0.8 mL/min and detection at 210 nm. The compounds were eluted from the column in less than 10 min. 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

KW - CYP2D6

KW - Debrisoquine

KW - HPLC

KW - Metabolic ratio

UR - http://www.scopus.com/inward/record.url?scp=17044382643&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=17044382643&partnerID=8YFLogxK

U2 - 10.1515/CCLM.2005.046

DO - 10.1515/CCLM.2005.046

M3 - Article

VL - 43

SP - 275

EP - 279

JO - Clinical Chemistry and Laboratory Medicine

JF - Clinical Chemistry and Laboratory Medicine

SN - 1434-6621

IS - 3

ER -