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Correlation of inter-individual variations of amitriptyline metabolism examined in hairs with CYP2C19 and CYP2D6 polymorphisms

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Abstract

The metabolite ratio of amitriptyline (AT), nortriptyline (NT) and its 10-hydroxy metabolites (E10-OHAT, Z10-OHAT, E10-OHNT and Z10-OHNT) was examined by liquid chromatography-mass spectrometry in hair samples of 23 white infants after long-term administration of AT. High inter-individual variation of the metabolite ratios were observed (e.g. NT/AT = 0.8–8.1, E10-OHNT/Z10-OHNT = 1.6–10.3). The significance of these variations was proven by confirmation of the temporary stability of these ratios within a hair fibre. Moreover, an association of the metabolic phenotype with genetic disposition was observed. The genotypes of CYP2C19 (alleles *2, *3 and *4) and of CYP2D6 (*3, *4, and *6) were examined by conventional polymerase chain reaction genotyping experiments. The relative amount of demethylation (NT/AT) is clearly affected by the number of functional alleles of CYP2C19. The demethylation capacity of CYP2C19 poor metabolizers (3 individuals, compared to 15 extensive metabolizers) was 4.3 times depleted. Moreover, the selectivity of hydroxylation (e.g. E10-OHNT/Z10-OHNT) is significantly correlated with CYP2C19.

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Authors and Affiliations

  1. Institute of Forensic Medicine, Nussbaumstrasse 26, 80336, Munich, Germany

    Detlef Thieme, Burkhard Rolf, Hans Sachs & Dagmar Schmid

  2. Forensic Toxicological Centre, Bayerstrasse 53, 80335, Munich, Germany

    Detlef Thieme & Hans Sachs

Authors
  1. Detlef Thieme
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  2. Burkhard Rolf
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  3. Hans Sachs
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  4. Dagmar Schmid
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Corresponding author

Correspondence to Detlef Thieme.

Electronic supplementary material

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Supplementary Fig. 1

Identification of amitriptyline (AT), nortriptyline (NT) and the hydroxymetabolites in a methanolic extract of hair by LC-MS-MS (DOC 28.5 KB)

Supplementary Table 1

Parameters for LC-MS detection of amitriptyline, nortriptyline, hydroxymetabolites and MPPH (internal standard), range and correlation coefficients of the calibration curves and limits of detection (LOD) (DOC 27.5)

Supplementary Table 2

Primers used for the multiplex PCR (DOC 24.0 KB)

Supplementary Table 3

Sequences of primer and the probe for CYP2D6 (DOC 19.5 KB)

Supplementary Table 4

Summary of gender, hair colour and hair concentrations of amitriptyline (AT), nortriptyline (NT), and corresponding hydroxymetabolites examined in hair bundles and alleles of CYP2D6 and CYP2D6 (DOC 63.0 KB)

Supplementary Table 5

Comparisons of mean values of metabolite ratios by variance analysis to examine significant correlations between demethylation and hydroxylation stereo specificity and the number of functional CYP2C19 and CYP2D6 alleles (DOC 23.5 KB)

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Thieme, D., Rolf, B., Sachs, H. et al. Correlation of inter-individual variations of amitriptyline metabolism examined in hairs with CYP2C19 and CYP2D6 polymorphisms. Int J Legal Med 122, 149–155 (2008). https://doi.org/10.1007/s00414-007-0184-4

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  • DOI: https://doi.org/10.1007/s00414-007-0184-4

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