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A liquid chromatographic method for quantitating amitriptyline in brain tissue.

S H Preskorn, R K Glotzbach

    Psychopharmacology
    |January 1, 1982
    PubMed
    Summary
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    This study adapted a plasma assay to measure tricyclic antidepressants (TCA), specifically amitriptyline and nortriptyline, in rat brain tissue. The modified method demonstrated high accuracy and precision for quantifying these crucial compounds in neurological samples.

    Area of Science:

    • Pharmacology
    • Neuroscience
    • Analytical Chemistry

    Background:

    • Tricyclic antidepressants (TCA) are vital in treating various neurological and psychiatric conditions.
    • Accurate quantification of TCAs in brain tissue is crucial for understanding their pharmacokinetics and pharmacodynamics.
    • Existing methods for TCA measurement often focus on plasma, necessitating adaptation for brain tissue analysis.

    Purpose of the Study:

    • To modify and validate a high-performance liquid chromatography (HPLC) method for measuring amitriptyline (AMI) and nortriptyline (NOR) in rat brain tissue.
    • To assess the accuracy, precision, and linearity of the adapted assay for TCA quantification in brain homogenates.

    Main Methods:

    • A previously established plasma assay for TCAs was adapted for rat brain tissue.

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  • Brains from drug-free and AMI-treated rats were extracted and analyzed using a Waters HPLC system.
  • Assay validation included drug recovery, intra-run and inter-run precision, and linearity assessments.

    • Main Results:

    • The modified HPLC method showed no interference from endogenous substances in drug-free brain tissue.
    • Average drug recovery was high, at 90 +/- 3.4% (mean +/- SEM).
    • Excellent precision was observed with low coefficients of variation for both AMI (intra-run: 2.7%, inter-run: 4.2%) and NOR (intra-run: 1.8%, inter-run: 3.5%).
    • Assay linearity was confirmed over a concentration range of 250--3,000 ng/g wet weight tissue (r = 0.96, P < 0.001).

    Conclusions:

    • The adapted HPLC method provides a reliable and precise means for quantifying amitriptyline and nortriptyline in rat brain tissue.
    • This validated assay facilitates further research into the central nervous system effects and distribution of these important TCAs.