A Simple High-Performance Liquid Chromatography Method for the Quantitation of Tricyclic Antidepressant Drugs in Human Plasma or Serum

Abstract

Objective: A simple reversed-phase high performance liquid chromatography method with a variable wavelength detection has been developed for determining the commonly used tricyclic antidepressant drugs in plasma. Methods: The assay procedure involves a liquid-liquid extraction with an initial extraction into hexane:ethyl acetate after basification of the homogenate. The column used was a Supelco Hypersil 5 μm, 150 × 3.2 mm. The mobile phase was acetonitrile, methanol, and phosphate buffer (0.01M, pH 7.4) at 12:3:5 ratio (v/v/v). Results: The recoveries ranged from 89 to 108% and the day-to-day precision was 1.1 to 13% CV depending on the compound. The method is sensitive to 15 ng/mL and linear to 400 ng/mL with a 25 μL injection. Conclusion: This method is simple, sensitive, precise, inexpensive, and is currently used in our laboratory for therapeutic monitoring of tricyclics. The data generated by this method were within the range outlined by the College of American Pathologists.