Analysis of benzodiazepines and tricyclic antidepressants in serum using a common solid-phase clean-up and a common mobile phase

Abstract

A single rapid and specific solid-phase clean-up procedure was developed for the analysis of benzodiazepines and tricyclic antidepressants using a carefully selected wash step and specific sequential elution. Benzodiazepines were eluted from the solid-phase column using a mixture of water-methanol-acetonitrile (2:3:3) followed by the elution of tricyclic antidepressants with methanol containing 0.6% diethylamine. A 30% solution of acetonitrile in phosphate buffer containing dimethylocylamine was used as a common isocratic mobile phase for the analysis of benzodiazepines and tricyclic antidepressants on a reversed-phase column and detection was carried out at 242 nm. The sensitivity limit of the assay for benzodiazepines and tricyclic antidepressants was 25 ng/ml in serum with recoveries of 95-105% for benzodiazepines and 76-95% for tricyclic antidepressants. The results were linear for benzodiazepines over the range 50-2000 ng/ml and for tricyclic antidepressants over the range 25-500 ng/ml. Analysis for benzodiazepines and tricyclic antidepressants gave good precision, with a coefficient of variation of less than 5.0%. The method described here will be suitable for use in a clinical setting, where there is a concomitnat use of benzodiazepines and tricyclic antidepressants.