Application of a novel form of solid-phase sorbent (Empore membrane) to the isolation of tricyclic antidepressant drugs from blood.
We employed a new form of solid-phase material, the Empore octyl (C8) extraction membrane (SPEM), for the efficient extraction of tricyclic drugs from patients' serum specimens. Both extraction and companion high-performance liquid chromatographic (HPLC) assay of doxepin (DOX), desmethyldoxepin (DDOX), imipramine (IMI), desmethylimipramine (DESI), amitriptyline (AMI), nortriptyline (NOR), clomipramine (CLO), and desmethylchlomipramine (DCLO) are presented here. Routinely, serum (1.0 mL or less) adjusted to pH 5.5 with phosphate buffer is passed through the SPEM secured in a MF-1 microfilter unit. Proteins and potential interferences retained on SPEM are removed with an acetonitrile-water wash. The tricyclic drugs are eluted with HPLC mobile phase and the eluate is injected directly on a Zorbax cyanopropyl (CN) HPLC column, thereby avoiding time consuming evaporation-concentration steps that can affect drug stability. Recovery for all drugs exceeds 90% and analytical responses are linear from a lower limit of sensitivity of 8 micrograms/L up to at least 1000 micrograms/L. Between-run coefficients of variation (CV) range from 2.9 to 8.3% through the concentration range of 75 to 300 micrograms/L. Performance characteristics of the SPEM are compared to those of conventional large particle silica- and polymeric-based sorbents. Within the requirements of this assay, the SPEM extraction requires less sample volume and reduces elution and solvent volumes.
Lensmeyer
G L
GL
Department of Pathology, University of Wisconsin Hospital & Clinics, Madison 53792.
Wiebe
D A
DA
Darcey
B A
BA
D000929:Antidepressive Agents, Tricyclic / Q000097:blood* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008567:Membranes, Artificial; D013056:Spectrophotometry, Ultraviolet
D000929:Antidepressive Agents, Tricyclic; D008567:Membranes, Artificial