Homogeneous Enzyme Immunoassay (EMIT) Protocol for Monitoring Tricyclic Antidepressants on the COBAS-BIO Centrifugal Analyzer

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Abstract
EMIT tests are available for quantitative determination of the tricyclic antidepressants amitryptyline (AMI), nortriptyline (NORT), imipramine (IMI), and desipramine (DMI). An extraction step before analysis eliminates cross-reacting polar metabolites. Excellent correlation between EMIT and high-performance liquid chromatography (HPLC) has been previously established. Most published protocols for using EMIT reagents on COBAS-BIO centrifugal analyzers were designed for analytes present at 1-50 mg/L in serum. AMI, NORT, IMI, and DMI are usually present at far less than 1 mg/L. We describe a COBAS-BIO EMIT protocol for assaying these analytes. Patient sample correlation beween COBAS-BIO EMIT and EMIT-AutoLab were > 0.99. Between-run precision for single-point determinations was comparable to SYVA AutoLab performance [.ltoreq. 11% at 40 .mu.g/L AMI, NORT, IMI (80 .mu.g/L DMI), and .ltoreq. 4% at 200 .mu.g/L AMI, NORT, IMI (400 .mu.g/L DMI)]. With stored-curve updating, working reagents were usable for at least 14 days (AMI) or 23 days (DMI).

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