Technical Note: Automated Homogeneous Immunoassay Analysis of Cotinine in Urine

Authors: Niedbala R.S.¹; Haley N.²; Kardos S.³; Kardos K.³

Source: Journal of Analytical Toxicology, Volume 26, Number 3, April 2002 , pp. 166-170(5)

Publisher: Preston Publications

Abstract:

A study was conducted to evaluate the performance comparison of a homogeneous enzyme immunoassay (EIA) designed to detect cotinine in urine and carbon monoxide (CO) breath measurements to determine smoking status. The clinical comparison was done using urine and breath specimens from 218 volunteers. Urine samples were analyzed by immunoassay and confirmed by gas chromatography–Mass spectrometry (GC–MS). Breath carbon monoxide was determined by a commercial analyzer. Using cutoffs of 10 ppm for CO and 500 ng/mL for urinary cotinine, the relative sensitivity/specificity was 93.6%/74.0%. The positive predictive value was 86.8%, and the negative predictive value was 86.5%. However, comparison of the EIA to GC–MS showed a sensitivity/specificity of 96.2%/98.4% and a positive predictive value of 99.3%. The EIA was also evaluated non-clinically for precision, stability, recovery, and interferences. In addition, the non-clinical evaluation demonstrated coefficients of variation from 0.37 to 1.09% across cotinine concentrations ranging from 0 to 5000 ng/mL. The assay was found to be highly specific for cotinine and cross-reacted to a limited degree with 3-hydroxycotinine. Finally, multiple freeze-thaw cycles of urines containing cotinine showed no degradation of the drug in the specimen when tested in the EIA. Thus, the EIA tested is a rapid, lab-based test that can reliably determine cotinine levels and their relation to smoking status.

Language: English

Document Type:

Affiliations: 1: OraSure Technologies, Inc. (OTI), 150 Webster Street, Bethlehem, Pennsylvania 18015-1389, Correspondence Address: E-mail:sniedbala@orasure.com 2: Metropolitan Life Insurance Company, Tarrytown, New York 3: OraSure Technologies, Inc. (OTI), 150 Webster Street, Bethlehem, Pennsylvania 18015-1389

Publication date: 2002-04-01

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  With an emphasis on practical application, JAT articles provide improved and novel techniques for use in clinical, forensic, workplace, sports testing (doping), and other toxicology laboratories. Articles describe newly developed methods in immunoassay testing, gas chromatography, liquid chromatography, mass spectrometry, atomic absorption spectrometry, solid and liquid phase extraction techniques, and other analytical approaches. Worldwide readership includes toxicologists, pathologists, chemists, clinicians, researchers, and educators working in medical examiner and law enforcement laboratories, hospitals, university and independent analytical laboratories, as well as the drug manufacturing industry.

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