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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 27, Number 8, November/December,
pp. 581-586
Validation of the Cozart® Microplate ELISA for Detection
of Opiates in Hair
Gail Cooper[1], Lisa Wilson[1], Claire Reid[1], Dene
Baldwin[1],
Chris Hand[2],
and Vina Spiehler[3]
[1]Cozart Bioscience Ltd, Abingdon, Oxfordshire OX14 4RU, United Kingdom;
[2]Cozart
Bioscience, Inc., Cupertino, California 95014; and
[3]Spiehler and Associates,
Newport Beach, California 92663
The purpose of this study was to determine the performance characteristics
of the Cozart Opiates Microplate ELISA assay for the detection of opiates in
hair specimens. One hundred and six hair specimens were collected from volunteers
and from drug-related deaths. The hair samples were extracted by sonication
followed by overnight extraction in methanol at 60°C. The methanol extract
was dried, reconstituted in ELISA negative calibrator, and then analyzed. For
gas chromatography–mass spectrometry (GC–MS) analysis, deuterated
internal standard mixture and 0.1M HCl were added to 20 mg of specimen or spiked
blank hair and sonicated for 1 h. The opiates were extracted by solid-phase
and derivatized with BSTFA + 1% TMS for GC–MS analysis. Fifty-one hair
specimens were confirmed positive by GC–MS. The true positives, true
negatives, false positives, and false negatives for different cutoffs with
the ELISA were determined by comparison of the ELISA response (normalized to
weight of hair extracted) to the GC–MS results as the reference method.
The optimum cutoff for the Cozart Opiate Microplate ELISA was determined to
be between 200 and 300 pg morphine equivalents/mg hair using a 20-mg hair sample.
The Cozart Opiates Microplate EIA for opiates in hair using a cutoff of 200
pg/mg hair with a 20-mg hair sample had a sensitivity of 98% ± 2% and
a specificity of 92.7% ± 3.5% versus GC–MS. Reproduction
of editorial content of this journal is prohibited without publisher’s
permission.
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