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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 27, Number 2, March 2003,
pp. 103-105
Enzyme Immunoassay Validation for the Detection of Buprenorphine
in Urine
V. Cirimele*, P. Kintz, S. Lohner, and B. Ludes
Institut de Médecine Légale, Laboratoire de Toxicologie, 11, rue
Humann, 67085 Strasbourg, France
A solid-phase enzyme immunoassay involving microtiter plates
was proposed by Microgenics to screen buprenorphine in urine. The intra-assay
precision at 10 ng/mL was 7.7% (coefficient of variation). The immunoassay
was determined to have no cross-reactivity with codeine, dihydrocodeine, morphine,
ethylmorphine, 6-monoacetylmorphine, methadone, pholcodine, propoxyphene, dextromoramide,
and dextromethorphan at 1 and 10 mg/L. A low cross-reactivity (3% at 1 ng/mL)
was observed at low concentrations of norbuprenorphine. After comparing this
new immunological test (Singlestep™ ELISA) for 76 urine specimens with
our validated high-performance liquid chromatography–electrospray mass
spectrometry (HPLC–ES-MS) procedure, an optimum cutoff concentration
of 2 ng/mL was determined for the kit. At this cutoff, the screening assay
was able to determine more than 90% of true results with 43.4% true positives
and 48.7% true negatives. Four positive urines (5.3%) were not confirmed by
HPLC–ES-MS. In only one case, the negative urine test was confirmed as
positive by HPLC–ES-MS (buprenorphine: 62.5 ng/mL). Buprenorphine concentrations
determined by HPLC–ES-MS ranged from 1.2 to 1052 ng/mL. Of the four potential
adulterants (hypochloride 50 mL/L, sodium nitrite 50 g/L, liquid soap 50 mL/L,
and sodium chloride 50 g/L) that might be added to a positive urine specimen,
none were able to cause a false-negative response by the immunoassay. The results
of this study support the concept that the Singlestep ELISA for buprenorphine
determination in urine should be considered as a new, valided screening procedure.
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