Bioluminescent Assay for Heroin and Its Metabolites

Illicit heroin is trafficked as a solid particulate drug, while heroin abuse is monitored by testing urine samples for its principal metabolites, morphine and morphine-3-glucuronide. Two novel bacterial enzymes were used in the development of a linked-enzyme assay for heroin and its metabolites:  he...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 1996-06, Vol.68 (11), p.1877-1882
Main Authors: Holt, Peter-John, Bruce, Neil C, Lowe, Christopher R
Format: Article
Language:English
Subjects:
Acetylesterase - analysis
Alcohol Oxidoreductases - analysis
Bacteria
Biological and medical sciences
Chemistry
Drug addictions
Enzymes
Enzymes, Immobilized
Heroin
Heroin - analysis
Heroin - pharmacokinetics
Humans
Luminescent Measurements
Medical sciences
Substance Abuse Detection
Toxicology
Vibrio - enzymology
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Summary:Illicit heroin is trafficked as a solid particulate drug, while heroin abuse is monitored by testing urine samples for its principal metabolites, morphine and morphine-3-glucuronide. Two novel bacterial enzymes were used in the development of a linked-enzyme assay for heroin and its metabolites:  heroin esterase, which converts heroin to morphine, and morphine dehydrogenase, which oxidizes morphine to morphinone with the concomitant reduction of NADP+. A bioluminescent assay involving heroin esterase, morphine dehydrogenase, and the bacterial luciferase from Vibrio harveyi was developed and shown to be sensitive to 89 ng/mL heroin and 2.0 ng/mL morphine. Excellent correlation with the results from 83 authentic samples submitted for urine drug screening at a hospital laboratory was obtained. The bioluminescent assay exhibited greater specificity and speed than current immunological screening methods. A novel format of the bioluminescent assay involving immobilized enzymes was sensitive to 101 ng (250 pmol) of heroin and responded well to particulate heroin. This form of the test was sensitive enough to respond to one or two typical particles of illicit heroin.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac951207r