| |
Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 45, Number 7, September,
pp.377-382
Development and GC–MS Validation of
a Highly Sensitive Recombinant G6PDH-Based Homogeneous Immunoassay
for the Detection of Buprenorphine and Norbuprenorphine in Urine
Guohong Wang[1], Michael Vincent[1],
Warren Rodrigues[1], Alpana Agrawal[1], Christine Moore[1], Rekha
Barhate[1], E. Abolencia[1], Cynthia Coulter[1], James Soares[1],
Yi-Feng Zheng[2], Celeste Taylor[2], and Nihmat Morjana[2]
[1]Immunalysis Corporation, 829 Towne Center Drive, Pomona, California
91767 and
[2]Dade Behring, Inc., Glasgow Research Laboratory,
Route 896, P.O. Box 6101, Newark, Delaware 19714
Buprenorphine is now increasingly prescribed as
an alternative to methadone for the treatment of heroin addiction.
Because of its potency (dosage usages from 0.2 mg to 8 mg), the
drug concentrations in body fluids are normally very low. Here,
we report the first recombinant glucose-6-phosphate dehydrogenase
(G6PDH)-based homogeneous immunoassay (EMIT-type assay) for free
buprenorphine and free norbuprenorphine in urine. The antibody
used in this assay cross-reacts nearly identically with buprenorphine
and norbuprenorphine and, at the same time, has less than 1% cross-reactivity
with a wide range of commonly prescribed opiates, particularly
those structurally related compounds such as morphine, codeine,
and dihydrocodeine. More importantly, this assay has a low detection
limit of 1 ng/mL for buprenorphine or norbuprenorphine. Further
evaluation of this technique using gas chromatography––mass
spectrometry (GC––MS) of authentic urine samples demonstrated
that the accuracy of the assay is greater than 95%. Because this
assay is designed to measure the free drugs in urine, it resulted
in simplification for GC––MS or liquid chromatography–MS
confirmation methods that did not require urine hydrolysis before
solid-phase or liquid––liquid extraction.
Reproduction
of editorial content of this journal is prohibited without publisher’s
permission.
This
article is available in its entirety by fax for $40.00 each.
Visa, MasterCard and AMEX accepted. To order electronically click here
or call: 847-647-2900 ext. 1323
or fax request to: 847-647-1155.
Please indicate JAT
volume and issue along with page numbers. |
|
Home | Subscribe
| Current Issue | Back Issues
| Search | Advertise | Other Publications
| |
