A Liquid Chromatographic-Electrospray Ionization-Tandem Mass Spectrometric Method for Determination of Buprenorphine, Its Metabolite, norBuprenorphine, and a Coformulant, Naloxone, That Is Suitable for in Vivo and in Vitro Metabolism Studies
Analytical Biochemistry, ISSN: 0003-2697, Vol: 306, Issue: 1, Page: 31-39
2002
- 125Citations
- 27Captures
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Metrics Details
- Citations125
- Citation Indexes125
- 125
- CrossRef98
- Captures27
- Readers27
- 27
Article Description
A liquid chromatographic-electrospray ionization-tandem mass spectrometric method has been developed and validated for determination of the antiabuse medication, buprenorphine, its primary metabolite, norbuprenorphine, and a proposed coformulant, naloxone. The method uses deuterated internal standards and a simple liquid–liquid extraction. Mass spectrometry employed selected reaction monitoring of the transitions of m/z 468 to 396 for buprenorphine, 472 to 400 for [ 2 H 4 ]buprenorphine, 414 to 101 for norbuprenorphine, 423 to 110 for [ 2 H 9 ]norbuprenorphine, 328 to 310 for naloxone, and 345 to 327 for its internal standard, [ 2 H 3 ]naltrexone. The method was accurate and precise across the dynamic range of 0.1 to 10 ng/ml. All analytes were stable in human plasma stored at room temperature for up to 24 h and after three freeze–thaw cycles. Reconstituted extracts were stable at −20°C for up to 3 days. In human subjects receiving a sublingual tablet of 8 mg buprenorphine and 2 mg naloxone, buprenorphine and norbuprenorphine were detected for up to 24 h with respective maximum concentrations at 1 and 1.5 h. Maximal concentrations ranged from 2.2 to 2.8 and 1.5 to 2.4 ng/ml for buprenorphine and norbuprenorphine, respectively (i.e., ≈6 nM). The method detected norbuprenorphine formation in human liver microsomes incubated with 5–82 nM buprenorphine, which encompasses the therapeutic plasma concentration range. When cDNA-expressed P450s were incubated with 21 nM buprenorphine, norbuprenorphine formation was detected for P450s 3A4, as previously described, but also for 3A5, 3A7, and 2C8. Buprenorphine utilization generally exceeded norbuprenorphine formation, suggesting that P450s 2C18, 2C19, 2D6, and 2E1 may also be involved in buprenorphine metabolism to other products. These results suggest this method is suitable for both in vivo and in vitro studies of buprenorphine metabolism to norbuprenorphine.
Bibliographic Details
http://www.sciencedirect.com/science/article/pii/S0003269702956733; http://dx.doi.org/10.1006/abio.2002.5673; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0036629194&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/12069411; https://linkinghub.elsevier.com/retrieve/pii/S0003269702956733; https://dx.doi.org/10.1006/abio.2002.5673
Elsevier BV
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