title: Disulfiram metabolite S-methyl-N,N-diethylthiocarbamate quantitation in human plasma with reverse phase ultra performance liquid chromatography and mass spectrometry.
abstract: Disulfiram has been used extensively for alcohol abuse and may have a role in treatment for cocaine addiction. Recent data suggest that disulfiram may also reactivate latent HIV in reservoirs. Disulfiram has complex pharmacokinetics with rapid metabolism to active metabolites, including S-methyl-N,N-diethylthiocarbamate (DET-Me) which is formed from cytochrome P450 (CYP450). Assessing disulfiram in HIV-infected individuals with a CYP450 inducing drug (e.g., efavirenz) or a CYP450 inhibiting drug (e.g., HIV-1 protease inhibitors) requires an assay that can measure a metabolite that is formed directly via CYP450 oxidation. Therefore, an assay to measure concentrations of DET-Me in human plasma was validated. DET-Me and the internal standard, S-ethyldipropylthiocarbamate (EPTC) were separated by isocratic ultra performance liquid chromatography using a Waters Acquity HSS T3 column (2.1 mm × 100 mm, 1.8 μm) and detection via electrospray coupled to a triple quadrupole mass spectrometer. Multiple reaction monitoring in positive mode was used with DET-Me at 148/100 and the internal standard at 190/128 with a linear range of 0.500-50.0 ng/mL with a 5 min run time. Human plasma (500 μL) was extracted using a solid phase procedure. The interassay variation ranged from 1.86 to 7.74% while the intra assay variation ranged from 3.38 to 5.94% over three days. Representative results are provided from samples collected from subjects receiving daily doses of disulfiram 62.5mg or 250 mg.
authors.affiliation: Translational Pharmacology Research Core, Center of Excellence in Bioinformatics and Life Sciences and School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY 14203, USA.
authors.forename: Jill
authors.initials: J
authors.lastname: Hochreiter
authors.forename: Elinore F
authors.initials: EF
authors.lastname: McCance-Katz
authors.forename: Jill
authors.initials: J
authors.lastname: Lapham
authors.forename: Qing
authors.initials: Q
authors.lastname: Ma
authors.forename: Gene D
authors.initials: GD
authors.lastname: Morse
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D004221:Disulfiram / Q000097:blood / Q000378:metabolism* / Q000493:pharmacokinetics; D004355:Drug Stability; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D015203:Reproducibility of Results; D013859:Thiocarbamates / Q000097:blood* / Q000378:metabolism / Q000493:pharmacokinetics
chemical_list: D013859:Thiocarbamates; C070631:S-methyl diethylthiocarbamate; D004221:Disulfiram