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 (2.1×100mm, 1.8μm) 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 to 50.0 ng/mL with a five minute 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 62.5 mg or 250 mg.

Interaction of disulfiram with antiretroviral medications: Efavirenz increases while atazanavir decreases disulfiram effect on enzymes of alcohol metabolism

“…DIS carbamate concentrations were determined using ultra performance liquid chromatography coupled to electrospray tandem mass spectrometry (27). S-Methyl-N-N-diethylthiocarbamate reference standard (lot number ELZ-125-3, 98% purity) was used to prepare calibration standards and quality controls and was supplied by Toronto Research Chemicals.…”

Degradation of amyloid beta species by multi-copper oxidases

“…Interestingly, EDTA and DDC could also shorten the lifetime of the transient state, suggesting the competition from EDTA and DDC may lead to more transient binding, which consequentially leads to more effective degradation. Since DDC is an active metabolite of disulfiram, an FDA-approved drug for alcoholism 42,59 , it may have the potential to utilize disulfiram to assist peripheral Aβ clearance. Nonetheless, more detailed studies are needed to clarify the degradation mechanism.…”

Disulfiram (Tetraethylthiuram Disulfide) in the Treatment of Lyme Disease and Babesiosis: Report of Experience in Three Cases

“…Disulfiram metabolism is complex [79,80,81,82,83,84]. The drug and its metabolites, including carbon disulfide and diethyldithiocarbamate, have a wide distribution in mammalian tissues, including lipids and the central nervous system, prolonged half-lives, and may require one to two weeks for complete elimination after a dose.…”