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.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 May 15;897:80-4. doi: 10.1016/j.jchromb.2012.03.035. Epub 2012 Apr 1.
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.
双硫仑被广泛用于治疗酒精滥用，并且可能在可卡因成瘾的治疗中发挥作用。最近的数据表明，双硫仑也可能使潜伏的 HIV 在储库中重新激活。双硫仑具有复杂的药代动力学特性，其代谢迅速，可生成包括 S-甲基-N,N-二乙基硫代氨基甲酸酯（DET-Me）在内的活性代谢物，该物质由细胞色素 P450（CYP450）形成。在接受 CYP450 诱导药物（例如依非韦伦）或 CYP450 抑制药物（例如 HIV-1 蛋白酶抑制剂）治疗的 HIV 感染者中评估双硫仑时，需要一种能够测量直接通过 CYP450 氧化形成的代谢物的检测方法。因此，建立了一种测量人血浆中 DET-Me 浓度的检测方法。使用 Waters Acquity HSS T3 柱（2.1mm×100mm，1.8μm）通过等度超高效液相色谱法分离 DET-Me 和内标 S-乙基二丙基硫代氨基甲酸酯（EPTC），通过电喷雾串联三重四极杆质谱仪进行检测。采用正离子模式下的多重反应监测，DET-Me 的检测离子对为 148/100，内标物的检测离子对为 190/128，线性范围为 0.500-50.0ng/mL，运行时间为 5 分钟。使用固相萃取法从 500μL 人血浆中提取样品。在三天内，通过检测三次，得到的结果显示，批内变异系数在 3.38%-5.94%之间，批间变异系数在 1.86%-7.74%之间。提供了来自接受双硫仑每日剂量 62.5mg 或 250mg 的受试者的样本的代表性结果。
