LC-MS/MS determination of carbamathione in microdialysis samples from rat brain and plasma.
LC-MS/MS determination of carbamathione in microdialysis samples from rat brain and plasma.
复制标题
LC-MS/MS 测定大鼠脑和血浆微透析样品中的氨基甲硫酮。
DOI：
10.1016/j.jpba.2009.07.026
复制
发表时间：
2010
影响因子：
3.4
通讯作者：
Faiman,MorrisD
中科院分区：
文献类型：
--
作者：
Kaul,Swetha;Williams,ToddD;Lunte,CraigE;Faiman,MorrisD
建立了一种选择性液相色谱-串联质谱（LC-MS/MS）测定大鼠脑和血浆微透析样品中S-（N，N-二乙基氨基甲酰基）谷胱甘肽（氨基甲酰）的方法。S-（N，N-二乙基氨基甲酰基）谷胱甘肽（氨甲酰谷胱甘肽）是双硫仑的代谢产物。这种代谢物可能是双硫仑治疗可卡因依赖有效性的原因。色谱分离在Alltech Altima C-18（50 mm长× 2.1 mm i.d.，溶剂A由10 mM甲酸铵、甲醇和甲酸（99：1：0.06，v/v/v）组成。溶剂B由甲醇、10 mM甲酸铵和甲酸（99：1：0.06，v/v/v）组成。使用从95%水溶液至95%有机溶液的20分钟线性梯度。在配备有ESI接口的Micromass Quattro Ultima“三重”四极杆质谱仪上获得串联质谱。在正离子模式选择反应监测（SRM）模式下进行定量质谱分析，观察氨甲喋呤的m/z 407-100和175跃迁以及内标物S-己基谷胱甘肽的m/z 392-263跃迁。同时收集血液和脑中的微透析液，用于监测中心和外周的卡巴肼浓度。在0.25- 10，000 nM的浓度范围内获得了良好的线性。最低定量限（LLOQ）确定为1 nM，最低检测限（LLOD）计算为0.25 nM。测定了日内和日间准确度和精密度，对于评价的所有样品，变异性均小于10%（R.S.D.）。
A selective liquid chromatography–tandem mass spectrometric (LC–MS/MS) method was developed for the determination of S-(N, N-diethylcarbamoyl) glutathione (carbamathione) in microdialysis samples from rat brain and plasma. S-(N, N-Diethylcarbamoyl) glutathione (carbamathione) is a metabolite of disulfiram. This metabolite may be responsible for disulfiram's effectiveness in the treatment of cocaine dependence. Chromatographic separations were carried out on an Alltech Altima C-18 (50mm long×2.1mm i.d., 3μm particles) analytical column at a flow rate of 0.3ml/min. Solvent A consisted of 10mM ammonium formate, methanol, and formic acid (99:1:0.06, v/v/v). Solvent B consisted of methanol, 10mM ammonium formate and formic acid (99:1:0.06, v/v/v). A 20min linear gradient from 95% aqueous to 95% organic was used. Tandem mass spectra were acquired on a Micromass Quattro Ultima “triple” quadrupole mass spectrometer equipped with an ESI interface. Quantitative mass spectrometric analysis was conducted in positive ion mode selected reaction monitoring (SRM) mode looking at the transition of m/z 407–100 and 175 for carbamathione and m/z 392–263 for the internal standard S-hexyl glutathione. The simultaneous collection of microdialysate from blood and brain was used to monitor carbamathione concentrations centrally and peripherally. Good linearity was obtained over a concentration range of 0.25–10,000nM. The lowest limit of quantification (LLOQ) was determined to be 1nM and the lowest limit of detection (LLOD) was calculated to be 0.25nM. Intra- and inter-day accuracy and precision were determined and for all the samples evaluated, the variability was less that 10% (R.S.D.).