Coles Rebecka, Kharasch Evan D
Division of Clinical and Translational Research, Department of Anesthesiology, Washington University, St Louis, MO 63110, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Sep 15;857(1):67-75. doi: 10.1016/j.jchromb.2007.07.007. Epub 2007 Jul 10.
A sensitive, stereoselective assay using solid phase extraction and LC-MS-MS was developed and validated for the analysis of (R)- and (S)-bupropion and its major metabolite (R,R)- and (S,S)-hydroxybupropion in human plasma and urine. Plasma or glucuronidase-hydrolyzed urine was acidified, then extracted using a Waters Oasis MCX solid phase 96-well plate. HPLC separation used an alpha(1)-acid glycoprotein column, a gradient mobile phase of methanol and aqueous ammonium formate, and analytes were detected by electrospray ionization and multiple reaction monitoring with an API 4000 Qtrap. The assay was linear in plasma from 0.5 to 200 ng/ml and 2.5 to 1000 ng/ml in each bupropion and hydroxybupropion enantiomer, respectively. The assay was linear in urine from 5 to 2000 ng/ml and 25 to 10,000 ng/ml in each bupropion and hydroxybupropion enantiomer, respectively. Intra- and inter-day accuracy was >98% and intra- and inter-day coefficients of variations were less than 10% for all analytes and concentrations. The assay was applied to a subject dosed with racemic bupropion. The predominant enantiomers in both urine and plasma were (R)-bupropion and (R,R)-hydroxybupropion. This is the first LC-MS/MS assay to analyze the enantiomers of both bupropion and hydroxybupropion in plasma and urine.
开发并验证了一种使用固相萃取和液相色谱 - 串联质谱的灵敏、立体选择性分析方法，用于分析人血浆和尿液中的（R）-和（S）-安非他酮及其主要代谢物（R，R）-和（S，S）-羟基安非他酮。将血浆或经葡萄糖醛酸酶水解的尿液酸化，然后使用沃特世Oasis MCX固相96孔板进行萃取。高效液相色谱分离使用α(1)-酸性糖蛋白柱、甲醇和甲酸铵水溶液的梯度流动相，通过电喷雾电离和API 4000 Qtrap进行多反应监测来检测分析物。该分析方法在血浆中，每种安非他酮和羟基安非他酮对映体的线性范围分别为0.5至200 ng/ml和2.5至1000 ng/ml。在尿液中，每种安非他酮和羟基安非他酮对映体的线性范围分别为5至2000 ng/ml和25至10,000 ng/ml。对于所有分析物和浓度，日内和日间准确度均>98%，日内和日间变异系数均小于10%。该分析方法应用于一名服用消旋安非他酮的受试者。尿液和血浆中的主要对映体均为（R）-安非他酮和（R，R）-羟基安非他酮。这是首个用于分析血浆和尿液中安非他酮和羟基安非他酮对映体的液相色谱 - 串联质谱分析方法。
