Department of Pharmacy, The First Affiliated Hospital of China Medical University, 155 Nanjing Street, Shenyang 110001, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 May 15;897:50-4. doi: 10.1016/j.jchromb.2012.04.007. Epub 2012 Apr 12.
A simple and rapid high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method has been developed and validated for the determination of 4-p-anisamidobutyric acid (ABA; or N-anysoyl-γ-aminobutiryc acid, N-anisoyl-GABA), a major active metabolite of aniracetam, in human plasma. After protein precipitation of plasma sample with methanol, ABA and the internal standard lisinopril were separated on a Venusil ASB C₁₈ column at 25 °C. The mobile phase consisted of methanol-ammonium acetate (10 mmol/L) (30:70, v/v). The detection was performed on a triple quadrupole tandem mass spectrometer with an ESI source in negative ion mode. Multiple reaction monitoring (MRM) using the precursor→product ion combinations of m/z 235.8→m/z 106.6, and m/z 403.8→m/z 113.6 was used to quantify ABA and lisinopril, respectively. This is the first LC-MS/MS method for ABA with advantages of short analysis time (4.5 min per sample run) and high selectivity attributable to the MRM detection and optimized HPLC conditions. The response was linear in a concentration range of 0.0485-19.4 μg/mL in plasma. The extraction recovery of ABA was between 89.1% and 100.7%. The precision (RSD) and accuracy (RE) of the method were evaluated to be within 7.3% and from 2.5% to 6.9%. The validated method has been applied to the pharmacokinetic study after a single oral administration of aniracetam dispersible tablets to human beings.
建立并验证了一种简单、快速的高效液相色谱-电喷雾串联质谱（LC-ESI-MS/MS）法，用于测定人血浆中作为茴拉西坦主要活性代谢物的 4-对茴香酰胺基丁酸（ABA；或 N-苯甲酰基-γ-氨基丁酸，N-苯甲酰基-GABA）。采用甲醇沉淀蛋白后，ABA 和内标赖诺普利在 25°C 下于 Venusil ASB C₁₈ 柱上进行分离。流动相由甲醇-乙酸铵（10mmol/L）（30:70，v/v）组成。检测采用电喷雾源在负离子模式下的三重四极杆串联质谱仪进行。采用母离子→子离子组合 m/z 235.8→m/z 106.6 和 m/z 403.8→m/z 113.6 的多重反应监测（MRM）分别定量 ABA 和赖诺普利。这是一种用于 ABA 的 LC-MS/MS 方法，具有分析时间短（每个样品运行 4.5 分钟）和高选择性的优点，这归因于 MRM 检测和优化的 HPLC 条件。在血浆中的浓度范围为 0.0485-19.4μg/mL 时，响应呈线性。ABA 的提取回收率在 89.1%至 100.7%之间。该方法的精密度（RSD）和准确度（RE）评价均在 7.3%和 2.5%至 6.9%范围内。该验证方法已应用于单次口服茴拉西坦分散片后人体的药代动力学研究。
