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目的:建立液相色谱-串联质谱法(LC-MS/MS)用于血浆中α-细辛脑浓度的测定。方法:取家兔血浆样品,加入适量内标(4-甲氧基苯甲酮),用甲醇沉淀蛋白,经高效液相色谱分离,色谱柱:ODS C_(18)分析柱(50mm×2.0mm,5μm),流动相:甲醇-1%醋酸水溶液(74:26),流速0.3mL·min~(-1),采用 APCI 离子化-串联质谱多离子反应监测(MRM)模式进行检测,放电电流(NC)=2,雾化气(GS1)=45.00,辅助气(GS2)=45.00,气帘气(CUR)=20.00,温度(TEM)=400.00,碰撞活化裂解(CAD)=Medium(以上单位均为仪器单位),离子选择通道分别为 m/z 209.40/194.10(α-细辛脑)和 m/z 213.40/105.10(内标)。结果:α-细辛脑血浆浓度在8～8000ng·mL~(-1)范围内线性关系良好(r=0.9987),日内和日间精密度试验的 RSD均小于16%,方法回收率为98.55%～103.4%,提取回收率为87.92%～93.22%,最低检测限为1ng·mL~(-1),血浆最低检测浓度为3ng·mL~(-1)。结论:建立的 LC-MS/MS 方法用于血浆中微量药物的浓度检测,具有专属性强、灵敏度高、准确性好的特点。
Objective: To establish liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the determination of α-asarone in plasma. METHODS: Rabbit plasma samples were taken, appropriate amount of internal standard (4-methoxybenzophenone) was added, protein was precipitated with methanol, and separated by high performance liquid chromatography. Column: ODS C 18 analytical column (50 mm × 2.0 mm) , 5μm), mobile phase: methanol-1% acetic acid aqueous solution (74:26), flow rate 0.3mL·min -1, detection by APCI ionization-tandem mass spectrometry and multi-ion reaction monitoring (MRM) mode, discharge current (NC)=2, atomizing gas (GS1)=45.00, auxiliary gas (GS2)=45.00, air curtain gas (CUR)=20.00, temperature (TEM)=400.00, collision activation cracking (CAD)=Medium (above units For the instrument unit), the ion-selection channels are m/z 209.40/194.10 (α-asarone) and m/z 213.40/105.10 (internal standard), respectively. Results: The linearity of α-asarone plasma concentration was in the range of 8-8000 ng·mL -1 (r=0.9987). The intra-day and inter-day precision RSD were less than 16%, and the method recovery was 98.55. %～103.4%, extraction recovery rate was 87.92%-93.22%, the lowest detection limit was 1 ng·mL -1, and the lowest plasma concentration was 3ng·mL -1 . Conclusion: The established LC-MS/MS method was used to detect the concentration of trace drugs in plasma. It was characterized by high specificity, high sensitivity, and good accuracy.