液相色谱串联质谱法测定人体血浆维拉帕米浓度及生物等效性评价研究

 目的 建立LC-MS/MS测定人体血浆中维拉帕米的浓度。方法 色谱柱为Thermo Hypurity C₁₈ (2.1 mm×150 mm， 5 μm)，流动相为10 mmol·L^-1甲酸铵(含0.1%甲酸)-乙腈（35∶65）；流速：0.3 mL·min^-1；进样量：10 μL；柱温：40 ℃，样品室温度为15 ℃。ESI源正离子扫描，维拉帕米和内标丙咪嗪离子监测分别为（m/z）455.0→165.0和（m/z）281.0→86.0。结果 维拉帕米线性范围为0.457 5～234.20 ng·mL^-1， 维拉帕米最低检测限为0.1 ng·mL^-1，方法灵敏、稳定、特异性高， 并成功地应用到人体维拉帕米药动学的研究。结论 该方法简便、准确、重复性好， 可以准确地定量人体血浆维拉帕米的浓度。

LC-MS/MS Determination of Verapamil in Human Plasma and Its Application in Bioequivalence Evaluation

OBJECTIVE To establish a LC-MS/MS method for determination of verapamil in human plasma.METHODS Thermo Hypurity C18 column(2.1 mm×150 mm,5 μm) was used in the experiment.The column temperature was set at 40 ℃.10 mmol·L-1 ammonium(including 0.1% ammonium formate)-acetonitrile(35∶65) was used as the mobile phase and the flow rate was 0.3 mL·min-1.The sample room temperature was set at 15 ℃ and the injection volume was 10 μL.ESI source and positive ion scan were used,verapamil and the internal standard imipramine were monitored at(m/z) 455.0→165.0 and(m/z) 281.0→86.0,respectively.RESULTS The calibration curve of verapamil was linear in the range of 0.457 5-234.20 ng·mL-1,and the detection limit for verapamil was about 0.1 ng·mL-1.The method had high sensitivity,stability and specificity and had been used for the pharmacokinetic study of verapamil in vivo successfully.CONCLUSION The method is simple,accurate and repetitive for the determination of verapamil in human plasma and suitable for pharmacokinetic study.