液相色谱-串联质谱法测定Beagle犬血浆中升麻苷H-1的浓度

目的: 建立液相色谱-串联质谱的方法测定Beagle犬血浆中升麻苷H-1的浓度。 方法: 采用20(S)-人参皂苷Rg3为内标,血浆样品经液-液萃取后,以甲醇和水为流动相,梯度洗脱进行分离。通过液相色谱-串联质谱法以多反应监测(MRM)方式检测。用于定量的离子反应分别为m/z 615.5→369.6(升麻苷H-1)和783.5→161.1(内标人参皂苷Rg3)。 结果: 血浆中升麻苷H-1的线性范围为0.5~1000 ng·mL^-1,定量下限为0.5 ng·mL^-1。在升麻苷H-1浓度为2.0,50,500 ng·mL^-1时的提取回收率分别为90.4%,94.0%,89.0%。方法的日内和日间精密度(RSD)分别为5.9%,4.7%,3.8%和10.8%,6.3%,7.8%。 结论: 该方法灵敏、准确,重复性好,可用于Beagle犬血浆中升麻苷H-1的浓度测定。

Objective: To establish a liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for the determination of cimicifugoside H-1 in Beagle dog plasma. Method: The structural analogues 20 (S)-ginsenoside Rg3 was used as the internal standard.After liquid-liquid extraction,samples were separated by a gradient elution procedure and the mobile phase consisted of methanol and water.The quantitation was performed on an API-4000 HPLC-MS/MS with negative electrospray ionization in multiple reaction monitoring (MRM) mode.The precursor to product ion transitions for cimicifugoside H-1 and 20 (S)-ginsenoside Rg3 (IS) were 615.5→369.6 and 783.5→161.1,respectively. Results: The assay was reproducible and linear for cimicifugoside H-1 in the range of 0.5-1000 ng·mL^-1.The lower limit of quantification (LLOQ) was 0.5 ng·mL^-1.The extract recovery was 90.4%,94.0% and 89.0% for cimicifugoside H-1 at concentrations of 2.0,50,500 ng·mL^-1,the intra-and inter-day precision of analysis was 5.9%,4.7%,3.8% and 10.8%,6.3%,7.8%,respectively. Conclusion: The method is sensitive,accurate and suitable for determination of cimicifugoside H-1 in Beagle dog plasma.