Wang Lin, Wen Yanqing, Meng Fanhao
School of Pharmacy, China Medical University, Shenyang, China.
Biomed Chromatogr. 2018 Jun;32(6):e4201. doi: 10.1002/bmc.4201. Epub 2018 Mar 2.
A simple, rapid and sensitive method using UPLC-MS/MS was established and validated for simultaneous determination of gelsemine and koumine in rat plasma after oral administration of Gelsemium elegans Benth extract. Plasma was performed with methanol precipitation and berberine was chosen as the internal standard. Plasma samples were separated on an Acquity UPLC® BEH C column (3.0 × 50 mm, 1.7 μm) with gradient elution using acetonitrile and 0.1% formic acid aqueous solution as the mobile phase at a flow rate of 0.4 mL/min. Multiple reaction monitoring mode in positive ion mode was utilized for detection. The calibration curves were linear over the range of 0.2-100 ng/mL for gelsemine and 0.1-50 ng/mL for koumine, with the lower limits of quantification 0.2 and 0.1 ng/mL, respectively. The intra- and inter-precision and accuracy were well within the acceptable ranges. The developed method was successfully applied to an in vivo pharmacokinetic study in rat after oral administration of 10 mg/kg Gelsemium elegans Benth extract.
建立并验证了一种使用超高效液相色谱-串联质谱法（UPLC-MS/MS）同时测定大鼠口服钩吻提取物后血浆中钩吻素甲和钩吻素子的简单、快速且灵敏的方法。血浆采用甲醇沉淀法处理，选择小檗碱作为内标。血浆样品在Acquity UPLC® BEH C18柱（3.0×50 mm，1.7 μm）上分离，以乙腈和0.1%甲酸水溶液作为流动相进行梯度洗脱，流速为0.4 mL/min。采用正离子模式下的多反应监测模式进行检测。钩吻素甲的校准曲线在0.2 - 100 ng/mL范围内呈线性，钩吻素子的校准曲线在0.1 - 50 ng/mL范围内呈线性，定量下限分别为0.2和0.1 ng/mL。批内和批间精密度及准确度均在可接受范围内。所建立的方法成功应用于大鼠口服10 mg/kg钩吻提取物后的体内药代动力学研究。
