Li Jianbo, Jin Yue, Fu Huiyan, Huang Yihui, Wang Xianqin, Zhou Yunfang
Yuhang Branch, Second Affiliated Hospital of Zhejiang University, Hangzhou, China.
Analytical and testing Centre, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, China.
Biomed Chromatogr. 2019 Mar;33(3):e4418. doi: 10.1002/bmc.4418. Epub 2018 Nov 22.
Gelsenicine is an indole alkaloid isolated from Gelsemium elegans Benth. In recent years, the role of G. elegans Benth preparations in anti-tumor, analgesic, dilatation and dermatological treatment has attracted attention, and it has been applied clinically, but it is easy to cause poisoning with its use. An UPLC-MS/MS method was established to determine the gelsenicine in mouse blood, and the pharmacokinetics of gelsenicine after intravenous (0.1 mg/kg) and intragastric (0.5 and 1 mg/kg) administration was studied. Deltalin was used as internal standard; a UPLC BEH C column was used for chromatographic separation. The mobile phase consisted of acetonitrile and 10 mmol/L ammonium acetate (0.1% formic acid) with a gradient elution flow rate of 0.4 mL/min. Multiple reaction monitoring mode was used for quantitative analysis of gelsenicine in electrospray ionization positive interface. Proteins from mouse blood were removed by acetonitrile precipitation. A validation of this method was performed in accordance with the US Food and Drug Administration guidelines. In the concentration range of 0.05-100 ng/mL, the gelsenicine in the mouse blood was linear (r > 0.995), and the lower limit of quantification was 0.05 ng/mL. In the mouse blood, the intra-day precision RSD was <12%, the inter-day precision RSD was <15%, the accuracy ranged from 89.8 to 112.3%, the average recovery was >76.8%, and the matrix effect was between 103.7 and 108.4%, which meet the pharmacokinetic research requirements of gelsenicine. The UPLC-MS/MS method is sensitive, rapid and selective, and has been successfully applied to the pharmacokinetic study of gelsenicine in mice. The absolute bioavailability of gelsenicine is 1.13%.
钩吻素甲是从断肠草中分离得到的一种吲哚生物碱。近年来，断肠草制剂在抗肿瘤、镇痛、扩张血管及皮肤科治疗方面的作用受到关注，并已应用于临床，但使用时易引起中毒。建立了一种超高效液相色谱-串联质谱法（UPLC-MS/MS）测定小鼠血液中的钩吻素甲，并研究了静脉注射（0.1 mg/kg）和灌胃（0.5和1 mg/kg）给药后钩吻素甲的药代动力学。以δ-生育三烯酚为内标；采用UPLC BEH C柱进行色谱分离。流动相由乙腈和10 mmol/L醋酸铵（0.1%甲酸）组成，梯度洗脱流速为0.4 mL/min。在电喷雾电离正离子模式下，采用多反应监测模式对钩吻素甲进行定量分析。通过乙腈沉淀去除小鼠血液中的蛋白质。按照美国食品药品监督管理局的指导原则对该方法进行了验证。在0.05-100 ng/mL的浓度范围内，小鼠血液中的钩吻素甲呈线性（r＞0.995），定量下限为0.05 ng/mL。在小鼠血液中，日内精密度RSD＜12%，日间精密度RSD＜15%，准确度在89.8%至112.3%之间，平均回收率＞76.8%，基质效应在103.7%至108.4%之间，符合钩吻素甲的药代动力学研究要求。该UPLC-MS/MS方法灵敏、快速且具有选择性，已成功应用于钩吻素甲在小鼠体内的药代动力学研究。钩吻素甲的绝对生物利用度为1.13%。
