Han Xiaoli, He Jinhua, Chen Qinyue, Sun Yali, Zhang Xiulei, Gu Zhengyi, Sha Xianyi
Key Laboratory of Smart Drug Delivery (Fudan University), Ministry of Education, Shanghai, China.
Department of Pharmaceutics, School of Pharmacy, Fudan University, Shanghai, China.
Biomed Chromatogr. 2018 Dec;32(12):e4356. doi: 10.1002/bmc.4356. Epub 2018 Aug 29.
Artemitin, a significant flavonol compound existing in Laggera pterodonta (DC.) Benth., Artemisia rupestris L, etc., is the subject of attention by researchers owing to its pharmacological activities (such as antioxidative, anti-inflammatory and antiviral). In this work, a highly sensitive and specific high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) assay combined with protein precipitation has been established and validated for determining artemitin concentration in rat plasma. Both artemitin and warfarin sodium (internal standard, IS) were separated on an Agela Venusil XBP Phenyl column through the isocratic elution mode of methanol-water containing 0.1% formic acid (80:20, v/v), at a flow rate of 0.4 mL/min. The MS/MS system was operated in a positive ion and ESI multiple reaction monitoring mode, and the multiple reaction monitoring transition was optimized as m/z 389.0 → 373.0 for artemitin and 309.2 → 163.0 for IS. The method showed good linearity in the range of 2.5-2000 ng/mL (R = 1.0000) and high sensitivity for artemitin with the lower limit of quantification of 2.5 ng/mL. The intra- and inter-day accuracies were 97.4-100.9 and 93.4-100.3%, respectively. The intra- and inter-day precisions were <4.8 and 6.5%, respectively. The extraction efficiency and absolute recovery were >66.5 and 71.3%, respectively. In addition, a good matrix effect of <9.5% was obtained. As a result, the method developed herein was successfully applied for the pharmacokinetic study of artemitin after an intravenous administration in rats.
青蒿素是存在于野马追（DC.）Benth.、岩蒿等植物中的一种重要黄酮醇化合物，因其药理活性（如抗氧化、抗炎和抗病毒）而受到研究人员的关注。在本研究中，建立了一种结合蛋白沉淀的高灵敏度、高特异性高效液相色谱-电喷雾电离-串联质谱（HPLC-ESI-MS/MS）测定法，并用于测定大鼠血浆中青蒿素的浓度。青蒿素和华法林钠（内标，IS）在Agela Venusil XBP苯基柱上通过含0.1%甲酸（80:20，v/v）的甲醇-水等度洗脱模式进行分离，流速为0.4 mL/min。MS/MS系统在正离子和ESI多反应监测模式下运行，青蒿素的多反应监测跃迁优化为m/z 389.0→373.0，内标的多反应监测跃迁优化为m/z 309.2→163.0。该方法在2.5 - 2000 ng/mL范围内线性良好（R = 1.0000），对青蒿素具有高灵敏度，定量下限为2.5 ng/mL。日内和日间准确度分别为97.4 - 100.9%和93.4 - 100.3%。日内和日间精密度分别<4.8%和6.5%。提取效率和绝对回收率分别>66.5%和71.3%。此外，获得了<9.5%的良好基质效应。因此，本文建立的方法成功应用于大鼠静脉注射青蒿素后的药代动力学研究。
