Yang Shengbao, Qu Ruiying, Sun Peilu, Xiong Shan, Yan Siyi, Deng Zhipeng
Jiaozhou People's Hospital, Qingdao, China.
Shandong Academy of Medical Sciences, Institute of Materia Medica, Jinan, China.
Biomed Chromatogr. 2018 Apr;32(4). doi: 10.1002/bmc.4138. Epub 2017 Dec 6.
Tubuloside B, a novel neuroprotective phenylethanoid, is a major active constituent of Cistanche tubulosa and Cistanche deserticola. A specific and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantification of tubuloside B in rat plasma. Sample preparation was conducted through a protein-precipitation extraction with methanol using tubuloside A as internal standard (IS). Chromatographic separation was achieved using a Capcell Pak C column (2.0 × 50 mm, 5 μm) with a mobile phase of methanol-10 mm ammonium acetate buffer (70:30, v/v) in an isocratic elution. Mass spectrometry analysis was performed in negative ionization mode with selected reaction monitoring transitions at m/z 665.1 → 160.9 for tubuloside B, and m/z 827.1 → 160.9 for IS. Calibration curves were linear over the range of 1.64-1640 ng/mL for plasma samples samples (R > 0.990). The lower limit of quantification (LLOQ) was 1.64 ng/mL. The intra- and inter-day accuracy was between 92.3 and 113.0% with the RSD <9.23% at all LLOQ and quality control levels. Finally, this method was successfully applied in the pharmacokinetics study of tubuloside B after intravenous administration.
管花苷B是一种新型神经保护苯乙醇苷类化合物，是管花肉苁蓉和荒漠肉苁蓉的主要活性成分。已开发并验证了一种特异性和灵敏的液相色谱串联质谱（LC-MS/MS）方法，用于定量大鼠血浆中的管花苷B。样品制备采用以管花苷A为内标（IS）的甲醇蛋白沉淀萃取法。使用Capcell Pak C柱（2.0×50 mm，5μm）进行色谱分离，流动相为甲醇-10 mM醋酸铵缓冲液（70:30，v/v），等度洗脱。质谱分析在负离子模式下进行，管花苷B的选择反应监测跃迁为m/z 665.1→160.9，内标的选择反应监测跃迁为m/z 827.1→160.9。血浆样品的校准曲线在1.64-1640 ng/mL范围内呈线性（R>0.990）。定量下限（LLOQ）为1.64 ng/mL。在所有LLOQ和质量控制水平下，日内和日间准确度在92.3%至113.0%之间，相对标准偏差（RSD）<9.23%。最后，该方法成功应用于管花苷B静脉给药后的药代动力学研究。
