School of Pharmacology, Henan University of Chinese Medicine, Zhengzhou, 450046, China; Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment & Chinese Medicine Development of Henan Province, Zhengzhou, 450046, China; Henan Key Laboratory of Chinese Medicine for Respiratory Disease, Zhengzhou 450046, China.
School of Pharmacology, Henan University of Chinese Medicine, Zhengzhou, 450046, China; Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment & Chinese Medicine Development of Henan Province, Zhengzhou, 450046, China.
J Pharm Biomed Anal. 2018 Jan 30;148:205-213. doi: 10.1016/j.jpba.2017.10.006. Epub 2017 Oct 13.
A rapid, sensitive and selective method based on high-performance liquid chromatography coupled with Q-Exactive mass spectrometry was developed and validated for simultaneous quantification of thirteen components in rat plasma, including chlorogenic acid, 5-caffeoylquinic acid, 4-caffeoylquinic acid, luteoloside, isochlorogenic acid A, B and C, scutellarin, baicalin, wogonin, baicalein, phillyrin and forsythoside A. After precipitating proteins from the plasma samples with methanol, chromatographic separation of the thirteen components was achieved by using an XBridge™ C column (2.1mm×150mm, 5μm) with the mobile phase consisting of methanol and 0.1% formic acid in water at a flow rate of 0.3mL/min. High-resolution MS quantification was adopted with detection on a Q-Exactive mass spectrometer in full-scan mode, and the results were obtained using a mass extraction window of 10ppm at a mass resolution of 70, 000. All the calibration curves exhibited good linearity (r>0.991) over the measured ranges. The lower limit of quantitation (LLOQ) was in the range of 1.05-8.13ng/mL. The intra- and inter-day precision (RSD) was less than 11.70% and the accuracy (RE) ranged from -5.58% to 12.29%. No significant matrix effect was observed and the extraction recoveries of all the analytes were more than 79.36%. The developed method was applied to a pharmacokinetic study of the thirteen ingredients in rats after intravenous administration of Tanreqing at three doses of 3, 6 and 12mL/kg. The results indicated that 8 of the 13 components, isochlorogenic acid A, B and C, chlorogenic acid, baicalin, wogonin, luteoloside and forsythoside A, had linear pharmacokinetic properties in the tested dosage range.
建立并验证了一种基于高效液相色谱-四极杆静电场轨道阱高分辨质谱联用的方法，用于同时定量测定大鼠血浆中的 13 种成分，包括绿原酸、5-咖啡酰奎宁酸、4-咖啡酰奎宁酸、芦丁、异绿原酸 A、B 和 C、汉黄芩苷、黄芩苷、木樨草素、黄芩素、汉黄芩素和连翘酯苷 A。用甲醇沉淀血浆样品中的蛋白质后，采用 XBridge™ C 柱（2.1mm×150mm，5μm），以甲醇和 0.1%甲酸水溶液为流动相，流速为 0.3mL/min，实现了 13 种成分的色谱分离。采用 Q-Exactive 质谱仪全扫描模式进行高分辨 MS 定量检测，在质量分辨率为 70,000 时，采用 10ppm 的质量提取窗口获得结果。所有校准曲线在测定范围内均具有良好的线性（r＞0.991）。定量下限（LLOQ）范围为 1.05-8.13ng/mL。日内和日间精密度（RSD）均小于 11.70%，准确度（RE）范围为-5.58%至 12.29%。未观察到明显的基质效应，所有分析物的提取回收率均大于 79.36%。该方法应用于三种剂量（3、6 和 12mL/kg）静脉注射痰热清后大鼠 13 种成分的药代动力学研究。结果表明，在测试剂量范围内，有 8 种成分（异绿原酸 A、B 和 C、绿原酸、黄芩苷、木樨草素、芦丁和连翘酯苷 A）具有线性药代动力学特性。
