School of Pharmaceutical Sciences, Shandong University of Traditional Chinese Medicine, No.4655 University Road, Changqing District, Jinan 250355, China and.
Department of Pharmacy, the First Hospital of Jilin University, No.71 Xinmin Street, Chaoyang District, Changchun 130021, China.
J Chromatogr Sci. 2020 Apr 22;58(3):234-240. doi: 10.1093/chromsci/bmz095.
Glaucocalyxin A, a novel potent negative Akt regulator, is a major active constituent of Rabdosia japonica. A simple, specific and sensitive ultra-high pressure liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the quantification of glaucocalyxin A in rat plasma, lung and brain tissues after intravenous administration. Sample preparation was carried out through a simple liqiud-liquid extraction with ethyl acetate using bullatine A as internal standard. The chromatographic separation was achieved by using an Agilent ZORBAX Eclipse Plus C18 column (3.0 mm × 100 mm, 1.8 μm) with a mobile phase of acetonitrile and water containing 0.1% formic acid in a gradient elution. Mass spectrometry analysis was conducted in positive ionization mode with multiple reaction monitoring transitions at m/z 333.2 → 157.1 for glaucocalyxin A and m/z 344.2 → 128.1 for IS. Calibration curves were linear over the ranges of 20.0-4000 ng/mL for both plasma and tissue samples (r2 > 0.99). The Lower limit of quantification (LLOQ) was 0.284 ng/mL. The intra-day and inter-day precision relative standard deviation (RSD%) were <14.9%, while the accuracy ranged from -12.5 to 17.0% for LLOQ and quality control samples. This UHPLC-MS/MS method was successfully applied in the pharmacokinetics, lung and brain tissue distributions of glaucocalyxin A after intravenous administration.
白杨素 A 是一种新型有效的 Akt 负调节剂，是罗布麻的主要活性成分。建立并验证了一种用于静脉给予后大鼠血浆、肺和脑组织中白杨素 A 定量分析的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法。样品制备采用乙酸乙酯进行简单的液液萃取，以 bullatine A 作为内标。采用 Agilent ZORBAX Eclipse Plus C18 柱（3.0mm×100mm，1.8μm），以含 0.1%甲酸的乙腈和水为流动相进行梯度洗脱，实现了色谱分离。质谱分析采用正离子化模式，在 m/z 333.2→157.1 处监测白杨素 A 的多重反应监测转换，在 m/z 344.2→128.1 处监测 IS。血浆和组织样品的定量范围均为 20.0-4000ng/mL（r2＞0.99）。定量下限（LLOQ）为 0.284ng/mL。日内和日间精密度相对标准偏差（RSD%）＜14.9%，而 LLOQ 和质控样品的准确度范围为-12.5%至 17.0%。该 UHPLC-MS/MS 方法成功应用于静脉给予后白杨素 A 的药代动力学、肺和脑组织分布研究。
