Wang Wen-Yu, Wang Wei, Wu Hong, Li Shu-Ping, Wang Rong, Dai Li, Zhang Zheng-Rong, Fu Jun, Deng Ran, Li Feng
College of Pharmacy, Anhui University of Chinese Medicine, Key Laboratory of Modernized Chinese Medicine in Anhui Province, Hefei, Anhui, China.
Rapid Commun Mass Spectrom. 2018 Mar 30;32(6):516-522. doi: 10.1002/rcm.8056.
Microdialysis has been used to detect the concentrations of drugs in tissues. Geniposide (GE), an iridoid glycoside compound, is the main bioactive component of Gardenia jasminoides Ellis fruit. We previously demonstrated that GE could control the activity of cytokines and reduce levels of inflammation in adjuvant arthritis (AA) rats, but the topic of concentration changes over time in the joint synovia of AA has rarely been studied.
In this study, a microdialysis technique combined with ultra-high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (UHPLC/ESI-MS/MS) was set up and confirmed to assay GE in the dialysate of the joint cavity in AA rats. Mass detection was conducted in multiple reaction monitoring (MRM) mode with negative ESI, and paeoniflorin (Pae) was used as an internal standard (IS).
A lower limit of quantitation (LLOQ) of 5 ng/mL was found using this method and with good linearity in the range of 5-4000 ng/mL. All the validation data including accuracy, precision, intra and inter-day repeatability and stability meet the requirements. The relative recoveries of GE were determined at approximately 40.01%.
The measurements based on microdialysis combined with UHPLC/ESI-MS/MS provide a method for sampling and rapid sensitive analysis of GE in the dialysate of the joint cavity in AA rats. This method should be considered for future pharmacokinetics studies.
微透析已被用于检测组织中的药物浓度。栀子苷(GE)是一种环烯醚萜苷类化合物,是栀子果实的主要生物活性成分。我们之前证明GE可以控制细胞因子的活性并降低佐剂性关节炎(AA)大鼠的炎症水平,但关于AA大鼠关节滑膜中GE浓度随时间变化的研究很少。
在本研究中,建立了一种结合超高效液相色谱/电喷雾电离串联质谱(UHPLC/ESI-MS/MS)的微透析技术,并经证实可用于测定AA大鼠关节腔透析液中的GE。采用负电喷雾电离在多反应监测(MRM)模式下进行质谱检测,芍药苷(Pae)用作内标(IS)。
使用该方法发现定量下限(LLOQ)为5 ng/mL,在5-4000 ng/mL范围内具有良好的线性。所有验证数据,包括准确度、精密度、日内和日间重复性以及稳定性均符合要求。GE的相对回收率约为40.01%。
基于微透析结合UHPLC/ESI-MS/MS的测量为AA大鼠关节腔透析液中GE的采样和快速灵敏分析提供了一种方法。未来的药代动力学研究应考虑使用该方法。