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Simultaneous UPLC–TQ-MS/MS determination of six active components in rat plasma: application in the pharmacokinetic study of Cyclocarya paliurus leaves

机译:同时用UPLC-TQ-MS / MS测定大鼠血浆中的六种活性成分:在青冈叶中药代动力学研究中的应用

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Cyclocarya paliurus (Batal.) Ijinskaja (CP) is a monotypic genus plant, also called sweet tea tree that belongs to the Juglandaceae family, which is mainly distributed in the subtropical highlands in China. Our previous work has verified that CP leaves exhibit a potent hyperglycemic effect by inhibiting pancreatic β cell apoptosis through the regulation of MPAK and Akt signaling pathways. However, the components that contribute to this potential health benefit remain undiscovered. A sensitive, reliable, and validated ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC–TQ-MS/MS) method was developed to simultaneously determine the presence of six active components (neochlorogenic acid, chlorogenic acid, quercetin-3-O-glucuronide, kaempferol-3-O-rhamnoside, quercetin, and kaempferol) in rat plasma after a single oral administration (in a dosage of 10.5?g/kg) of an extract of CP leaves to rats. The separation was performed on a Waters ACQUITY BEH C18 column (50?mm?×?2.1?mm, 1.7?μm). The detection was conducted by multiple reaction monitoring (MRM) in negative ionization mode. The two highest abundant MRM transitions without interference were optimized for each analyte. Acetonitrile and formic acid aqueous solution (0.1%) was used as the mobile phase at a flow rate of 0.3?ml/min. The precision, accuracy, and recovery all satisfied the criteria of international guidance (Bioanalytical Method Validation Guidance for Industry, Food and Drug Administration), and the analytes were stable in plasma for all tested conditions. The main pharmacokinetic parameters were calculated by plasma concentration versus time profiles using the pharmacokinetics program. The pharmacokinetic parameters of each compound can facilitate future clinical studies.
机译:Cyclocarya paliurus(Batal。)Ijinskaja(CP)是单型属植物,也称为甜茶树,属于胡桃木科,主要分布于中国的亚热带高原。我们以前的工作已经证实,CP叶片通过调节MPAK和Akt信号通路抑制胰腺β细胞凋亡,从而显示出有效的高血糖作用。但是,仍未发现有助于这种潜在健康益处的成分。开发了灵敏,可靠且经过验证的超高效液相色谱结合三重四极杆串联质谱(UPLC–TQ-MS / MS)方法,可同时测定六个活性成分(新绿原酸,绿原酸,槲皮素-在大鼠单次口服CP提取物(剂量为10.5?g / kg)后,在大鼠血浆中产生3-O-葡萄糖醛酸苷,山奈酚3-O-鼠李糖苷,槲皮素和山奈酚。分离是在Waters ACQUITY BEH C18柱上进行的(50?mm?×?2.1?mm,1.7?μm)。通过负电离模式下的多反应监测(MRM)进行检测。对于每种分析物,优化了两个最高的无干扰的MRM跃迁。乙腈和甲酸水溶液(0.1%)用作流动相,流速为0.3?ml / min。精密度,准确性和回收率均满足国际指南(工业,食品和药物管理局的生物分析方法验证指南)的标准,并且在所有测试条件下,分析物在血浆中均稳定。使用药代动力学程序通过血浆浓度与时间的关系曲线计算出主要的药代动力学参数。每种化合物的药代动力学参数可以促进将来的临床研究。

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