Simultaneous Determination of Seven Components in Rat Plasma by the UPLC-MS/MS Method and Application of Pharmacokinetic Studies to SimiaoYong'an Decoction.

Simultaneous Determination of Seven Components in Rat Plasma by the UPLC-MS/MS Method and Application of Pharmacokinetic Studies to SimiaoYong'an Decoction.
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DOI:
10.3390/molecules22111937
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发表时间:
2017-11-09
期刊:
Molecules (Basel, Switzerland)
影响因子:
--
通讯作者:
Liu B
Liu B
中科院分区:
其他
文献类型:
--
作者:
Liu Y;Chi S;Wang W;Su L;Liu B

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四妙永安汤(SYD)是治疗坏疽、清热解毒、止痛的经典方剂。我们开发了一种灵敏的超高效液相色谱-串联质谱(UPLC-MS/MS)方法,用于同时测定大鼠血浆中SYD提取物的七种主要活性成分(即哈巴苷、绿原酸、苦菜苷、马钱素、甘草苷、安果苷C和哈巴苷)。血浆分析的初步步骤是添加内标(例如 linarin),然后用甲醇沉淀蛋白质。采用 ACQUITY UPLC® BEH C18 色谱柱(100 mm × 2.1 mm,1.7 μm)以 0.2 mL/min 的流速,以甲醇/水 0.1% 甲酸水溶液 (v/v) 作为流动相,进行活性成分的分离。在多反应监测 (MRM) 模式下通过负离子电喷雾电离在三重四极杆串联 MS (QqQ-MS) 上进行检测。所有校准曲线在浓度范围内均表现出良好的线性 (r > 0.99),定量下限在 0.029 至 5.813 ng/mL 之间。通过日内和日间测定评估精密度,RSD百分比均在8.1%以内。提取效率和基质效应分别为80.6-113.6%和82.9-99.5%。经验证的方法已成功应用于大鼠口服SYD提取物和相应的单味和组合中药(TCM)后的药代动力学研究。由于不同共存成分之间的相互作用,七种成分的药代动力学特性呈现出动态变化。事实证明,所建立的方法对于中药活性成分的研究是有用的。
SimiaoYong’an Decoction (SYD) is a classical traditional Chinese prescription that is used for the treatment of gangrene, heat-clearing, detoxification and pain alleviation. We developed a sensitive ultra-performance liquid chromatography-tandem mass spectrum (UPLC-MS/MS) method for the simultaneous determination of seven major active ingredients of SYD extract (i.e., harpagide, chlorogenic acid, sweroside, loganin, liquiritin, angoroside C and harpagoside) in rat plasma. The preliminary steps in the plasma analysis were the addition of an internal standard such as linarin, followed by protein precipitation with methanol. Separation of the active ingredients was performed on an ACQUITY UPLC® BEH C18 column (100 mm × 2.1 mm, 1.7 μm) at a flow rate of 0.2 mL/min with methanol/water 0.1% formic acid aqueous (v/v) as the mobile phase. Detection was performed on a triple quadrupole tandem MS (QqQ-MS) via negative ion electrospray ionization in multiple reactions monitoring (MRM) mode. All calibration curves showed good linearity (r > 0.99) over the concentration range with a low limit of quantification between 0.029 and 5.813 ng/mL. Precision was evaluated by intra-day and inter-day assays, and the percentages of the RSD were all within 8.1%. The extraction efficiency and matrix effect were 80.6–113.6% and 82.9–99.5%, respectively. The validated method was successfully applied to a pharmacokinetic study in rats after oral administration of SYD extract and the corresponding single and combined traditional Chinese medicines (TCMs). The pharmacokinetic properties of the seven ingredients showed dynamic changes due to counteraction among the different coexisting components. The established approach has proven useful in the study of the active constituents in a traditional Chinese prescription.
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