基于UPLC-LTQ-Orbitrap-MS及网络药理学的复方黄柏液治疗糖尿病足活性成分研究

该研究采用超高效液相色谱-线性离子阱-静电场轨道阱质谱(UPLC-LTQ-Orbitrap-MS)对复方黄柏液化学成分进行定性分析,色谱柱为Waters HSS T3-C18色谱柱(2. 1 mm×100 mm,1. 8μm),流动相为0. 1%甲酸水(A)-乙腈(B),梯度洗脱,流速0. 3 m L·min^-1。根据化合物的母离子、子离子的质谱信息,对照品比对、文献信息进行成分鉴定。在此基础上,利用网络药理学研究方法,对复方黄柏液化学成分进行靶标预测、功能富集以及分子对接验证,筛选出与糖尿病足相关的活性成分、阐释其作用机制。结果表明,从复方黄柏液中共鉴定了138种化合物,包括:生物碱28种,黄酮类16种,苯乙醇苷类11种,环烯醚类9种,环已基乙醇衍生物11种,酚酸及衍生物28种,木脂素类3种,萜类4种,挥发油及其他28种。从中共筛选出36种与糖尿病足相关的活性成分。靶标功能富集结果表明复方黄柏液主要从调控炎症反应、生长因子等7个功能方面实现对糖尿病足的治疗。该研究结合LTQ-Orbitrap-MS质谱分析与网络药理学研究方法,为复方黄柏液的有效成分的识别、质量标志物的筛选以及解析其作用机制提供参考。

Study on active components of Fufang Huangbai Ye for diabetic foot treatment by UPLC-LTQ-Orbitrap-MS and network pharmacology

Chemical constituents of the Fufang Huangbai Ye( FFHB) were analyzed and identified by UPLC-ESI-LTQ-OrbitrapMS. The analysis was performed on an Waters HSS T3 reverse phase column( 2. 1 mm×100 mm,1. 8 μm). The mobile phase consisting of 0. 1% aqueous formic acid( A) and acetonitrile( B) was used with gradient elution,and the flow rate was 0. 3 mL·min^-1.Based on the information of the accurate mass,the multistage fragment ions,the mass spectrometric data of the standard substance and the relative reference literature,the structure of the chemical constituents in FFHB were identified. Based on the identified compounds,network pharmacology study,including target prediction,functional enrichment,and molecular docking was applied to screen out the main active substances for treatment of diabetes foot and explore the potential mechanism. The results showed that a total of 138 compounds were identified,including 28 alkaloids,16 flavonoids,11 phenylethanoid glycosides,9 cycloolefins,11 cyclohexylethanol derivatives,28 phenolic acids and derivatives,3 lignans,4 terpenes,28 volatile oils and the others. Further,36 active substances for diabetes foot were screened out,and the functional enrichment showed the potential mechanism of FFHB were mainly seven functional items including inflammatory response,growth factor activity. This study combining the UPLC-LTQ-Orbitrap-MS technology and the network pharmacology provide a useful reference and basis for active compounds,quality control markers and the pharmacological mechanism of FFHB for diabetic foot treatment.