UPLC-Q-TOF-MS筛选穿心莲中亲和Caco-2细胞的活性成分

目的采用超高效液相色谱-四级杆飞行时间质谱(UPLC-Q-TOF-MS)技术筛选穿心莲与Caco-2细胞亲和的活性成分。方法利用Caco-2细胞进行药物摄取结合药物吸收转运实验共同筛选穿心莲的亲和活性成分,采用UPLC-Q-TOF-MS测定亲和前后的细胞样品。根据各化学成分的保留时间、质谱信息,并结合提取离子流图与对照品、相关文献质谱数据对比鉴定各亲和活性成分。结果从穿心莲中共筛选出10种能与Caco-2细胞亲和的活性成分,分别为12,13-双氢穿心莲内酯(1)、黄芩黄酮I-2′-O-葡萄糖苷(2)、穿心莲内酯(3)、14-脱氧-11,12-二脱氢穿心莲内酯苷(4)、7-羟基脱水穿心莲内酯(5)、新穿心莲内酯(6)、3-去氧脱水穿心莲内酯苷(7)、去氧穿心莲内酯(8)和脱水穿心莲内酯(9)和1种未知成分。结论结合UPLC-Q-TOF-MS技术建立的筛选穿心莲中与Caco-2细胞亲和活性成分的方法,可用于筛选中药复杂体系中的活性成分,为进一步研究活性成分群间的协同配伍效应奠定基础。

Active components from Andrographis paniculata with affinity to Caco-2 cells screened by UPLC-Q-TOF-MS

Objective To identify the active components of Andrographis paniculata coupled with Caco-2 cells screened by UPLC-Q-TOF-MS. Methods The cell-combining compounds from A. paniculata were screened based on drug uptake and transport absorption experiment using Caco-2 cells, then the cell dissociation solution was detected before and after treating with A. paniculata by using UPLC-Q-TOF-MS method. The cell-combining compounds from the ethanol extracts of A. paniculata were identified based on the retention time and mass spectrometry information of each chemical composition combined with comparing the extracted ion chromatograms and mass spectrometry datas of reference substances and the related articles. Results A total of 10 compounds combined with Caco-2 cells in ethanol extract of A. paniculata, which were detected and identified as 12,13-dihydroandrographolide (1), skullcapflavone I-2''-O-glucoside (2), andrographolide (3), 14-deoxy-11,12-didehydro andrographiside (4), 7-hydroxy dehydroandrographolide (5), neoandrographolide (6), 3-deoxy dehydroandrographiside (7), deoxyandrographolide (8), dehydroandrographolide (9), and one unidentified component. Conclusion This research establishes a method of screening the active components of A. paniculata coupled with Caco-2 cells using UPLC-Q-TOF-MS analysis, which can be used to screen out the active components of the complex system on traditional Chinese medicines, as well as to lay the foundation for further study of the synergetic compatibility effects among active ingredients.