College of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan, Gyeonggi-do 15588, Korea.
Department of Pharmacognosy, Faculty of Pharmacy, University of Sindh, Jamshoro 76088, Pakistan.
Molecules. 2020 Jul 6;25(13):3077. doi: 10.3390/molecules25133077.
A comprehensive linear gradient solvent system for centrifugal partition chromatography (CPC) was developed for the bioassay-guided isolation of natural compounds. The gradient solvent system consisted of three different ternary biphasic solvents types: -hexane-acetonitrile-water (10:2:8, /), ethyl acetate-acetonitrile-water (10:2:8, /), and water-saturated -butanol-acetonitrile-water (10:2:8, /). The lower phase of the -hexane-acetonitrile-water (10:2:8, /) was used as the stationary phase, while its upper phase, as well as ethyl acetate-acetonitrile-water (10:2:8), and water-saturated -butanol-acetonitrile-water (10:2:8, /) were pumped to generate a linear gradient elution, increasing the mobile phase polarity. We used the gradient CPC to identify antioxidant response elements (AREs), inducing compounds from , using an ARE-luciferase assay in HepG2 cells, which led to the purification of the active molecules 3-methoxyquercetin and brevilin A. The developed CPC solvent systems allow the separation and isolation of compounds with a wide polarity range, allowing active molecule identification in the complex crude extract of natural products.
开发了一种用于离心分配色谱（CPC）的综合线性梯度溶剂系统，用于基于生物测定的天然化合物的分离。梯度溶剂系统由三种不同的三元双相溶剂类型组成：-己烷-乙腈-水（10:2:8，/）、乙酸乙酯-乙腈-水（10:2:8，/）和水饱和正丁醇-乙腈-水（10:2:8，/）。-己烷-乙腈-水（10:2:8，/）的下相用作固定相，而上相以及乙酸乙酯-乙腈-水（10:2:8）和水饱和正丁醇-乙腈-水（10:2:8，/）被泵送到产生线性梯度洗脱，增加流动相的极性。我们使用梯度 CPC 通过 HepG2 细胞中的 ARE-荧光素酶测定法从 中鉴定抗氧化反应元件（ARE）诱导化合物，从而纯化了活性分子 3-甲氧基槲皮素和布瑞维林 A。开发的 CPC 溶剂系统允许分离和分离具有广泛极性范围的化合物，允许在天然产物的复杂粗提取物中鉴定活性分子。
