Cui Yulei, Shao Yun, Wang Qilan, Mei Lijuan, Tao Yanduo
Northwest Institute of Plateau Biology, College of Life Science, Chinese Academy of Sciences, Xining, Qinghai, P.R. China.
Key Laboratory of Tibetan Medicine Research, College of Life Science, Chinese Academy of Sciences, Xining, Qinghai, P.R. China.
J Chromatogr Sci. 2019 May 1;57(5):418-425. doi: 10.1093/chromsci/bmz010.
Herbal plants are significant for the reason that they have a great potential in discovering drug precursors. However, how to purify compounds with higher purity from them is a question which needs to be discussed. In present study, an offline 2D reversed-phase (RP) preparative liquid chromatography coupled with solid-phase extraction (SPE) method was successfully developed for the separation of flavonolignan diastereoisomers from Arenaria kansuensis. Based on the analysis of results, the major conclusion that we have drawn from it is a RP-SPE was selected for enriching target flavonolignan sample from A. kansuensis. After that, an ODS preparative column was used for 1D preparation, and the target sample (4.6 g) was divided into five fractions with a recovery of 83.9%. Then, a C18HCE preparative column, a polar-modified RP (polar-copolymerized) type, was used for isolating flavonolignan diastereoisomers in the 2D preparation. By establishing optimal 2D chromatography, hydrophilic interaction chromatography (HILIC) columns and normal-phase (NP) columns were tested simultaneously, and the result showed that diastereoisomers are not suitable for HILIC and NP chromatography mode. Our study resulted in a tricin and five analogous derivative flavonolignans with purity >98% were successfully purified from A. kansuensis. This is the initial report of Salcolin C, Salcolin B, Tricin 4'-O-(C-veratroylglycol) ether and 5'-methoxyhydnocarpin D from A. kansuensis. In addition, it tended to be the first time that Tricin 4'-O-(C-veratroylglycol) ether is isolated from natural resource. This method has great potential for efficiently isolating flavonolignan diastereoisomers from A. kansuensis, and it shows a great prospect for the separation of flavonolignans from complex samples.
草药植物意义重大，因为它们在发现药物前体方面具有巨大潜力。然而，如何从它们中纯化出更高纯度的化合物是一个需要讨论的问题。在本研究中，成功开发了一种离线二维反相（RP）制备液相色谱与固相萃取（SPE）联用的方法，用于从甘肃雪灵芝中分离黄酮木脂素非对映异构体。基于结果分析，我们得出的主要结论是选择RP-SPE从甘肃雪灵芝中富集目标黄酮木脂素样品。之后，使用ODS制备柱进行一维制备，将目标样品（4.6 g）分成五个馏分，回收率为83.9%。然后，使用C18HCE制备柱，一种极性改性的RP（极性共聚）类型，在二维制备中分离黄酮木脂素非对映异构体。通过建立最佳二维色谱，同时测试了亲水相互作用色谱（HILIC）柱和正相（NP）柱，结果表明非对映异构体不适用于HILIC和NP色谱模式。我们的研究成功地从甘肃雪灵芝中纯化出了纯度>98%的小麦黄素和五种类似的衍生黄酮木脂素。这是关于甘肃雪灵芝中Salcolin C、Salcolin B、小麦黄素4'-O-（C-藜芦酰二醇）醚和5'-甲氧基非洲防己碱D的首次报道。此外，小麦黄素4'-O-（C-藜芦酰二醇）醚首次从自然资源中分离出来。该方法在从甘肃雪灵芝中高效分离黄酮木脂素非对映异构体方面具有巨大潜力，并且在从复杂样品中分离黄酮木脂素方面显示出广阔前景。
