Enrichment and separation of high‐polar compounds from Saussurea obvallata using solid‐phase extraction combining with offline two‐dimensional liquid chromatography

Abstract The high‐polar compounds from natural products are often used as medicines due to their good bioactivities. However, owing to the complexity and diversity of their structure, the separation of high‐polar compounds is still a challenging work. For this, an efficient method for enrichment and separation of the high‐polar compounds from Saussurea obvallata was developed. First, the target compounds were enriched from the total extract using a solid‐phase extraction method. An offline two‐dimensional liquid chromatography method was used for the separation of pure compounds from the enriched sample. After optimization of chromatographic conditions, high separation selectivity of target compounds was obtained on a polar‐modified C18 column and a HILIC XAmide column. Hence, a two‐dimensional reversed‐phase × hydrophilic interaction liquid chromatography system was constructed and enlarged from the analytical level to the preparative level. In the first dimension, four fractions were obtained on the XCharge C18 column with a recovery rate of 71.2%. In the second‐dimension preparation on the XAmide column, eight high‐polar compounds with more than 96% purity were isolated. All compounds were isolated from Saussurea obvallata for the first time. The results demonstrated that this developed strategy is effective for preparative‐scale isolation of high‐polar compounds from natural products.