title: Comparison of ultra-high performance supercritical fluid chromatography and ultra-high performance liquid chromatography for the separation of spirostanol saponins.
abstract: Spirostanol saponins are important active components of some herb medicines, and their isolation and purification are crucial for the research and development of traditional Chinese medicines. We aimed to compare the separation of spirostanol saponins by ultra-high performance supercritical fluid chromatography (UHPSFC) and ultra-high performance liquid chromatography (UHPLC). Four groups of spirostanol saponins were separated respectively by UHPSFC and UHPLC. After optimization, UHPSFC was performed with a HSS C18 SB column or a Diol column and with methanol as the co-solvent. A BEH C18 column and mobile phase containing water (with 0.1% formic acid) and acetonitrile were used in UHPLC. We found that UHPSFC could be performed automatically and quickly. It is effective in separating the spirostanol saponins which share the same aglycone and vary in sugar chains, and is very sensitive to the number and the position of hydroxyl groups in aglycones. However, the resolution of spirostanol saponins with different aglycones and the same sugar moiety by UHPSFC was not ideal and could be resolved by UHPLC instead. UHPLC is good at differentiating the variation in aglycones, and is influenced by double bonds in aglycones. Therefore, UHPLC and UHPSFC are complementary in separating spirostanol saponins. Considering the naturally produced spirostanol saponins in herb medicines are different both in aglycones and in sugar chains, a better separation can be achieved by combination of UHPLC and UHPSFC. UHPSFC is a powerful technique for improving the resolution when UHPLC cannot resolve a mixture of spirostanol saponins and vice versa.
authors.affiliation: Beijing Institute of Radiation Medicine, No. 27, Taiping Road, Beijing 100850, PR China.
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Zhu
authors.affiliation: Beijing Institute of Radiation Medicine, No. 27, Taiping Road, Beijing 100850, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Waters Corporation (Shanghai), Shanghai 201206, PR China.
authors.forename: Yong-Wei
authors.initials: YW
authors.lastname: Xu
authors.affiliation: Waters Corporation (Shanghai), Shanghai 201206, PR China.
authors.forename: Qing-Long
authors.initials: QL
authors.lastname: Sun
authors.affiliation: Beijing Institute of Radiation Medicine, No. 27, Taiping Road, Beijing 100850, PR China.
authors.forename: Xin-Guang
authors.initials: XG
authors.lastname: Sun
authors.affiliation: Beijing Institute of Radiation Medicine, No. 27, Taiping Road, Beijing 100850, PR China.
authors.forename: Li-Ping
authors.initials: LP
authors.lastname: Kang
authors.affiliation: Ovation Health Science and Technology Co., Ltd., ENN Group, Langfang 065001, PR China.
authors.forename: Ren-Yi
authors.initials: RY
authors.lastname: Yan
authors.affiliation: Ovation Health Science and Technology Co., Ltd., ENN Group, Langfang 065001, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Beijing Institute of Radiation Medicine, No. 27, Taiping Road, Beijing 100850, PR China. Electronic address: liuchao9588@sina.com.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.affiliation: Beijing Institute of Radiation Medicine, No. 27, Taiping Road, Beijing 100850, PR China. Electronic address: mabaiping@sina.com.
authors.forename: Bai-Ping
authors.initials: BP
authors.lastname: Ma
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D025924:Chromatography, Supercritical Fluid / Q000379:methods; D000432:Methanol / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry; D012503:Saponins / Q000737:chemistry*; D013150:Spirostans / Q000737:chemistry*; D014867:Water / Q000737:chemistry
chemical_list: D012503:Saponins; D013150:Spirostans; D014867:Water; D000432:Methanol
keywords: Chromatographic behavior; Natural products; Spirostanol saponins; Ultra-high performance liquid chromatography; Ultra-high performance supercritical fluid chromatography