Comparison of ultra-high performance supercritical fluidchromatography and ultra-high performance liquid chromatographyfor the separation of spirostanol saponins
Comparison of ultra-high performance supercritical fluidchromatography and ultra-high performance liquid chromatographyfor the separation of spirostanol saponins
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超高效超临界流体色谱法与超高效液相色谱法分离螺甾烷皂苷的比较
DOI：
10.1016/j.jpba.2015.12.002
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发表时间：
2016
影响因子：
3.4
通讯作者：
mabaiping
中科院分区：
文献类型：
--
作者：
zhu lingling;zhao yang;xu yongwei;sun qinglong;sun xinguang;kang liping;yan renyi;zhang jie;liu chao;mabaiping
螺甾烷醇皂苷是一些中药材的重要活性成分，其分离纯化对于中药的研发至关重要。我们的目的是比较超高效超临界流体色谱（UHPSFC）和超高效液相色谱（UHPLC）对螺甾烷皂苷的分离。采用UHPSFC和UHPLC分别分离了四组螺甾烷皂苷。优化后，采用 HSS C18 SB 柱或 Diol 柱，以甲醇为共溶剂进行 UHPSFC。 UHPLC 中使用 BEH C18 柱和含有水（含 0.1% 甲酸）和乙腈的流动相。我们发现UHPSFC可以自动且快速地执行。它可以有效分离具有相同苷元但糖链不同的螺甾烷皂苷，并且对苷元中羟基的数量和位置非常敏感。然而，UHPSFC对不同苷元和相同糖部分的螺甾烷皂苷的分离效果并不理想，可以用UHPLC来分离。 UHPLC 擅长区分糖苷配基的变异，并且受糖苷配基中双键的影响。因此，UHPLC和UHPSFC在分离螺甾烷皂苷方面是互补的。考虑到草药中天然产生的螺甾烷皂苷在苷元和糖链方面都不同，因此UHPLC和UHPSFC的组合可以实现更好的分离。当 UHPLC 无法分离螺甾烷皂苷混合物时，UHPSFC 是一种提高分离度的强大技术，反之亦然。
tSpirostanol saponins are important active components of some herb medicines, and their isolation andpurification are crucial for the research and development of traditional Chinese medicines. We aimed tocompare the separation of spirostanol saponins by ultra-high performance supercritical fluid chromatog-raphy (UHPSFC) and ultra-high performance liquid chromatography (UHPLC). Four groups of spirostanolsaponins were separated respectively by UHPSFC and UHPLC. After optimization, UHPSFC was performedwith a HSS C18 SB column or a Diol column and with methanol as the co-solvent. A BEH C18 column andmobile phase containing water (with 0.1% formic acid) and acetonitrile were used in UHPLC. We foundthat UHPSFC could be performed automatically and quickly. It is effective in separating the spirostanolsaponins which share the same aglycone and vary in sugar chains, and is very sensitive to the numberand the position of hydroxyl groups in aglycones. However, the resolution of spirostanol saponins withdifferent aglycones and the same sugar moiety by UHPSFC was not ideal and could be resolved by UHPLCinstead. UHPLC is good at differentiating the variation in aglycones, and is influenced by double bonds inaglycones. Therefore, UHPLC and UHPSFC are complementary in separating spirostanol saponins. Con-sidering the naturally produced spirostanol saponins in herb medicines are different both in aglyconesand in sugar chains, a better separation can be achieved by combination of UHPLC and UHPSFC. UHPSFC isa powerful technique for improving the resolution when UHPLC cannot resolve a mixture of spirostanolsaponins and vice versa.