Li Zhenyu, Fu Qing, Li Kuiyong, Liang Tu, Jin Yu
Se Pu. 2014 May;32(5):506-12. doi: 10.3724/sp.j.1123.2013.12035.
A fast chromatographic separation of indole alkaloids from Evodiae fructus was developed by supercritical fluid chromatography (SFC). The initial screening of four stationary phases was investigated with a standard mixture of evodiamine and rutaecarpine, and a complex sample of indole alkaloids prepared from Evodiae fructus as probes. Later, the effects of chromatographic parameters on separation were studied including injection volume, organic modifier, additive, temperature and back pressure. The injection volume had significant impact on the peak shape. With the additives in the mobile phase, slight changes in peak shape and retention time were observed in separation. Variation in organic modifier led to dramatic change in chromatographic behavior. Both decreased temperature and increased back pressure shortened the retention time. Finally, a fast analytical method using SFC, on a Waters ACQUITY UPC2 BEH column, methanol as modifier, under 35 degrees C and 2.07 x 10(7) Pa, was developed to separate a complex sample of indole alkaloids in less than 15 min. Another rapid approach for the separation of a complex sample of indole alkaloids was developed by using ultra-high performance liquid chromatography (UHPLC). As a result, SFC can be used in the separation of natural products, giving high performance, good resolution and fast analysis speed. The difference in selectivity with UHPLC can be used to the development of natural product separation.
采用超临界流体色谱法(SFC)建立了吴茱萸中吲哚生物碱的快速色谱分离方法。以吴茱萸碱和吴茱萸次碱的标准混合物以及从吴茱萸中制备的吲哚生物碱复杂样品为探针,对四种固定相进行了初步筛选。随后,研究了色谱参数对分离的影响,包括进样体积、有机改性剂、添加剂、温度和背压。进样体积对峰形有显著影响。在流动相中加入添加剂后,分离过程中峰形和保留时间有轻微变化。有机改性剂的变化导致色谱行为发生显著变化。降低温度和增加背压均会缩短保留时间。最后,建立了一种快速分析方法,使用SFC法,在Waters ACQUITY UPC2 BEH柱上,以甲醇为改性剂,在35℃和2.07×10⁷Pa条件下,在不到15分钟内分离吲哚生物碱复杂样品。还开发了另一种使用超高效液相色谱法(UHPLC)分离吲哚生物碱复杂样品的快速方法。结果表明,SFC可用于天然产物的分离,具有高性能、良好的分离度和快速的分析速度。与UHPLC选择性的差异可用于天然产物分离方法的开发。