Simultaneous determination of saikosaponins in Bupleurum Radix from different locations by high performance liquid chromatography- charged aerosol detection (HPLC-CAD) method and its immunomodulation effects on mouse splenocytes

A high performance liquid chromatography (HPLC)-Charged Aerosol Detection (CAD) was employed to simultaneously determine saikosaponins which include saikosaponin a (SSa), saikosaponin b2 (SSb2) and saikosaponin d (SSd) in Bupleurum Radix from different locations. Meanwhile, MTT assay was used to compare the proliferation effect of SSa, SS and SSb2 on mouse splenocytes, and enzyme linked immunosorbent assay (ELISA) method was used to determined cytokines which include interleukin (IL)-4, IL-10, interfernon (IFN)-γ, TNF-α. The results show that there existed significant differences of the three saikosaponins from different locations, and SSa, SSd, and SSb2 all presented immunomodulation effects on mouse splenocytes via promoting splenocyte proliferation and regulating the balance of Th1/Th2 factors.   Key words: HPLC-CAD, Bupleurum Radix, saikosaponin a (SSa), saikosaponin b2 (SSb2), saikosaponin d (SSd).