ABSTRACT
Hollow
fibre cell fishing with HPLC (HFCF-HPLC) based on the human ovarian cancer cell
line SKOV-3, human renal tubular cell line ACHN or hepatoma cell line HepG-2
was employed to screen active groups of coumarin and volatile oil in Radix angelicae sinensis, Radix angelicae dahuricae and Fructus citri sarcodactylis. Simultaneously,
hollow fibre liquid phase microextraction with HPLC (HFLPME-HPLC) was conducted
to enrich and determine the contents of active components in the same sample
solution. Before application, for HFCF-HPLC, cells growth states and survival
rates on the fibre, effect of ethanol concentration in the extract of samples
on cell survival rates, non-specific binding between fibre active centres and
the target components, positive and negative controls and repeatabilities were
validated; for HFLPME, extraction solvent, sample phase pH, agitation speed,
extraction time and sample phase volume were investigated. Many active components were screened from three medicines. Some
of them, such as scoparone, psoralen, bergapten, oxypeucedanin, imperatorin,
ligustilide, were identified by MS. The target fishing factors of active components
and the cell apoptosis rates of three cells under the medicines effect were researched.
The binding sites of active groups on HepG-2 cells were preliminarily determined.
The results demonstrated that HFCF-HPLC, coupled with HFLPME-HPLC, is a simple
and universal approach to find bioactive components at the cellular level,
determine their content and research traditional Chinese medicines (TCMs) entirety
effect of multi-component and multi-target. The approach may provide us a new
and good solution to clarify the material basis of anti-cancer effect and
conduct personalized quality control for the components associated with
efficacy in TCMs.