Multichannel microperfusion cell-capture system for screening anticancer active ingredients in traditional Chinese medicine formulas†

Abstract

The screening of effective active ingredients from traditional Chinese medicine (TCM) formulas is currently a key focus of research. In this study, a multichannel microperfusion cell-capture system utilizing human hepatocellular carcinoma HepG2 cells was established and used to screen for anticancer active ingredients in the ancient formula Huangqisan. A polypropylene solid film carrying HepG2 cells was used as the cell-capture platform, and a multichannel microperfusion system was utilized for perfusion, enabling the screening of multiple ingredients' activities. This study optimized the main factors influencing the screening results and conducted a methodological examination under the optimal screening conditions. The method was coupled with high-performance liquid chromatography (HPLC) to analyze and identify the anticancer ingredients in Huangqisan. The optimal conditions were determined as follows: use of a polypropylene film as the cell support, a 7 mL sample phase solution (3 μg mL⁻¹) for perfusion, and a screening duration of 30 min. After screening, eight anticancer active ingredients in Huangqisan, namely, calycosin, calycosin glycoside, formononetin, ononin, genistein, isorhamnetin, puerarin, and morin, were preliminarily determined by comparing their chromatographic retention times and peak areas with standard substances. The cell fishing factor (CFF) of these ingredients ranged from 1.1 to 19.8. In summary, the use of a multichannel microperfusion cell-capture system combined with HPLC enabled the screening of the anticancer active ingredients in Huangqisan. This method thus provides a new approach for screening complex TCM matrices and holds promising prospects for future development.