Phytochemical identification of SH003 solvent fractions and evaluation of their anti-cancer effects across various cancer types

SH003 is a natural anti-cancer agent derived from the traditional herbal formula Danggui Buxue Tang. It is an extract prepared by mixing Astragalus membranaceus, Angelica gigas , and Trichosanthes kirilowii at a ratio of 1:1:1. In previous studies, the research team reported anti-tumor, anti-cancer, anti-inflammatory, and immune-enhancing effects of SH003. However, research identifying which specific phytochemicals are responsible for these anti-cancer effects is lacking. To address this gap of knowledge, the present study performed solvent fractionation of SH003 using solvents with different polarities to identify active components. Qualitative and quantitative phytochemical analyses of each solvent fraction were conducted using high performance liquid chromatography (HPLC). In addition, whether anti-cancer effect of each solvent fraction was enhanced compared to SH003. Through HPLC qualitative analysis and liquid chromatography mass spectrometry analysis (LC-MS/MS), the presence of eight phytochemicals bergapten, isopimpinellin, methylnissolin, osthenol, nodakenin, marmesin, calycosin-7-O-β-D-glucoside, and cucurbitacin D was confirmed in ethyl acetate (EA) and chloroform (CHCl ₃ ) fractions. Quantitative analysis showed that these compounds were the most abundant in EA and CHCl ₃ fractions. In vitro results revealed that EA and CHCl ₃ fractions had significantly higher anti-cancer efficacies than the original SH003 extract, with the CHCl ₃ fraction exhibiting the highest anti-cancer activity. Based on results of HPLC qualitative/quantitative analysis and the in vitro model, the list of candidate active compounds was narrowed down.