Bioactive assay and hyphenated chromatography detection for complex supercritical CO2 extract from Chaihu Shugan San using an experimental design approach

In this study, Super Critical CO2 (SCC) was applied to extract the active ingredients from Multi-herbal formula Chaihu Shugan San (CSS). Response Surface Methodology (RSM) coupled with an improved chaotic Gray Wolf Optimization (ACGWO) was employed to the experimental design. After gas chromatography-mass spectrometer (GC-CI/EI-MS) detection, liquid chromatography-mass spectrometry- ion trap-time of flight (LC-MS-IT-ToF) further indicated that several phthalides are mainly responsible for the SCC extract; the next is α-Cyperone in Cyperus rotundus; and Meranzin hydrate, Isomeranzin in Citrus reticulata Blanco again. Further pharmacological tests were also performed for the SCC extracts. THP-1 macrophages were used to observe the anti-inflammatory effects. The expression of IL-6 was obviously suppressed by the SCC extract from CSS, through a positive correlation with the dose. Then, ROS content in a cell model was used to detect the antioxidant capacities. The results showed that the SCC extract from CSS, exert dose-dependent inhibitory effects on the oxidative stress of the cells.