Abstract
OBJECTIVE: This study aimed to optimize the green extraction of phenolic compounds from TSH and evaluate their antioxidant and anti-liver cancer activities. METHODS: TSH was extracted using 60% ethanol and distilled water in different temperatures using a 2-level factorial design as the variables, followed by tray drying. Extracts were analyzed for total phenolic content (TPC) using Folin-Ciocalteu reagent as the response and DPPH radical scavenging assay. Oligomeric proanthocyanidins were characterized by high performance liquid chromatography (HPLC) and liquid chromatography tandem mass spectrometry (LC-ESI-QTOF-MS/MS). Cytotoxicity and antimigratory effects on HepG2 cells were assessed using sulforhodamine B (SRB) and wound healing assays. RESULT: The optimal model equation was TPC = 0.9 - 0.0064A + 0.0059B + (2.61 × 10-5)AB. Although the ethanolic extract had higher total phenolic content, but the water extract showed stronger antioxidant activity and exhibited significantly stronger cytotoxic activity (IC50 at 72 hours: 44.1±0.9 µg/mL) than ethanolic extract (408.3±9.8 µg/mL) in a time- and dose-dependent manner. Migration of HepG2 cells was also significantly inhibited by the water extract in a dose-dependent manner. CONCLUSION: The water extract of TSH may serve as a promising candidate for functional ingredient development and natural anti-liver cancer applications.