Comparative LC-MS Analysis and In Vitro Evaluation of Antioxidant, Antimicrobial, and Enzyme Inhibitory Properties of Ultrasound-Assisted and Soxhlet Extracts of Cachrys cristata DC

Abstract

This study evaluated the extraction efficiency, phenolic composition, and bioactivity of Cachrys cristata using different methods. Ultrasound-assisted extraction (US) yielded higher extract amounts and phenolic contents, including flavonoids and flavonols, compared to Soxhlet extraction (S). While US 70% ethanol gave the highest extract yield, the US methanol (MeOH) extract contained the highest total phenolic content (105.47 mg gallic acid equivalents/g dry extract) and was selected for LC-MS profiling. Seventeen compounds were identified, with gentisic acid (39.03/50.73 mg/L), ferulic acid (27.00/25.68 mg/L), and rutin (13.28/11.91 mg/L) as dominant in Soxhlet and US extracts, respectively. The US MeOH extract showed strong antioxidant activity in DPPH and ABTS assays and eightfold higher anti-tyrosinase activity than kojic acid, while no acetylcholinesterase inhibition was observed. The hexane extract exhibited potent antibacterial effects against Staphylococcus aureus, Enterococcus faecalis, and Listeria monocytogenes, inhibited biofilm formation and pyocyanin in Pseudomonas aeruginosa, and had limited antifungal activity. It also strongly inhibited bovine serum albumin denaturation (88.88% vs. 95.60% for diclofenac). For antidiabetic potential, the US methylene chloride extract inhibited α-amylase better than acarbose (IC50 0.267 vs. 0.832 mg/mL) and moderately reduced α-glucosidase activity. These results provide novel insights into the phenolic profile and multifunctional bioactivity of C. cristata extracts.