Optimizing the Operational Conditions of Simultaneous Ultrasound-Assisted Enzymatic Aqueous Two-Phase Extraction for Strobilanthes crispus
An innovative extraction method, incorporating simultaneous ultrasonic, enzymatic, and two-phase aqueous extraction, was employed to enhance the ethanol extract of phenolics and flavonoids from Strobilanthes crispus leaves. The study investigated the correlation between extraction method parameters and the total phenolic content (TPC), total flavonoids content (TFC), partition coefficient (k), and recovery (R) of the ethanol extract from leaves. The cellulase enzyme from Aspergillus niger was utilized, and a two-phase solvent system (ethanol-ammonium sulfate, 33:14 %w/w) was employed. Isolation of the crude extract was carried out using the eluent ethyl acetate:methanol (1:1). Subsequently, an inhibition test on the HMG CoA reductase enzyme was conducted. Optimal extraction conditions included a solvent-to-solid ratio of 35 mL/g, a temperature of 60°C, an enzyme loading of 30 mg/g, and an extraction time of 45 minutes. The analysis results revealed a TPC content of 5.30 ± 0.34 mg GAE/g DLP, TFC of 1.16 ± 0.05 mg QE/g DLP, a partition coefficient of 19.91, and a recovery of 98.01%. Regression analysis revealed significant correlations (p<0.05) between extraction parameters and TPC, TFC, k, and R. Fraction I demonstrated competitiveness with pravastatin, highlighting its pharmaceutical potential. Characterization of the crude extract revealed the presence of bioactive compounds, including genistein, umbelliferone, and pinocembrin, known for their potential anticholesterol properties, as supported by existing literature. This research introduces an effective extraction technique for increasing phenolic and flavonoid bioactive compounds with anticholesterol activity, contributing to advancements in the pharmaceutical field.