Effects of Extraction Solvents on Antioxidant Activity and Chemical Composition of Kae Mussakaya Thatu Atisan Formulation

Abstract

This research aimed to study the effect of solvents on the antioxidant activity of the Kae Mussakaya Thatu Atisan formulation, a traditional Thai medicine used to treat diarrhea. The study employed three solvents which are hexane, ethyl acetate (EtOAc), and ethanol (EtOH) for extracting active compounds from the formulation. Antioxidant properties were evaluated using DPPH, ABTS, and FRAP assays. Total phenolic and flavonoid contents were determined by Folin-Ciocalteu and Aluminum Chloride Colorimetric Method, respectively. Chemical analysis was performed using HPLC technique. Results showed
that ethanol has the highest yielded extract (7.08%), followed by ethyl acetate (4.75%) and hexane (3.18%). The DPPH antioxidant assay revealed that the ethanol extract had the highest antioxidant activity with an IC₅₀ value of 90.01±5.55 µg/ml, while the ABTS assay showed that the hexane extract had the highest activity with an IC₅₀ value of 17.74±1.42 µg/ml and FRAP value in rang from 23.45 - 59.57 mmol/100 g extract. However, the antioxidant activity of all extracts showed lower than that of Trolox. In the chemical composition analysis, the ethanol extract had the highest total phenolic content (33.64±2.99 mg GAE/g extract), while the hexane extract had the highest total flavonoid content (70.19±4.79 mg QE/g extract). For cannabinoid analysis by HPLC showed the highest ∆9-THC content in EtOAc extract as 60.42±0.04 mg/g extract and the highest CBN content was found in both hexane and EtOH extracts, with values of 59.17±0.07 and 39.58±0.04 %w/w, respectively. In addition, GC-MS analysis identified 59 compounds with the main active compounds being THC, CBD and (+)-2-bornanone.
This research demonstrates that the type of solvent significantly affects the extract yield, antioxidant activity, and chemical composition of the Kae Mussakaya Thatu Atisan formulation. The findings from this study can be used as a guideline for developing appropriate extraction methods to obtain extracts with maximum antioxidant activity and may lead to the development of pharmaceutical products from the traditional Thai formulation in the future.