Efficient recovery of allylpyrocatechol from Piper betle using alcohol/salt aqueous two-phase system (ATPS)

The emergence of antibiotic -resistant bacteria presents significant public health threat, triggering a growing demand for natural compounds possessing antibacterial properties within the realm of Complementary and Alternative Medicine (CAM). Our previous research identified allylpyrocatechol from Piper betle as having strong antibacterial and QS inhibition properties. However, isolating phenolic compounds is time-consuming and relies on significant organic solvent use through methods like chromatography. The aqueous two-phase system (ATPS) emerges as an innovative, efficient, and eco-friendly approach for biomolecule recovery. This study assesses ATPS efficiency in allylpyrocatechol recovery from P. betle , optimizing parameters such as phase component, composition, crude load, and pH. Optimal conditions include 22% 1-propanol, 24% ammonium sulfate at pH 7, and a 10% sample load, resulting in a higher yield (5.82 +/- 0.04 g/kg plant material) and purity (14.09 +/- 0.10%) compared to hydrodistillation (yield: 0.29 +/- 0.01 g/kg plant material, purity: 7.17 +/- 0.17%). The ATPS extract demonstrates greater antibacterial potency against Staphylococcus aureus (MIC: 500 mu g/mL) and Micrococcus luteus (MIC: 500 mu g/mL) than hydrodistillation (1000 mu g/mL respectively). Additionally, it possesses stronger inhibition effect on violacein (48.00 +/- 0.83%) and biofilm production (43.70 +/- 1.22%) while hydrodistillation only inhibits by 35.44 +/- 0.60% and 33.15 +/- 0.66%, respectively. This study underscores ATPS as rapid and effective method for phenolic compound recovery, exemplified by allylpyrocatechol from P. betle .