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The traditional ways of protein extraction is disadvantageous from both economic and environment perspective. In this study, ultrasonic assisted extraction, UAE technique was used for the first time for the optimization process of extracting proteins from Eurycoma longifolia roots. The experiments demonstrated the successful effect of applying eccentric agitation with UAE to provide a combination of agitation and sonication to extract the proteins in short extraction times. Central composite design (CCD) was used to optimize the Ultrasonic-assisted extraction of proteins from roots of Eurycoma longifolia by water. Response surface methodology (RSM) was employed to investigate the effects of five independent variables; particle sizes ( A ), extraction temperatures ( B ), agitation speeds ( C ), amplitude ( D ) and duty cycle ( E ). The second enhanced model ( Y 2 ) prevailed to be more efficient in protein recovery with optimum conditions of A : 0.022 ± 0.022 mm, B : 49 °C, C : 1314 rpm, D : 9 W and E : 63% UI : 2.94 W·cm −2 was 9.543 ± 0. 946%. High agitation speed didn’t only affect the solvent properties, the diffusion of the solutes from the particles, cooling the extraction temperature, but influenced the distribution of cavitation bubbles and their impact on the solid particles. UAE was found to minimize the extraction of the same amount by conventional extraction 7-fold times. The results obtained in this study have exposed the capability of ultrasonic assisted extraction, UAE technology in extraction of protein from E. longifolia roots. Further works are nevertheless required to provide deeper understanding of the mechanisms involved to facilitate the development of optimum system applicable to the industry.

Central composite designs (CCDs) for UAE and WE were established by seven and five extraction factors, respectively, to determine their optimum conditions that maximized the saponins yields. These optimum conditions were employed successively to establish a sequential extraction (SE) of saponins from Eurycoma longifolia roots. Results identified the agitation speed as an influencing factor in altering the interactive effects of the extraction factors in UAE and WE. Optimized conditions illustrated that higher agitation speeds increased the yields of saponins in WE while UAE preferred lower agitation speed ranges. This implied that the high agitation speeds decreased the collision of microjets and particles and hence the the decrease of sonication efficiency. The SE extraction was proposed by initial sonication of the plant sample for a duration of 5 minutes and 29 seconds to 10 minutes followed by conventional WE for 20-25 minutes.