Acoustic and hydrodynamic cavitation assisted hydrolysis and valorisation of waste human hair for the enrichment of amino acids

Hair waste in large amount is produced in India from temples and saloons, India alone exported approximately 1 million kg of hair in 2010. Incineration and degradation of waste human hair leads to environmental concerns. The hydrothermal process is a conventional method for the production of hair hydrolysate. The hydrothermal process is carried out at a very high temperature and pressure, which causes the degradation of heat-sensitive essential amino acids, thereby depleting the nutritional value. This work deals with alkaline hydrolysis of human hair using acoustic and hydrodynamic cavitation, and comparison with the conventional method. The optimal operating conditions for highest efficiency was observed, for the hydrolysis of 1 g of sample hairs in 100 mL of solution, at 4:1 (KOH: hair) ratio, soaking time of 24 h, the ultrasonic power density of 600 W dm⁻³ (20 KHz frequency and input power 200 W) or hydrodynamic cavitation inlet pressure of 4 or 7 bars. Cavitation results in rupture of disulfide linkages in proteins and mechanical effects lead to cleavage of several hydrogen bonds breaking the keratin sheet structure in hair. Breakdown of bonds leads to a decrease in viscosity of the solution. 10% and 6% reduction in viscosity is obtained at optimal conditions for ultrasonic and hydrodynamic cavitation treatment, respectively. FTIR analysis of produced hair hydrolysate confirmed that the disulfide bonds in hair proteins are broken down during cavitation. The amino acid of hair hydrolysate, prepared using cavitation, has a relatively higher digestibility and nutritional value due to the enhancement of amino-acid content, confirmed using amino acid analysis. Cavitation assisted hair hydrolysate has a potential application in agricultural engineering as a fertilizer for improvement of the quality of the soil and land. Cavitation based hair hydrolysate can also be used as an environmentally friendly and economical source of essential amino acids and digestibles for animal or poultry feed.     

Degradation of organic wastewater by hydrodynamic cavitation combined with acoustic cavitation

In this paper, the decomposition of Rhodamine B (RhB) by hydrodynamic cavitation (HC), acoustic cavitation (AC) and the combination of these individual methods (HAC) have been investigated. The degradation of 20 L RhB aqueous solution was carried out in a self-designed HAC reactor, where hydrodynamic cavitation and acoustic cavitation could take place in the same space simultaneously. The effects of initial concentration, inlet pressure, solution temperature and ultrasonic power were studied and discussed. Obvious synergies were found in the HAC process. The combined method achieved the best conversion, and the synergistic effect in HAC was even up to 119% with the ultrasonic power of 220 W in a treatment time of 30 min. The time-independent synergistic factor based on rate constant was introduced and the maximum value reached 40% in the HAC system. Besides, the hybrid HAC method showed great superiority in energy efficiency at lower ultrasonic power (88–176 W). Therefore, HAC technology can be visualized as a promising method for wastewater treatment with good scale-up possibilities.     

Intensification of biogas production using pretreatment based on hydrodynamic cavitation

The present work investigates the application of hydrodynamic cavitation (HC) for the pretreatment of wheat straw with an objective of enhancing the biogas production. The hydrodynamic cavitation reactor is based on a stator and rotor assembly. The effect of three different speeds of rotor (2300, 2500, 2700 rpm), wheat straw to water ratios (0.5%, 1% and 1.5% wt/wt) and also treatment times as 2, 4 and 6 min have been investigated in the work using the design of experiments (DOE) approach. It was observed that the methane yield of 31.8 ml was obtained with untreated wheat straw whereas 77.9 ml was obtained with HC pre-treated wheat straw confirming the favourable changes during the pre-treatment. The combined pre-treatment using KOH and HC gave maximum yield of biogas as 172.3 ml. Overall, it has been established that significant enhancement in the biogas production can be obtained due to the pretreatment using HC which can also be further intensified by combination with chemical treatment.     

Degradation of imidacloprid using combined advanced oxidation processes based on hydrodynamic cavitation

The harmful effects of wastewaters containing pesticides or insecticides on human and aquatic life impart the need of effectively treating the wastewater streams containing these contaminants. In the present work, hydrodynamic cavitation reactors have been applied for the degradation of imidacloprid with process intensification studies based on different additives and combination with other similar processes. Effect of different operating parameters viz. concentration (20–60 ppm), pressure (1–8 bar), temperature (34 °C, 39 °C and 42 °C) and initial pH (2.5–8.3) has been investigated initially using orifice plate as cavitating device. It has been observed that 23.85% degradation of imidacloprid is obtained at optimized set of operating parameters. The efficacy of different process intensifying approaches based on the use of hydrogen peroxide (20–80 ppm), Fenton’s reagent (H₂O₂:FeSO₄ ratio as 1:1, 1:2, 2:1, 2:2, 4:1 and 4:2), advanced Fenton process (H₂O₂:Iron Powder ratio as 1:1, 2:1 and 4:1) and combination of Na₂S₂O₈ and FeSO₄ (FeSO₄:Na₂S₂O₈ ratio as 1:1, 1:2, 1:3 and 1:4) on the extent of degradation has been investigated. It was observed that near complete degradation of imidacloprid was achieved in all the cases at optimized values of process intensifying parameters. The time required for complete degradation of imidacloprid for approach based on hydrogen peroxide was 120 min where as for the Fenton and advance Fenton process, the required time was only 60 min. To check the effectiveness of hydrodynamic cavitation with different cavitating devices, few experiments were also performed with the help of slit venturi as a cavitating device at already optimized values of parameters. The present work has conclusively established that combined processes based on hydrodynamic cavitation can be effectively used for complete degradation of imidacloprid.     

Oxidation of Sulphur pollutants in model and real fuels using hydrodynamic cavitation

Hydrodynamic Cavitation (HC) offers an attractive platform for intensifying oxidative desulphurization of fuels. In the first part of this work, we present new results on oxidising single ring thiophene in a model fuel over the extended range of volume fraction of organic phase from 2.5 to 80 v/v %. We also present influence of type and scale of HC device on performance of oxidative desulphurization. Further experiments revealed that oxidising radicals generated in-situ by HC alone were not able to oxidise dual ring thiophenes. External catalyst (formic acid) and oxidising agents (hydrogen peroxide, H₂O₂) were therefore used with HC. Based on our prior work with acoustic cavitation (AC), the volumetric ratios for H₂O₂ and formic acid were identified as 0.95 v/v % and 6.25 v/v % respectively. The data of oxidation of dual ring thiophenes with n-dodecane and n-hexane as model fuels and typical transport fuels (diesel, kerosene, and petrol) using these oxidant and catalyst is presented. The observed performance with HC was compared with results obtained from a stirred tank and AC set-up. The presented data indicates that HC is able to intensify oxidation of sulphur species. The presented results provide a sound basis for further developments on HC based oxidative desulphurization processes.     

乳清分离蛋白-葡聚糖接枝物性质的荧光光谱法分析

乳清分离蛋白与葡聚糖的混合物在干热处理条件下,发生了以褐变为特征的美拉德反应。当葡聚糖分子量由67kD增至150kD时,游离氨基含量分别下降了35.77%和30.53%,糖链越长,其接入到蛋白质肽链的难度越大。采用荧光光谱对乳清分离蛋白-葡聚糖接枝物的性质进行分析。内源荧光光谱图显示,接枝产物在405nm的最大荧光强度显著提高,且350～500nm范围内的荧光强度顺序为:G67>G150,这说明接枝物中有Maillard反应体系所特有的荧光物质生成;由外援荧光光谱图得出,接枝产物在470nm的最大荧光强度均有明显降低,各溶液体系中荧光强度高低顺序依次为:WPI>G150>G67。疏水性指数的测定进一步说明两种不同分子量的葡聚糖接入到蛋白质肽链中,对乳清分离蛋白的疏水性均有一定的屏蔽作用。     

Degradation of reactive orange 4 dye using hydrodynamic cavitation based hybrid techniques

In the present work, degradation of reactive orange 4 dye (RO4) has been investigated using hydrodynamic cavitation (HC) and in combination with other AOP’s. In the hybrid techniques, combination of hydrodynamic cavitation and other oxidizing agents such as H₂O₂ and ozone have been used to get the enhanced degradation efficiency through HC device. The hydrodynamic cavitation was first optimized in terms of different operating parameters such as operating inlet pressure, cavitation number and pH of the operating medium to get the maximum degradation of RO4. Following the optimization of HC parameters, the degradation of RO4 was carried out using the combination of HC with H₂O₂ and ozone. It has been found that the efficiency of the HC can be improved significantly by combining it with H₂O₂ and ozone. The mineralization rate of RO4 increases considerably with 14.67% mineralization taking place using HC alone increases to 31.90% by combining it with H₂O₂ and further increases to 76.25% through the combination of HC and ozone. The synergetic coefficient of greater than one for the hybrid processes of HC + H₂O₂ and HC + Ozone has suggested that the combination of HC with other oxidizing agents is better than the individual processes for the degradation of dye effluent containing RO4. The combination of HC with ozone proves to be the most energy efficient method for the degradation of RO4 as compared to HC alone and the hybrid process of HC and H₂O₂.     

Degradation of dichlorvos using hydrodynamic cavitation based treatment strategies

The degradation of an aqueous solution of dichlorvos, a commonly used pesticide in India, has been systematically investigated using hydrodynamic cavitation reactor. All the experiments have been carried out using a 20 ppm solution of commercially available dichlorvos. The effect of important operating parameters such as inlet pressure (over a range 3-6 bar), temperature (31 °C, 36 °C and 39 °C) and pH (natural pH = 5.7 and acidic pH = 3) on the extent of degradation has been investigated initially. It has been observed that an optimum value of pressure gives maximum degradation whereas low temperature and pH of 3 are favorable. Intensification studies have been carried out using different additives such as hydrogen peroxide, carbon tetrachloride, and Fenton’s reagent. Use of hydrogen peroxide and carbon tetrachloride resulted in the enhancement of the extent of degradation at optimized conditions but significant enhancement was obtained with the combined use of hydrodynamic cavitation and Fenton’s chemistry. The maximum extent of degradation as obtained by using a combination of hydrodynamic cavitation and Fenton’s chemistry was 91.5% in 1 h of treatment time. The present work has conclusively established that hydrodynamic cavitation in combination with Fenton’s chemistry can be effectively used for the degradation of dichlorvos.     

Biodiesel production process intensification using a rotor-stator type generator of hydrodynamic cavitation

Triglyceride transesterification for biodiesel production is a model reaction which is used to compare the conversion efficiency, yield, reaction time, energy consumption, scalability and cost estimation of different reactor technology and energy source. This work describes an efficient, fast and cost-effective procedure for biodiesel preparation using a rotating generator of hydrodynamic cavitation (HC). The base-catalyzed transesterification (methanol/sodium hydroxide) has been carried out using refined and bleached palm oil and waste vegetable cooking oil. The novel HC unit is a continuous rotor-stator type reactor in which reagents are directly fed into the controlled cavitation chamber. The high-speed rotation of the reactor creates micron-sized droplets of the immiscible reacting mixture leading to outstanding mass and heat transfer and enhancing the kinetics of the transesterification reaction which completes much more quickly than traditional methods. All the biodiesel samples obtained respect the ASTM standard and present fatty acid methyl ester contents of >99% m/m in both feedstocks. The electrical energy consumption of the HC reactor is 0.030 kW h per L of produced crude biodiesel, making this innovative technology really quite competitive. The reactor can be easily scaled-up, from producing a few hundred to thousands of liters of biodiesel per hour while avoiding the risk of orifices clogging with oil impurities, which may occur in conventional HC reactors. Furthermore it requires minimal installation space due to its compact design, which enhances overall security.     

Degradation of diclofenac sodium using combined processes based on hydrodynamic cavitation and heterogeneous photocatalysis

Diclofenac sodium, a widely detected pharmaceutical drug in wastewater samples, has been selected as a model pollutant for degradation using novel combined approach of hydrodynamic cavitation and heterogeneous photocatalysis. A slit venturi has been used as cavitating device in the hydrodynamic cavitation reactor. The effect of various operating parameters such as inlet fluid pressure (2–4 bar) and initial pH of the solution (4–7.5) on the extent of degradation have been studied. The maximum extent of degradation of diclofenac sodium was obtained at inlet fluid pressure of 3 bar and initial pH as 4 using hydrodynamic cavitation alone. The loadings of TiO₂ and H₂O₂ have been optimised to maximise the extent of degradation of diclofenac sodium. Kinetic study revealed that the degradation of diclofenac sodium fitted first order kinetics over the selected range of operating protocols. It has been observed that combination of hydrodynamic cavitation with UV, UV/TiO₂ and UV/TiO₂/H₂O₂ results in enhanced extents of degradation as compared to the individual schemes. The maximum extent of degradation as 95% with 76% reduction in TOC has been observed using hydrodynamic cavitation in conjunction with UV/TiO₂/H₂O₂ under the optimised operating conditions. The diclofenac sodium degradation byproducts have been identified using LC/MS analysis.     

Effect of ultrasound treatment on interactions of whey protein isolate with rutin

Rutin is a biologically active polyphenol, but its poor water solubility and low bioavailability limit its application to the food industry. We investigated the effect of ultrasound treatment on the properties of rutin (R) and whey protein isolate (WPI) using spectral and physicochemical analysis. The results revealed that there was covalent interaction between whey protein isolate with rutin, and the binding degree of whey isolate protein with rutin increased with ultrasound treatment. Additionally, solubility and surface hydrophobicity of WPI-R complex improved with ultrasonic treatment, and a maximum solubility of 81.9 % at 300 W ultrasonic power. The ultrasound treatment caused the complex to develop a more ordered secondary structure, resulting in a three-dimensional network structure with small and uniform pore sizes. This research could provide a theoretical reference for studying protein–polyphenol interactions and their applications in food delivery systems.     

Decolorization of azo dyes Orange G using hydrodynamic cavitation coupled with heterogeneous Fenton process

The present work demonstrates the application of the combination of hydrodynamic cavitation (HC) and the heterogeneous Fenton process (HF, Fe⁰/H₂O₂) for the decolorization of azo dye Orange G (OG). The effects of main affecting operation conditions such as the inlet fluid pressure, initial concentration of OG, H₂O₂ and zero valent iron (ZVI), the fixed position of ZVI, and medium pH on decolorization efficiency were discussed with guidelines for selection of optimum parameters. The results revealed that the acidic conditions are preferred for OG decolorizaiton. The decolorization rate increased with increasing H₂O₂ and ZVI concentration and decreased with increasing OG initial concentration. Besides, the decolorization rate was strongly dependent on the fixed position of ZVI. The analysis results of degradation products using liquid chromatography–ESI–TOF mass spectrometry revealed that the degradation mechanism of OG proceeds mainly via reductive cleavage of the azo linkage due to the attack of hydroxyl radical. The present work has conclusively established that the combination of HC and HF can be more energy efficient and gives higher decolorization rate of OG as compared with HC and HF alone.     

Continuous production of nanoemulsion for skincare product using a 3D-printed rotor-stator hydrodynamic cavitation reactor

In this study, nanoemulsions for skincare products were continuously produced using a hydrodynamic cavitation reactor (HCR) designed with a rotor and stator. The key component of this research is the utilization of a 3D-printed rotor in a HCR for the production of an oil-in-water nanoemulsion. Response surface methodology was used to determine the process conditions, such as speed of the rotor, flow rate, as well as, Span60, Tween60, and mineral oil concentrations, for generating the optimal droplet size in the nanoemulsion. The results showed that a droplet size of 366.4 nm was achieved under the recommended conditions of rotor speed of 3500 rpm, flow rate of 3.3 L/h, Span60 concentration of 2.36 wt%, Tween60 concentration of 3.00 wt%, and mineral oil concentration of 1.76 wt%. Moreover, the important characteristics for consideration in skincare products, such as polydispersity index, pH, zeta potential, viscosity, stability, and niacin released from formulations, were also assessed. For the niacin release profile of emulsion and nanoemulsion formulations, different methods, such as magnetic stirring, ultrasound, and hydrodynamic cavitation, were compared. The nanoemulsion formulations provided a greater cumulative release from the formulation than the emulsion. Particularly, the nanoemulsion generated using the HCR provided the largest cumulative release from the formulation after 12 h. Therefore, the present study suggests that nanoemulsions can be created by means of hydrodynamic cavitation, which reduces the droplet size, as compared to that generated using other techniques. The satisfactory results of this study indicate that the rotor-stator-type HCR is a potentially cost-effective technology for nanoemulsion production.     

Degradation of benzene present in wastewater using hydrodynamic cavitation in combination with air

The degradation of benzene present in wastewater using hydrodynamic cavitation (HC) alone as well as in combination with air has been studied using nozzles as cavitating device of HC reactor. Initially, the energy efficiency of the HC reactor operated at different inlet pressures was determined using the calorimetric studies. Maximum energy efficiency of 53.4% was obtained at an inlet pressure of 3.9 bar. The treatment processes were compared under adiabatic as well as isothermal conditions and it was observed that under the adiabatic condition, the extent of degradation is higher as compared to isothermal condition. Studies related to the understanding the effect of inlet pressure (range of 1.8–3.9 bar) revealed that the maximum degradation as 98.9% was obtained at 2.4 bar pressure using the individual operation of HC under adiabatic conditions and in 70 min of treatment. The combination of HC with air was investigated at different air flow rates with best results for maximum degradation of benzene achieved at air flow rate of 60 mL/sec. A novel approach of using cavitation for a limited fraction of total treatment time was also demonstrated to be beneficial in terms of the extent of degradation as well as energy requirements and cost of operation. Based on the cavitational intensity, the resonant radius of aggregates of cavitation bubbles was also determined for distilled water as well as for aqueous solution of benzene. Overall, significant benefits of using HC combined with air have been demonstrated for degradation of benzene along with fundamental understanding into cavitation effects.     

The effects of waste-activated sludge pretreatment using hydrodynamic cavitation for methane production

Disintegration of waste-activated sludge (WAS) is regarded as a prerequisite of the anaerobic digestion (AD) process to reduce sludge volume and increase methane yield. Hydrodynamic cavitation (HC), which shares a similar underlying principle with ultrasonication but is energy-efficient, was employed as a physical means to break up WAS. Compared with ultrasonic (180–3600 kJ/kg TS) and thermal methods (72,000 kJ/kg TS), HC (60–1200 kJ/kg TS) found to consume significantly low power. A synergetic effect was observed when HC was combined with alkaline treatment in which NaOH, KOH, and Ca(OH)₂ were used as alkaline catalysts at pH ranging from 8 to 13. As expected, the production yield of CH₄ gas increased proportionally as WAS disintegration proceeded. HC, when combined with alkaline pretreatment, was found to be a cost-effective substitute to conventional methods for WAS pretreatment.     

High-intensity ultrasound pretreatment influence on whey protein isolate and its use on complex coacervation with kappa carrageenan: Evaluation of selected functional properties

The aim of this work was to evaluate the influence of high-intensity ultrasound (HIUS) treatment on whey protein isolate (WPI) molecular structure as a previous step for complex coacervation (CC) with kappa-carrageenan (KC) and its influence on CC functional properties. Protein suspension of WPI (1% w/w) was treated with an ultrasound probe (24 kHz, 2 and 4 min, at 50 and 100% amplitude), non HIUS pretreated WPI was used as a control. Coacervation was achieved by mixing WPI and KC dispersions (10 min). Time and amplitude of the sonication treatment had a direct effect on the molecular structure of the protein, FTIR-ATR analysis detected changes on pretreated WPI secondary structure (1600–1700 cm⁻¹) after sonication. CC electrostatic interactions were detected between WPI positive regions, KC sulfate group (1200–1260 cm⁻¹), and the anhydrous oxygen of the 3,6 anhydro-D-galactose (940–1066 cm⁻¹) with a partial negative charge. After ultrasound treatment, a progressive decrease in WPI particle size (nm) was detected. Rheology results showed pseudoplastic behavior for both, KC and CC, with a significant change on the viscosity level. Further, volume increment, stability, and expansion percentages of CC foams were improved using WPI sonicated. Besides, HIUS treatment had a positive effect on the emulsifying properties of the CC, increasing the time emulsion stability percentage. HIUS proved to be an efficient tool to improve functional properties in WPI-KC CC.     

Degradation of 2,4-dinitrophenol using a combination of hydrodynamic cavitation,chemical and advanced oxidation processes

In the present work, degradation of 2,4-dinitrophenol (DNP), a persistent organic contaminant with high toxicity and very low biodegradability has been investigated using combination of hydrodynamic cavitation (HC) and chemical/advanced oxidation. The cavitating conditions have been generated using orifice plate as a cavitating device. Initially, the optimization of basic operating parameters have been done by performing experiments over varying inlet pressure (over the range of 3–6 bar), temperature (30 °C, 35 °C and 40 °C) and solution pH (over the range of 3–11). Subsequently, combined treatment strategies have been investigated for process intensification of the degradation process. The effect of HC combined with chemical oxidation processes such as hydrogen peroxide (HC/H₂O₂), ferrous activated persulfate (HC/Na₂S₂O₈/FeSO₄) and HC coupled with advanced oxidation processes such as conventional Fenton (HC/FeSO₄/H₂O₂), advanced Fenton (HC/Fe/H₂O₂) and Fenton-like process (HC/CuO/H₂O₂) on the extent of degradation of DNP have also been investigated at optimized conditions of pH 4, temperature of 35 °C and inlet pressure of 4 bar. Kinetic study revealed that degradation of DNP fitted first order kinetics for all the approaches under investigation. Complete degradation with maximum rate of DNP degradation has been observed for the combined HC/Fenton process. The energy consumption analysis for hydrodynamic cavitation based process has been done on the basis of cavitational yield. Degradation intermediates have also been identified and quantified in the current work. The synergistic index calculated for all the combined processes indicates HC/Fenton process is more feasible than the combination of HC with other Fenton like processes.     

Effects of ultrasound synergized with microwave on structure and functional properties of transglutaminase-crosslinked whey protein isolate

In the present study, ultrasound (400 W, U), microwave heating (75 ℃ for 15 min, M) and ultrasound synergized with microwave heating (UM) pretreatments of whey protein isolate (WPI) were applied to investigate and compare their influence on structure, physicochemical and functional characteristic of transglutaminase (TGase)-induced WPI. From the results of size exclusion chromatography, it could be seen that all three physical pretreatments could promote the formation of polymers in TGase cross-linked WPI, whose polymer amounts were increased by the order of U, UM and M pretreatment. Among three physical methods, M pretreatment had the strongest effect on structure and functional characteristics of TGase-induced WPI. Furthermore, compared with TGase-induced WPI, α-helix and β-turn of M−treated TGase-induced WPI (M−WPI−TGase) were reduced by 7.86% and 2.93%, whereas its β-sheet and irregular curl were increased by 15.37% and 7.23%. Zeta potential, emulsion stability and foaming stability of M−WPI−TGase were increased by 7.8%, 59.27% and 28.95%, respectively. This experiment exhibited that M was a more effective pretreatment method than U, UM for WPI, which could promote its reaction with TGase and improve its functional properties.     

Intensification of Red-G dye degradation used in the dyeing of alpaca wool by advanced oxidation processes assisted by hydrodynamic cavitation

Red-G dye is one of the main dyes used in the textile industry to dye alpaca wool. Therefore, considering the large volume of processed wool in Perú, the development of efficient technologies for its removal is a present scientific issue. In this study, an integrated system based on hydrodynamic cavitation (HC) and photo-Fenton process was evaluated to remove the Red-G dye. Using a hybrid cavitation device (venturi + orifice plate), the effect of pH was evaluated, achieving 21 % of removal at pH 2 which was more than 80 % higher compared to pH 4 and 6. The effect of temperature was also evaluated in HC-system at pH 2, where percentage of dye degradation increased at lower temperatures (around 20 °C). Then, 50.7 % of dye was removed under optimized condition of HC-assisted Fenton process (FeSO₄:H₂O₂ of 1:30), that value was improved strongly by UV-light incorporation in the HC-system, increasing to 99 % removal efficiency with respect to HC-assisted Fenton process and reducing the time to 15 min. Finally, the developed cavitation device in combination with photo-Fenton process removed efficiently the dye and thus could be considered an interesting option for application to real wastewater.     

Continuous double-step acid catalyzed esterification production of sludge palm oil using 3D-printed rotational hydrodynamic cavitation reactor

Sludge palm oil (SPO) with high free fatty acid (FFA) content was processed using a continuous and double-step esterification production process in a rotor-stator-type hydrodynamic cavitation reactor. Three-dimensional printed rotor was made of plastic filament and acted as a major element in minimizing the FFA content in SPO. To evaluate the reduced level of FFAs using both methods, five independent factors were varied: methanol content, sulphuric acid content (H₂SO₄), hole diameter, hole depth, and rotor speed. The first-step conditions for the esterification process included 60.8 vol% methanol content, 7.2 vol% H₂SO₄ content, 5.0 mm diameter of the hole, 6.1 mm depth of the hole, and 3000 rpm speed of the rotor. The initial free fatty acid content decreased from 89.16 wt% to 35.00 wt% by the predictive model, while 36.69 wt% FFA level and 94.4 vol% washed first-esterified oil yield were obtained from an actual experiment. In the second-step, 1.0 wt% FFA was achieved under the following conditions: 44.5 vol% methanol content, 3.0 vol% H₂SO₄ content, 4.6 mm hole diameter, 5.8 mm hole depth, and 3000 rpm rotor speed. The actual experiment produced 0.94 wt% FFA content and 93.9 vol% washed second-esterified oil yield. The entire process required an average electricity of 0.137 kWh/L to reduce the FFA level in the SPO below 1 wt%.