Ultrasonic-assisted preparation of eucalyptus oil nanoemulsion: Process optimization,in vitro digestive stability,and anti-Escherichia coli activity

Eucalyptus oil (EO) is a natural and effective antimicrobial agent; however, it has disadvantages such as poor water solubility and instability. The aim of this study was to investigate the effect of process vessels and preparation process parameters on the particle size of the emulsion droplets using ultrasonic technique and response surface methodology to prepare eucalyptus oil nanoemulsion (EONE). The optimal sonication process parameters in conical centrifuge tubes were confirmed: sonication distance of 0.9 cm, sonication amplitude of 18%, and sonication time of 2 min. Under these conditions, the particle size of EONE was 18.96 ± 4.66 nm, the polydispersity index was 0.39 ± 0.09, and the zeta potential was −31.17 ± 2.15 mV. In addition, the changes in particle size, potential, micromorphology, and anti-Escherichia coli activity of EONE during digestion were investigated by in vitro simulated digestion. The emulsion was stable in simulated salivary fluid, tended to aggregate in simulated gastric fluid, and increased in particle size and potential value in simulated intestinal fluid. EONE showed higher anti-E. coli activity than EO by simulated digestion. These results provide a useful reference for the in vivo antimicrobial application of the essential oil.     

Process optimization and stability of D-limonene-in-water nanoemulsions prepared by ultrasonic emulsification using response surface methodology

d-limonene in water nanoemulsion was prepared by ultrasonic emulsification using mixed surfactants of sorbitane trioleate and polyoxyethylene (20) oleyl ether. Investigation using response surface methodology revealed that 10% d-limonene nanoemulsions formed at S₀ ratio (d-limonene concentration to mixed surfactant concentration) 0.6-0.7 and applied power 18 W for 120 s had droplet size below 100 nm. The zeta potential of the nanoemulsion was approximately −20 mV at original pH 6.4, closed to zero around pH 4.0, and around −30 mV at pH 12.0. The main destabilization mechanism of the systems is Ostwald ripening. The ripening rate at 25 °C (0.39 m³ s⁻¹ × 10²⁹) was lower than that at 4 °C (1.44 m³ s⁻¹ × 10²⁹), which was in agreement with the Lifshitz-Slezov-Wagner (LSW) theory. Despite of Ostwald ripening, the droplet size of d-limonene nanoemulsion remained stable after 8 weeks of storage.     

Optimization of ultrasound-assisted extraction of bioactive compounds from coffee pulp using propylene glycol as a solvent and their antioxidant activities

In the cosmetic and pharmaceutical industries, it has been increasingly popular to use alternative solvents in the extraction of bioactive compounds from plants. Coffee pulp, a by-product of coffee production, contains different phenolic compounds with antioxidant properties. The effects of polyols, amplitude, extraction time, solvent concentration, and liquid–solid ratio on total phenolic content (TPC) using ultrasound-assisted extraction (UAE) were examined by single-factor studies. Three main factors that impact TPC were selected to optimize the extraction conditions for total phenolic content (TPC), total flavonoid content (TFC), total tannin content (TTC), and their antioxidant activities using the Box-Behnken design. Different extraction methods were compared, the bioactive compounds were identified and quantified by liquid chromatography triple quadrupole mass spectrometer (LC-QQQ), and the cytotoxicity and cellular antioxidant activities of the extract were studied. According to the response model, the optimal conditions for the extraction of antioxidants from coffee pulp were as follows: extraction time of 7.65 min, liquid–solid ratio of 22.22 mL/g, and solvent concentration of 46.71 %. Under optimized conditions, the values of TPC, TFC, TTC, 1,1-diphenyl-2-picryl-hydrazil (DPPH) radical scavenging assay, 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical scavenging assay, and Ferric reducing antioxidant power assay (FRAP) were 9.29 ± 0.02 mg GAE/g sample, 58.82 ± 1.38 mg QE/g sample, 8.69 ± 0.25 mg TAE/g sample, 7.56 ± 0.27 mg TEAC/g sample, 13.59 ± 0.25 mg TEAC/g sample, and 10.90 ± 0.24 mg FeSO₄/g sample, respectively. Compared with other extraction conditions, UAE with propylene glycol extract (PG-UAE) was significantly higher in TPC, TFC, TTC, DPPH, ABTS, and FRAP response values than UAE with ethanol (EtOH-UAE), maceration with propylene glycol (PG-maceration), and maceration with ethanol (EtOH -maceration) (p < 0.05). Major bioactive compounds detected by LC-QQQ included chlorogenic acid, caffeine, and trigonelline. At higher concentrations starting from 5 mg/ml, PG-UAE extract showed higher cell viability than EtOH-UAE in both cytotoxicity and cellular antioxidant assays. The researcher expects that this new extraction technique developed in this work could produce a higher yield of bioactive compounds with higher biological activity. Therefore, they can be used as active ingredients in cosmetics (anti-aging products) and pharmaceutical applications (food supplements, treatment for oxidative stress-related diseases) with minimal use of chemicals and energy.     

Experimental investigation and metallographic characterization of fiber laser beam welding of Ti-6Al-4V alloy using response surface method

In the present study, experimental investigations of fiber-laser-beam-welding of 5 mm thick Ti-6Al-4V alloy are carried out based on statistical design of experiments. The relationship between the process parameters such as welding power, welding speed, and defocused position of the laser beam with the output responses such as width of the fusion zone, size of the heat affected zone, and fusion zone area are established in terms of regression models. Also, the most significant process parameters and their optimum ranges are identified and their percentage contributions on output responses are calculated. It is observed that welding power and speed plays the major role for full penetration welding. Also, welding power shows direct effect whereas welding speed shows the inverse effect on the output responses. The bead geometry is influenced by the defocused position of the laser beam due to the change in power density on the workpiece surface. However, overall fusion zone area is unaffected. Mechanical characterization of the welded samples such as microstructural analysis, hardness, and tensile tests are conducted. It is noticed that the hardness value of the FZ is higher than the HAZ and BM zone due to the difference in cooling rate during welding which promotes the formation of α′ martensitic phase in the FZ. Also, an average hardness value in the FZ is compared for two different defocusing positions (i.e. 1 and 2 mm). It is found that hardness value is higher for 1 mm defocused position than 2 mm due the decrement in grain size below a critical range at 2 mm defocused position. The ultimate tensile strength and % elongation of the welded samples are degraded as compared to BM which can be further improved by post heat treatment.     

Artificial neural network model- and response surface methodology-based optimization of Atractylodis Macrocephalae Rhizoma polysaccharide extraction,kinetic modelling and structural characterization

Atractylodis Macrocephalae Rhizoma (AMR) is the dried rhizome of Atractylodes macrocephala Koidz, which is widely used in the development of health products. AMR contains a large number of polysaccharides, but at present there are fewer applications for these polysaccharides. In this study, the effects of different extraction methods on the Atractylodis Macrocephalae Rhizoma polysaccharide (AMRP) yield were investigated, and the conditions for ultrasound-assisted extraction were optimized by response surface methodology (RSM) and three neural network models (BP neural network, GA-BP neural network and ACO-GA-BP neural network). The best conditions were a liquid-to-solid ratio of 17 mL/g, ultrasonic power of 400 W, extraction temperature of 72 °C, and extraction time of 40 min, which yielded 31.31% AMRP. The kinetic equation of AMRP was determined and compared with the results predicted by three neural network models. It was finally determined that the extraction conditions, kinetic processes and kinetic equation predicted by the GA-ACO-BP neural network were optimal. In addition, AMRP was characterized using SEM, FTIR, HPLC, UV, XRD, and NMR, and the structural study revealed that AMRP has a rough exterior and a porous interior; moreover, it contains high levels of glucose (5.07%), arabinose (0.80%), and galactose (0.74%). AMRP has three crystal structures, consisting of two β-type monosaccharides and one α-type monosaccharide. Additionally, the effectiveness of AMRP as an antioxidant was demonstrated in an in vitro experiment.     

Structural-borne acoustics analysis and multi-objective optimization by using panel acoustic participation and response surface methodology

This paper is aimed to investigate the structural-borne acoustics analysis and multi-objective optimization of an enclosed box structure by using the panel acoustic participation (PAP) and response surface methodology (RSM). The acoustic frequency response function is applied to achieve the critical frequency of interest under each excitation. The PAP analysis is then carried out at all critical frequencies and the remarkable acoustic panels are identified. The correlation coefficient matrix method is proposed for reselecting and grouping the positions of acoustic panels identified to paste damping layer to control noise. With the help of faced central composite design, an efficient set of sample points are generated and then the second-order polynomial functions of sound pressure response at each critical frequency are computed and verified by the adjusted coefficient of multiple determination. The functional relationships between sound pressure responses and the thicknesses of damping layers are investigated, and multi-objective optimization of the thicknesses of damping layers is developed. The results indicate that, by using the PAP and RSM, the structural-borne acoustics at critical frequencies are calculated conveniently and controlled effectively. The optimization process of the explicit optimization model proposed in this paper is simple and the computational time is saved.     

Study on the emulsification and oxidative stability of ovalbumin-pectin-pumpkin seed oil emulsions using ovalbumin solution prepared by ultrasound

Pumpkin seed oil (PSO), which is a valuable compound with high nutritional value used for the prevention of various chronic diseases, is prone to oxidation. In this work, small and uniform (su) ovalbumin (OVA) and pectin (PEC) were used to stabilize PSO in the form of an emulsion. The results showed that suOVA-PEC-PSO emulsion with a droplet size of 9.82 ± 0.05 μm was successfully self-assembled from PSO, PEC, and suOVA solution (with a droplet size of 230.13 ± 14.10 nm) treated with 300 W ultrasound, owing to the formation of a more stable interfacial film on the surface of droplets. The interfacial, rheological, emulsifying, and antioxidant properties of the suOVA-PES-PSO emulsions were excellent, owing to the synergistic effects between PEC and suOVA solution. Moreover, the physical stability of the suOVA-PEC-PSO emulsions to salt stress, a freeze-thaw cycle, and heat treatment was also increased and the oxidation of linolenic acid was notably delayed. These results have extended the food-related applications of OVA and PSO, and provide a promising foundation for further exploration of the self-assembly of composite emulsions by small and uniform proteins.     

Formulation development and optimization of a novel Cremophore EL-based nanoemulsion using ultrasound cavitation

In the present study, response surface methodology (RSM) based on central composite design (CCD) was employed to investigate the influence of main emulsion composition variables, namely drug loading, oil content, emulsifier content as well as the effect of the ultrasonic operating parameters such as pre-mixing time, ultrasonic amplitude, and irradiation time on the properties of aspirin-loaded nanoemulsions. The two main emulsion properties studied as response variables were: mean droplet size and polydispersity index. The ultimate goal of the present work was to determine the optimum level of the six independent variables in which an optimal aspirin nanoemulsion with desirable properties could be produced. The response surface analysis results clearly showed that the variability of two responses could be depicted as a linear function of the content of main emulsion compositions and ultrasonic processing variables. In the present investigation, it is evidently shown that ultrasound cavitation is a powerful yet promising approach in the controlled production of aspirin nanoemulsions with smaller average droplet size in a range of 200-300 nm and with a polydispersity index (PDI) of about 0.30. This study proved that the use of low frequency ultrasound is of considerable importance in the controlled production of pharmaceutical nanoemulsions in the drug delivery system.     

Effect of the lactose source on the ultrasound-assisted enzymatic production of galactooligosaccharides and gluconic acid

It is well known that one of the main problems in galactooligosaccharide production (GOS) via tranglycosylation of lactose is the presence of monosaccharides that contribute to increasing the glycaemic index, as is the case of glucose. In this work, as well as studying the effect of ultrasound (US) on glucose oxidase (Gox) activation during gluconic acid (GA) production, we have carried out an investigation into the selective oxidation of glucose to gluconic acid in multienzymatic reactions (β-galactosidase (β-gal) and Gox) assisted by power US using different sources of lactose as substrate (lactose solution, whey permeate, cheese whey). In terms of the influence of matrix on GOS and GA production, lactose solution gave the best results, followed by cheese whey and whey permeate, salt composition being the most influential factor. The highest yields of GOS production with the lowest glucose concentration and highest GA production were obtained with lactose solution in multienzymatic systems in the presence of ultrasound (30% amplitude) when Gox was added after 1 h of treatment with β-gal. This work demonstrates the ability of US to enhance efficiently the obtainment of prebiotic mixtures of low glycaemic index.     

Nanoemulsification of soybean oil using ultrasonic microreactor: Process optimization,scale-up and numbering-up in series

Ultrasonically-induced nanoemulsions have been widely investigated for the development of functional food, cosmetics, and pharmaceuticals due to ideal droplet sizes (DS), low polydispersity index (PDI), and superior physical stability. However, a series of frequently-used ultrasonic set-ups mainly suffered from a low ultrasonic energy efficiency caused by the large acoustic impedance and energy consumption, subordinately confronted with a low throughput, complicated fabrication with complex structure and weak ultrasonic cavitation. Herein, we employed a typical ultrasonic microreactor (USMR) that ensured the high-efficient energy input and generated intense cavitation behavior for efficient breakage of droplets and continuous production of unified oil-in-water (O/W) nanoemulsions in a single cycle and without any pre-emulsification treatment. The emulsification was optimized by tuning the formula indexes, technological parameters, and numerical analysis using Response Surface Methodology (RSM), followed by a comparison with the emulsification by a traditional ultrasonic probe. The USMR exhibited superior emulsification efficiency and easy scale-up with remarkable uniformity by series mode. In addition, concurrent and uniform nanoemulsions with high throughput could also be achieved by a larger USMR with high ultrasonic power. Based on RSM analysis, uniform DS and PDI of 96.4 nm and 0.195 were observed under the optimal conditions, respectively, well consistent with the predicted values. Impressively, the optimal nanoemulsions have a uniform spherical morphology and exhibited superior stability, which held well in 45 days at 4℃ and 25℃. The results in the present work may provide a typical paradigm for the preparation of functional nanomaterials based on the novel and efficient emulsification tools.     

Composition and antioxidant activity of anthocyanins from Aronia melanocarpa extracted using an ultrasonic-microwave-assisted natural deep eutectic solvent extraction method

A time-saving, efficient, and environmentally friendly ultrasonic-microwave-assisted natural deep eutectic solvent (UMAE-NADES) extraction method was developed for the extraction of anthocyanins from Aronia melanocarpa. Eight different natural eutectic solvents were screened initially, and choline chloride-glycerol was selected as the extraction solvent. The extraction conditions were optimized using the response surface methodology, and the extraction rate of anthocyanins was higher than those achieved using the traditional ethanol method, natural deep eutectic solvent extraction method, and ultrasonic-microwave-assisted ethanol method. Six anthocyanins, including cyanidin-3-O-galactoside, cyanidin-3-O-glucoside, cyanidin-3-O-arabinoside, cyanidin-3-O-xyloside, cyanidin-3,5-O-dihexoside, and the dimer of cyanidin-hexoside were identified and extracted at a purity of 448.873 mg/g using high performance liquid chromatography-mass spectrometry (HPLC-MS). The compounds extracted using UMAE-NADES had higher antioxidant capacities than those extracted by the other three methods. The UMAE-NADES demonstrated significant efficiency toward the extraction of bioactive substances and has potential utility in the food and pharmaceutical industries.     

Prediction of weld strength and seam width for laser transmission welding of thermoplastic using response surface methodology

In the present work, a study is made to investigate the effects of process parameters, namely, laser power, welding speed, size of the laser beam and clamp pressure, on the lap-shear strength and weld-seam width for laser transmission welding of acrylic (polymethyl methacrylate), using a diode laser system. Response surface methodology (RSM) is employed to develop mathematical relationships between the welding process parameters and the output variables of the weld joint to determine the welding input parameters that lead to the desired weld quality. In addition, using response surface plots, the interaction effects of process parameters on the responses are analyzed and discussed. The statistical software Design-Expert v7 is used to establish the design matrix and to obtain the regression equations. The developed mathematical models are tested by analysis-of-variance (ANOVA) method to check their adequacy. Finally, a comparison is made between measured and calculated results, which are in good agreement. This indicates that the developed models can predict the responses adequately within the limits of welding parameters being used.     

An ammonium sulfate/ethanol aqueous two-phase system combined with ultrasonication for the separation and purification of lithospermic acid B from Salvia miltiorrhiza Bunge

We studied the effect of ultrasonication extraction technology combined with ammonium sulfate/ethanol aqueous two-phase system (ATPS) for the separation of lithospermic acid B (LAB) from Salvia miltiorrhiza Bunge. According to the literature and preliminary studies, ammonium sulfate concentration, ethanol concentration, pH, ultrasonication power, ultrasonication time and the ratio of solvent-to-solid were investigated using a single factor design to identify the factors affecting separation. Taking into consideration a simultaneous increase in LAB recovery (R (%)) and partition coefficient (K), the best performance of the ATPS was obtained at 25 °C and pH 2 using ammonium sulfate 22% (w/w) and ethanol 30% (w/w). To keep the solvent-to-solid ratio at 10, response surface methodology was used to find the optimal ultrasonication power and ultrasonication time. Quadratic models were predicted for LAB yield in the upper phase. Optimal conditions of 572.1 W ultrasonication power and 42.2 min produced a maximum yield of LAB of 42.16 mg g⁻¹ sample. There was no obvious degradation of LAB with ultrasound under the applied conditions, and the experimental yield of LAB was 42.49 mg g⁻¹ sample and the purity was 55.28% (w/w), which was much higher than that obtained using conventional extraction. The present study demonstrated that ultrasound coupled with aqueous two-phase systems is very efficient tool for the extraction and purification of LAB from Salvia miltiorrhiza Bunge.     

Simultaneous ultrasound-assisted removal of sunset yellow and erythrosine by ZnS:Ni nanoparticles loaded on activated carbon: Optimization by central composite design

The present study focused on the simultaneous ultrasound-assisted removal of sunset yellow and erythrosine dyes from aqueous solutions using ZnS:Ni nanoparticles loaded on activated carbon (ZnS:Ni-NP-AC) as an adsorbent. ZnS:Ni nanoparticles were synthesized and characterized using different techniques such as FESEM, XRD and TEM. The effects of various parameters such as sonication time, pH, initial dye concentrations and adsorbent dose on the percentage of dye removal were investigated. Parameters were optimized by central composite design (CCD) combined with response surface methodology (RSM) and desirability function (DF). A good agreement between experimental and predicted values was observed. The ultrasound-assisted adsorbent (0.04 g) was capable of high percentage removal (98.7% and 99.6%) of sunset yellow and erythrosine in short time (3.8 min).     

Spectroscopic, energetic and metallographic investigations of the laser lap welding of AISI 304 using the response surface methodology

Spectroscopic signals originated by the laser-induced plasma optical emission have been simultaneously investigated together with energetic and metallographic analyses of CO₂ laser welded stainless steel lap joint, using the Response Surface Methodology. This statistical approach allowed us to study the influence of the laser beam power and the laser welding speed on the following response parameters: plasma plume electron temperature, joint penetration depth and melted area. A clear correlation has been found between all the investigated response parameters. The results have been shown to be consistent with quantitative considerations on the energy supplied to the workpiece as far as the laser power and travel speed were varied. The regression model obtained in this way could be a valuable starting point to develop a closed loop control of the weld penetration depth and the melted area in the investigated process window.     

A new direction in design and manufacture of co-sensitized dye solar cells: Toward concurrent optimization of power conversion efficiency and durability

A novel methodology was implemented in the present study to concurrently control power conversion efficiency (η) and durability (D) of co-sensitized dye solar cells. Applying response surface methodology (RSM) and Desirability Function (DF), the main influential assembling (dye volume ratio and anti-aggregation agent concentration) and operational (performance temperature) parameters were systematically changed to probe their main and interactive effects on the η and D responses. Individual optimization based on RSM elucidated that D can be solely controlled by changing the ratio of vat-based organic photosensitizers, whereas η takes both effects of dye volume ratio and anti-aggregation concentration into account. Among the studied factors, the performance temperature played the most vital role in η and D regulation. In particular, however, multi-objective optimization by DF explored the degree to which one should be careful about manipulation of assembling and operational parameters in the way maximization of performance of a co-sensitized dye solar cell.     

Zirconia supported 12-molybdophosphoric acid: Physico-chemical characterization and non-solvent liquid phase oxidation of styrene

A series of solid catalysts comprising 12-molybdophosphoric acid (PM) and hydrous zirconia (Z) have been synthesized by wet impregnation and characterized by various physico-chemical techniques. The structure of PM onto the surface of the support has been confirmed by ³¹P MAS NMR and XPS. Non-solvent liquid phase oxidation of styrene has been carried out with 30% aqueous H₂O₂ as an oxidant by varying different parameters over fresh catalysts. The catalytic activity of calcinated catalysts has also been evaluated by carrying out same oxidation reaction under optimized conditions. As solid catalysts, they show high conversion of styrene with good selectivity towards benzaldehyde and styrene epoxide.     

Statistical optimization of process parameters for improvement of phycobiliproteins (PBPs) yield using ultrasound-assisted extraction and its kinetic study

The present study is mainly concerned for the development of an optimal ultrasound-assisted extraction (UAE) condition for phycobiliproteins (PBPs) from Oscillatoria sp. (BTA 170) using Taguchi methodology. Four process parameters viz. solid to liquid ratio, duty cycle, electrical acoustic intensity, and pH, for UAE were optimized using Taguchi methodology for enhanced PBPs extraction. The ratio of signal to noise (S/N) was used to compute the optimized condition required to attain a higher yield of PBPs, the average performance of individual parameter and corresponding interactive effects. The statistically significant parameters with their contribution were assessed using Analysis of variance (ANOVA). Results showed that duty cycle contributed the maximum influence (30.81%) on phycocyanin (PC) extraction followed by a solid liquid ratio (28.62%), pH (22.46%) and electrical acoustic intensity (18.10%). The highest contribution on the extraction of phycoerythrin (PE) was found from pH (33.16%), followed by duty cycle (31.57%), solid to liquid ratio (22.83%) and electrical acoustic intensity (12.45%). For extraction of allophycocyanin (APC), the duty cycle, solid to liquid ratio, pH and electrical acoustic intensity contributed 29.47, 29.07, 29.03, and 12.43% respectively. Results obtained from Taguchi methodology indicated that enhanced PC (94.10%), PE (95.20%) and APC (90.54%) can be achieved with solid–liquid ratio (0.2 g/ml), electrical acoustic intensity (16.99 w/cm²), duty cycle (75%), and pH 7 than the yield of PBPs obtained under unoptimized condition. In the present study, higher yield of PC (38.99%), PE (20.84%), and APC (11.93%) were attained with UAE compared to yield obtained from homogenized Oscillatoria sp. BTA 170 using 0.05 M phosphate buffer. Batch extraction data of PBPs under UAE was fitted well with the second order model. The values of second-order rate constant (k) were computed as 6.66 × 10⁻⁴, 64.09 × 10⁻⁴ and 1.49 × 10⁻⁴ L/mg/min for extraction of PC, PE and APC respectively. The PBPs exhibited significant antioxidant property and hydrogen peroxide scavenging activity, which were increased with the enhancement of PBPs concentration.     

Synthesis of nanocomposites of iron oxide/gold (Fe3O4/Au) loaded on activated carbon and their application in water treatment by using sonochemistry: Optimization study

This paper focuses on the finding best operational conditions using response surface methodology (RSM) for Rhodamine123 (R123) and Disulfine blue (DSB) dyes removal by ultrasound assisted adsorption onto Au-Fe₃O₄ nanoparticles loaded on activated carbon (Au-Fe₃O₄ NPs-AC). The influences of variables such as initial R123 (X₁) and DSB concentration (X₂), pH (X₃), adsorbent mass (X₄) and sonication time (X₅) on their removal were investigated by small central composite design (CCD) under response surface methodology. The significant variables and the possible interactions among variables were investigated and estimated accordingly. The best conditions were set as: 4 min, 4.0, 0.025 g, 13.5 and 26.5 mg L⁻¹ for sonication time, pH, adsorbent weight, initial R123 and DSB concentration, respectively. At above conditions, the adsorption equilibrium and kinetic follow the Langmuir isotherm and pseudo-second-order kinetic model, respectively. The maximum monolayer capacity (Q_max) of 71.46 and 76.38 mg g⁻¹ for R123 and DSB show sufficiency of model for well presentation of experimental data.     

Process optimization studies of 10-Hydroxycamptothecin (HCPT)-loaded folate-conjugated chitosan nanoparticles by SAS-ionic crosslink combination using response surface methodology (RSM)

10-Hydroxycamptothecin (HCPT) is a well-established topoisomerase I inhibitor of a broad spectrum of cancers. However, poor aqueous solubility, low instability, and toxicity to normal tissues have limited its clinical development. A novel HCPT-containing drug carrier system was developed to overcome these disadvantages. The response surface methodology was used to optimize the process of preparing HCPT-chitosan nanoparticles (HCPT-CSNPs) by the SAS-ionic crosslink (supercritical antisolvent SAS) combination method; the resulting HCPT-CSNPs were then conjugated with folate for specific targeting. A central composite design, composed of four independent variables, namely, chitosan concentration, TPP concentration, HCPT nanoparticle concentration, and crosslink time, was applied in the modeling process. The mean particle size and drug entrapment efficiency (DEE) of HCPT-CSNPs were chosen as response variables. The interactive effects of the four independent variables on the response variables were also studied. Nanoparticle characteristics such as morphology, DEE, and mean particle size were investigated. The optimum conditions for preparing HCPT-CSNPs were determined as follows: folate-coupled chitosan concentration 2.46 mg/ml, TPP concentration 7.73 mg/ml, HCPT nanoparticle concentration 0.48 mg/ml, and crosslinking time 47.4 min. Optimum conditions for preparing desired HCPT-CSNPs with a mean particle size of 173.5 nm and entrapment efficiency of 77.3% were obtained. The resulting folate-conjugated HCPT-CSNPs (FA-HCPT-CSNPs) reveal that the amount of folate conjugation was 197.64 mg/g CS. FA-HCPT-CSNPs used in drug carrier systems could have potential value in HCPT-sensitive tumors.