Box-Behnken Experimental Design Research Articles

Photocatalytic Degradation of Tartrazine and Naphthol Blue Black Binary Mixture with the TiO2 Nanosphere under Visible Light: Box-Behnken Experimental Design Optimization and Salt Effect

In this study, TiO2 nanospheres (TiO2-NS) were synthesized by the solvothermal method. Firstly, the synthesized nanomaterial was characterized by X-ray diffraction (XRD), Fourier Transformed Infrared (FTIR), scanning electron microscopy (SEM) and UV-Vis Diffuse Reflectance Spectroscopy (DRS). To study the photocatalytic degradation of Tartrazine (TTZ) and Naphthol Blue Black (NBB) in a binary mixture, the influence of some key parameters such as pH, pollutant concentration and catalyst dose was taken into account under visible and UV light. The results show a 100% degradation efficiency for TTZ after 150 min of UV irradiation and 57% under visible irradiation at 180 min. The kinetic study showed a good pseudo-first-order fit to the Langmuir–Hinshelwood model. Furthermore, in order to get closer to the real conditions of textile wastewater, the influence of the presence of salt on TiO2-NS’s photocatalytic performance was explored by employing NaCl as an inorganic ion. The optimum conditions provided by the Response Surface Methodology (RSM) were low concentrations of TTZ (2 ppm) and NBB (2.33 ppm) and negligible salt (NaCl) interference. The percentage of photodegradation was high at low pollutant and NaCl concentrations. However, this yield became very low as NaCl concentrations increased. The photocatalytic treatment leads to 31% and 53% of mineralization yield after 1 and 3 h of visible light irradiation. The synthesis of TiO2-NS provides new insights that will help to develop an efficient photocatalysts for the remediation of contaminated water.

Abrasive waterjet cutting of r‐GO infused magnesium fiber metal laminates: Experimental investigations and optimization through gorilla troops algorithm

AbstractLaminated metal‐composite structures, known as fiber metal laminates (FMLs), are modern lightweight materials extensively utilized in diverse industries such as aerospace and automotive manufacturing. These materials offer enhanced impact and fatigue resistance, making them invaluable for various applications. However, machining FMLs poses a challenge due to the occurrence of delamination and heterogenous in nature during conventional methods. Therefore, this study aims to investigate the quality characteristics such as kerf width, surface roughness and kerf taper of Magnesium based FMLs processed with an unconventional machining process namely abrasive water jet cutting (AWJC). These FMLs comprise alternately stacked kevlar/carbon fibers adhesively bonded with epoxy resin matrix filled with varying wt% of reduced graphene oxide (r‐GO), combined with AZ31B alloy sheet as the skin material. AWJC experiments were performed by varying the process parameters including waterjet pressure (275–325 MPa), stand‐off distance (2.5–3.5 mm), and cutting speed (600–800 mm/min) based on the box–behnken experimental design. The findings from the statistical analysis underscore the significant influences of stand‐off distance and waterjet pressure on kerf characteristics, while the inclusion of r‐GO is observed to notably affect the surface quality. In pursuit of enhancing cut quality, a multi‐response optimization was conducted employing the Gorilla Troops Optimization (GTO) algorithm. Comparative analysis with established metaheuristics like the gray wolf algorithm, dragonfly algorithm, and harmony search algorithm reveals GTO's superior performance across various metrics, including rapid convergence, diversity, and spacing values. Further, the cutting‐induced damages such as matrix plowing, fiber‐metal debonding and composite delamination were observed through microstructure analysis.Highlights Magnesium (AZ31B) fiber metal laminate comprising with various wt% of reduced graphene oxide was fabricated. Abrasive waterjet cutting of FMLs were performed based on box–behnken design experimental approach. Statistical analysis and influence of process parameters on the kerf width, surface roughness and kerf taper were investigated. Multi‐response optimization was carried out using metaheuristic‐based gorilla troops algorithm.

Optimization of biomass durian peel as a heterogeneous catalyst in biodiesel production using microwave irradiation

Abstract The present study investigated biodiesel production from the transesterification of palm oil with methanol using calcined biomass durian peel (BDP) as a heterogeneous catalyst assisted by microwave irradiation. Characterization of the calcined BDP showed that K2O is the main compound with a concentration of 86.15 wt%. The effect of three independent variables of catalyst weight (3–12 wt%), reaction time (1–10 min), and power of microwave (180–900 W) was used to determine the optimum condition on biodiesel production using the response surface method-based on the Box–Behnken design experiment. The optimum biodiesel conversion of 97.3% was achieved under experimental parameters of catalyst concentration of 12 wt%, reaction time of 9 min, and microwave power of 180 W. The catalyst concentration and reaction time have significant effects on biodiesel conversion.

Design of experiments using box behnken design in the development, characterization, mathematical modeling, and evaluation of lung targeted nebulized antiviral camostat mesylate loaded pegylated nanosuspension product

Box Behnken Design is a factorial statistical approach that helps to overcome limitations associated with time-consuming full factorial experimental design. This study used the Box Behnken design and Design of Experiments (DoE) to develop the nebulized antiviral Camostat Mesylate loaded pegylated nanoliposomal suspension product with the target profile specifications of particle size <250 nm, polydispersity index (PDI) between 0.1 and 0.3, and zeta potential (ZP) of 0±5 mV, % entrapment efficiency (%EE) between 40%w/v to 70 % w/v and % drug loading between 20%w/w to 50%w/w. Box Behnken design with a robust standard least squares model was employed to develop the pharma product. Camostat Mesylate loaded pegylated nanoliposomal suspension was developed using the scalable ethanol injection method. The three independent factors at three levels were concentrations of the lipids employed to develop the formulation. Applying holistic QbD during formulation development using Design of Experiments (DoE) ensures a science-based, risk-managed approach to developing high-quality pharmaceutical products. The novelty in this investigation is that, for the first time, nebulized antiviral Camostat mesylate-loaded Nanoliposomes with a size of <250 nm, size <250 nm, polydispersity index (PDI) between 0.1 and 0.3, and zeta potential (ZP) of 0±5 mV, % entrapment efficiency (%EE) between 40%w/v to 70 % w/v and % drug loading between 20%w/w to 50%w/w were developed using DoE, Box Behnken Design, mathematical modeling and Response Surface Methodology.Furthermore, the Camostat mesylate-loaded Nanoliposomes were evaluated for inhibiting SARS-CoV-2 injection. Based on our findings, we kept other concentrations, i.e., Camostat Mesylate concentration at 2.25 mg/mL, 0.9%w/v NaCl, and 90%v/v ethanol and process parameters, namely, stirring speed, stirring time, ethanol addition rate, syringe diameter constant. From the mathematical modeling and Response Surface Methodology, we observed that the particle size of the nanosuspension increased with an increase in concentrations of DPPC and cholesterol. At the same time, increasing concentrations of DPPC increased the % entrapment efficiency and % drug loading; an increase in the concentration of Cholesterol led to a decrease in both those parameters. DoE successfully developed Camostat Mesylate loaded pegylated nanoliposomes with a particle size of 188 ± 0.75 nm, 75.47 ± 5.87 % entrapment of Camostat Mesylate and 49.65 ± 3.86 % drug load of Camostat Mesylate in liposomal product using the ethanol-injection method meeting target profile. The in vitro extended release of Camo from the Camo-pegNLs was observed in 24 h. The nebulized Camo-pegNL product showed MMAD of 4.01 ± 0.21 μm, GSD of 1.34 ± 0.03, and %FPF of 29.53 ± 6.5 %. In conclusion, integrating nebulized product development, Box Behnken Design, enabled the successful development of nebulized Camostat Mesylate loaded pegylated nanoliposomal suspension product, demonstrating their suitability for lung delivery of the antiviral drug.

Bead-Free Electrospun Nanofibrous Scaffold Made of PVOH/Keratin/Chitosan Using a Box–Behnken Experimental Design and In Vitro Studies

Bead-free nanofibrous scaffolds composed of PVOH/keratin/chitosan were prepared using electrospinning after optimising the process parameters using a Box–Behnken experimental design. Two quadratic models were developed to optimise the fibre diameter and the diameter of fibre beads considering the voltage, flow rate, spinning distance, and amount of biopolymer as independent variables. All independent variables were found to be significant in determining responses, although not all interactions among these were significant. The models were highly effective in describing responses, with an R2 of 98.58 and 99.67%. The optimum conditions were determined to be 15.82 kV voltage, 0.25 mL/h flow rate, 105 mm spinning distance, and 30% biopolymers. The accuracy of the models was verified and found to be within an acceptable range. The bead-free nanofibrous scaffold exhibited no cytotoxicity to Human Aneuploid Immortal Keratinocyte (HaCaT) and Normal Human Dermal Fibroblast (NHDF) cell lines, enabling cell adhesion and proliferation. Both cell lines remained attached with perfect cell morphology when co-cultured on the scaffold for 30 days, indicating the scaffold’s potential for biomedical applications.

Study on temperature and hardness behaviors of Al-6060 alloy during magnetic abrasive finishing process using artificial neural networks

Magnetic Abrasive Finishing (MAF) revolutionizes surface finishing by utilizing magnetic fields to apply pressure, ensuring exceptionally high-quality surfaces. Its potential for ultra-precise finishing with fine magnetic abrasives is particularly noteworthy. This study analyzes subsurface temperature dynamics during the MAF process on Al-6060, recognizing temperature's pivotal role in shaping surface texture and mechanical-chemical properties. Unbonded magnetic abrasives containing SiC were used without lubrication or coolants in experimentation. Experimental parameters, including machining gap, abrasive weight, voltage, and rotational speed, were systematically varied using a Box-Behnken Design of Experiments. An artificial neural network facilitated comprehensive data modelling, enabling a detailed parametric study. Post-processing surface characterization provided crucial insights into the impact of surface temperature rise on surface finish and hardness. This holistic approach enhances understanding of temperature's influence on MAF efficacy and its outcomes on Al-6060 surfaces. Experimental findings demonstrate MAF's effectiveness in achieving low-temperature finishing of Al-6060, with a maximum surface temperature of 36 °C. ANOVA analysis quantitatively determined that voltage has greatest influence (55.56%), followed by abrasive weight (20.00%) and machining gap (13.89%), with rotational speed having the least impact (2.22%). Qualitatively, the developed ANN model accurately predicted changes in surface roughness (ΔRa), temperature (ΔT), and hardness (ΔH), with maximum errors of 4.107%, 5.588%, and 6.680%, respectively. XRD analysis provided quantitative evidence of SiC diffusion into the surface, resulting in a hardness increase from 2.7 HV to 5.6 HV. Additionally, chemical substrates deposition, such as silicon carbide, kaolinite, iron silicide, and cristobalite, elucidated chemical-mechanical interactions during the process at such low temperatures.

Optimization use of watermelon rind in the coagulation-flocculation process by Box Behnken design for copper, zinc, and turbidity removal

Watermelon rinds were investigated as a bio-coagulant for treating water contaminated by metals and turbidity, owing to their biodegradability and greater environmental friendliness compared to chemical coagulants. Fourier transform infrared spectroscopy, scanning electron microscopy paired with energy dispersive X-ray analysis and X-ray diffraction characterized the watermelon rinds before and after use. A Box-Behnken experimental design optimized the most influential parameters of initial pH, coagulant dose, and particle size based on response surface methodology. This analysis revealed the experimental data fit quadratic polynomial models, achieving maximum removal efficiencies of 97.51 % for zinc, 99.88 % for copper, and 99.21 % for turbidity under optimal conditions. Statistical analysis confirmed the models effectively captured the experimental data. Analysis of variance denoted the high significance of the quadratic effects of dose and pH. Removal of metal ions Zn2+ and Cu2+ was significantly impacted by these factors. The watermelon rind powder retained its coagulation efficiency after five cycles of reuse, with removal rates of 80.04 % for Zn, 88.33 % for Cu and 86.24 % for turbidity. These results demonstrate the potential of watermelon rind as an alternative coagulant for wastewater treatment. Further testing on real industrial effluents at larger scales would help assess their feasibility for real-world applications.

Investigation of the effects of corn flour, Spirulina powder, and buffalo yogurt on the quality characteristics of gluten-free muffins.

This study aimed to investigate the properties of buckwheat flour-based gluten-free muffins formulated by using corn flour (CF), Spirulina powder (SP), and buffalo yogurt (BY) levels as the factors in the Box-Behnken experimental design. The color values, total soluble polyphenol content (TPC), radical scavenging activity (RSA), specific volume, moisture content, and baking loss results were evaluated as the response variables in the design. The buckwheat flour in the control sample was substituted with CF and SP. Cow yogurt was used in the control muffin instead of milk and was substituted with BY at different levels in other samples. Among the response variables, TPC and RSA values were selected for the optimization studies. Decreasing the CF and SP levels in the formulations resulted in observing higher TPC and RSA values. Three optimal formulations were obtained by response surface methodology. Both cow and buffalo yogurt did not change the properties of the muffins adversely compared to milk. Increasing the SP content made the muffins firmer and caused a decrease in the L*, a*, b*, and browning index values of the muffins. However, sensory scores of SP-containing muffins were also satisfactory.

Sustainable Utilization of Mushroom By-Products Processed with a Combined Osmotic Dehydration Pretreatment and a Hot-Air-Drying Step.

Mushroom production and consumption are gaining increased interest due to their unique flavor and nutritional value. However, in the mushroom industry, large amounts of by-products are generated, which have a high negative environmental and economic impact. In this study, an osmotic dehydration process followed by hot-air-drying was applied to mushroom stems to produce dried mushrooms as the end product. The osmotic dehydration conditions (concentration of hypertonic solution, specifically, 10-30% maltodextrin and 20-40% oligofructose; a treatment time of 40-80 min; and a temperature range of 30-50 °C) were optimized using response surface methodology (RSM). The results showed that a four-factor three-level Box-Behnken experimental design was effectively implemented to evaluate the effect of the process parameters and identify the optimal osmotic dehydration conditions for producing osmotically dehydrated mushrooms. The main factor affecting mass transfer was the osmosis temperature, and the optimal conditions were found to be 38 °C, 40% oligofructose and 19.3% maltodextrin as the osmotic agents, and 80 min of immersion time. Moreover, the results showed that osmotic pretreatment, in the optimal conditions, significantly reduced the required drying time of the by-products compared to traditional hot-air-drying, especially at milder drying temperatures. Consequently, the required energy was also reduced by at least 40% at 50 °C.

Elucidating co-conditioning strategies of aluminium-based sludge using natural biopolymeric magnetite composite for leveraging dewaterability

Significant co-conditioning of aluminum-based sludge using alternative conditioner (natural biopolymer-magnetite/H2O2) as a Fenton conditioner for improving the sludge dewaterability is investigated in the current study. Firstly, natural biopolymer/magnetite (NBP-M) were prepared in various ratios and the samples labeled as (NBP-M-(1:1); NBP-M-(1:3) and NBP-M-(2:1). Focuses were placed on factors influencing such novel Fenton conditioner. Capillary suction time (CST) was utilized to assess sludge dewaterability. Experiments have exhibited that significant development of alum sludge dewaterability could be attained at minimum reaction time and using low concentrations of the applied catalyst, i.e. natural biopolymer/magnetite and H2O2. It was found that sludge conditioning and dewaterability became better with increasing both reagents concentrations. A Box–Behnken experimental design based on the response surface methodology was investigated to explore the optimum of the influencing variables, i.e. catalyst (NBP-M) concentration, H2O2 loading and pH. The optimized values for NBP-M, H2O2, and pH are 46 and 380 mg/L, respectively at pH 3.0, at which the CST reduction efficiency of 58 ± 3% can be attained, this approved with that predicted by an established polynomial model in the current investigation.

Optimization of 3D printing formulation of shrimp surimi based on response surface method

The Box-Behnken experimental design is an optimization statistical tool. In this study, shrimp surimi was taken as the research object, Box-Behnken experiment design and regression analysis were used, and the addition of cross-linked starch (CLS), soy protein isolate (SPI) and lard oil (LO) were used as experimental factors. The purpose was to explore the effects of adding different mixed ingredients on the properties of shrimp surimi. The study found that in order to improve the accuracy and stability of 3D printing of shrimp surimi, the material characteristics should meet two conditions at the same time: (i) Water holding capacity is controlled between 89% and 92%; (ii) AF value is controlled between 3500 and 4500 Pa·S1/z. When the addition amount of 3.00% LO, 4.50% CLS, and SPI was controlled between 1.58% and 6.53%, or when the addition amount of 3.00% LO, 4.50% SPI, and CLS was controlled between 1.80% and 6.75%, the above two conditions can be met at the same time, making the material properties of shrimp surimi conducive to 3D printing. This study determined the optimization range of the added amount of mixed ingredients to improve the adaptability of 3D printed shrimp surimi, which could meet the needs of consumers for personalized nutritious diets and improve the added value of shrimp surimi products.

Waste-derived activated carbons for effective adsorptive removal of strontium, barium, and binary pollutants: A response surface methodology study

A double-pronged approach to pollution management is proposed by sustainably managing solid wastes and converting them to activated carbons that are then utilized for water treatment. In this study, gas-to-liquids (GTL) derived biosolids, carboard and their mixed samples are used to produce the activated carbons. In a laboratory batch study, a Box-Behnken experimental design was used with four factors and three levels to optimize the removal of single component strontium and barium, and the binary system of pollutants from an aqueous solution. The design incorporated response surface modeling (RSM) techniques with a total of 29 different experimental data points collected and analyzed in this study. The study was conducted considering four parameters: the initial pH of the solution (ranging from 4.0 to 8.0), temperature (ranging from 20 to 40 ℃), the percentage of cardboard (fixed at 0 %, 50 %, and 100 %), and the amount of adsorbent (between 0.05 and 0.5 g). These factors were assigned three levels, represented as −1, 0, and 1. A second-order polynomial regression equation was then developed to estimate the responses. The statistical significance of the independent variables and their interactions was assessed using analysis of variance (ANOVA) with a 95 % confidence level (α = 0.05). The results revealed that only temperature and dosage show significant effects on the responses and that optimum values of the selected parameters were obtained by solving the prediction equations, which were validated with less than 4 % error %. In an attempt to optimize the factors, a pH of 5.5, temperature of 40 ℃ and dose of 0.3 g is found for all three samples. Validation results for optimization also proved that varying the percentage cardboard showed little difference in the percentage removal of all the pollutants. The results from this study can be directly applied for any such systems trying to optimize these parameters and the prediction equations can be utilized effectively.

A ropivacaine-eluting poly(lactide-co-caprolactone) wound dressing provided enhanced analgesia in partial-thickness porcine injuries.

Partial-thickness skin wounds are some of the most painful injuries due to large areas of exposed nerve endings. These injuries often require systemic opioid treatments to manage pain adequately. However, in 2021 alone, the CDC reported nearly 17,000 prescription opioid-related deaths in the USA, highlighting the ongoing need for non-opioid treatment strategies. In this manuscript, we developed a novel single-application ropivacaine-eluting primary wound dressing that could provide sustained ropivacaine delivery to partial-thickness wounds and assessed its in vivo feasibility for prolonged non-opioid analgesia. Sustained release of ropivacaine from a poly(lactide-co-e-caprolactone) matrix was first optimized in vitro using dissolution testing and a Box Behnken design of experiments. The optimized dressing was then tested against a clinical control silicone dressing in a porcine partial-thickness wound study to assess analgesic effect, pharmacokinetics, and wound healing. The ropivacaine-eluting dressing showed a moderate analgesic effect in vivo, where normalized single pinprick scores significantly improved pain over the testing period (4-168h) (control vs treatment: 232±25% vs 145±16%, p<0.0003). Ropivacaine blood plasma levels peaked at 8 hours post-treatment, with a maximum concentration of 246 ± 74 ng/mL. No significant differences in wound healing were found when compared to control. The ropivacaine-loaded poly(lactide-co-e-caprolactone)-based wound dressing provided sustained delivery of ropivacaine to partial-thickness skin wounds and enhanced analgesic effect compared to a clinical standard control dressing.

Optimization of the fermentation media and growth conditions of Bacillus velezensis BHZ-29 using a Plackett-Burman design experiment combined with response surface methodology.

Bacillus velezensis occurs extensively in the soil environment. It produces a range of antimicrobial compounds that play an important role in the field of biological control. However, during the actual application process it is often affected by factors such as the medium formulation and fermentation conditions, and therefore biocontrol measures often do not achieve their expected outcomes. In this study, the B. velezensis BHZ-29 strain was used as the research object. The carbon and nitrogen sources, and inorganic salts that affect the number of viable bacteria and antibacterial potency of B. velezensis BHZ-29, were screened by a single factor test. A Plackett-Burman design experiment was conducted to determine the significant factors affecting the number of viable bacteria and antibacterial potency, and a Box-Behnken design experiment was used to obtain the optimal growth of B. velezensis BHZ-29. The medium formula that produced the highest number of viable bacteria and most antibacterial substances was determined. The initial pH, temperature, amount of inoculant, liquid volume, shaking speed, and culture time were determined by a single factor test. The factors that had a significant influence on the number of viable bacteria of B. velezensis BHZ-29 were selected by an orthogonal test. A Box-Behnken design experiment was conducted to obtain the optimal fermentation conditions, and highest number of viable bacteria and antibacterial titer. Molasses, peptone, and magnesium sulfate had significant effects on the viable count and antibacterial titer of B. velezensis BHZ-29. The viable count of B. velezensis BHZ-29 increased from 7.83 × 109 to 2.17 × 1010 CFU/mL, and the antibacterial titer increased from 111.67 to 153.13 mm/mL when the optimal media were used. The optimal fermentation conditions for B. velezensis BHZ-29 were as follows: temperature 25.57°C, pH 7.23, culture time 95.90 h, rotation speed 160 rpm, amount of inoculant 2%, and liquid volume 100 ml. After the optimization of fermentation conditions, the number of viable bacteria increased to 3.39 × 1010 CFU/mL, and the bacteriostatic titer increased to 158.85 mm/ml.The plant height and leaf number of cotton plants treated with BHZ-29 fermentation broth were higher than those of cotton inoculated with Verticillium dahliae. The number of bacteria was 1.15 × 107 CFU/g, and the number of fungi was 1.60 × 105 spores/g. The disease index of the cotton seedlings treated with the optimized fermentation broth was 2.2, and a control effect of 93.8% was achieved. B. velezensis BHZ-29 could reduce the disease index of cotton Verticillium wilt and had a controlling effect on the disease. The best effect was achieved in the treatment group with an inoculation concentration of 2 × 108 CFU/ml, the disease index was 14.50, and a control effect of 84.18% was achieved. The fermentation process parameters of the number of viable bacteria and antibacterial titer by strain B. velezensis BHZ-29 were optimized to lay a foundation for the practical production and application of strain B. velezensis BHZ-29 in agriculture.

Response surface experimental design for simultaneous chromatographic determination of two antiviral agents “Favipiravir and Remdesivir” in pharmaceuticals and spiked plasma samples

AbstractThe antiviral agents, Favipiravir (FAV) and Remdesivir (REM), were introduced in the last few years alone or as combination regimen for successful management of the rapidly spreading CORONA virus pandemic. A newly developed rapid and sensitive high performance liquid chromatographic method (HPLC) has been developed for the simultaneous determination of their mixture. Firstly, one factor at a time optimization (OFAT) has been applied. Afterwards, quality by design approach (QbD) has been utilized using Box Behnken experimental design (BBD) for the development of an experimental design of four independent and nine dependent variables for much better refining of the optimized parameters. The established model has given an optimum resolution at; acetonitrile percentage of 52.66, mobile phase of pH 2.91, percentage of triethylamine of 0.15 and 1.30 mL/min flow rate. The proposed method has been validated according to the USP 31 NF 26 guidelines. Good linearity ranges have been obtained from 5.00 up to 50.00 μg/mL for FAV and from 2.00 up to 60.00 μg/mL for FAV and REM, respectively. Excellent relative standard deviation values (not more than 1.40) were obtained upon investigation of accuracy, precision and robustness. The developed method has succeeded in analysis of investigated drugs in their pharmaceutical formulations and spiked plasma samples with good recoveries of 99.00 and up to 106.00%. The proposed method is considered eligible for the quality control laboratories as well as in‐vivo determinations of both analytes.

Harvesting optimization and Omega-3 recovery improvement from Schizochytrium DT3 using surfactant-aided dispersed air flotation: Response surface methodology

Microalgae's potential to produce high value bioactives is contingent on the cost-effective harvesting of algal biomass. The use of CTAB (cetyl trimethylammonium bromide) assisted dispersed air flotation as a harvesting technique for Schizochytrium biomass, was optimized as a cost-effective means for recovery (R) and concentration factor (CF) using Response Surface Methodology. To the best of our knowledge, this is the first study to employ surfactant assisted dispersed air flotation (SDAF) in the recovery of a heterotrophic thraustochytrid strain. A Box-Behnken design of experiment investigating the operating parameters of CTAB amount, air flow rate and volume on Schizochytrium was employed. Initially, both responses were analyzed individually and then used to simultaneously maximize both variables. The optimized conditions of CTAB (500 mg/L), air flow of 2 L/min and volume of 600 mL resulted in a R of 91 % and CF of 19 times. Although a secondary step is necessary for further concentration, this technique utilizes 70 times less energy as compared to conventional centrifugation techniques which are used to recover Schizochytrium sp. This is significant as this technique can be easily adapted to existing bioreactors, as they are already equipped with gas spargers. In addition, the presence of surfactant carried through demonstrated an improvement in the recovery of long chain poly unsaturated fatty acid (PUFA) by 6 %, particularly in DHA and DPA which was not observed in washed biomass.

Supercritical CO2 Extraction of Fatty Acids, Phytosterols, and Volatiles from Myrtle (Myrtus communis L.) Fruit.

Myrtle (Myrtus communis L.) is a coastal Mediterranean aromatic medicinal plant rich in essential oil components, flavonoids, and phenolic acids. Studies highlight the potential health benefits of myrtle bioactive compounds with antioxidant and antiproliferative properties. Since limited research exists on myrtle fruit's lipid fraction, the aim of this study was to apply supercritical CO2 extraction to obtain bioactive compounds from myrtle berries focusing on the fatty acids, sterols, and essential oils. The optimization of the supercritical CO2 extraction of myrtle fruit using CO2 as solvent was carried out using the response surface methodology with Box-Behnken experimental design. The following conditions were tested: temperature (40, 50, and 60 °C), pressure (200, 300, and 400 bar), and flow rate (20, 30, and 40 g min-1) on the yield of lipid extract as well as on the yield of fatty acids, phytosterols, and volatiles present in the extract and constituting its bioactive potential. In the extracts examined, 36 fatty acids, 7 phytosterols, and 13 volatiles were identified. The average yield of the extract was 5.20%, the most abundant identified fatty acid was essential cis-linolenic acid (76.83%), almost 90% of the total phytosterols were β-sitosterol (12,465 mg kg-1), while myrtenyl acetate (4297 mg kg-1) was the most represented volatile compound. The optimal process conditions obtained allow the formulation of extracts with specific compositions.

Optimization of preparation conditions for Salsola laricifolia protoplasts using response surface methodology and artificial neural network modeling

BackgroundSalsola laricifolia is a typical C3–C4 typical desert plant, belonging to the family Amaranthaceae. An efficient single-cell system is crucial to study the gene function of this plant. In this study, we optimized the experimental conditions by using Box-Behnken experimental design and Response Surface Methodology (RSM)-Artificial Neural Network (ANN) model based on the previous studies.ResultsAmong the 17 experiment groups designed by Box-Behnken experimental design, the maximum yield (1.566 × 106/100 mg) and the maximum number of viable cells (1.367 × 106/100 mg) were obtained in group 12, and the maximum viability (90.81%) was obtained in group 5. Based on these results, both the RSM and ANN models were employed for evaluating the impact of experimental factors. By RSM model, cellulase R-10 content was the most influential factor on protoplast yield, followed by macerozyme R-10 content and mannitol concentration. For protoplast viability, the macerozyme R-10 content had the highest influence, followed by cellulase R-10 content and mannitol concentration. The RSM model performed better than the ANN model in predicting yield and viability. However, the ANN model showed significant improvement in predicting the number of viable cells. After comprehensive evaluation of the protoplast yield, the viability and number of viable cells, the optimal results was predicted by ANN yield model and tested. The amount of protoplast yield was 1.550 × 106/100 mg, with viability of 90.65% and the number of viable cells of 1.405 × 106/100 mg. The corresponding conditions were 1.98% cellulase R-10, 1.00% macerozyme R-10, and 0.50 mol L−1 mannitol. Using the obtained protoplasts, the reference genes (18SrRNA, β-actin and EF1-α) were screened for expression, and transformed with PEG-mediated pBI121-SaNADP-ME2-GFP plasmid vector. There was no significant difference in the expression of β-actin and EF1-α before and after treatment, suggesting that they can be used as internal reference genes in protoplast experiments. And SaNADP-ME2 localized in chloroplasts.ConclusionThe current study validated and evaluated the effectiveness and results of RSM and ANN in optimizing the conditions for protoplast preparation using S. laricifolia as materials. These two methods can be used independently of experimental materials, making them suitable for isolating protoplasts from other plant materials. The selection of the number of viable cells as an evaluation index for protoplast experiments is based on its ability to consider both protoplast yield and viability. The findings of this study provide an efficient single-cell system for future genetic experiments in S. laricifolia and can serve as a reference method for preparing protoplasts from other materials.

Formulation and Evaluation of Gambier (Uncaria gambir)-Chitosan Microparticle Intranasal Delivery for Alzheimer’s Diseases

Alzheimer’s Disease (AD), the most common form of dementia continues to be the deadliest neuro degenerative disease in recent years. Despite significant efforts to mitigate the progression of the disease, there is no known cure and development towards a more effective treatment is still lacking. AD is marked by exceptionally low amount of acetylcholine in the brain, formation of tau protein, and amyloid beta plaque. Current drugs of choice for treating AD, namely donepezil and memantine, are acetylcholinesterase (AChE) inhibitors which focused on delaying the onset of cognitive decline by maintaining acetylcholine concentration. Gambier water extract (GWE) contains high level of polyphenols which act as an antioxidant, exhibit strong correlation with AChE inhibitor. The aim of this research is to formulate and encapsulate GWE inside a microparticle system composed of chitosan and different crosslinkers, STPP (IMGS) and CaCl2 (IMGC), which were then characterized as AChE inhibitor using Ellman’s method. Variations of the formula were designed following Box-Behnken experimental design with chitosan and crosslinker concentration, crosslinker type, and stirring speed as variables. Initial activity of GWE, IMGS and IMGC as antioxidant were confirmed with DPPH method, obtaining a strong activity of 88.01, 82.11, and 84.99% DPPH inhibition at 100 ppm respectively. Promisingly, at concentration of 100 ppm GWE demonstrated AChE inhibition of 30.36%. However, this activity reduced after encapsulation into IMGS and IMGC, with 14.63% and 18.65% AChE inhibition, which can be linked to the relatively sustained diffusion of GWE from the polymer matrix. IMGS and IMGC diffusion profile showed release of 23.24% and 21.89% after 6 hours, with significant increase in diffusion after 24 hours with 74.92% and 71.19% respectively. Despite showing sustained release behaviour, both IMGS and IMGC ex-vivo diffusion significantly improved when compared to GWE which only diffused 51.84% after 24 hours. This result indicates encapsulation of GWE into a polymeric carrier could increase gambier diffusion through the nasal mucous membrane, significantly improving the potential to penetrate into the brain systemic circulation. Combined with desirable intranasal delivery characteristics, this research was able to demonstrate the promising potential of gambier water extract polymeric system as AChE inhibitors for AD therapy.

Optimization of a high through-flow design turbine using response surface method

Enhancing the through-flow capability of the turbine facilitates the potential to decrease the external dimension of the engine, leading to a reduction in weight and an increase in the thrust-to-weight ratio. The optimization of blade profile in terms of aerodynamic design is an essential strategy to improve the overall performance of the high through-flow design turbine. This study aims to develop an automated optimization technique, utilizing the response surface method (RSM), to enable reliable prediction and rapid optimization of aerodynamic performance. A parametric modeling method is devised to generate blade profiles with continuous curvature throughout the automated optimization process. The Box–Behnken experimental design, in combination with Reynolds-averaged Navier–Stokes numerical calculation, is employed to construct a second-order polynomial RSM approximation model. The optimization process comprises two levels: improving the through-flow capability and reducing blade profile loss. The optimized profile, Opt1, achieves a substantial 13.19% improvement in through-flow capability at the cost of a 12.27% increase in blade profile loss. Thus, further optimization is performed to minimize blade profile loss based on the Opt1 scheme. Geometric constraints are applied to the most influential parameters affecting through-flow capability to mitigate their impacts. Compared to the Opt1, the final optimized profile, Opt2, achieves a substantial 31.83% decrease in blade profile loss with a negligible sacrifice of 0.23% in through-flow capability.