Box-Behnken Experimental Design Research Articles

Optimization of the Extraction System of Total Phenolic from Garcia mangostana using Response Surface Methodology

Mangosteen (Garcia mangostana) consist of 40% flesh and 60% pericarp. Mangosteen pericarp has unpleasant taste of bitterness and usually disposed as waste. However, mangosteen pericarp contains xanthones which are polyphenolic compound that proven has high antioxidant activity were found abundantly in mangosteen pericarp. This study aims to investigate water extraction optimum operating conditions to be applied in industry by using RAM (response surface methodology) for the recovery of total phenolic extract from the pericarp of mangosteen. Box-Behnken experimental design used 3 factors. Solid-to-liquid ratio (g/ml), temperature (°C) and extraction time (hour) were analysed to compare the response of total phenolic content (TPC). Under the operating conditions, the highest TPC is 0.906 (g/l) at the factors of 1:10 (g/ml), 65 °C, and 3-hour. The optimum parameters determined by using RSM are at 1:10 (g/ml), 59.74 °C, and 2.87-hour with TPC 0.835 (g/l).

Experimental investigation of micro EDM drilling in the CFRP using response surface methodology

Recently, most of the conventional (industrial) materials such as steel, aluminum, and copper are being replaced by composite materials. Among the various composite materials, CFRP material is one of the most popular materials in many industries due to its excellent mechanical properties like-high strength, high modulus, compressive strength, etc. However, the micro hole in micro EDM drilling in the CFRP composite material is challenging due to fiber orientation, uneven distribution of conducting fiber, and delamination affecting the material removal rate (MRR) and micro hole quality performance. The objective of the article is to optimize the process parameters for optimum hole quality in the micro EDM drilling of CFRP composites. For this purpose, response surface methodology is applied to investigate the effect of the input process parameters which are voltage, capacitance, and tool rotation speed on material removal rate, roundness error, and tapper of the micro-hole. The experiments have been conducted according to the box behnken design of experiments. Regression equations, and response surfaces are developed. Optimum responses are determined by the desirability function approach. The experimental investigation shows that capacitance is the highly significant factor among the input process parameters affecting hole quality output responses such as material removal rate, roundness error, and tapper. The MRR increases with increasing the capacitance and tool rotation speed initially but its value decreases on further increasing the capacitance at a particular voltage. Roundness error and the tapper initially decrease on increasing capacitance and tool rotation speed then increases. The optimum material removal rate and hole quality are obtained in a range of 150–250 nF capacitance, and 1400–1600 rpm tool rotation speed, at 100 voltage.

Response surface optimization of poly-β-hydroxybutyrate synthesized by Bacillus cereus L17 using acetic acid as carbon source

A strain of Bacillus that can tolerate 10 g/L acetic acid and use the volatile fatty acids produced by the hydrolysis and acidification of activated sludge to produce polyhydroxyalkanoate was screened from the activated sludge of propylene oxide saponification wastewater. The strain was identified by 16S rRNA sequencing and phylogenetic tree analysis and was named Bacillus cereus L17. Various characterization methods showed that the polymer synthesized by strain L17 is poly-β-hydroxybutyrate, which has low crystallinity, good ductility and toughness, high thermal stability and a low polydispersity coefficient. It has wide thermoplastic material operating space as well as industrial and medicinal applications. The optimal fermentation conditions were determined by single factor optimization. Then, Plackett-Burman and Box-Behnken design experiments were carried out according to the single factor optimization results, and the response surface optimization was completed. The final results were: initial pH 6.7, temperature 25 °C, and loading volume 124 mL. The verification experiment showed that the yield of poly-β-hydroxybutyrate after optimization increased by 35.2 % compared to that before optimization.

A HPLC method for simultaneous quantification of chlorhexidine and thymol using Box-Behnken design for robustness of the method assessment

The antimicrobial chlorhexidine is considered the gold standard in dentistry. Thymol is a phenol found in the essential oils of various plant species and also stands out for its antimicrobial potential. Synergistic effects can be promoted by applying these two active pharmaceutical ingredients together in technological products, for example in controlled release microparticles. The objective of this work was to develop and validate an analytical methodology applying a Box-Behnken experimental design and using High Performance Liquid Chromatography capable of quantifying chlorhexidine and thymol simultaneously in a matrix including pharmaceutical excipients. After optimization, the mobile phase consisted of methanol and 0.03 M monobasic sodium phosphate buffer (60:40), with 0.4% triethylamine and octylsilane as the applied stationary phase. The method proved selective, even in the presence of chlorhexidine and thymol degradation products. For chlorhexidine, the method was linear from 4.8 to 19.2 µg/mL, and for thymol from 8.0 to 32.0 µg/mL. Accuracy was close to 100%, and the precision assessment yielded coefficient variation values of <5%. Being based on the Box-Behnken design, the method was robust and therefore validated for assisting in quality control processes involving these active pharmaceutical ingredients.

Optimization of Supercritical Carbon Dioxide Fluid Extraction of Medicinal Cannabis from Quebec

Research on cannabis oil has evolved to encompass the pharmaceutical industry for the therapeutic potential of the active compounds for pathologies such as Alzheimer, auto-immune disorders, and cancer. These debilitating diseases are best treated with cannabinoids such as tetrahydrocannabinol (∆9-THC), cannabigerol (CBG), and cannabinol (CBN), which relieve neuropathic pain and stimulate the immune system. We extracted cannabinoids from plants with supercritical CO2 and produced an extract with a total yield close to 26%. The three-level Box–Behnken experimental design considered four factors: Temperature, pressure, CO2 flow rate, and processing time, with predetermined parameters at low, medium, and high levels. The mathematical model was evaluated by regression analysis. The yield of ∆9-THC and CBG reached a maximum after 2 h and 15 g/min of CO2, 235 bar, 55 °C (64.3 g THC/100 g of raw material and 4.6 g CBG/100 g of raw material). After another 2 h of extraction time, the yield of CBN reached 2.4 g/100 g. The regression analysis identified pressure and time as the only significant factors for total yield while pressure was the only significant factor for ∆9-THC and CBG. Time, temperature, pressure, and flow rate were all significant factors for CBN.

Global Sensitivity Analysis Method for Embankment Dam Slope Stability Considering Seepage–Stress Coupling under Changing Reservoir Water Levels

Ensuring the long-term, efficient, and safe operation of reservoir dams relies on the slope stability of embankment dams. Periodic fluctuations of the reservoir water level due to reservoir scheduling operations make the slope of the reservoir bank vulnerable to instability. To investigate the influence of various factors and their interactions with embankment dam slope stability under changing reservoir water levels, a global sensitivity analysis method is proposed that accounts for seepage–stress coupling. An embankment dam in Shaanxi Province, China, is studied as an example, with COMSOL Multiphysics software simulating the seepage and slope stability of the dam under fluctuating reservoir water level conditions and seepage–stress coupling. The global sensitivity analysis of factors affecting dam slope stability is accomplished by combining Plackett–Burman and Box–Behnken experimental designs, with ANOVA determining the sensitivity of each factor and interaction term. The results demonstrate that during the impoundment period of the reservoir, the saturation line is concave, and the overall stability safety of the dam slope increases first and then tends to be stable, according to the coefficient. The internal friction angle φ, cohesion c, and soil density ρs represent the three most sensitive factors affecting the stability and safety of the dam slope, while c × ρs is a second-order interaction term with significant sensitivity to the stability and safety coefficient of the dam slope. The reservoir drainage period infiltration line is convex, and dam slope stability first reduced and then increased. The magnitude of water level change H, internal friction angle φ, cohesion c, and soil density ρs are the four most sensitive factors for the coefficient of safety of dam slope stability, while c × ρs, H × ρs, and φ × ρs are the second-order interaction terms with significant sensitivity to the coefficient of safety of dam slope stability. These research findings and methods can offer valuable technical support and reference for the investigation and evaluation of the stability of embankment dam slopes.

Acoustic Characteristics of Jute and Coir Non-woven Fabric

Effective noise reduction is essential because it can influence the health and well-being of living organisms significantly, both physically and psychologically. This necessitates the use of appropriate absorbing materials to minimize the negative impact of noise. In recent years, there has been growing interest in using jute, coir and natural fibre-based acoustic materials in architectural and interior applications due to their exceptional acoustic properties, eco-friendliness, cost-effectiveness and widespread availability. The study focuses on analysing the effects of some scrutinised testing parameters of jute, coir and jute-coir blended non-woven fabric samples on noise reduction coefficient through observing sound absorption coefficient. The sound absorption coefficient was measured using the impedance tube method as per the ASTM E-1050 standard. The mean value of the sound absorption coefficient observed at frequencies 250 Hz, 500 Hz, 1000 Hz, and 2500 Hz was used to calculate the noise reduction coefficient. A three-factor &amp; three-level Box–Behnken experimental design was used for designing research and non-woven jute, coir and jute coir blended fabric sample planning. The experimental work showed that with an increase in non-woven jute fabric sample thickness from 15 mm to 45 mm, blending jute 50 % with coir 50% and non-woven fabric samples tested by creating air gaps (15 mm to 30 mm) resulted in higher sound absorption coefficient and improved noise reduction coefficient. The experimental findings and statistical analysis revealed that blending jute with coir and increasing the thickness of non-woven fabric led to a significant improvement in the acoustic performance of the tested samples. However, increasing the air gap resulted in only a marginal improvement in acoustic performance.

Ultrasound-assisted extraction of phenolic compounds from blueberry leaves using natural deep eutectic solvents (NADES) for the valorization of agrifood wastes

The food industry demands novel green solvents for the extraction of bioactive compounds, particularly from residues of the agrifood industry. Herein, an ultrasound-assisted method has been developed for the environmentally friendly extraction of phenolic compounds from blueberry leaves using natural deep eutectic solvents (NADES). After the screening of multiple NADES, the best extraction efficiencies in terms of total phenol content and antioxidant activity were provided by NADES composed of lactate, sodium acetate, and water (3:1:2), and of choline chloride and oxalic acid (1:1). Using a Box–Behnken experimental design, the optimal extraction conditions were achieved by sonicating for 45 min at 65 °C and using solvent:sample ratios of 15 and 75 (v/w) for the NADES based on lactic acid and choline, respectively. Compared with conventional organic solvents, the use of these NADES composed of lactic acid and choline provided superior performance for the recovery of phenolic compounds (1.6-fold and 2.2-fold greater efficacy, respectively) and antioxidant compounds (1.6-fold and 2.8-fold greater efficacy, respectively). The chromatographic characterization of the extracts obtained under these optimized conditions evidenced that the lactic-based NADES enabled the extraction of a wide range of hydroxycinnamic acids and flavonol derivatives, whereas the choline-based NADES was selective towards the extraction of anthocyanins. These results indicate that the proposed method could represent an excellent green alternative for the recovery of phenolic compounds from plant materials and agrifood wastes, with improved extraction efficacy and/or selectivity compared to that provided by traditional organic solvents.

Evaluation of pilocarpine in situ forming buccal gels as potential formulations for the management of xerostomia

Buccally administrated pilocarpine (PIL) can be a unique formulation approach for the management of xerostomia. In this way the severity of systemically induced side effects can be effectively controlled, while maintaining a high level of therapeutic efficacy. The aim of the present study was to evaluate the sol–gel and mucoadhesive properties of an in situ thermoresponsive hydrogel designed for PIL's buccal administration. For this purpose, poloxamer 407 (P407) and 188 (P188) were used as in situ gel forming agents, while xanthan gum (XG) was added as a mucoadhesion enhancer. Implementation of a Box-Behnken experimental design, combined with multi-linear regression (MLR) fitting, unraveled the role of formulation factors on several critical product attributes (such as gel transition temperature, gelation time, solution viscosity and gel viscosity). Artificial neural networks (ANNs) showed excellent generalizing ability, with verification experiments showing good agreement between the ANN- predicted and the experimentally-obtained results. The optimum sol-gel formulations were sprayable at room/storage temperature, while forming a strong gel at temperatures prevailing in the buccal cavity. In vitro dissolution profiles of all optimum formulations exhibited a sustained release pattern (releasing almost 80% of the total dose in 360 min) characterized by an initial rapid release followed by a steady state plateau. The evaluation of drug release kinetics, by applying different release kinetic models, suggested that the drug release from the in situ gels predominantly follows Fick's law and is primarily diffusion-controlled. The in vitro studies were complemented with ex vivo studies, performed on a freshly excised porcine buccal mucosa, showing good mucoadhesion properties with approximately 27% of the total drug permeating the buccal porcine mucosa, while no damage to the epithelium was observed based on the histological evaluation.

Optimization of Process Parameters for Pellet Production from Corn Stalk Rinds Using Box–Behnken Design

In the current study, corn stalk rinds were used as feedstock for the production of solid-fuel pellets. In an effort to comprehensively analyze the effects of different operational parameters on the physical properties of pellets, response surface methodology (RSM) was employed in conjunction with a Box–Behnken experimental design (BBD). By assessing multiple variables simultaneously and examining their interactions, BBD facilitates the development of a reliable response model that can predict how changes in independent variables will impact response variables. The recorded responses included relaxed density, mechanical durability, and compressive strength. Based on the results, greater R2 values of 0.9467, 0.8669, and 0.9196, could be, respectively, attained for the quadratic regression models. The analysis of variance revealed that all independent variables had significant effects on the responses. The optimal processing condition for the pellets was established by determining the ideal combination of operational parameters. The process entailed the choice of a particle dimension measuring 0.5 mm, a moisture level of 11.35%, the application of heat at 125.7 °C on the die, and the utilization of a molding pressure of 154.2 MPa. Based on these factors, the predicted response values were determined to be 1639.61 kg/m3 for relaxed density, 97.95% for mechanical durability, and 10.18 MPa for compressive strength. The values obtained experimentally under the optimized conditions were similar to the predicted values with a desirability value of 1.00.

Development and Assessment of Acyclovir Gel Plaster Containing Sponge Spicules

Acyclovir is an acyclic purine nucleoside analog that is highly effective in inhibiting the herpes simplex virus. However, topical acyclovir has poor efficacy because of its low skin permeability. This study aimed to develop an acyclovir gel plaster containing sponge spicules (AGP-SS) to achieve synergistic improvements in skin absorption and deposition of acyclovir. The process of preparing the gel plaster was optimized by orthogonal experiments, while the composition of the formulation was optimized using the Plackett-Burman and Box–Behnken experimental designs. The selected formula was tested for physical properties, in vitro release, stability, ex vivo permeation, skin irritation, and pharmacokinetics. The optimized formulation exhibited good physical characteristics. In vitro release and ex vivo permeation studies showed that acyclovir release from AGP-SS was dominated by diffusion with significantly higher skin permeation (20.00 ± 1.07 μg/cm2) than that of the controls (p < 0.05). Dermatopharmacokinetic analyses revealed that the maximum concentration (78.74 ± 11.12 μg/g), area under the curve (1091.81 ± 29.05 μg/g/h) and relative bioavailability (197.12) of AGP-SS were higher than those of the controls. Therefore, gel plaster containing sponge spicules show potential for development as transdermal delivery systems to achieve higher skin absorption and deposition of acyclovir, especially in deep skin layers.

Box Behnken design application for optimization of bio-oil yield from catalytic pyrolysis of agro-residue

A Response Surface Methodology (RSM) was used to investigate the effect of reaction temperature and biomass to catalyst (b/c) ratio on the catalytic pyrolysis of three wood sawdust samples in a fixed bed reactor using Green zeolite-Y catalyst synthesized from Ficus exasperate (L.) leaf particles. Temperature (400–700 °C), biomass percentage (60–100%), and catalyst (0–40%) were the independent variables with a total of 15 experimental runs, including 3 center runs, were generated via the Box-Behnken experimental design. The results reveal that biomass/ catalyst (b/c) ratio of 80/20% at 550 °C yielded optimum pyrolytic liquid for Melicia excelsa (Me), Diospyros crassiflora (Dc) and Entada Africana (Ea) as 31 wt.%, 31 wt.%, 30 wt.%, respectively while with the attendance of catalysts at 20% increased the yield of pyrolytic liquid for Me (45 wt.%), Dc (42 wt.%), and Ea (43 wt.%). GCMS analysis of Me (80.81 wt.%), Dc (73.96 wt.%), and Ea (70.26 wt.%) pyrolytic oil reveals the dominance of phenols, ethers, alcohols, ketones, alkanes, and acids. Reduction in acidity, decrease in oxygen content, increase in viscosity of the bio-oil were noticed in biomass/catalyst (b/c) ratio of 80/20 at 550 °C. These measurements show enhanced pyrolysis oil characteristics, which is a boost to its bioenergy potential.

Integrated extraction-purification and anti-inflammatory activity of berberine-rich extracts from Coptis chinensis Franch

In this study, an innovative integrated extraction-purification process (IEPP) was developed to obtain berberine-rich extracts (BRE) from Coptis chinensis Franch. (C. chinensis). Cation exchange resins were screened, single-factor experiments and Box-Behnken design experiments (BBD) were performed to optimize the IEPP. D151 resin was selected as adsorbent and the optimal extraction-purification conditions were 80% ethanol aqueous solution,16 mL/g ratio of liquid to solid, 70 °C extraction temperature, 1.5 h extraction time, 2.44 ratio of resin to raw material, acetic acid concentration of 10%, ultrasonic power of 480 W and ratio of eluent to resin of 2:1 mL/g. The purity of berberine obtained by the IEPP method was 5.57 times than that of the reflux method, the elution solvent consumption was about 1/3–1/7 of that of the previous resin desorption method. The new method allowed high purity and minor solvent consumption compared to previous approaches. The results indicated that the IEPP was energy efficient, green process for extracting BRE from C. chinensis. Meanwhile, the anti-inflammatory property of BRE was evaluated in RAW264.7 macrophages induced with lipopolysaccharide (LPS). The results showed that BRE could significantly prevent the elevation of inflammatory factors, including nitricoxide (NO), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1 β (IL-1β), prostaglandin E2 (PGE2) and promote the expression of anti-inflammatory factors, including interleukin-10 (IL-10). In addition, the BRE could effectively inhibit reactive oxygen species (ROS) in LPS-induced RAW264.7 cells.

Optimisation of β-Glucosidase Production in a Crude Aspergillus japonicus VIT-SB1 Cellulase Cocktail Using One Variable at a Time and Statistical Methods and its Application in Cellulose Hydrolysis.

Pulp and paper mill sludge (PPMS) is currently disposed of into landfills which are reaching their maximum capacity. Valorisation of PPMS by enzymatic hydrolysis using cellulases is an alternative strategy. Existing commercial cellulases are expensive and contain low titres of β-glucosidases. In this study, β-glucosidase production was optimised by Aspergillus japonicus VIT-SB1 to obtain higher β-glucosidase titres using the One Variable at a Time (OVAT), Plackett Burman (PBD), and Box Behnken design (BBD)of experiments and the efficiency of the optimised cellulase cocktail to hydrolyse cellulose was tested. β-Glucosidase production was enhanced from 0.4 to 10.13 U/mL, representing a 25.3-fold increase in production levels after optimisation. The optimal BBD production conditions were 6 days of fermentation at 20 °C, 125 rpm, 1.75% soy peptone, and 1.25% wheat bran in (pH 6.0) buffer. The optimal pH for β-glucosidase activity in the crude cellulase cocktail was (pH 5.0) at 50 °C. Optimal cellulose hydrolysis using the crude cellulase cocktail occurred at longer incubation times, and higher substrate loads and enzyme doses. Cellulose hydrolysis with the A. japonicus VIT-SB1 cellulase cocktail and commercial cellulase cocktails resulted in glucose yields of 15.12 and 12.33 µmol/mL glucose, respectively. Supplementation of the commercial cellulase cocktail with 0.25 U/mg of β-glucosidase resulted in a 19.8% increase in glucose yield.

Evaluation of the Survival of Lactobacillus fermentum K73 during the Production of High-Oleic Palm Oil Macroemulsion Powders Using Rotor-Stator Homogenizer and Spray-Drying Technique.

This study aimed to evaluate the survival of the probiotic Lactobacillus fermentum when it is encapsulated in powdered macroemulsions to develop a probiotic product with low water activity. For this purpose, the effect of the rotational speed of the rotor-stator and the spray-drying process was assessed on the microorganism survival and physical properties of probiotic high-oleic palm oil (HOPO) emulsions and powders. Two Box-Behnken experimental designs were carried out: in the first one, for the effect of the macro emulsification process, the numerical factors were the amount of HOPO, the velocity of the rotor-stator, and time, while the factors for the second one, the drying process, were the amount of HOPO, inoculum, and the inlet temperature. It was found that the droplet size (ADS) and polydispersity index (PdI) were influenced by HOPO concentration and time, ζ-potential by HOPO concentration and velocity, and creaming index (CI) by speed and time of homogenization. Additionally, HOPO concentration affected bacterial survival; the viability was between 78-99% after emulsion preparation and 83-107% after seven days. The spray-drying process showed a similar viable cell count before and after the drying process, a reduction between 0.04 and 0.8 Log10 CFUg-1; the moisture varied between 2.4% and 3.7%, values highly acceptable for probiotic products. We concluded that encapsulation of L. fermentum in powdered macroemulsions at the conditions studied is effective in obtaining a functional food from HOPO with optimal physical and probiotic properties according to national legislation (>106 CFU mL-1 or g-1).

Simultaneous microwave-assisted extraction of bioactive compounds from aged garlic

In this work, the simultaneous extraction of bioactives (organosulfur compounds, such as S-allyl-L-cysteine (SAC), carbohydrates, such as neokestose and neonystose, and total phenolic compounds) from aged garlic has been optimized for the first time to obtain multifunctional extracts for further application as food ingredients. Analytical methods using liquid chromatography coupled to mass spectrometry (HPLC-MS) and by hydrophilic interaction liquid chromatography with evaporative light scattering detection (HILIC-ELSD) were also previously optimized. High sensitivity (limits of detection between 0.013 and 0.77 µg mL−1) and appropriate repeatability (< 12%) and accuracy (> 92%) for the analysis of bioactives were achieved. After selecting water as the extraction solvent and microwave-assisted extraction (MAE) as the most efficient technique, operation conditions were optimized using a Box-Behnken experimental design (60 min; 120 °C; 0.05 g mL−1; 1 cycle) to maximize the content of bioactives from different aged garlic samples. Regarding organosulfur compounds, only SAC (traces-2.32 mg g−1 dry sample) and cycloalliin (1.23–3.01 mg g−1 dry sample) were detected in all samples, while amino acids such as arginine (0.24–3.45 mg g−1 dry sample) and proline (0.43–3.91 mg g−1 dry sample) were, in general, the most abundant. Bioactive carbohydrates (from trisaccharides to nonasaccharides) were only detected in fresh garlic and aged garlic processed under mild conditions, whereas all garlic extracts showed antioxidant activity. The developed MAE methodology is shown as a successful alternative to other procedures for the simultaneous extraction of aged garlic bioactives intended by the food and nutraceutical industries, among others.

Box-Behnken Experimental Design for Electrochemical Aptasensor Optimization on Screen Printed Carbon Electrode/Silica-Ceria

This study aims to optimize the epithelial sodium channel (ENaC) electrochemical aptasensor with the Box-Behnken experimental design. ENaC is a protein that plays a role in sodium ion transport in several epithelial tissues and is associated with hypertension. The ENaC protein aptamer is held in place in the electrochemical aptasensor by a modified screen-printed carbon electrode (SPCE) of silica-ceria composite (SiO2-CeO2). The unique structure of a silica matrix with high biocompatibility can form composites through a hydrothermal process. The Box-Behnken (BBD) experimental design is an efficient optimization method of factors that affect the experiment at three levels. The FTIR results of the silica-ceria composites were 549.35 cm-1 (Ce-O), 1095.3 cm-1 (Si-O-Si), and 491.28 cm-1 (Si-O). Meanwhile, SPCE/silica-ceria characterized by differential pulse voltammetry (DPV) showed an increase in peak current [Fe(CN)6]3-/4- from 3.190 μA to 9.073 μA. Three experimental factors, aptamer concentration, streptavidin incubation time, and aptamer incubation time, were optimized with BBD and obtained at 0.5 μg.mL-1, 30 minutes, and 1 hour. The optimum conditions observed resulted in a selective current response for ENaC protein detection. The optimization results can be applied to aptamer-based ENaC protein detection in samples.

Preparation, Characterization and Optimization of Sustained Release Matrix Tablets of Repaglinide using Box–Behnken Design

The aim of this investigation was to develop and optimize the Sustained release Matrix formulations of Repaglinide (RLD) using response surface methodology by employing a 3-factor, 3-level Box–Behnken statistical design. The independent variables studied were the amount of hydroxypropyl methylcellulose (HPMC K4M), Ethyl cellulose (EC) and PVP K30. The Swelling index (Y1), drug release at 8 hr and 12hr were the target responses. The response surface methodology and multiple response optimizations utilizing a polynomial equation were used to search for the optimal formulation with a specific release rate at different time intervals. The results showed that the effect of combination of HPMC K4M and EC was the most influencing factor on the drug release from ER matrix tablets. The mechanism of drug release from RLD Matrix tablets was dependent on the added amount of EC. Validation of the optimization technique demonstrated the reliability of the model. The optimized formulation containing 50mg of HPMC K4, 35mg of EC, and 30mg of PVP K30 was prepared according to the software determined levels. DSC and FTIR studies combined with the stability study of the optimized formulation proved the integrity of the developed formulation. The Box–Behnken experimental design facilitated the formulation and optimization of extended release hydrophilic matrix systems of RLD in a short period of time and with the fewest number of experiments. The optimized Matrix tablet of Repaglinide showed good pharmacokinetic result over conventional formulation.

Effect of Nitrogen, Salinity, and Light Intensity on the Biomass Composition of Nephroselmis sp.: Optimization of Lipids Accumulation (Including EPA).

Microalgal biomass is characterized by high protein, carbohydrates, and lipids concentrations. However, their qualitative and quantitative compositions depend not only on the cultivated species but also on the cultivation conditions. Focusing on the microalgae's ability to accumulate significant fatty acids (FAs) amounts, they can be valorized either as dietary supplements or for biofuel production, depending on the accumulated biomolecules. In this study, a local isolate (Nephroselmis sp.) was precultured under autotrophic conditions, while the Box-Behnken experimental design followed using the parameters of nitrogen (0-250 mg/L), salinity (30-70 ppt) and illuminance (40-260 μmol m-2 s-1) to evaluate the accumulated biomolecules, with an emphasis on the amount of FAs and its profile. Regardless of the cultivation conditions, the FAs of C14:0, C16:0, and C18:0 were found in all samples (up to 8% w/w in total), while the unsaturated C16:1 and C18:1 were also characterized by their high accumulations. Additionally, the polyunsaturated FAs, including the valuable C20:5n3 (EPA), had accumulated when the nitrogen concentration was sufficient, and the salinity levels remained low (30 ppt). Specifically, EPA approached 30% of the total FAs. Therefore, Nephroselmis sp. could be considered as an alternative EPA source compared to the already-known species used in food supplementation.

Ultrasound-Assisted Extraction of Hydroxytyrosol from Lactiplantibacillus plantarum Fermented Olive Leaves: Process Optimization and Bioactivity Assessment

Olive leaves are important by-products for the recovery of phenolic compounds and extracts with high phenolic content using lactic acid bacteria during fermentation. Lactiplantibacillus plantarum (L. plantarum) strains as glucosidase-positive strains are starter cultures used to control the fermentation process. The main objective of the present work is to determine the most effective strain for the biodegradation of oleuropein to hydroxytyrosol using two L. plantarum strains for the fermentation of olive leaves. Box–Behnken experimental design was applied to determine the optimum ultrasound-assisted extraction (UAE) conditions to obtain hydroxytyrosol-rich extract using the brine of the fermented leaves. L. plantarum ATCC 14917 (hydroxytyrosol; 126.89 ± 1.59 mg/L) strain showed higher oleuropeinolytic activity than L. plantarum ATCC-BAA 793 (85.93 ± 0.70 mg/L) in olive leaf brine. When the UAE method was applied, it was seen that the hydroxytyrosol concentration of samples that were inoculated with L. plantarum ATCC 14917 (362.29 ± 2.31 mg/L) compared to L. plantarum ATCC-BAA 793 (248.79 ± 4.14) increased. The optimum UAE conditions were determined as 30% amplitude-5 min-30% ethanol for ATCC 14917 and 45% amplitude-9 min-10% ethanol for BAA 793 strain. This study showed that the brines of fermented olive leaves with oleuropeinolytic strains can be considered high added value products.