Search

IntroductionToxic heavy metal pollution has been considered a major ecosystem pollution source. Unceasing or rare performance of Pb2+ to the surrounding environment causes damage to the kidney, nervous, and liver systems. Microbial remediation has acquired prominence in recent decades due to its high efficiency, environment-friendliness, and cost-effectiveness.MethodsThe lead biosorption by Bacillus subtilis was optimized by two successive paradigms, namely, a definitive screening design (DSD) and an artificial neural network (ANN), to maximize the sorption process.ResultsFive physicochemical variables showed a significant influence (p < 0.05) on the Pb2+ biosorption with optimal levels of pH 6.1, temperature 30°C, glucose 1.5%, yeast extract 1.7%, and MgSO4.7H2O 0.2, resulting in a 96.12% removal rate. The Pb2+ biosorption mechanism using B. subtilis biomass was investigated by performing several analyses before and after Pb2+ biosorption. The maximum Pb2+ biosorption capacity of B. subtilis was 61.8 mg/g at a 0.3 g biosorbent dose, pH 6.0, temperature 30°C, and contact time 60 min. Langmuir’s isotherm and pseudo-second-order model with R2 of 0.991 and 0.999 were suitable for the biosorption data, predicting a monolayer adsorption and chemisorption mechanism, respectively.DiscussionThe outcome of the present research seems to be a first attempt to apply intelligence paradigms in the optimization of low-cost Pb2+ biosorption using B. subtilis biomass, justifying their promising application for enhancing the removal efficiency of heavy metal ions using biosorbents from contaminated aqueous systems.

Environmentally friendly and economically viable methods are essential in the selection of materials and techniques for the synthesis of nano-zero-valent iron. Plants, with their high polyphenol content and antioxidant capacity, have found application in eco-friendly synthesis processes. The definitive screening design (DSD) monitored four key process parameters for the concurrent removal of copper and phosphate: copper concentration (ranging from 1 to 9 mg L-1), phosphate concentration (ranging from 1 to 9 mg L-1), initial pH values (ranging from 2 to 10), and the dosage of nano-zero-valent iron (ranging from 2 to 16 mL). The analysis results provide valuable insights into the significant individual factors influencing the process, along with the potential for their interactions. The model also proposes process optimization to attain maximum removal efficiency, and subsequent verification confirmed its superiority among the alternatives. Mechanisms such as sorption, reduction, complexation, electrostatic attraction, and ligand exchange play pivotal roles in the effective removal of copper and phosphate using nano-zero-valent iron. In summary, this research yields several benefits: the utilization of environmentally sustainable materials, a substantial reduction in experimental complexity, coupled with the ease of the entire procedure, simultaneous and highly efficient copper and phosphate removal, favorable pH levels and, notably, no requirement for additional treatment.

Background/Objectives: Patients with schizophrenia have significant challenges in adhering to and complying with oral medicines, resulting in adverse consequences such as symptom worsening and psychotic relapse. Methods: This study aimed to develop clove oil-based bilosomes using definitive screening design (DSD) to maximize the anti-schizophrenic action of clozapine and promote its nose-to-brain delivery. The target was to optimize the physicochemical properties of bilosomes and incorporate them into mucoadhesive intranasal in situ gels, searching for augmented ex vivo and in vivo clozapine delivery. Results: The bilosomes’ particle size was decreased by increasing the span, SDC, and clove oil amounts. In addition to using a high lipid amount, the aforementioned components also helped increase the entrapment efficiency values. Increased zeta potential was only observed by increasing surfactant amount and reducing clozapine concentration. After incorporation of optimized liquid clove oil-based bilosomes, which had a spherical nano-sized vesicular shape, into P 407-dependent gels, an HPMC (2% w/w)/P 407 (20% w/w)-containing formulation (G6) was selected as an optimized gel owing to its acceptable gelation time (13.28 s), gel strength (27.72 s), viscosity (12,766.67 cP), and mucoadhesive strength (4273.93 dyne/cm2). The optimized G6 exhibited higher Jss (50.86 μg/cm2·h−1) through the nasal mucosa compared to the control gel (23.03 μg/cm2·h−1). Compared to the control gel, G6 displayed higher relative bioavailability (491.37%) than a commercial tablet (264.46%). Following ELISA analysis, dopamine and serotonin were significantly reduced, while BDNF was remarkably increased after administration of optimized G6 into schizophrenic rats. Conclusion: Our study indicates the potential of intranasal bilosomal gels in upgrading the anti-schizophrenic and neuroprotective activity of clozapine.

The Analytical Quality by Design (AQbD) concept was adopted to establish a quantitative analysis of multi-components with a single marker (QAMS) method for industrial lanolin alcohol, targeting cholesterol, lanosterol, and 24,25-dihydrolanosterol. The potential critical method parameters (CMPs) were identified as column temperature, flow rate, and gradient. Definitive screening design and statistical modeling were employed to optimize the gradient conditions of the mobile phase, column temperature, and flow rate. The Method Operable Design Region (MODR) was determined using a risk-based quantification approach. The robustness was assessed using a Plackett–Burman experimental design, followed by methodological validation. Optimal analytical conditions were as follows: acetonitrile (B)—water (A) mobile phase system; flow rate of 1.58 mL/min; detection wavelength of 205 nm; injection volume of 10 µL; and column temperature of 37 °C. A gradient elution program was implemented as follows: 0–19.0 min, 90.5% B; 19.0–25.0 min, 90.5–100% B; and 25.0–55.0 min, 100% B. Cholesterol served as an internal standard for quantifying lanosterol and 24,25-dihydrolanosterol, with relative correction factors of 0.4227 and 0.8228, respectively. This analytical method utilized only the cholesterol reference substance as an internal standard to quantify the content of cholesterol, lanosterol, and 24,25-dihydrolanosterol in industrial lanolin alcohol. It reduced the testing costs and enhanced efficiency, making it potentially suitable for widespread adoption in lanolin alcohol processing industries.

This study showcases a promising approach to sustainably unlocking plant biomass residues by combining biodegradation with artificial intelligence to optimize the process. Specifically, we utilized the definitive screening design (DSD) and artificial neural networks (ANNs) to optimize the degradation of common bean biomass by the endophytic fungus Trichoderma asperellum WNZ-21. The optimized process yielded a fungal hydrolysate rich in 12 essential and non-essential amino acids, totaling 18,298.14 μg/g biomass. GC-MS analysis revealed four potential novel components not previously reported in microbial filtrates or plants and seven components exclusive to plant sources but not reported in microbial filtrates. The hydrolysate contained phenolic, flavonoid, and tannin compounds, as confirmed by FT-IR analysis. High-resolution transmission electron microscopy depicted structures resembling amino acid micelles and potential protein aggregates. The hydrolysate exhibited antioxidant, antibacterial, and anticancer properties and innovatively induced apoptotic modulation in the MCF7 cancer cell line. These findings underscore the potential of ANN-optimized fermentation for various applications, particularly in anticancer medicine due to its unique composition and bioactivities. The integration of the DSD and ANNs presents a novel technique for biomass biodegradation, warranting the valorization of plant biomass and suggesting a further exploration of the new components in the fungal hydrolysate. This approach represents the basic concept for exploring other biomass sources and in vivo studies.

In the present work, biodiesel was produced from watermelon waste oil by the transesterification process. A definitive screening design was used to investigate the effect of transesterification parameters such as catalyst concentration (4−10%), methanol-to-oil ratio (2−10mol/mol)and reaction temperature (46-60°C). The quadratic regression analysis was used to predict the yield of biodiesel. It is found that the optimum catalyst concentration, methanol-to-oil molar ratio and reaction temperature are 5.51%, 5.77:1 and 60°C, respectively, resulting in a 96.763% yield of methyl esters, which is in excellent confirmation with the predicted value. According to the results, all variables had a significant effect on the yield of biodiesel. The physicochemical property of the extracted watermelon waste oil and produced biodiesel using optimum process parameters was analyzed. The results revealed that the extracted oil and produced biodiesel met the requirements of ASTM and EN14214 (European committee for standardization) standard methods. Oleic acid and Linoleic acid were detected to be the dominant unsaturated fatty acids; while palmitic acid was found to be the highest amount of saturated fatty acid. It can be concluded that a definitive screening design for optimization of transesterification reaction parameters is an effective way to boost the yield of biodiesel. In addition, this method significantly reduces the number of experiments, which is important for a limited amount of raw materials. In general, the finding of this work proposes that the watermelon waste oils seem to be a potential and alternative feedstock for the production of biodiesel from waste materials.

Conversion of nonfood natural products to value-added products is an important work, which is going on worldwide. In addition, to obtain a product with better quality, a sufficient separation method is helpful to improve its utilization value by using effective, rapid and environmentally friendly techniques. Thus in this work, the microwave-assisted hydrodistillation (MAHD) process of essential oils from important natural plant Rumex Crispus leaves was investigated. The experimental design and extraction conditions were optimized using a definitive screening design. The yield of essential oils of 4.67 ± 0.02 % was investigated under the optimum conditions; 534.89 W of microwave power, 23.48 min of hydrodistillation time and 4.5 mL/g of the volume of water to plant mass ratio. Under these conditions, the essential oils were analyzed using gas chromatography-mass spectrometry (GC–MS). Results revealed that the essential oil extracted by MAHD possessed high quality as proved by its higher percentage of oxygenated compounds. In addition, α-santol (29.63 %) and β-santol (25.60 %) are the primary components of oxygenated compounds in essential oils. In conclusion, the MAHD was successfully employed to obtain mainly Oxygenated compounds-rich essential oil that may be used in several industrial applications. Subsequently, a definitive screening design may be regarded as an alternative and reliable method for the prediction of experimental parameters. It was concluded that the weeds plant (Rumex Crispus) contains a reliable quantity of oils that is extremely feasible to use and recommended as good feedstock for possible use in industrial applications.

The huge amount of solid waste produced during the hydrodistillation of Pelargonium graveolens L’Hér. is a serious environmental hazard. The aim of this research is to reduce this environmental pollution and to valorize this waste by using its extract as a natural dye. Extracts were used for dyeing wool samples. The dyed fabrics showed shades ranging between brown and beige. By using the Minitab 19 Software, a definitive screening design was used to optimize the dye extraction process. The studied extraction process parameters were pH, temperature, and extraction time. The main studied responses were the color strength and the polyphenol, flavonoid, and condensed tannin contents (mg.g−1). Based on the photochemical study and the UV–vis absorption spectra, it was found that the polyphenol content varies between 4.144 and 7.567 mg.g−1. For the tannin content, the highest value was equal to 120.967 mg.g−1. To optimize the colorant extraction process, the K/S was also measured by the spectrocolorimeter. Process optimum conditions were of 11, 100°C, and 65 min, respectively, for pH, temperature, and extraction time. The optimum K/S value was equal to 115.15. Therefore, the solid waste hydrodistillation of Pelargonium graveolens can be an interesting choice for the natural dyeing of textile fabrics.

A definitive screening design was used in order to evaluate the effects of starch, glycerol and montmorillonite (MMT) concentrations, as well as the drying temperature, drying tray type and starch species, on packaging film’s functional properties. Optimization showed that in order to obtain films with the minimum possible thickness, the maximum elongation at break, the maximum tensile strength, as well as reduced water vapor permeability and low opacity, a combination of factors should be used as follows: 5.5% wt starch concentration, 30% wt glycerol concentration on a dry starch basis, 10.5% wt MMT concentration on a dry starch basis, 45 °C drying temperature, chickpea as the starch species and plexiglass as the drying tray type. Based on these results, starch films were prepared, and fresh minced meat was stored in them for 3 days. It was shown that the incorporation of MMT at 10.5% wt on a dry starch basis in the packaging films led to a decreased mesophilic and psychrotrophic bacteria growth factor compared to commercial packaging. When assessed for their biodegradability, the starch films disintegrated after 10 days of thermophilic incubation under simulated composting conditions. Finally, to prove their handling capability during industrial production, the starch films were rewound in a paper cylinder using an industrial-scale rewinding machine.

The definitive screening design (DSD) and artificial neural network (ANN) were conducted for modeling the biosorption of Co(II) by Pseudomonas alcaliphila NEWG-2. Factors such as peptone, incubation time, pH, glycerol, glucose, K2HPO4, and initial cobalt had a significant effect on the biosorption process. MgSO4 was the only insignificant factor. The DSD model was invalid and could not forecast the prediction of Co(II) removal, owing to the significant lack-of-fit (P < 0.0001). Decisively, the prediction ability of ANN was accurate with a prominent response for training (R2 = 0.9779) and validation (R2 = 0.9773) and lower errors. Applying the optimal levels of the tested variables obtained by the ANN model led to 96.32 ± 2.1% of cobalt bioremoval. During the biosorption process, Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray spectroscopy, and scanning electron microscopy confirmed the sorption of Co(II) ions by P. alcaliphila. FTIR indicated the appearance of a new stretching vibration band formed with Co(II) ions at wavenumbers of 562, 530, and 531 cm–1. The symmetric amino (NH2) binding was also formed due to Co(II) sorption. Interestingly, throughout the revision of publications so far, no attempt has been conducted to optimize the biosorption of Co(II) by P. alcaliphila via DSD or ANN paradigm.

Background: Xiaochaihu capsule is composed of seven traditional Chinese medicines. The pharmacopoeia only focuses on the quantitative detection of baicalin, which cannot fully reflect the quality of the preparation. Some medium polar components were used to establish the fingerprint of Xiaochaihu capsule, but there was no report on the strong polar components. Methods: A high performance liquid chromatography-corona charged aerosol detection technology was used to establish a fingerprint analysis method for Xiaochaihu capsules following an analytical quality by design approach. Definitive screening designed experiments were used to optimize the method parameters. A stepwise regression method was used to build quantitative models. The method operable design region was calculated using the experimental error simulation method. Plackett–Burman designed experiments were carried out to test robustness. Results: The contents of four components were simultaneously determined. There were seven common peaks in the fingerprint. The common peak area accounted for 91.72%. Both fingerprint and quantitative analysis methods were validated as applicable in the methodology study. The quantitative fingerprint analysis method for sugar components can fill the gap in the detection of strong polar components in the existing methods. It provides a new technology for the comprehensive overall evaluation of Xiaochaihu capsule.

Abstract Background Quality by design (QbD) is an advanced drug quality control concept that has been gradually implemented in the optimization of manufacturing processes of Chinese medicines. However, the variation of Chinese medicinal material quality has rarely been considered in published works. Because manufacturing processes may lower the variation introduced through different batches of materials, a material quality control strategy should be developed considering the influences of manufacturing processes. Methods In this work, the processes of extraction, concentration, water precipitation, and chromatography for notoginseng total saponin (NTS) production were investigated while considering Panax notoginseng quality variation as a sample. Ten process parameters were studied simultaneously using a definitive screening design. After the process critical quality attributes (CQAs) were determined, critical process parameters (CPPs) and critical material attributes (CMAs) were identified simultaneously. Then, models utilizing the CMAs, CPPs, and process CQAs were developed. The design space was then calculated using a Monte Carlo simulation method with an acceptable probability of 0.90. A material quality control strategy considering the influences of manufacturing processes was proposed. Results The ginsenoside Rd purity and total saponin purity in the eluate were identified as process CQAs. The ethanol solution concentration used for extraction, the ethanol solution concentration used for elution, and elution time were identified as CPPs. The extractable dry matter content of Panax notoginseng was one of the CMAs. The extractable contents of notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1, and ginsenoside Rd were the other CMAs. The inequalities implemented to discriminate the high quality and low quality of Panax notoginseng were developed according to the NTS standard of the Xuesaitong injection. Low quality Panax notoginseng should not be released for NTS production. High quality Panax notoginseng can be treated with feasible manufacturing processing parameters. Verification experiments were carried out successfully for 2 batches of high quality Panax notoginseng. Conclusions In this work, a quality control strategy for herbal materials was developed considering the matching of process characteristics and material quality attributes. This strategy is promising for application to other Chinese medicines.

Heterogeneous Fenton catalyst Fe2(MoO4)3 was prepared and the process efficiency was evaluated for oxidation of water-based Cyan flexo dye in synthetic aqueous solution and printing wastewater. The removal process of printing dye was analyzed by UV/VIS spectrophotometry, while dye mineralization was evaluated by the determination of total organic carbon content and chemical oxygen demand. Four determinants of the heterogeneous Fenton system, including initial dye concentration, iron concentration, pH and hydrogen peroxide concentration were investigated. Statistical method, definitive screening design was applied to generate optimal operational conditions of the four variables, which maximizes the process of dye removal. The initial dye concentration of 20 mgL-1, catalyst dosage of 0.75 mgL-1, pH of 2 and H2O2 concentration of 11 mM were chosen as the best operational conditions, contributing to 82% of the process efficiency. The Fenton process efficiency of 79% was achieved within the treatment of printing wastewater under optimal conditions for a 90 minute reaction time. The maximum COD removal efficiency was 61.1%, while 67% mineralization was achieved. The obtained results confirmed synergistic effect of Fe3+ and MoO4 2− which contributed to high catalytic activity and high heterogeneous Fenton efficiency. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR 34014 and Grant no. III43005]

Plasmid DNA is a vital biological tool for molecular cloning and transgene expression of recombinant proteins; however, decades ago, it has become an exceptionally appealing as a potential biopharmaceutical product as genetic immunization for animal and human use. The demand for large-quantity production of DNA vaccines also increases. Thus, we, herein, presented a systematic approach for process characterization of fed-batch Escherichia coli DH5α fermentation producing a porcine DNA vaccine. Design of Experiments (DoE) was employed to determine process parameters that have impacts on a critical quality attribute of the product, which is the active form of plasmid DNA referred as supercoiled plasmid DNA content, as well as the performance attributes, which are volumetric yield and specific yield from fermentation. The parameters of interest were temperature, pH, dissolved oxygen, cultivation time, and feed rate. Using the definitive-screening design, there were 16 runs, including 3 additional center points to create the predictive model, which then was used to simulate the operational ranges for capability analysis.

Abstract Low grade high ash coals with difficult physical washability characteristics require chemical cleaning techniques such as solvent extraction to obtain super clean coal with lower ash contents. Kedla, an Indian low grade coal has ash content around 44%–50% and there is a need to maximize its solvent extraction yield so as to make this power grade coal industrially useful. This paper highlights the use of definitive screening design and response surface methodology to extract the maximum organic matter from Kedla coal using ethylenediammine (EDA) and N-methyl pyrollidone (NMP) as solvents under mild atmospheric pressure conditions. The coal was pre-treated with acetic acid before extractions were carried out. The parameters such as the acid pre-treatment time, acid concentration, particle size, the co-solvent concentration and the solvent concentration were optimized in this paper. With this design of experiments technique about 40% of the organic matter with minimal ash (less than 2%) content was rendered extractable unlike the already postulated designer solvent system—e, N system (i.e. NMP containing small amount of EDA) that uses coal to solvent ratio as 1:17 (wt/vol) and coal to co-solvent ratio as 1:1(wt/vol) thus only giving 19% extraction yield. Therefore, the present design of experiments technique helps in conserving the number of experiments for process intensification.

CO2 microbubbles have recently been used in enhanced oil recovery for blocking the high permeability zone in heterogeneous reservoirs. Microbubbles are colloidal gas aphrons stabilized by thick shells of polymer and surfactant. The stability of CO2 microbubbles plays an important role in improving the performance of enhanced oil recovery. In this study, a new class of design of experiment (DOE)—definitive screening design (DSD) was employed to investigate the effect of five quantitative parameters: xanthan gum polymer concentration, sodium dodecyl sulfate surfactant concentration, salinity, stirring time, and stirring rate. This is a three-level design that required only 11 experimental runs. The results suggest that DSD successfully evaluated how various parameters contribute to CO2 microbubble stability. The definitive screening design revealed a polynomial regression model has ability to estimate the main effect factor, two-factor interactions and pure-quadratic effect of factors with high determination coefficients for its smaller number of experiments compared to traditional design of experiment approach. The experimental results showed that the stability depend primarily on xanthan gum polymer concentration. It was also found that the stability of CO2 microbubbles increases at a higher sodium dodecyl sulfate surfactant concentration and stirring rate, but decreases with increasing salinity. In addition, several interactions are presented to be significant including the polymer–salinity interaction, surfactant–salinity interaction and stirring rate–salinity interaction.

β-glucosidases are a class of enzyme that are widely distributed in the living world, with examples noted in plants, fungi, animals and bacteria. They offer both hydrolysis and synthesis capacity for a wide range of biotechnological processes. However, the availability of native, or the production of recombinant β-glucosidases, is currently a bottleneck in the widespread industrial application of this enzyme. In this present work, the production of recombinant β-glucosidase from Streptomyces griseus was optimised using a Design of Experiments strategy, comprising a two-stage, multi-model design. Three screening models were comparatively employed: Fractional Factorial, Plackett-Burman and Definitive Screening Design. Four variables (temperature, incubation time, tryptone, and OD600 nm) were experimentally identified as having statistically significant effects on the production of S.griseus recombinant β-glucosidase in E. coli BL21 (DE3). The four most influential variables were subsequently used to optimise recombinant β-glucosidase production, employing Central Composite Design under Response Surface Methodology. Optimal levels were identified as: OD600 nm, 0.55; temperature, 26 °C; incubation time, 12 h; and tryptone, 15 g/L. This yielded a 2.62-fold increase in recombinant β-glucosidase production, in comparison to the pre-optimised process. Affinity chromatography resulted in homogeneous, purified β-glucosidase that was characterised in terms of pH stability, metal ion compatibility and kinetic rates for p-nitrophenyl-β-D-glucopyranoside (pNPG) and cellobiose catalysis.

The metabolism of seaweeds depends on environmental parameters, the availability of nutrients, and biotic/abiotic stresses; therefore, their chemical composition fluctuates throughout the year. This study investigated seasonal variations in the metabolome of the Baltic Sea brown alga Fucus vesiculosus and its potential relation to the bioactivity profile. By using a definitive screening design (DSD) combined with pressurised liquid extraction (PLE), an optimised protocol was developed to extract algal biomass monthly for a full calendar year. An untargeted metabolomics approach using ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MSn)-based molecular networking and manual dereplication was employed. The extracts were simultaneously screened for their in vitro antimicrobial, anticancer/apoptotic, and free radical scavenging activities. 44 compounds were putatively dereplicated in the metabolome. Many compounds were found to vary with the sampling month; phlorotannin total ion count (TIC) was highest in summer, whilst chlorophylls, lipids, and carotenoids peaked in winter and spring. The greatest radical scavenging and apoptotic activities against pancreas cancer cells observed in the summer months were attributed to high phlorotannin TIC. Methicillin-resistant Staphylococcus aureus (MRSA) inhibitory activity was produced year-round without a clear seasonal trend. This is the first study applying DSD-based optimised PLE extraction combined with a metabolome analysis of F. vesiculosus for the identification of seasonal variations in both metabolome and bioactivity.