Acoustic resonance technology and quality by design approach facilitate the development of the robust tetrandrine nano-delivery system.

The aim of this study was to develop a sufficiently robust tetrandrine (Tet) nano-delivery system using acoustic resonance (AR) technology and freeze-drying technology. This system can effectively improve the solubility and dissolution properties of Tet, along with high stability and scale-up adaptability. Firstly, 54 stabilizers were screened simultaneously in a high-throughput manner with the help of AR technology to fully explore the optimal prescription space of tetrandrine nanosuspension (Tet-NS). The Plackett-Burman design was used to screen for critical variables severely affecting the quality of Tet-NS. The Box-Behnken design was used to investigate and optimize critical variables to obtain optimal nanosuspensions. The optimal prescription was successfully scaled up by 100 times, which was the initial exploration of its commercial scale production. Solidification studies have shown that formulations with 2.44% fructose as the cryoprotectant have excellent redispersibility. Compared with pure Tet, Tet in Tet-NS showed a significant increase in solubility and dissolution rate in water. Fourier transform infrared (FT-IR) demonstrated that no significant interactions occurred between the drug and excipients in Tet-NS. Powder x-ray diffraction analysis (PXRD) indicated that some of the Tet transformed into amorphous state during the preparation process. In short-term stability study, Tet-NS successfully maintained its physical stability. In summary, under the guidance of the QbD concept, this study rapidly developed Tet-NS using acoustic resonance technology, which can effectively improve the solubility and dissolution properties of Tet. During the development of Tet-NS, AR technology has demonstrated high particle size reduction capability, the ability to process multiple sets of formulations in parallel, and excellent scale-up capability. Meanwhile, the method and concept of this study are not limited to Tet, but also applicable to other poorly water-soluble drugs.

Mandarin Peels-Derived Carbon Dots: A Multifaceted Fluorescent Probe for Cu(II) Detection in Tap and Drinking Water Samples.

Carbon dots (CDs) derived from mandarin peel biochar (MBC) at different pyrolysis temperatures (200, 400, 600, and 800 °C) have been synthesized and characterized. This high-value transformation of waste materials into fluorescent nanoprobes for environmental monitoring represents a step forward towards a circular economy. In this itinerary, CDs produced via one-pot hydrothermal synthesis were utilized for the detection of copper (II) ions. The study looked at the spectroscopic features of biochar-derived CDs. The selectivity of CDs obtained from biochar following carbonization at 400 °C (MBC400-CDs towards various heavy metal ions resulted in considerable fluorescence quenching with copper (II) ions, showcasing their potential as selective detectors. Transmission electron microscopic (TEM) analysis validated the MBC-CDs' consistent spherical shape, with a particle size of <3 nm. The Plackett-Burman Design (PBD) was used to study three elements that influence the F0/F ratio, with the best ratio obtained with a pH of 10, for 10 min, and an aqueous reaction medium. Cu (II) was detected over a dynamic range of 4.9-197.5 µM and limit of detection (LOD) of 0.01 µM. Validation testing proved the accuracy and precision for evaluating tap and mountain waters with great selectivity and no interference from coexisting metal ions.

Unlocking sustainable solutions: wood waste as a novel substrate for polyhydroxybutyrate (PHB) production to combat plastic pollution.

Polyhydroxybutyrate (PHB) is considered as a hope for bioplastic production, which can serve as a sustainable alternative. Utilizing feedstock as substrate is widely explored for the production but wood waste, which is abundant in cellulose, hemicellulose and lignocellulose, has limited studies for PHB production. Herein, wood waste is used as a biobased feedstock Hydrolyses of wood waste was done using sulphuric acid (H2SO4) to break down of cellulose and hemicellulose into simple carbon forms. The hydrolysed product was analysed for sugar presence by quantitative and qualitative methods. Pseudomonas fluorescens bacterial strain was used for the production purpose using hydrolysed wood waste as substrate media. The Plackett-Burman design (PBD) and response surface methodology (RSM) were applied to optimize the growth media. The results of PBD were used to identify significant factors influencing PHB production, which were then further optimized using RSM. The work's results conclusively demonstrated that P. fluorescens possesses the capability to effectively utilize wood waste and wastewater as substrate media up to production rate of 13-14 mg mL-1 of PHB. Fourier Transformed Infra-Red (FTIR) spectroscopic peaks confirm the produced product is PHB, which is a type of polyhydroxyalkanoate (PHA), classified within the polyester family highlighting wood waste potential as a sustainable solution to address plastic pollution.

Statistical Modelling of Thermostable Cellulase Production Conditions of Thermophilic Geobacillus sp. TP-1 Isolated from Tapovan Hot Springs of the Garhwal Himalayan Mountain Ranges, India.

A thermo-alkali stable cellulase from Geobacillus sp. TP-1 was isolated from Tapovan hot spring soil sample. The BLASTn sequence analysis of 16S rRNA sequence revealed that the isolate belonged to the Geobacillus genus and shared the highest degree of sequence similarity (99.43%) with the different strains of Geobacillus subterraneus. The neighbour joining method of multiple sequence alignment revealed that the 16S rRNA sequence of Geobacillus sp. TP-1 shows maximum similarity with Geobacillus stearothermophilus strain S_YE6-1017-022. One-Factor-At-a-Time analysis was used to optimize the carbon source, nitrogen source, pH, temperature, inoculum size and growth profile with respect to cellulase production. When compared to un-optimized basal media, optimised medium increased cellulase production by around 3.6 times. The Plackett Burman factorial design was employed to identify the critical medium components influencing cellulase activity and temperature was determined to have a significant effect on overall cellulase production. The current strain was capable of utilising lignocellulosic waste as an alternative carbon source. The use of sugarcane molasses and wheat bran as carbon sources resulted in a significant increase (~ 7.2 fold) in cellulase production in the current study, indicating the bacterium's potential for valorising lignocellulosic biomass into value-added products, which encourages its use in lignocellulosic-based bio refineries. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01258-x.

Novel Green Strategy to Recover Bioactive Compounds with Different Polarities from Horned Melon Peel.

Around 20-30% of the horned melon's weight is peel. This peel is often discarded or underutilized despite containing valuable bioactive compounds. Conventional methods for extracting polyphenols and carotenoids from horned melon peel are typically inefficient, environmentally harmful, or require significant time and energy. The potential of green cloud point extraction (CPE) or green surfactant-based extraction for recovering bioactives with different polarities from this kind of by-product has not been thoroughly investigated. Therefore, this study focused on optimizing CPE process parameters using a one-variable-at-a-time (OVAT) approach. Optimal CPE demonstrated superior yields compared to conventional, ultrasound, microwave, ultrasound-assisted CPE, and microwave-assisted CPE methods. Further, a Plackett-Burman design identified key factors influencing optimal CPE conditions, while artificial neural network (ANN) analysis assessed each input variable's impact on outcomes. Maximum extraction efficiency for total phenolics (352.49 mg GAE/100 g), total carotenoids (16.59 mg ß-carotene/100 g), and antioxidant activity (989.02 µmol TE/100 g) was achieved under conditions of: surfactant type = Tween 80, surfactant concentration = 2%; solid:liquid ratio = 1:100; pH = 6612; equilibration temperature = 35 °C; equilibration time = 60 min; salt type = NaCl; salt concentration = 16.4%; centrifugation speed = 7906× g ; centrifugation time = 13.358 min; and No. of CPE steps = Step 1. This comprehensive approach aimed to enhance the understanding and optimization of CPE for maximizing the recovery of bioactives from the horned melon peel, addressing the inefficiencies of traditional extraction methods.

Oxidative weathering of pyrite in acidic environments: Data-driven experimental evaluation coupled with Raman hyperspectral imaging.

This study deals with the identification of the factors that affect pyrite oxidation in acid mine drainage conditions. For this scope, weathering experiments have been carried at laboratory scale based on the design of experiments methodology to evaluate the effect of factors such as major ion concentrations, crystal size, and humic acids presence over the amount of elemental sulfur produced due to the involved weathering reactions. In particular, metal and anionic concentrations in solution were quantified by inductively coupled plasma-atomic emission spectroscopy and ion-chromatography techniques, respectively, whereas the amount of elemental sulfur was quantified with a high-performance liquid chromatography with diode-array detection technique after proper extraction procedure. A partial least squares regression was calculated to establish a quantitative relationship between the considered factors and the amount of elemental sulfur. After evaluation of the model, ferric iron, crystal size and the presence of humic acids were identified as the relevant factors for pyrite oxidation under acidic conditions. In addition, the surface of the samples was characterized by Raman imaging spectroscopy and subsequently analyzed by explorative hyperspectral analysis methods to assess the spatial distribution of the elemental sulfur as the main weathering product, resulting in a homogenous distribution.

Effective remediation of mercury (II) from aqueous solutions using CS-MnFe2O4-CoS-MWCNTs: Box-Behnken design optimization and adsorption isotherm, kinetics, and thermodynamics evaluation.

Mercury (Hg) is a hazardous heavy metal, non-biodegradable and toxic, posing a serious threat to aquatic life and human health. Therefore, the removal of Hg ions from contaminated water using effective and eco-friendly adsorbents is necessary. In the present study, three magnetic chitosan-based organic-inorganic nanocomposites, such as CS-MnFe2O4, CS-MnFe2O4-CoS, and CS-MnFe2O4-CoS-MWCNTs, were designed and constructed to investigate their capacity for adsorbing Hg ions from aqueous solutions. The physicochemical properties of prepared composites were characterized by various analyses. The BET analyses indicated their high surface area and porous structure, and the N2 adsorption-desorption showed that the modification of CS in three stages by MnFe2O4 and crosslinking reaction, CoS preparation, and MWCNT incorporation resulted in increased N2 adsorption. The XRD confirms the synthesis of MnFe2O4 and CoS in the CS matrix and also the distinct peaks of MWCNTs. The CS-MnFe2O4-CoS-MWCNTs showed acceptable thermal stability with 45% char yields and superparamagnetic properties with magnetic saturation (Ms) of 16 emu g-1. The interactive impacts of independent variables (pH, contact time, and adsorbent dosage) on the removal percentage of Hg(II) onto three prepared adsorbents, as well as the process optimization, were assessed by the Box-Behnken design. The optimum conditions were identified, and the data from the analysis of variance showed that the three independent factors (pH, contact time, and adsorbent dosage) significantly influenced the adsorption of Hg(II). The adsorption isotherm and thermodynamics analysis investigation showed that at low concentrations of Hg(II), the adsorption process was both endothermic and spontaneous for the studied adsorbents.

Polysaccharides from Hericium erinaceus and its immunomodulatory effects on RAW 264.7 macrophages.

This study aimed to optimize the extraction of Hericium erinaceus polysaccharides (HEP) using ultrasound-assisted enzymatic extraction combined with Plackett-Burman design (PBD) and response surface methodology (RSM). The optimal extraction conditions were identified as: 33 min extraction time, 30:1 liquid to material ratio, 38 °C extraction temperature, 9 g/kg cellulase amount, pH 4, and 20 % ethanol concentration. Under these conditions, the extraction yield of HEP was 5.87 ± 0.16 %, consistent with the predicted results. Additionally, the potential immunomodulatory activity of HEP on RAW 264.7 macrophage was evaluated. The results revealed that HEP improved the immunostimulatory activity of RAW264.7 cells, evident from increased production of IL-6 and TNF-α. These findings suggest that HEP is capable of enhancing the immune activity of RAW 264.7 macrophage.

A statistical optimization for almost-complete methylene blue biosorption by Gracilaria bursa-pastoris.

In this study, the dried biomass of four marine algae, namely Porphyra sp., Gracilaria bursa-pastoris, Undaria pinnatifida and Laminaria sp., were screened for their ability to remove methylene blue (MB) dye from aqueous solutions. Statistical approaches of the Plackett-Burman Design (PBD) and Box-Behnken Design (BBD) were applied to optimize different environmental conditions in order to achieve the maximum MB removal percentage by Gracilaria bursa-pastoris. The biosorbent was characterized before and after adsorption process using FTIR, XRD and SEM analysis. Additionally, isotherms, kinetics and thermodynamics studies were conducted to investigate the adsorption behavior of the adsorbent. The results showed that Gracilaria bursa-pastoris achieved the highest dye removal efficiency (98.5 %) compared to 96.5 %, 93.5 % and 93.9 % for Undaria pinnatifida, Porphyra sp. and Laminaria sp., respectively. PBD analysis revealed that the agitation speed, pH, and biomass dose were found to be the significant parameters affecting MB removal onto Gracilaria dried biomass. According to the BBD results, the maximum dye removal percentage (99.68 %) was obtained at agitation speed of 132 rpm, pH 7 and biomass dose of 7.5 g/L. FTIR, XRD and SEM analysis demonstrated the participation of several functional groups in the adsorption process and changes in the cell surface morphology of the adsorbent following the dye adsorption. The adsorption isotherms showed better fit to Freundlich model (R2 = 0.9891) than the Langmuir, Temkin, and Dubinin-Radushkevich models. The adsorption kinetics were best described by the pseudo-second-order model (R2 = 0.9999), suggesting the chemical interactions between dye ions and the algal biomass. The thermodynamic parameters indicated that the adsorption of MB onto Gracilaria dried biomass was spontaneous, feasible, endothermic and random. These results indicate that dried biomass of Gracilaria bursa-pastoris is an attractive, environmentally friendly, cheap and effective agent for MB dye removal from environmental discharges.

Optimized phenol degradation and lipid production by Rhodosporidium toruloides using response surface methodology and genetic algorithm-optimized artificial neural network.

Oleaginous yeast can produce lipids while degrading phenol in wastewater treatment. In this study, a Plackett-Burman Design (PBD) was adopted to identify key factors of phenol degradation and lipid production using R toruloides 9564T. While temperature, inoculum size, and agitation were significant for both the processes (p < 0.05), pH and incubation were significant for lipid production, and phenol removal, respectively. Results from four factors (pH, temperature, inoculum size, and incubation period) central composite design (CCD) experiment were used to formulate quadratic and genetic algorithm-optimized ANN models. The reduced quadratic model for phenol degradation (R2: 0.993) and lipid production (R2: 0.958) were marginally inferior to ANN models (R2: 0.999, 0.982, respectively) on training sets. Multi-objective optimization with equal importance suggests phenol degradation between 106.4 and 108.76%, and lipid production of 0.864-0.903 g/L, by polynomial and ANN models. Complete phenol degradation (100%) and 3.35-fold increment (0.918 g/L) in lipid production were obtained at pH 6.07, inoculum size 14.68% v/v, at 29.5 °C in 92.17 h experimentally.

Pomegranate seed as a novel source of plant protein: Optimization of protein extraction and evaluation of in vitro digestibility, functional, and thermal properties.

This research was carried out to optimize the extraction process of proteins from pomegranate seeds and characterize their in vitro digestibility as well as their thermal and functional properties. For this purpose, the study screened five parameters (liquid/solid ratio, pH, temperature, NaCl concentration, and time) that could potentially influence the extraction process. This screening was conducted using a two-level Placket-Burman design (PBD). The significant parameters (pH and NaCl concentration) were subsequently optimized using a three-level face-centered central composite design (FCCD) to determine the optimum extraction conditions. A maximum protein recovery of 83.8% was obtained at pH 11.0 and NaCl concentration of 0.0 M. Pomegranate seed protein isolate (PSPI) with a protein content of 92.4% (w/w) was obtained through the isoelectric precipitation of pomegranate seed protein extracted under the optimized conditions. An emulsifying activity index of 14.1 m2 g-1 was observed at the isoelectric pH, where the emulsion stability index was at 8.2%. PSPI also showed high water- and oil-holding capacities (3.7 and 4.3 g g-1, respectively). The essential amino acid levels in PSPI (except for valine and isoleucine) exceeded the recommended amounts set by WHO/FAO/UNU for adults, highlighting its high nutritional value. Based on thermal analysis data, denaturation of PSPI could occur at 89.5°C. The in vitro digestibility of PSPI was found to be 74.3%. PSPI shows a potential as a novel ingredient for substituting animal-based proteins in various food applications.

Optimizing Polyhydroxyalkanoate production using a novel Bacillus paranthracis isolate: A response surface methodology approach.

Microorganisms have emerged as promising resources for producing economical and sustainable bioproducts like Polyhydroxyalkanoate (PHA), a biodegradable polymer that can replace synthetic plastics. In this study, we screened a novel isolate, Bacillus paranthracis RSKS-3 strain, to produce PHA from sewage water, identifying it using Whole Genome Sequence. This study represents the first report on optimizing PHA production using B. paranthracis RSKS-3, employing Design Expert 12.0 software. Our findings reveal that four factors (temperature, inoculum size, potassium dihydrogen phosphate, and magnesium sulfate) significantly affect PHA production in the Plackett-Burman design experiment. Through Response Surface Methodology, we optimized PHA production to 0.647 g/L with specific values for potassium dihydrogen phosphate (0.55 %), inoculum size (3 %), magnesium sulfate (0.055 %), and a temperature of 35 °C, in agreement with the predicted value of 0.630 g/L. This optimization resulted in a substantial 13.29-fold increase in PHA production from 0.34 g/L to 4.52 g/L, underscoring the promising role of B. paranthracis RSKS-3 in eco-friendly PHA production and advancing sustainable bioproduct development.

Ammonia recovery via direct contact membrane distillation: Modeling and performance optimization.

Ammonia recovery from wastewater has positive environmental benefits, avoiding eutrophication and reducing production energy consumption, which is one of the most effective ways to manage nutrients in wastewater. Specifically, ammonia recovery by membrane distillation has been gradually adopted due to its excellent separation properties for volatile substances. However, the global optimization of direct contact membrane distillation (DCMD) operating parameters to maximize ammonia recovery efficiency (ARE) has not been attempted. In this work, three key operating factors affecting ammonia recovery, i.e., feed ammonia concentration, feed pH, and DCMD running time, were identified from eight factors, by a two-level Plackett-Burman Design (PBD). Subsequently, Box-Behnken design (BBD) under the response surface methodology (RSM) was used to model and optimize the significant operating parameters affecting the recovery of ammonia though DCMD identified by PBD and statistically verified by analysis of variance (ANOVA). Results showed that the model had a high coefficient of determination value (R2 = 0.99), and the interaction between NH4Cl concentration and feed pH had a significant effect on ARE. The optimal operating parameters of DCMD as follows: NH4Cl concentration of 0.46 g/L, feed pH of 10.6, DCMD running time of 11.3 h, and the maximum value of ARE was 98.46%. Under the optimized conditions, ARE reached up to 98.72%, which matched the predicted value and verified the validity and reliability of the model for the optimization of ammonia recovery by DCMD process.

Optimization of the chromium (Cr+6) reduction from waterways using chemically and bacterially treated agro-waste.

Heavy metals, a major source of pollution in the environment, pose a substantial threat due to their non-biodegradability and ability to accumulate in living organisms, causing health problems. Recently, researchers have been searching for cost-effective and safe ways to remove heavy metals from polluted waterways using agricultural waste substitutes. The present study focused on the low-cost treatments for the reduction of chromium Cr+6 metal from the effluent, wherein it has been found that chemically and bacterially treated agro-waste had increased heavy metal ion adsorption capabilities. A sequential optimization of the process parameters was attempted using Plackett-Burman design (PBD) and central composite design of response surface methodology (CCD-RSM) for the maximum reduction of the chromium metal from the effluent. A total of eight parameters were screened out using a 12-run PBD experiment. Out of the eight parameters, time, HCl, NaOH, and bacterial treatments were found to be significantly affecting the maximum reduction of Cr+6 from the effluent. To investigate the interactions' effects of the chosen parameters, they were evaluated using CCD-RSM. Maximum 74% Cr+6 reduction was achieved under the optimum treatment to rice husk of HCl 4.52 N, NaOH 3.53 N, bacterial suspension 7.41%, and with an interaction time 14.32 min using 30 run CCD-RSM experiment. A scanning electron microscope was used to confirm the effects of selected variables on the agro-waste for the Cr+6 reductions, as well as a Fourier transform infrared spectrometer.

Screening factors to affect ultrasound-assisted extraction of (poly)phenols from date palm seeds.

The aim of the current work was to compare the (poly)phenol profile (free, soluble-conjugate, and insoluble-bound) and antioxidant activity of date palm seed flour using different extraction methods (conventional vs. ultrasound-assisted extraction [UAE]) and to determine the most critical variables in the extraction of (poly)phenols through UAE using the Plackett-Burman design experiment. Using the Plackett-Burman design, seven factors, namely, ethanol concentration, liquid:solid ratio (mL/g), sonotrode, amplitude (%), extraction time, extractant pH, and extraction cycle, were studied. After the factors were studied using conventional extraction methods, 23 compounds were quantified, with protocatechuic acid and catechin being the predominant (poly)phenols. Furthermore, the distribution of (poly)phenols within the cell varied, with glycosylated quercetins and caffeoyl shikimic acids predominantly found in free forms. Ultrasound-assisted extraction demonstrated efficiency in extracting free and soluble-conjugate (poly)phenols. However, it showed limitations in extracting insoluble-bound (poly)phenols. Nevertheless, similar amounts of total (poly)phenols were shown after conventional extraction and UAE, that is, 259.69 ± 43.54 and 189.00 ± 3.08 mg/100 g date seed flour, respectively. The Plackett-Burman design revealed the liquid-solid ratio as a crucial factor affecting (poly)phenol extraction, with higher ratios yielding better results. The sonotrode choice also influenced the extraction efficiency, highlighting that the sonotrode with a smaller diameter but higher displacement amplitude showed the best polyphenol recovery and antioxidant activity values. The nature of (poly)phenols influenced the studied extraction variables differently, emphasizing the complexity of the extraction process. In this line, pure water was sufficient to extract flavan-3-ols after UAE, whereas ethanol was a crucial factor in extracting quercetin. These findings underscore the importance of optimizing extraction methods for maximizing (poly)phenol recovery from date palm seed flour for various applications in food and pharmacology industries.

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

Introduction: 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. Methods: 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. Results: 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. Discussion: 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.

Determination of pesticide residues in soybeans using QuEChERS followed by deep eutectic solvent-based DLLME preconcentration prior to gas chromatography-mass spectrometry analysis.

A reliable and greener alternative to the usual extraction methods is reported for the determination of pesticide residues in soybeans. This novel approach combines the classical QuEChERS extraction method with a DLLME (dispersive liquid-liquid microextraction) step, utilizing a deep eutectic solvent (DES) - camphor: hexanoic acid (1:1 molar ratio) - as the microextraction solvent. This DES has never been employed in pesticide analysis by gas chromatography-mass spectrometry of complex matrices like soybeans. A Plackett-Burman screening design was employed to optimize sample preparation variables of QuEChERS (amount of sodium chloride and magnesium sulfate, and amount of PSA and C18 sorbents) and DLLME (pH of medium, amount of sodium chloride, and volume of microextraction solvent). This design allowed for a systematic evaluation of the impact of each parameter on the method's performance. The optimized method was evaluated using a certified reference material and commercial samples of soybeans. The method exhibited high accuracy and precision for most of the analytes under study, demonstrating its applicability for pesticide residue analysis in soybeans. To assess the greenness and practicality of the developed method, the Analytical Greenness (AGREE) and Blue Applicability Grade Index (BAGI) metric systems were employed, respectively. Overall, the proposed QuEChERS-DLLME method using a DES solvent is a reliable and greener alternative to conventional extraction methods for the determination of pesticide residues in soybeans. Its high performance, coupled with its environmental friendliness, makes it a promising tool for food safety analysis.

Cloning, expression and purification of cellobiohydrolase gene from Caldicellulosiruptor bescii for efficient saccharification of plant biomass.

Anthropogenic activities have led to a drastic shift from natural fuels to alternative renewable energy reserves that demand heat-stable cellulases. Cellobiohydrolase is an indispensable member of cellulases that play a critical role in the degradation of cellulosic biomass. This article details the process of cloning the cellobiohydrolase gene from the thermophilic bacterium Caldicellulosiruptor bescii and expressing it in Escherichia coli (BL21) CondonPlus DE3-(RIPL) using the pET-21a(+) expression vector. Multi-alignments and structural modeling studies reveal that recombinant CbCBH contained a conserved cellulose binding domain III. The enzyme's catalytic site included Asp-372 and Glu-620, which are either involved in substrate or metal binding. The purified CbCBH, with a molecular weight of 91.8 kDa, displayed peak activity against pNPC (167.93 U/mg) at 65°C and pH 6.0. Moreover, it demonstrated remarkable stability across a broad temperature range (60-80°C) for 8 h. Additionally, the Plackett-Burman experimental model was employed to assess the saccharification of pretreated sugarcane bagasse with CbCBH, aiming to evaluate the cultivation conditions. The optimized parameters, including a pH of 6.0, a temperature of 55°C, a 24-hour incubation period, a substrate concentration of 1.5% (w/v), and enzyme activity of 120 U, resulted in an observed saccharification efficiency of 28.45%. This discovery indicates that the recombinant CbCBH holds promising potential for biofuel sector.

Utilizing experimental design and desirability function in optimizing RP-HPLC method for simultaneous determination of some skeletal muscle relaxants and analgesics.

An experimental design and response surface methodologies using Plackett-Burman and Box-Behnken designs were applied for selecting and optimizing the most appropriate parameters which significantly affect the separation and quantitative estimation of five skeletal muscle relaxants and four analgesic drugs (baclofen, methocarbamol, dantrolene sodium, orphenadrine citrate, cyclobenzaprine hydrochloride, ketoprofen, etoricoxib, ibuprofen, and mefenamic acid) with a relatively short duration of analysis in a single run. For the separation of the nine drugs, an INERTSIL ODS-V3-5 µm C18 column (250 × 4.6 mm I.D.) was used with the optimum mobile phase conditions (45.15 mM ammonium acetate buffer pH 5.56 adjusted with acetic acid, acetonitrile, and methanol in a ratio of 30.5:29.5:40, v/v/v with a flow rate of 1.5 mL/min) and UV-detection at 220 nm. The optimized method was successfully subjected to the validation steps as described in ICH guidelines for linearity, precision, accuracy, robustness, and sensitivity. The optimized and validated method was effectively applied to determine the content of the studied drugs in their pharmaceutical preparations and to expand its applicability to the counterfeit estimation of etoricoxib in different brands of tablet dosage forms.

Characterization and optimization of exopolysaccharide extracted from a newly isolated halotolerant cyanobacterium, Acaryochloris Al-Azhar MNE ON864448.1 with antiviral activity.

Several antiviral agents lost their efficacy due to their severe side effects and virus mutations. This study aimed to identify and optimize the conditions for exopolysaccharide (EPS) production from a newly isolated cyanobacterium, Acaryochloris Al-Azhar MNE ON864448.1, besides exploring its antiviral activity. The cyanobacterial EPS was purified through DEAE-52 cellulose column with a final yield of 83.75%. Different analysis instruments were applied for EPS identification, including Fourier-transform infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), and gas chromatographic-mass spectrometry (GC-MS). Plackett-Burman's design demonstrated that working volume (X1), EDTA (X2), inoculum size (X3), CaCl2 (X4), and NaCl (X5) are the most important variables influencing EPS production. Central composite design (CCD) exhibited maximum EPS yield (9.27 mg/mL) at a working volume of 300 mL in a 1 L volumetric flask, EDTA 0.002 g/L, inoculum size 7%, CaCl2 0.046 g/L, and NaCl 20 g/L were applied. EPS showed potent antiviral activities at different stages of herpes simplex virus type-1 and 2 (HSV-1, HSV-2), adenovirus (ADV) and coxsackievirus (A16) infections. The highest half-maximal inhibitory concentration (IC50) (6.477 µg/mL) was recorded during HSV-1 internalization mechanism, while the lowest IC50 (0.005669 µg/mL) was recorded during coxsackievirus neutralization mechanism.