Eco-friendly engineering of micro composite-based hydroxyapatite bio crystal and polyaniline for high removal of OG dye from wastewater: Adsorption mechanism and RSM@BBD optimization.

The presence of harmful substances such as dyes in water systems poses a direct threat to the quality of people's lives and other organisms living in the ecosystem. Orange G (OG) is considered a hazardous dye. The existing paper attempts to evaluate a low-cost adsorbent for the effective removal of OG dye. The developed adsorbent Polyaniline@Hydroxyapatite extracted from Cilus Gilberti fish Scale (PANI@FHAP) was elaborated through the application of the in situ chemical polymerization method to incorporate PANI on the surface of naturally extracted hydroxyapatite FHAP. The good synthesis of PANI@FHAP was evaluated through multiple techniques including X-ray diffraction (XRD), Scanning electron microscopy coupled with energy dispersive X-ray spectrometry (SEM/EDS), Fourier Transforms Infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA) coupled with thermal differential analysis (DTA) analysis. The results reveal a highly ordered disposition of PANI chains on FHAP, resulting in a well-coated FHAP in the PANI matrix. Furthermore, the presence of functional groups on the surface of PANI such as amine (-NH2) and imine (=NH) groups would facilitate the removal of OG dye from contaminated water. The adsorption of OG onto PANI@FHAP was conducted in batch mode and optimized through response surface methodology coupled with box-Behnken design (RSM/BBD) to investigate the effect of time, adsorbent dose, and initial concentration. The outcomes proved that OG adsorption follows a quadratic model (R2 = 0.989). The kinetic study revealed that the adsorption of OG fits the pseudo-second-order model. On the other hand, the isotherm study declared that the Freundlich model is best suited to the description of OG adsorption. For thermodynamic study, the adsorption of OG is spontaneous in nature and exothermic. Furthermore, the regeneration-reusability study indicates that PANI@FHAP could be regenerated and reused up to five successive cycles. Based on the FTIR spectrum of PANI@FHAP after OG adsorption, the mechanism governing OG adsorption is predominantly driven by π-π interaction, electrostatic interaction, and hydrogen bonding interactions. The obtained results suppose that PANI@FHAP adsorbent can be a competitive material in large-scale applications.

Nanosponge hydrogel of octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propanoate of Alcaligenes faecalis.

Octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propanoate (ODHP) was extracted in a previous study from the culture broth of soil isolate Alcaligenes faecalis MT332429 and showed a promising antimycotic activity. This study was aimed to formulate ODHP loaded ß-cyclodextrins (CD) nanosponge (NS) hydrogel (HG) to control skin fungal ailments since nanosponges augment the retention of tested agents in the skin. Box-Behnken design was used to produce the optimized NS formulation, where entrapment efficiency percent (EE%), polydispersity index (PDI), and particle size (PS) were assigned as dependent parameters, while the independent process parameters were polyvinyl alcohol % (w/v %), polymer-linker ratio, homogenization time, and speed. The carbopol 940 hydrogel was then created by incorporating the nanosponges. The hydrogel fit Higuchi's kinetic release model the best, according to in vitro drug release. Stability and photodegradation studies revealed that the NS-HG remained stable under tested conditions. The formulation also showed higher in vitro antifungal activity against Candida albicans compared to the control fluconazole. In vivo study showed that ODHP-NS-HG increased survival rates, wound contraction, and healing of wound gap and inhibited the inflammation process compared to the other control groups. The histopathological examinations and Masson's trichrome staining showed improved healing and higher records of collagen deposition. Moreover, the permeability of ODHP-NS-HG was higher through rats' skin by 1.5-folds compared to the control isoconazole 1%. Therefore, based on these results, NS-HG formulation is a potential carrier for enhanced and improved topical delivery of ODHP. Our study is a pioneering research on the development of a formulation for ODHP produced naturally from soil bacteria. KEY POINTS: • Octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propanoate was successfully formulated as a nanosponge hydrogel and statistically optimized. • The new formula exhibited in vitro good stability, drug release, and higher antifungal activity against C. albicans as compared to the fluconazole. • Ex vivo showed enhanced skin permeability, and in vivo analysis showed high antifungal activity as evidenced by measurement of various biochemical parameters and histopathological examination.

Response surface methodology for optimizing methylene blue dye removal by mesoporous activated carbon derived from renewable woody Bambusoideae waste.

In this study, the focus was on utilizing tropical plant biomass waste, specifically bamboo (BB), as a sustainable precursor for the production of activated carbon (BBAC) via pyrolysis-induced K2CO3 activation. The potential application of BBAC as an effective adsorbent for the removal of methylene blue (MB) dye from aqueous solutions was investigated. Response surface methodology (RSM) was employed to evaluate key adsorption characteristics, which included BBAC dosage (A: 0.02-0.08 g/L), pH (B: 4-10), and time (C: 2-8 min). The adsorption isotherm analysis revealed that the adsorption of MB followed the Freundlich model. Moreover, the kinetic data were well-described by the pseudo-second-order model, suggesting the role of a chemisorption process. The BBAC demonstrated a notable MB adsorption capacity of 195.8 mg/g, highlighting its effectiveness as an adsorbent. Multiple mechanisms were identified as controlling factors in MB adsorption by BBAC, including electrostatic forces, π-π stacking, and H-bonding interactions. The findings of this study indicate that BBAC derived from bamboo has the potential to be a promising adsorbent for the treatment of wastewater containing organic dyes. The employment of sustainable precursors like bamboo for activated carbon production contributes to environmentally friendly waste management practices and offers a solution for the remediation of dye-contaminated wastewater.

This works introduces a renewable and woody Bambusoideae waste as promising and low-cost precursor for producing mesoporous activated carbon via microwave assisted K₂CO₃ activation. The effectiveness of the prepared activated carbon was tested toward removal of a toxic cationic dye, namely; methylene blue from aqueous environment.

Optimization of ultrasound- and enzymatic-assisted extractions of polyphenols from dried red onion peels and evaluation of their antioxidant activities.

There is a recognized need for exploring new natural antioxidants that have high antioxidant potential. Thus, the aim of this study was to optimize the extraction process of onion peels using ultrasound- and enzymatic-assisted extractions (UAE and EAE) methods to utilize the optimal extracts as natural antioxidants. Box-Behnken Design (BBD) was implemented to reach the optimal extracts with the highest simultaneous content of total phenolic content (TPC) and total flavonoid content (TFC). The optimal UAE and EAE extracts were subjected to High Performance Liquid Chromatography (HPLC) analysis to identify their chemical profile. The antioxidant activities of optimal extracts obtained by UAE and EAE were evaluated in vitro using DPPH and FRAP assays and their abilities to increase the oxidative stability of sunflower oil were studied using Rancimat test. The optimal conditions for UAE were 34.02 W, 26.87 mL/g solid, and 45.43 min, meanwhile they were 30.00 mL/g solid, 45.43 min at enzyme concentration of 0.52% for EAE method. DPPH and FRAP assays results revealed that EAE optimal extract show superior antioxidant activity over UAE optimal extract. The protection factor of optimal EAE extract against sunflower oil oxidation was close to that of butylated hydroxytoluene (BHT).

Statistical optimization and sequential scale-up of α-galactosidase production by Actinoplanes utahensis B1 from shake flask to pilot scale.

α-Galactosidase (α-GAL) is a class of hydrolase that releases galactose from galacto-oligosaccharides and synthetic substrates such as pNPG. In this study, the production of α-GAL by Actinoplanes utahensis B1 in submerged fermentation was enhanced by using statistical methods. The effects of temperature, pH, and inoculum percentage on enzyme secretion were optimized using BBD of RSM. The optimized process was scaled up from the shake flask to the laboratory scale (5 L) and to pilot scale (30 L) using KLa based scale-up strategy. By using BBD, a maximum yield of 62.5 U/mL was obtained at a temperature of 28 °C, a pH of 6.9, and an inoculum of 6.4%. Scale-up was performed successfully and achieved a yield of 74.4 U/mL and 76.8 U/mL in laboratory scale and pilot scale fermenters. The TOST was performed to validate the scale-up strategy and the results showed a confidence level of 95% for both scales indicating the perfect execution of scale-up procedure. Through the implementation of BBD and scale-up strategy, the overall enzyme yield has been significantly increased to 76%. This is the first article to explore the scale-up of α-GAL from the A. utahensis B1 strain and provide valuable insights for industrial applications.

Novel route to enhance the thermo-optical performance of bicyclic diene photoswitches for solar thermal batteries.

Harnessing solar energy by employing chemical photoswitches in molecular solar thermal (MOST) energy storage systems is a topic of appealing research interest. However, incorporating all the features desired for an ideal MOST system in a single photoswitching couple is challenging. Inspired by experimental synthesis, herein we report our attempt to enhance both the thermochemical and photophysical properties in a single-bridged bicyclic diene (BBD)-based photoswitch by elongating the unsaturated bridge with different heteroatomic units. To elucidate the best elongation unit, the energy storage capacity and the TBR barriers were accounted using the DLPNO-CCSD(T) and (8,8)-CASPT2 methods, respectively. The photophysical properties including the absorption onset, excitation wavelengths, and the absorption intensities were extensively investigated with the time-dependent calculations. The result provides information on the most versatile solvent to exhibit the best photoswitching behaviour which is beneficial for real-life energy storage applications. Additionally, the stability and reversibility of the photoswitching system with elongated unsaturated bridges have also been assessed. By means of the studied modification, the storage energy of 158.57 kJ/mol, energy storage density of 1.48 MJ/kg, TBR barrier of 136.36 kJ/mol, and the absorption onset of 305.00 nm is achieved in acetonitrile. These values are substantially higher when compared with the storage energy (96.06 kJ/mol), energy storage density (1.04 MJ/kg), and TBR barrier (121.76 kJ/mol) of prototype NBD/QC in the gas phase. The outcomes render useful insights into the stability and properties of bicyclic diene-based photoswitches having elongated unsaturated bridges and indeed paves the way for the rational design of practical MOST systems.

Vancouver Symptom Score for Dysfunctional Elimination Syndrome: Hindi translation and validation.

BACKGROUND: Bladder and bowel dysfunction (BBD) is a common problem in school-aged children and is evaluated using questionnaires. Among available questionnaires, "Vancouver Symptom Score for Dysfunctional Elimination Syndrome" (VSSDES) is considered superior in validity and reliability. We translated VSSDES into Hindi and performed validity and reliability testing of Hindi-VSSDES in children aged 4-16 years with lower urinary tract dysfunction (LUTD). METHODS: In this cross-sectional study, we followed Consensus-based Standards for the selection of health Measurement Instruments recommendation to develop Hindi-VSSDES. Forward-translation-harmonization-back-translation-expert committee review-prefinal version sequence of translation was followed by cognitive debriefing. One hundred and sixteen children with symptoms suggestive of LUTD and 106 healthy controls were recruited. Construct validity of Hindi-VSSDES was assessed via structural validity (factor analysis) and hypotheses testing. In addition, test-retest reliability and internal-consistency reliability were assessed. RESULTS: Hindi-VSSDES had acceptable face validity. Factor analysis identified four factors representing three domains. All a priori hypotheses were met. The questionnaire displayed excellent convergent validity and discriminative ability (area under receiver operating characteristic curve 0.83). The cutoff scores for screening and diagnosis were 7 and 11, respectively. Scores were comparable whether children self-filled the questionnaire or parents were proxy. Scores in girls were significantly higher. It had an excellent test-retest reliability (intraclass correlation coefficient = 0.96) and Cronbach's ɑ was 0.62. The smallest detectable change was 4.1 points. Median time to complete the questionnaire was 7 min and majority (91%) rated it as easy. CONCLUSIONS: Hindi-VSSDES is the first validated and reliable Hindi questionnaire for children with BBD. Clinimetric properties of responsiveness and interpretability are yet to be tested.

Exploration of polymethacrylate and Hypromellose for the development of a non-sulfhydryl ACE inhibitor mucoadhesive system using Box-Behnken design: in-vitro and ex-vivo evaluation.

PURPOSE: To counteract early morning pathology like hypertension a time-dependent release of the drug is required. This study is focused to formulate a pulsatile and mucoadhesive drug delivery system of an ACE inhibitor Perindopril Erbumine. METHOD: Two matrix tablets were punched with Eudragit RSPO, Eudragit RLPO, and HPMC K15M using a 3-3-3 Box-Behnken Design of Response Surface Methodology. Based on the design-optimized formulation P1T3 and P2T8 were coated for a lag time with compression coating of HPMC K4M and a blend of 1:1 ratio of ethylcellulose and carbopol polymer and further encapsulated in a Eudracap™ capsule to provide gastric resistance. RESULT: The in-vitro release data confirmed an initial pause phase of 4.5 h then release of the drug for 5.2 ± 0.3 h to cope with the early morning rush in blood pressure. After that, a gap of 6 h and then sustained release of the drug for 10.5 ± 0.5 h. From the ex-vivo study, mucoadhesive strength was obtained as 55.13 ± 0.03 gm and 56.39 ± 0.02 gm for P1T3 and P2T8 respectively. The lag time for coated tablet P1T3 came to 2.15 ± 0.15 h and for P2T8 11.9 ± 0.10 h proving the coating efficiency of polymers. CONCLUSION: The current study strongly suggests that perindopril Erbumine in association with Eudragit and Hypromellose polymer can open a path for the time-regulated release of the drug for hypertension chronotherapy with less risk of dose dumping.

Quality by Design (QbD) Based Method for Estimation of Xanthohumol in Bulk and Solid Lipid Nanoparticles and Validation.

The analytical quality by design (AQbD) approach is utilized for developing and validating the simple, sensitive, cost-effective reverse-phase high performance liquid chromatographic method for the estimation of xanthohumol (XH) in bulk and nanoformulations. The Box-Behnken design (BBD) is applied for method optimization. The mobile phase ratio, pH and flow rate were selected as independent variables, whereas retention time, peak area, peak height, tailing factor, and theoretical plates were selected as dependent variables. The chromatogram of XH obtained under optimized conditions has given optimum conditions such as retention time (5.392 min), peak area (1,226,737 mAU), peak height (90,121 AU), tailing factor (0.991) and theoretical plates (4446.667), which are contoured in the predicted values shown by BBD. Validation of the method has been performed according to ICH Q2(R1) recommendations, using optimized conditions for linearity, limit of detection (LOD) and limit of quantification (LOQ), accuracy, precision, robustness and system suitability. All the values of validation parameters lie within the acceptable limits prescribed by ICH. Therefore, the developed and validated method of XH by the AQbD approach can be applied for the estimation of XH in bulk and various nanoformulations.

Integrated Supercritical Fluid Extraction and Pre-Formulation Process of Punica granatum L. Pericarp Polar Compounds.

Pomegranate (Punica granatum L.) is a widely used fruit in the dietary supplement industry due to its richness in bioactive compounds. In this study, an experimental design was applied to optimize supercritical fluid extraction (SFE) of polar compounds of interest (ellagic acid and punicalagins), known for antioxidant and skin care properties from pomegranate's pericarp. The effects of temperature, modifier percentage, and water additive percentage added in the modifier were explored through a Box-Behnken design, followed by a study of the extraction kinetics. The results indicated that 40 °C, 20% EtOH:H2O 80:20 v:v, with an extraction duration of 60 min allowed for the highest recovery of the above-mentioned molecules (19.59 mg/g). Due to solubilization issues encountered by the extract, a screening of cosmetic solvents was carried out to solubilize SFE pomegranate extracts and a composition of Gly:H2O 80:20 v:v was selected. Furthermore, an integrated SFE pre-formulation process of pomegranate pericarp extract (PPE) was elaborated. This allowed for the recovery of the extracts in cosmetic solvent, avoiding a full evaporation. Finally, the stability of the pre-formulated extracts was evaluated and showed high stability for over 3 months at 5 °C.

Ultrasound-Assisted Extraction of Antioxidants from Melastoma malabathricum Linn.: Modeling and Optimization Using Box-Behnken Design.

This study presents modeling and optimization of ultrasound-assisted extraction (UAE) of Melastoma malabathricum with the objective of evaluating its phytochemical properties. This one-factor-at-a-time (OFAT) procedure was conducted to screen for optimization variables whose domains included extraction temperature (XET), ultrasonic time (XUT), solvent concentration (XSC), and sample-to-liquid ratio (XSLR). Response surface methodology (RSM) coupled with Box-Behnken design (BBD) was applied to establish optimum conditions for maximum antioxidant extraction. Modeling and optimization conditions of UAE at 37 kHz, XET 32 °C for XUT 16 min and dissolved in an XSC 70% ethanol concentration at a XSLR 1:10 ratio yielded scavenging effects on 2,2-diphenyl-1-picryl-hydrazyl (DPPH) at 96% ± 1.48 and recorded values of total phenolic content (TPC) and total flavonoid content (TFC) at 803.456 ± 32.48 mg GAE (gallic acid equivalents)/g, and 102.972 ± 2.51 mg QE (quercetin equivalents)/g, respectively. The presence of high flavonoid compounds was verified using TWIMS-QTOFMS. Chromatic evaluation of phytochemicals using gas chromatography-mass spectrometry (GC-MS) revealed the presence of 14 phytocompounds widely documented to play significant roles in human health. This study provides a comparative evaluation with other studies and may be used for validation of the species' potential for its much-acclaimed medicinal and cosmeceutical uses.

Mathematical Modelling and Optimization for Facile Synthesis of Structured Activated Carbon (ACs) from Adansonia kilima (Baobab) Wood Chips Integrating Microwave-Assisted Pyrolysis for the Elimination of Lead (II) Cations from Wastewater Effluents.

In this research, activated carbon (AC) was synthesized from ligno-cellulosic residues of Adansonia kilima (Baobab) wood chips (AKTW) using two-step semi-carbonization and subsequent pyrolysis using microwave-induced heating (MWP) in the presence of a mild activating agent of K2CO3. The influence of process input variables of microwave power (x1), residence time (y1), and amount of K2CO3 (z1) were analysed to yield superior quality carbon having maximum removal efficiencies (R1) for lead (II) cations from waste effluents, fixed carbon percentages (R2), and carbon yield percentages (R3). Analysis of variance (ANOVA) was used to develop relevant mathematical models, with an appropriate statistical assessment of errors. Level factorial response surface methodology (RSM) relying on the Box-Behnken design (BBD) was implemented for the experimental design. The surface area and porous texture of the samples were determined using Brunauer, Emmett, and Teller (BET) adsorption/desorption curves based on the N2 isotherm. Surface morphological structure was observed using field emission scanning electron microscopic (FESEM) analysis. Thermogravimetric analysis (TGA) was carried out to observe the thermal stability of the sample. Change in the carbon content of the samples was determined using ultimate analysis. X-ray diffraction (XRD) analysis was performed to observe the crystalline and amorphous texture of the samples. The retention of a higher proportion of fixed carbon (80.01%) ensures that the synthesized adsorbent (AKTWAC) will have a greater adsorption capacity while avoiding unwanted catalytic activity for our synthesized final sample.

Quality-by-Design Approach for Investigating the Efficacy of Tacrolimus and Hyaluronic Acid-Loaded Ethosomal Gel in Dermal Management of Psoriasis: In Vitro, Ex Vivo, and In Vivo Evaluation.

Psoriasis is an auto-immune condition with high keratinocyte hyperproliferation due to lower p53 and p22 levels. Tacrolimus, an immune suppressor, is considered one of the most effective drugs in suppressing psoriasis. Systematic administration of tacrolimus often leads to challenging side effects, namely increased infection risk, renal toxicity, neurological symptoms such as tremors and headaches, gastrointestinal disturbances, hypertension, skin-related problems, etc. To address this, a nanocarrier-based formulation of tacrolimus along with inclusion of hyaluronic acid was developed. The optimization and formulation of ethosomes via the ethanol injection technique were done based on the Box-Behnken experimental design. The results revealed hyaluronic acid-based tacrolimus ethosomes (HA-TAC-ETH) had nanometric vesicle size (315.7 ± 2.2 nm), polydispersity index (PDI) (0.472 ± 0.07), and high entrapment efficiency (88.3 ± 2.52%). The findings of drug release and skin permeation showed sustained drug release with increased dermal flux and enhancement ratio. The effectiveness of HA-TAC-ETH was confirmed in an imiquimod (5%)-prompted psoriasis model. The skin irritation score and Psoriasis Area and Severity Index (PASI) score indicated that HA-TAC-ETH gel has validated a decline in the entire factors (erythema, edema, and thickness) in the imiquimod-induced psoriasis model in contrast with TAC-ETH gel and TAC ointment. The fabricated HA-TAC-ETH opt gel proved to be safe and effective in in vivo studies and could be employed to treat psoriasis further.

Detection rate is not higher for women with BBD history in breast cancer screening.

BACKGROUND: To investigate whether women with benign breast disease (BBD) history have higher breast cancer detection rate in screening. METHODS: We reviewed data for 33 001 female participants in Multi-modality Independent Screening Trial (MIST). Corresponding data for 6823 breast cancer patients were retrieved from the Tianjin Breast Cancer Cases Cohort (TBCCC) and analyzed for comparison. RESULTS: The breast cancer detection rate was 2.83‰ among women with BBD history and 3.28‰ in women without. Moreover, the proportion of carcinoma in situ (CIS) was also lower in women with BBD history than women without (7.69 versus 20.31%). In contrast, analysis of TBCCC data revealed a higher proportion of CIS in patients with BBD history (5.05%) than patients without (3.26%). Our data showed that a larger proportion of women with BBD history had undergone previous breast examinations. Additionally, among participants diagnosed with both breast cancer and BBD in MIST, we found a lower proportion of CIS in women with BBD history (11.76%) compared to women without (32.14%). CONCLUSIONS: Women with BBD history were not found to have higher detection rate in breast cancer screening. Women with BBD history were more likely to be proactive in seeking breast examinations and to have breast cancer be diagnosed in clinic.

Impact of redox-mediators in the degradation of olsalazine by marine-derived fungus, Aspergillus aculeatus strain bpo2: Response surface methodology, laccase stability and kinetics.

The indiscriminate disposal of olsalazine in the environment poses a threat to human health and natural ecosystems because of its cytotoxic and genotoxic nature. In the present study, degradation efficiency of olsalazine by the marine-derived fungus, Aspergillus aculeatus (MT492456) was investigated. Optimization of physicochemical parameters (pH. Temperature, Dry weight) and redox mediators {(2,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), p-Coumaric acid and 1-hydroxybenzotriazole (HOBT)} was achieved with Response Surface Methodology (RSM)-Box-Behnken Design (BBD) resulting in 89.43% removal of olsalazine on 7th day. The second-order polynomial regression model was found to be statistically significant, adequate and fit with p < 0.0001, F value=41.87 and correlation coefficient (R2=0.9826). Biotransformation was enhanced in the redox mediator-laccase systems resulting in 99.5% degradation of olsalazine. The efficiency of ABTS in the removal of olsalazine was more pronounced than HOBT and p-Coumaric acid in the laccase-mediator system. This is attributed to the potent nature of the electron transfer mechanism deployed during oxidation of olsalazine. The pseudo-second-order kinetics revealed that the average half-life (t1/2) and removal rates (k1) increases with increasing concentrations of olsalazine. Michaelis-Menten kinetics affirmed the interaction between laccase and olsalazine under optimized conditions with maximum removal rate, Vmax=111.11 hr-1 and half-saturation constant, Km=1537 mg L-1. At the highest drug concentration (2 mM); 98%, 95% and 93% laccase was remarkably stabilized in the enzyme-drug degradation system by HOBT, ABTS and p-Coumaric acid respectively. This study further revealed that the deactivation of laccase by the redox mediators is adequately compensated with enhanced removal of olsalazine.

Formulation design and optimization of cationic-charged liposomes of brimonidine tartrate for effective ocular drug delivery by design of experiment (DoE) approach.

OBJECTIVE: The present study was aimed to design and optimize brimonidine tartrate (BRT) loaded cationic-charged liposome formulation with enhanced trans-corneal drug permeation, prolonged corneal residence, and sustained drug release for effective ocular delivery. METHODS: Design of experiment (DoE) based formulation optimization was done by three-factor, three-level Box-Behnken design selecting lipid, cholesterol, and drug content as independent variables and particle size (PS), polydispersity index (PDI), zeta potential (ZP), entrapment efficiency (EE%), and cumulative % drug release (CDR) as response variables. The optimized formulation consisting of 79.2 mM lipid, 36.2 mM cholesterol, and 15.8 mg/mL drug was prepared by thin film hydration-sonication method using EPCS:DOTAP (1:1) as lipid component and characterized for all desired critical quality attributes (CQAs), drug release kinetics, TEM, DSC, XRD analysis, ex vivo trans-corneal drug permeation, and physical stability studies. RESULTS: The optimized liposome formulation exhibited experimentally observed responses close to predicted values having 150.4 nm (PS), 0.203 (PDI), 30.62 mV (ZP), and 55.17% (EE). The observed CDR (%) was 36.15% at 1 h and 91.13% at 12 h exhibiting sustained drug release profile and followed Higuchi drug release kinetics. The TEM, DSC, and XRD studies revealed spherical, nanosized, small unilamellar vesicles effectively entrapping BRT in liposomes. The ex vivo permeation study across goat cornea recorded apparent permeability (Papp) 1.011 ± 0.07 cm.min-1 and steady-state flux (Jss) 17.63 ± 1.22 µg.cm-2.min-1 showing >2-fold enhanced drug permeation as compared to BRT solution. CONCLUSION: The developed liposomal formulation possessed all recommended CQAs in optimal range with enhanced trans-corneal drug permeation and remained physically stable in 3 months stability study.

Optimization of Ultrasonic Cellulase-Assisted Extraction and Antioxidant Activity of Natural Polyphenols from Passion Fruit.

In this paper, ultrasonic cellulase extraction (UCE) was applied to extract polyphenols from passion fruit. The extraction conditions for total phenol content (TPC) and antioxidant activity were optimized using response surface methodology (RSM) coupled with a Box-Behnken design (BBD). The results showed that the liquid-to-solid ratio (X2) was the most significant single factor and had a positive effect on all responses. The ANOVA analysis indicated quadratic models fitted well as TPC with R2 = 0.903, DPPH scavenging activity with R2 = 0.979, and ABTS scavenging activity with R2 = 0.981. The optimal extraction parameters of passion fruit were as follows: pH value of 5 at 30 °C for extraction temperature, 50:1 (w/v) liquid-to-solid ratio with extraction time for 47 min, the experimental values were found matched with those predicted. Infrared spectroscopy suggested that the extract contained the structure of polyphenols. Furthermore, three main polyphenols were identified and quantified by HPLC. The results showed the content of phenolic compounds and antioxidant activity of the optimized UCE were 1.5~2 times higher than that determined by the single extraction method and the Soxhlet extraction method, which indicates UCE is a competitive and effective extraction technique for natural passion fruit polyphenols.

Optimization of purification conditions for papain in a polyethylene glycol-phosphate aqueous two-phase system using quaternary ammonium ionic liquids as adjuvants by BBD-RSM.

This work attempts to study and optimize the conditions for separating and purifying papain in aqueous two-phase systems (ATPSs). Quaternary ammonium ionic liquids (ILs, 4 wt%) were added as adjuvants to a PEG-phosphate ATPS. On the basis of single-factor experiments, a Box-Behnken design with response surface methodology (BBD-RSM) was used to optimize the purification conditions of papain in the ATPS by setting the NaH2PO4·2H2O concentration, PEG concentration and pH as independent variables and the overall desirability (OD) of the recovery rate of papain, the protein recovery rate and the purification factor as dependent variables. The following optimum conditions were determined: PEG4000 16.4 wt%, NaH2PO4·2H2O 13.7 wt%, pH 6.22, temperature 60 °C and enzyme concentration 12.0 mg/ml. Under the optimized conditions, the purification factor for the ATPS supplemented with commercial enzyme increased from 1.331 (no ILs) to 3.380 (containing 4 wt% [N2222]BF4). The total evaluation OD was 0.9979, the maximum predicted OD was 0.9994, and the deviation rate was -0.15%. Therefore, the model established in this experiment could predict the experimental value well. To verify the practical effect of the model, papain obtained from fresh papaya latex (papain crude extract) was applied to the same ATPS. The results showed that the purification factor of the ATPS with papain crude extract increased from 3.517 (no ILs) to 12.04 (containing 4 wt% [N2222]BF4). In summary, the addition of 4 wt% ILs to partially replace PEG greatly improved the purification factor for crude papain extract enriched in the phosphate phase, providing a potential method for the large-scale industrial production of papain.

Efficient racemization of N-phenylacetyl-D-glufosinate for L-glufosinate production.

Most amino acids contain chiral centres and exist as both D-enantiomer and L-enantiomer. The optically pure enantiomer is often more valuable than the racemate. Enzymatic resolution provides an effective strategy to obtain optically pure amino acids but often results in large amounts of unwanted isomer. In this study, optically pure L-glufosinate (L-PPT) was obtained by coupling amidase-mediated hydrolysis of N-phenylacetyl-D,L-glufosinate with racemization of N-phenylacetyl-D-glufosinate (NPDG), which exclusively exhibits effective herbicidal properties compared with its D-enantiomer. To improve the yield of L-PPT, the racemization reaction conditions were optimized, and through single-factor experiments, the optimal reaction temperature, reaction time, and mole ratio of phenylacetic acid to NPDG were determined to be 150°C, 30 minutes, and 1.5, respectively. The response surface methodology was applied to further optimize the racemization conditions, and the final yield of L-PPT reached 96.13% with optimum reaction temperature of 154°C, reaction time of 23 minutes, and phenylacetic acid/NPDG mole ratio of 1.7, respectively. Moreover, adding a small amount of acetic anhydride further raised the yield of L-PPT to 97.02%.

Optimization of the Cellulase-Ultrasonic Synergistic Extraction Conditions of Polysaccharides from Lenzites betulina.

Lenzites betulina has been recognized as a rich source of chemical components, including polysaccharides, sterides and sugar alcohols. In this study, cellulase-ultrasonic synergistic extraction method was applied to extract polysaccharides from L. betulina, and the response surface methodology was used to optimize the extraction conditions. The eight basic factors affecting extraction yield were evaluated by Plackett-Burman design (PBD). Then, the four important factors significantly affecting the yield of polysaccharides from L. betulina, including enzymolysis temperature, enzymolysis time, ultrasound time and ultrasound temperature, were optimized by Box-Behnken design (BBD). Maximum extraction yield of L. betulina polysaccharides was 13.64±0.09 % at a cellulase dosage of 0.8 %, enzymolysis temperature of 60 °C, enzymolysis time of 180 min, pH of 4.5, liquid-solid ratio of 45 ml/g, ultrasound power of 300 W, ultrasound time of 20 min and ultrasound temperature of 45 °C. Subsequently, the characteristic structure of crude polysaccharides was determined by FT-IR. Results indicated that cellulase-ultrasonic synergistic treatment is suitable for L. betulina polysaccharides extraction, and it has good prospect for development and utilization.