Screening and optimization of the formulation of human recombinant neutrophil inhibitory factor and hirulog hybrid for injection / 中国生物制品学杂志

@#Objective To screen and optimize the formulation and technology of human recombinant neutrophil inhibitory factor and hirulog hybrid(TNHH)for injection,and investigate its stability.Methods Based on the results of the single factor experiment,with the pH range,mannitol dosage and povidone K30 dosage as independent variables,and the content of high molecular protein as response value,the response surface design(CCF)test was used to analyze the effects of the respective variables and their interaction on the content of high molecular protein in TNHH for injection to screen out the optimal formulation. In order to facilitate the operation,the optimal formulation was adjusted to prepare three batches of samples in pilot scale,which were placed at 40 ℃,75% relative humidity(RH)for 2,4 weeks and 2 ～ 8 ℃ for 3,6 months,respectively. The samples were taken and the appearance,pH,purity of reversed phase-high performance liquid chromatography(RP-HPLC)and purity of size exclusion chromatography-high performance liquid chromatography(SECHPLC)were detected to verify the stability of this formulation and process.Results The optimal formulation was pH 4. 982 6,mannitol 7. 986 4% and povidone K30 1. 902 7%,which was finally adjusted to pH 5. 0,mannitol 8. 0% and povidone K302. 0%. The TNHH preparation for injection prepared by the optimized prescription and process were stable in quality and met the clinical medication requirements.Conclusion The optimum formulation of TNHH preparation for injection is reasonable in the process and suitable for industrial production.

Trametes versicolor (L.) Lloyd as a source of thermostable serine protease: production and characterization

Proteases are ubiquitously present and are among the largest groups of commercially important enzymes. Here, we investigated a wood-rot basidiomycete Trametes versicolor (L.) Lloyd [Syn. Coriolus versicolor (L.) Quél.; Polyporus versicolor (L.) Fr.] as a source of the enzyme serine protease, its production, and optimized to obtain a higher yield of the enzyme.. The significant variables with optimized values for maximum production of the enzyme were temperature (30?C), incubation time (120 h) and wheat bran (10 g). The yield increased by 30.76% by statistically optimizing the media. The optimized temperature and pH for the maximum protease activity was 50?C and pH 7.0, respectively. The enzyme was purified through ion exchange (using DEAE cellulose 52 resin) and gel filtration chromatography (using Superdex 200 column). The purified enzyme had a retention time of 7 min in RP-HPLC. The enzyme was stable at a broad range of temperature (30-60?C) and pH (5.0-8.0) with a half-life of 58.72 min, Vmax of 37.17 ?M min/mL and Km of 0.657 mg/mL. Its activity was enhanced by Na+, Ca2+, Mg2+ ions and SDS surfactant. These properties make this enzyme a valuable candidate for industrial applications

Bioconversion of glycerol waste to ethanol by Escherichia coli and optimisation of process parameters

Biofuel is one of the best ways to reduce our dependence on fossil fuels. Ever since commercial biodiesel production began, waste glycerol, the biodiesel byproduct, has gained researchers’ interest, especially its recycling. Here, we explored using glycerol residue (carbon source) as a substrate in the fermentation process for ethanol production by Escherichia coli K12 in anaerobic conditions. The factors affecting the ethanol production was optimised by response surface methodology (RSM). Significant variables that impact the ethanol concentration were pH, temperature and the substrate, with a statistically significant effect (P <0.05) on ethanol formation. The significant factor was analyzed by the Box-Behnken design. The optimum conditions for bioethanol formation using glycerol as substrate was obtained at pH 7 and temperature 37°C. The ethanol productivity was 0.77 g/L/h. The ethanol concentration of 9.2 g/L achieved from glycerol residue was close to the theoretical value with the fermentation achieved at optimised terms.

Quality evaluation of Saposhnikoviae Radix based on QAMS combined with information entropy-response surface method / 中国中药杂志

This research was used with high performance liquid chromatography(HPLC), combined with information entropy-response surface method(RSM) to investigate the ethanol concentration, extraction time, liquid-to-material ratio. Taking the content of four chromogens as evaluation indexes, the weight coefficients of each index were given, and the comprehensive score was calculated to optimize the extraction process. Then, prim-O-glucosylcimifugin was used as the reference, the relative calibration factors(RCFs) of cimifugin, 4'-O-β-D-glucosyl-5-O-methylvisamminol and sec-O-glucosylhamaudo to prim-O-glucosylcimifugin were calculated respectively. The contents of four components in Saposhnikoviae Radix were determined by both external standard method(ESM)and quantitative analysis of multi-components by single marker(QAMS) method, and the results were compared. At last, combined with principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) to evaluate the quality of the Saposhnikoviae Radix in different production areas. The optimal extraction process parameter of the Saposhnikoviae Radix was as follows: liquid-to-material ratio is 60∶1(mL·g~(-1)), extraction time is 35 min, and ethanol concentration is 70%. The repeatability of the RCFs was perfect, and the results calculated by the QAMS were consistent with the results from the ESM. The stoichiometric results indicate that there are obvious differences in the distribution of Saposhnikoviae Radix in different production areas, and cimifugin and prim-O-glucosylcimifugin are the characteristic compounds that cause this difference. In this study, the optimal extraction process is stable and feasible, and the method of QAMS is accurate and reliable. From the perspective of four chromogens, there are differences in the quality of the Saposhnikoviae Radix in different production areas. Therefore, the established extraction process combined with the method of QAMS can be used to evaluate the quality of Saposhnikoviae Radix and provide a scientific basis for the quality control of Saposhnikoviae Radix.

Microencapsulation of immunoglobulin Y: optimization with response surface morphology and controlled release during simulated gastrointestinal digestion / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Immunoglobulin Y (IgY) is an effective orally administered antibody used to protect against various intestinal pathogens, but which cannot tolerate the acidic gastric environment. In this study, IgY was microencapsulated by alginate (ALG) and coated with chitooligosaccharide (COS). A response surface methodology was used to optimize the formulation, and a simulated gastrointestinal (GI) digestion (SGID) system to evaluate the controlled release of microencapsulated IgY. The microcapsule formulation was optimized as an ALG concentration of 1.56% (15.6 g/L), COS level of 0.61% (6.1 g/L), and IgY/ALG ratio of 62.44% (mass ratio). The microcapsules prepared following this formulation had an encapsulation efficiency of 65.19%, a loading capacity of 33.75%, and an average particle size of 588.75 µm. Under this optimum formulation, the coating of COS provided a less porous and more continuous microstructure by filling the cracks on the surface, and thus the GI release rate of encapsulated IgY was significantly reduced. The release of encapsulated IgY during simulated gastric and intestinal digestion well fitted the zero-order and first-order kinetics functions, respectively. The microcapsule also allowed the IgY to retain 84.37% immune-activity after 4 h simulated GI digestion, significantly higher than that for unprotected IgY (5.33%). This approach could provide an efficient way to preserve IgY and improve its performance in the GI tract.

Optimization of Formulation of Econazole Solid Lipid Nanoparticles Based on Pseudo-ternary Phase Diagrams and Central Composite Design-Response Surface Methodology / 中国药学杂志

OBJECTIVE: To optimize the formulation of econazole solid lipid nanoparticles(E-SLN) by combining pseudo-ternary phase diagrams and central composite design-response surface methodology (CCD-RSM). METHODS: Econazole solubility in different solid lipids and the capacity of lipid emulsion were tested. The microemulsion region was obtained by the pseudo-ternary phase diagrams. Then the E-SLN were prepared by microemulsion method. Drug/lipid (X1), lipid/surfactant (X2) and surfactant/cosurfactant (X3) were taken as individual factors, the encapsulation efficiency (Y1), particle size (Y2), Zeta potential (Y3) were taken as the dependent factors. The possible optimum formulation was predicted by CCD-RSM and validated. RESULTS: Econazole could be dissolved in tripalmitic acid glyceride (TAG), monostearic acid glyceride, stearic acid and lauric acid glyceride. TAG had a good capacity of emulsion. The optimized formulation was econazole 0.06 g, glyceryl palmitate 0.48 g, Tween-80 1.194 g, glycerol 0.274 g and added water to 30 mL by CCD-RSM. According to the optimized formulation, the encapsulation efficiency, particle size and Zeta potential were (94.06±1.54)%, (18.88±0.38)nm and (3.53±0.01)mV, respectively. The deviation was less than 5%. CONCLUSION: The stable and ultra-small size E-SLN with high encapsulation efficiency could be obtained by combining pseudo-ternary phase diagrams and CCD-RSM.

Enhanced biobutanol production by optimization of the medium from Clostridium acetobutylicum MTCC 11274 using macroalgae Gracilaria edulis

Clostridial strain Clostridium acetobutylicum MTCC 11274 was employed for producing biobutanol inbatch culture fermentation. The effects of various carbon sources, i.e., xylose, starch, dextrin, glucose, andmannose as well as nitrogen sources, i.e., yeast extract, peptone, beef extract, and soya protein were studiedconventionally (one-factor-at-a-time). It was found that the maximum amount of biobutanol, i.e., 6.27 and 7.40g/l was obtained from 60 g/l glucose and 5 g/l yeast extract, respectively. In addition to this, the interactionsbetween pH, temperature, and glucose concentration were also taken into consideration for the optimization ofbiobutanol production with the help of Central Composite Design (CCD) of Response Surface Methodology.CCD design was used for the optimization of the above-mentioned parameters and low and high values ofvariables were chosen by performing the steepest ascent experiment. The analysis of variance (ANOVA)model was used for estimating the significance of the model coefficients. ANOVA revealed that the modelwas significant (p < 0.05) and the effects of the glucose concentration, pH, and temperature on biobutanolproduction were significant. It was found that 8.56 g/l biobutanol was produced under optimum fermentationconditions with 40 g/l Gracilaria edulis supplemented with 20 g/l glucose as a carbon source

Development of andrographolide colon-targeting release pellets based on quality by design (QbD) approach / 中草药

Objective: To optimize andrographolide pellet by using a quality by design (QbD) approach. Methods: Usingthe cumulative in vitro release as evaluation index, the single-factor investigation and hazard analysis were used to study the core and coating process of andrographolide colon-targeting pellets, and the central composite design-response surface method was used to optimize and predict the three key factors of plasticizer dosage, aging time, and coating weight gain. Results: The optimum coating process parameters were as following: plasticizer dosage of 3 g, coating weight gain of 20%, and aging time of 1 h. The process had been verified that the optimal formulation process had a cumulative in vitro release of 6.9% in the acid phase (0.1 mol/L HCl)-buffered salt phase (pH 6.0) and a cumulative in vitro release of more than 90% in the pH 7.2 buffered salt phase. Conclusion: It is feasible to apply the QbD concept optimization in the study of andrographolide colon-targeting pellets.

Quality Characteristics of Ginger Extract Candy with Salicornia herbacea L. for Calming Effect on Morning Sickness

The primary objective of this study was to develop an optimal composite recipe for ginger extract candy with Salicornia herbacea L., for consumption during the first trimester of pregnancy. The secondary objective was to examine quality characteristics of the candy. The physical and mechanical properties and sensory properties for pregnant women in were measured, and these values were applied to mathematical models. Time of stirring water solution, saltiness, pH, and redness of the candy increased as concentrations of ginger juice did, but variations in pH were not significant. The hardness values of the candy ranged from 3,063.90 to 5,681.65 dyne/cm³. The average values of sweetness and time stirring the water solution were 5.36% and 14.1 minutes, respectively. However, hardness and sweetness stirring water solution were not significant. The range of sensory values of color (P < 0.01), flavor (P < 0.05), sweetness, saltiness, spiciness, and overall quality (P < 0.05) ranged from were 3.73~5.32, 4.05~5.05, 3.67~5.14, 3.59~5.09, 3.55~5.15, and 3.32~5.45, respectively. Results suggest that ginger extract candy with Salicornia herbacea L. should be comprised of 7.37 g of ginger juice and 1.77 g of salt. Consequently, it could be a functional candy for pregnant women.

Technology research and quality assessment on prescription of imperatorin lipid microspheres by central composite design-response surface method / 中草药

Objective To optimize prescription and evaluate the quality of imperatorin lipid microspheres (ILM) by central composite design-response surface methodology (CCD-RSM), therefore to use the mean particle size, particle size distribution (polydispersity index, PDI) and Zeta potential as the primary indicator. Methods Mean particle size, particle size distribution and Zeta potential of lipid microspheres were measured to investigate the effect of the concentration of egg yolk lecithin, poloxamer and percentage of soybean oil in oil phase on the properties of ILM preparation. Mathematic relation between indicator and factor was constructed by binominal fitting. Then the response surface method according to the best mathematical model of evaluation indicators was used to make predictive analysis of the best prescription. Optimized prescription was used to prepare ILM and evaluate its quality. Results Optimized prescription was egg yolk lecithin 1.50 g, F68 0.35 g, and MCT 10 g. All items of optimized prescription were similar to target values. According to the optimized prescription, mean particle size of ILM was (165.00 ± 0.22) nm, particle size distribution was 0.046 ± 0.070, Zeta potential was (−30.30 ± 0.13) mV, encapsulation efficiency was about 90.09%, drug-loading rate was 1.0 mg/mL. Conclusion Due to the better predictability of constructed mathematical model, CCD-RSM can be applied to optimize prescription of lipid microspheres and the optimized ILM meet pharmacy requirements.

Optimization of purification process of saponins in steamed Panax notoginseng by failure mode and effects analysis and central composite design-response surface methodology / 中草药

Objective: To optimize the purification technology of saponins in steamed Panax notoginseng with macroporous resin. Methods: The main factors affecting the purification process were screened by failure mode and effects analysis (FMEA). The purification method with macroporous resin was optimized by central combination design-response surface method (CCD-RSM) based on the recovery and purity of saponins. In this experiment, the concentration of sample solution, loading volume, washing volume, ethanol concentration, and ethanol elution volume were used to investigate the purification of saponins in steamed P. notoginseng. Results: The optimized purification process with macroporous resin was as follows: maximum recovery (82.81%) and purity (77.24%) of saponins were obtained with the concentration of saponin solution of 11.22 mg/mL, loading volume of 4.97 BV, washing volume of 2 BV, ethanol concentration of 70%, and ethanol elution volume of 3.31 BV. Conclusion: The optimized purification process based on FMEA and CCD-RSM is convenient and stable, with high recovery and purity of saponins, which has a certain practical value.

Optimization of polyhydroxyalkanoate (PHA) production by Burkholderia cepacia BPT1213 utilizing waste glycerol as the sole carbon source

@#Aims:This study was carried out to optimize the fermentation conditions using statistical approach for polyhydroxyalkanoate(PHA) production by a local isolate, Burkholderia cepaciaBPT1213, in the shake flask system.Methodology and results:Throughout this study, B. cepaciaBPT1213 was grown in minimal salt medium (MSM) supplemented with 2% of waste glycerol (86.70% purity).The strain can produce up to 1.33 g/L cell dry weight (CDW) with 22.21% of PHA content, thus giving a total PHA concentration 0.30 g/L before optimization. A factorial design experiment that was carried out showed all parameters KH2PO4, Na2HPO4·2H2O, carbon-to-nitrogen ratio (C/N), initial pH of medium, and temperature significantly affected the growth (cell dry weight, CDW) and PHA content. Response surface methodology (RSM) using central composite design (CCD) was then applied to optimize these parameters. The optimum conditions suggested were at 2.5 g/L KH2PO4, 4.5 g/L Na2HPO4·2H2O, 30 (g/g) C/N ratio, initial medium pH of 8.5 and 37 °C cultivation temperature, with a predicted CDW of 3.43 g/L and PHA content of 45.71% contributing to 1.57 g/L total PHA concentration. The verification experiment resulted in 3.60 g/L of CDW with 48.08% of PHA content contributing to 1.73 g/L total PHA concentration.Conclusion, significance and impact of study:The statistical approach using factorial design and RSM have succeeded in increasing the production of PHA by B. cepaciaBPT1213 using waste glycerol as the sole carbon source which is a promising renewable and cheaper feedsto

Optimization of an Activity Assay of Coprinus Cinereus Peroxidase

ABSTRACT To seek a simple, rapid and sensitive Coprinus cinereus Peroxidase (CIP) activity assay, a convenient one-factor-at-a-time (OFAT) method and a response surface methodology (RSM) were used. The recombinant CIP expressed in Pichia pastoris was purified with the Ni-NTA spin column. Based on the results of catalytic efficiency (kcat/Km) analysis, 2,2'-azinobis (ethylbenzthiazoline -6-sulfonate) (ABTS) was selected as the optimal enzyme substrate. Results of the OFAT method showed that enzymatic reaction performed in 0.1 mol/L sodium acetate (pH 5.0) buffer in a 200-µl reaction mixture containing 0.5 mmol/L ABTS, 10 mmol/L hydrogen peroxide (H2O2), 49.7 ng CIP at 25°C gave an average CIP activity of 88 U/mL. The ABTS and H2O2 concentrations were then further optimized to improve the sensitivity of the assay. To do that, RSM was conducted through central composite design, and a reduced quadratic model with good fit regression equation was generated. ANOVA analysis of this model indicated that the concentrations of ABTS and H2O2 and their interaction had significant impact on the assay sensitivity. The optimal reaction mixture was determined to include an initial ABTS concentration of 0.82 mmol/L 49.7 ng CIP and 16.36 mmol/L H2O2, and the activity under this condition was determined to be 138.89 U/mL.

Design of Experiments (DoE) applied to Pharmaceutical and Analytical Quality by Design (QbD)

According to Quality by Design (QbD) concept, quality should be built into product/method during pharmaceutical/analytical development. Usually, there are many input factors that may affect quality of product and methods. Recently, Design of Experiments (DoE) have been widely used to understand the effects of multidimensional and interactions of input factors on the output responses of pharmaceutical products and analytical methods. This paper provides theoretical and practical considerations for implementation of Design of Experiments (DoE) in pharmaceutical and/or analytical Quality by Design (QbD). This review illustrates the principles and applications of the most common screening designs, such as two-level full factorial, fractionate factorial, and Plackett-Burman designs; and optimization designs, such as three-level full factorial, central composite designs (CCD), and Box-Behnken designs. In addition, the main aspects related to multiple regression model adjustment were discussed, including the analysis of variance (ANOVA), regression significance, residuals analysis, determination coefficients (R2, R2-adj, and R2-pred), and lack-of-fit of regression model. Therefore, DoE was presented in detail since it is the main component of pharmaceutical and analytical QbD.

Microwave-assisted extraction in dry fruit of andean species Vaccinium meridionale: Experimental conditions on the recovery of total polyphenols / Extração assistida por microondas para frutos da espécie andina Vaccinium meridionale: Condições experimentais na remoção de polifenóis totais

ABSTRACT Due to their nutraceutical properties, the fruits of the species Vaccinium meridionale, which inhabit the Andean region, are of scientific interest. Microwave-assisted extraction has been applied to different vegetal matrices for to extract efficiently polyphenolic compounds. In this work we study in microwave assisted extraction processes, the effect on the extraction of total polyphenols in dried fruits of Vaccinium meridionale, using response surface methodology. The main objective of this research was to analyze the relationships between power, temperature, solid-liquid ratio, time and ethanol concentration in aqueous media on the effectiveness of total polyphenols recovery in dry fruits of Vaccinium meridionale. A central composite design face-centered with three levels for each variable was used: P = 300 - 900 W; T = 70 -110 °C; L:Srat. = 30:1 - 70:1 w/w; t = 5 - 15 min; [EtOH] = 0 - 80%. In this sense, the effect of the input variables on the recovery of total polyphenols, the optimization process for maximum extraction, and the comparison to other solid-liquid extractions in terms of quantity of total polyphenols, are reported. It was found that the significant variables in the extraction process were ethanol concentration, temperature and time; the best yields were obtained in the range [EtOH] = 44 - 49%, T = 110 °C, and t = 10 - 15 min. In terms of quantity, time and consumption of energy, microwave-assisted extraction technique is more efficient than other solid-liquid extraction processes for the extraction of total polyphenols.

RESUMO Devido a suas propriedades nutracêuticas em termos do conteúdo de polifenóis totais, os frutos da espécie andina Vaccinium meridionale tem associado um grande interesse cientifico. A extração assistida por microondas tem sido utilizada em diferentes matrizes vegetais para a remoção de forma eficiente de compostos polifenólicos que apresentam uma atividade biológica. Neste artigo são apresentados os principais resultados experimentais obtidos em diferentes processos de extração assistida por microondas, com respeito à quantia de polifenóis totais removidos em frutos secos de Vaccinium meridionale por meio da metodologia da superfície de resposta. O principal objetivo desta pesquisa foi analisar as relações entre as variáveis independentes da potência, a temperatura, a relação sólido-líquido, tempo e concentração de etanol com respeito à quantidade polifenóis totais extraídos de frutos secos de Vaccinium meridionale. Foi utilizado um desenho composto centrado nas caras com três diferentes níveis para cada variável: P = 300 - 900 W; T = 70 -110 °C; L:Srat. = 30:1 - 70:1 w/w; t = 5 - 15 min; [EtOH] = 0 - 80%. Foi utilizado o método de Folin-Ciocalteu para a quantificação de polifenóis totais com ácido gálico como molécula de referência. Neste sentido, foi reportado neste trabalho o efeito dos fatores de entrada com respeito a quantidade de polifenóis totais removidos, o processo de optimização para obter a máxima remoção possível, e também uma comparação dos resultados experimentais com outros processos de extração sólido-líquido. Foi achado que a concentração de etanol, a temperatura e o tempo foram a variáveis mais importantes no processo de extração. A máxima quantidade de recuperação achada neste trabalho foi obtida no intervalo [EtOH] = 44 - 49%, T = 110 °C e t = 10 - 15 min. Em termos de quantidade, tempo e gasto de energia, a extração assistida por microondas é uma técnica eficiente para a remoção de polifenóis totais em frutos secos de Vaccinium meridionale.

Optimizing culture conditions for production of intra and extracellular inulinase and invertase from Aspergillus niger ATCC 20611 by response surface methodology (RSM)

Abstract The aim of this study was obtain a model that maximizes growth and production of inulinase and invertase by Aspergillus niger ATCC 20611, employing response surface methodology (RSM). The RSM with a five-variable and three-level central composite design (CCD) was employed to optimize the medium composition. Results showed that the experimental data could be appropriately fitted into a second-order polynomial model with a coefficient of determination (R2) more than 0.90 for all responses. This model adequately explained the data variation and represented the actual relationships between the parameters and responses. The pH and temperature value of the cultivation medium were the most significant variables and the effects of inoculum size and agitation speed were slightly lower. The intra-extracellular inulinase, invertase production and biomass content increased 10-32 fold in the optimized medium condition (pH 6.5, temperature 30 °C, 6% (v/v), inoculum size and 150 rpm agitation speed) by RSM compared with medium optimized through the one-factor-at-a-time method. The process development and intensification for simultaneous production of intra-extracellular inulinase (exo and endo inulinase) and invertase from A. niger could be used for industrial applications.

Optimization of processing technology of Hemsleya omeiensis by orthogonal design and central-composite design-response surface methodology / 中草药

Objective: To optimize the processing technology of Hemsleya omeiensis processed by licorice juice. Methods: HPLC was employed to determine the content of hemslecin A.Taking content of hemslecin A as index, the orthogonal test was adopted to optimize the covered moistening time, drying-time, and processing temperature. And the central composite design-response surface methodology (CCD-RSM) was adapted to optimize moistening time and drying-time further. Results: The optimum processing technology of H. omeiensis by the orthogonal test was covered moistening time of 7 h, drying-time of 12 h, and processing temperature at 80℃. The optimum processing technology by CCD-RSM was covered moistening time of 7.48-8.56 h, drying-time of 12.06-13.12 h, and processing temperature at 80℃. Conclusion: The experimental design method is precise and the data are reliable with the model. It is the first time that H. omeiensis is processed with licorice juice. Besides, it establishes the processing technology of H. omeiensis and provides a theoretical basis for the processing technology of H. omeiensis with licorice juice.

Development and validation of stability indicating liquid chromatographic (RP-HPLC) method for estimation of ubidecarenone in bulk drug and formulations using quality by design (QBD) approach

ABSTRACT A novel, accurate, precise and economical stability indicating Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) method, was developed and validated for the quantitative determination of ubidecarenone (UDC) in bulk drug, UDC marketed formulation and UDC loaded cubosomes (CBMs) nanocarriers through Response surface methodology (RSM) design with three factors and three levels was performed to optimize the chromatographic variables followed by forced degradation studies of UDC were performed to detect degradation peak. RP-HPLC separation was achieved using mobile phase consisting of Acetonitrile:Tetrahydrofuran:Deionised water in the ratio 55:42:3 and a flow rate of 1.0 mL/min was optimized with a standard retention time (Rt) of 2.15 min, through experiment. The method was found linear in the concentration range of 5-100 µg/mL with a regression coefficient of 0.999. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 3.04 µg/mL and 9.11 µg/mL, respectively.

Intranasal mucoadhesivemicroemulsion for neuroprotective effect of curcuminin mptp induced Parkinson model

ABSTRACT This study was to investigate the neuroprotective effect of curcumin against inflammation-mediated dopaminergic neurodegeneration in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mice model of Parkinson's disease (PD). Curcumin loaded sodium hyaluronate based mucoadhesive microemulsion (CMME) was developed by using Box Behnken design of Response surface method (RSM) and was characterized. Male C57BL/6 mice were first treated with four intraperitoneal injections of MPTP (20 mg/kg of body weight) at 2 h intervals followed CMME intranasal administration for 14 days at 2.86 mg of curcumin/kg of body weight per once a day. Optimal CMME containing 3% Capmul MCM as oil phase, 37 % of Accenon CC and Transcutol HP at 2.5:1 ratio and 0.5% sodium hyaluronate was stable, non-ciliotoxic with 57.66 nm±3.46 as average globule size. PdI value (0.190 ± 0.19) and TEM result depicted the narrow size distribution of CMME.All three independent variables had a significant effect (p<0.05) on the responses and the designed model was significant for all taken responses. In-vivo results revealed significant reduction of MPTP-mediated dopamine depletion after nasal administration of CMME. MPTP intoxication significantly decreased striatal DA content to 21.29 % which was then elevated to 55.37% after intranasal curcumin treatment. Significant improvement in motor performance as well as gross behavioural activity of mice was observed from rota-rod and open field test findings. Findings of the investigation revealed the symptomatic neuroprotection of curcumin against MPTP-induced neurodegradation in the striatum and hence could be considered as a promising approach to treat PD.

Quality Characteristics of Rice Cookies Prepared with Stevia rebaudiana Leaf

The purpose of this study was to determine the optimal recipe of rice cookies with two different amounts of Stevia rebaudiana leaf and grape seed oil, using a central composite design (CCD). In addition, mixing conditions of rice cookies were optimized by sensory evaluation and mechanical and physicochemical analysis using response surface methodology (RSM). RSM was used to obtain 10 experimental points (including two replicates of Stevia rebaudiana leaf and Grape seed oil), and the formulation of Stevia rebaudiana leaf added rice cookies was optimized using rheology. The results of mechanical and physicochemical analysis showed significant values for lightness, redness, yellowness, hardness, spread factor, loss rate, leavening rate, sweetness, moisture, pH, and density (P<0.001), results of the sensory evaluation showed significant values for color, flavor, taste, texture, appearance, and overall quality (P<0.05). As a results, optimal sensory ratio was found to be 1.98 g of Stevia rebaudiana leaf and 37.94 g of Grape seed oil.