Extraction of effective components of Psoralea corylifolia and efficacy evaluation in the treatment of vitiligo / 中国药房

OBJECTIVE To extract the effective components of Psoralea corylifolia and evaluate its efficacy in the treatment of vitiligo. METHODS The concentrations of psoralen， isopsoralen， neobavaisoflavone， corylin， psoralidin， corylifolinin， and bakuchiol in P. corylifolia extract were determined by ultra-performance liquid chromatography. Based on the analytic hierarchy process （AHP） and Plackett-Burman design， with the concentrations of the 7 components as evaluation indexes and the crushing degree， ethanol concentration， and soaking time as factors， the extraction process of P. corylifolia was optimized by Box-Behnken response surface methodology and the validation test was conducted. Zebrafish were divided into blank control group， positive control group （8-methoxypsoralen， 10.8 μg/mL）， and low-， medium-， and high-concentration groups of P. corylifolia extract （500， 1 000， 2 000 μg/mL）， with 6 fish in each group. The effects of P. corylifolia extract on the melanin production of zebrafish were studied by density analysis. RESULTS The best extraction process was P. corylifolia powder over 60 meshes and soaked in 80% ethanol for 72 hours. The average comprehensive score of three validation experiments was 98.27， with an RSD of 1.36%， and the relative error was 1.02% compared with the predicted value of the fitting equation （97.28）. Compared with the blank control group， the melanin pigmentation of zebrafish in the low-， medium-， and high-concentration groups of P. corylifolia extract was significantly increased （P＜0.01）. CONCLUSIONS The optimized extraction process of P. corylifolia is reasonable and feasible， and the obtained P. corylifolia extract can significantly promote the production of melanin in zebrafish.

Optimization of the medium and fermentation condition for the Penicillium aurantiocandidum Z12 strain with molluscicidal actions against Oncomelania hupensis / 中国血吸虫病防治杂志

Objective To optimize the culture and fermentation conditions of the Penicillium aurantiocandidum Z12 strain, a fungal strain with molluscicidal actions against Oncomelania hupensis, so as to provide the basis for the research and development of molluscicidal active substances from the P. aurantiocandidum Z12 strain and its fermentation broth and large-scale fermentation. Methods The carbon source, nitrogen source and mineral salts were identified in the optimal culture medium for the P. aurantiocandidum Z12 strain with a single-factor experiment to determine the best fermentation condition for the P. aurantiocandidum Z12 strain. Factors that significantly affected the growth of the P. aurantiocandidum Z12 strain were identified using the Plackett-Burman design, and the best range of each factor was determined using the steepest climb test. Response surface analyses of temperature, pH value, seeding amount and liquid-filling quantity were performed using the Box-Behnken design to create a regression model for fermentation of the P. aurantiocandidum Z12 strain to identify the optimal culture medium. Results Single-factor experiment preliminarily identified the best culture medium and conditions for the P. aurantiocandidum Z12 strain as follows: sucrose as the carbon source at approximately 20 g/L, tryptone as the nitrogen source at approximately 5 g/L, K2HPO4 as the mineral salt at approximately 5 g/L, initial pH at approximately 8, temperature at approximately 28 °C, seeding amount at approximately 6%, and liquid-filling quantity at approximately 50 mL/100 mL. Plackett-Burman design showed that factors that significantly affected the growth of the P. aurantiocandidum Z12 strain included temperature (t = −5.28, P < 0.05), seeding amount (t = 5.22, P < 0.05), pH (t = −4.30, P < 0.05) and liquid-filling quantity (t = −4.39, P < 0.05). Steepest climb test showed the highest mycelial growth at pH of 7.5, seeding amount of 8%, and liquid-filling quantity of 40 mL/100 mL, and this condition was selected as the central point of response surface analysis for the subsequent optimization of fermentation conditions. Response surface analyses using the Box-Behnken design showed that the optimal conditions for fermentation of the P. aurantiocandidum Z12 strain included sucrose at 15 g/L, tryptone at 5 g/L, K2HPO4 at 5 g/L, temperature at 28.2 °C, pH at 7.5, seeding amount at 10%, and liquid-filling quantity at 35.8 mL/100.0 mL, resulting in 0.132 g yield of the P. aurantiocandidum Z12 strain. Conclusion The optimal culture condition for the P. aurantiocandidum Z12 strain has been identified, and the optimized culture medium and fermentation condition may effectively improve the fermentation yield of the P. aurantiocandidum Z12 strain.

Optimization of extraction process of polysaccharide from Baihe dihuang decoction and study on its anti-anxiety and anti-depression efficacy / 中国药房

OBJECTIVE To optimize the extraction process and to primarily evaluate the anti-anxiety and anti-depression efficacy of polysaccharide from Baihe dihuang decoction. METHODS Based on Plackett-Burman experimental design， using the comprehensive score of yield and content of polysaccharide as indicators， with extraction time， water amount， alcohol precipitation concentration as factors， Box-Behnken response surface methodology was used to optimize the extraction process of polysaccharide from Baihe dihuang decoction； and the validation test was conducted. Forty ICR mice were divided into control group， venlafaxine group [positive control， 13.5 mg/（kg·d）]， Baihe dihuang polysaccharide high-dose and low-dose groups [5.28， 2.64 g/（kg·d），by raw material]， with 10 mice in each group （half male and half female）. Administration groups were given corresponding drug solution intragastrically， and control group was given water 10 mL/kg intragastrically， once a day， for 7 test， forced swimming test and tail suspension test were used to evaluate the effects of the extract prepared by the optimal process on the anxiety-like and depression-like behavior of mice； enzyme-linked immunosorbent assay was used to detect the effects of the extract on the levels of neurotransmitter in cerebral tissue of mice. RESULTS The optimal extraction process of Baihe dihuang decoction was： the water amount of 25 times， extract time of 1.5 hours， and alcohol precipitation concentration of 70%. In 3 times of validation test， the average yield and content of polysaccharide were 33.10% and 0.62 mg/mg， the relative deviations of which from the predicted values （36.14% and 0.65 mg/mg） were 8.40% and 4.62% respectively （RSD＜2%， n＝3）. The polysaccharide extract of Baihe dihuang decoction could effectively increase the percentages of open-arms entry， the percentages of open-arms time， the total distance of voluntary activities and the activity distance in central area， and significantly shortened the immobility time of forced swimming test and tail suspension test （P＜0.05 or P＜0.01）. The polysaccharide extract could significantly increase the levels of 5-hydroxytryptamine， norepinephrine （except for the Baihe dihuang polysaccharide low-dose group） and gamma-aminobutyric acid in cerebral tissue of mice， while significantly decrease the levels of glutamic acid （except for the Baihe dihuang polysaccharide low-dose group） （P＜0.05 or P＜ 0.01）. CONCLUSIONS The optimized extraction process of polysaccharide from Baihe dihuang decoction is stable and feasible， and the obtained polysaccharide extract has obvious anti-anxiety and anti-depression effect in vivo.

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

A method for measuring the ADCP potency of anti-CD38 monoclonal antibody / 药学学报

A method to measure the antibody-dependent cell-mediated phagocytosis (ADCP) potency of anti-CD38 mAb was developed based on design of experiment (DoE) with a Jurkat/NFAT/CD32a-FcεRIγ transgenic cell line as the effector cell, the Daudi cell line as the target cells, and luciferase as the detection system. The DoE method was used for optimization of experimental parameters and methodological validation. The results show that anti-CD38 mAb exhibits a dose-response relationship with the following four-parameter equation: y = (A - D) / [1 + (x / C)B] + D. Several experimental parameters were optimized by statistical experimental design and determined as follows: the working concentration of anti-CD38 mAb was 800-20.81 ng·mL-1, the density of the target cells was 7.5×104 per well, and the density of effector cells was 2.5×104 per well, with an induction time of 6 h. The method showed good specificity. The recovery rate for samples from 5 different groups showed that the relative potencies of anti-CD38 mAb were (59.97 ± 4.74) %, (82.44 ± 5.15) %, (110.69 ± 11.71) %, (129.23 ± 5.22)% and (162.15 ± 3.66) %. The recoveries ranged from 103% to 120% and the RSDs of the above results were all less than 11%. The linear detection range was 50%-150%. Based on DoE design, this method for measuring ADCP potency of anti-CD38 mAb was optimized and validated with good specificity, repeatability and accuracy. This method can be used for evaluation of ADCP biological activity of anti-CD38 mAbs.

Identification of critical influencing factors in dropping process of Xuesaitong Dropping Pills / 中国中药杂志

With the dropping process of Xuesaitong Dropping Pills(XDP) as the study object, critical factors affecting the quality indicators of pill pass rate, average weight of drop pills and roundness were screened out, so as to deepen the understanding of the dropping process. The critical process units, critical quality attributes and potential critical process influencing factors of XDP were determined by risk analysis and prior knowledge, and then the critical influencing factors were screened out by Plackett-Burman design. First, according to the risk assessment, the critical technique of XDP preparation process was dropping, and then the critical quality attributes of dropping process were pill pass rate, average weight of drop pills and roundness. Then, according to fishbone diagram and failure mode and effects analysis(FMEA), potential critical influencing factors were determined as flow rate, matrix ratio, solid-liquid ratio, feed-liquid temperature, top temperature of condensate, bottom temperature of condensate and dropping distance. Finally, among these seven potential factors, the critical influencing factors were determined as material liquid ratio, dropping distance, top temperature of condensate, bottom temperature of condensate. This study revealed the potential of Plackett-Burman design in screening and understanding the influence of selected factors on XDP dropping process, which could provide a reference for studying the dropping process.

To Study The Significance Of Processing Variables Using Quality By Design For Optimization Of Nanoparticulate System Of Cilnidipine

This investigation aimed to prepare Cilnidipine Nanoparticles by nanoprecipitation ultrasonication method and to study the significance of processing variables by applying quality by design. Cilnidipine is fourth-generation dual L/N-type Ca2+ channel blocker used for the management of hypertension. It is BCS class-II drug exhibiting lower aqueous solubility, which tends to lower bioavailability. The combination of Poloxamer 188 and Tween 80 was used as a stabilizer. The design of the experiment is one of the tools of Quality by design. Plackett -Burman design was applied for the screening of processing variables, which are significant for the method. The processing variables screened were stirring speed, antisolvent ratio, drug concentration, polymer concentration, stabilizer concentration. The effect of each parameter evaluated by particle size, entrapment efficiency, and drug release at 10 minutes of prepared Nanoparticles of Cilnidipine. Analysis of variance and Pareto-plot of Plackett-Burman design were utilized to find the significance of the factor and extent of the effect. The surface morphology of Cilnidipine Nanoparticles was studied by SEM. The Pareto plot, as well as statistical analysis of design, had shown that the Concentration of drug, solvent: antisolvent ratio and concentration of poloxamer 188 were the significant parameters for the method. The stabilizer concentration, the stirring speed, and the antisolvent ratio had a negative effect of while the concentration of drug has a positive effect on the particle size of Nanoparticles and drug release at 10 minutes and positive effect of entrapment efficiency of Cilnidipine Nanoparticles. The Cilnidipine Nanoparticles were characterized by FTIR and DSC analysis.

Aislamiento y caracterización de una bacteria marina degradadora de testosterona: Vibrio sp. N3 / Isolation and characterization of a marine testosterone-degrading bacterium: Vibrio sp. N3

Steroids, including testosterone, estrone, 17β-estradiol, estriol and 17β-ethinyl estradiol, are harmful not only to the population dynamics of aquatic life forms but also to public health. In this study, a marine testosterone-degrading bacterium (strain N3) was isolated from Nanao Island in the South China Sea. In addition, the strain could also use 17β-estradiol (E2), 17β-ethinyl estradiol (EE2), estriol (E3) or cholesterol as a sole carbon source. According to the 16S rRNA gene sequence analysis, strain N3 was identified as Vibrio sp. Further characterization showed that the strain is aerobic, gram-negative, and mobile and exhibits resistance to ampicillin, carbenicillin, penicillin and spectinomycin. For enhancing its capacity of testosterone degradation, the Plackett-Burman factorial design and the central composite design were used to optimize the culture condition. Under optimal conditions, 92% of testosterone was degraded by Vibrio sp. N3 in 48 h.

Los esferoides-que incluyen la testosterona, la estrona, el 17 β-estradiol, el estriol y el 17 p-etinilestradiol-son nocivos no solo para la población dinámica de las formas de vida acuática, sino también para la salud pública. En este estudio se aisló una bacteria marina degradadora de testosterona de la isla de Nanao, en el Mar del Sur de China, a la que se denominó cepa N3. Se determinó que esta cepa también podría usar 17 β-estradiol (E2), 17 p-etinilestradiol (EE2), estriol (E3) o colesterol como únicas fuentes de carbono. De acuerdo con el análisis de la secuencia del gen 16S rRNA, la cepa N3 se identificó como Vibrio sp. La caracterización adicional mostró que dicha bacteria es un organismo aerobio, gram negativo y móvil, y que presenta resistencia a ampicilina, carbenicilina, penicilina y espectinomicina. Para optimizar la condición de cultivo en relación con su capacidad de degradar la testosterona, se utilizaron el diseño factorial Plackett-Burman y el diseno compuesto central. En condiciones óptimas, el 92% de la testosterona fue degradada por Vibrio sp. N3 en 48 h.

Colombian propolis as starting material for the preparation of nanostructured lipid carriers

ABSTRACT The value of propolis is scientifically and commercially measured through the content of biologically active molecules as phenolic compounds and flavonoids; on the other hand, a high percentage of waxes in the propolis composition makes it a substandard beekeeping product. Colombian propolis is characterized by a high content of waxes; however, this drawback turns into an advantage when this material is used for preparing lipid nanocarriers. Accordingly, in this research work, a propolis-extracted material obtained by Randall method is characterized by differential scanning calorimetry, infrared spectroscopy, X-ray diffraction, and 1H-Nuclear Magnetic Resonance. Then, it is used for obtaining nanostructured lipid carriers by the emulsification-diffusion technique, whose recipe and operating work conditions were established by a Plackett-Burman statistical screening design. The obtained particles exhibit sizes less than 300 nm, polydispersity indices around 0.1, zeta potential values less than ±2 mV, good physical stability and they show to be safe in the in vitro irritation test. Thus, Colombian propolis arises as an attractive natural source for obtaining lipid carriers that could be used in pharmaceutical or cosmetic industries for developing innovative products.

Biosurfactant from endophytic Bacillus cereus: Optimization, characterization and cytotoxicity study

Aims@#The increased importance of biosurfactant in the recent past is mainly due to their applications in various industries ranging from petroleum to pharmaceuticals. Their biodegradability and environmental compatibility with low toxicity makes it even more interesting. Microbial production of biosurfactant is found to be a viable option as they are diverse, eco-friendly, facilitate large scale production, able to perform under extreme conditions etc. One class of microbes that is endophytes are known to show great potential in producing different varieties of medically and industrially significant biological compounds. The present study focuses on the screening and production of biosurfactant from endophytic bacteria. @*Methodology and results@#Of all the isolates tested, one endophyte identified as Bacillus cereus HM998898 was found to produce maximum biosurfactant. Statistical method Plackett burman was used to optimize the media for the maximum production and the ideal composition was found to be KNO3 (1 g/L), Gingley oil (2 mL), K2HPO4 (2.5 g/L), KH2PO4 (0.75 g/L), MgSO4·5H2O (0.5 g/L), FeSO4.7H2O (0.005 g/L) and NaCl (0.025 g/L). The extracted biosurfactant was characterized and was identified to be glycolipid. This was further tested for biocompatibility against Fibroblast (3T3) cells and was evaluated for their anti tumor activity against Hep2 cells. @*Conclusion, significance and impact of study@#The biosurfactant produced was found to induce toxicity to cancer cells at appreciable levels while they remained non-toxic to normal cells supporting the possible applications of biosurfactant in medical field.

Optimal extraction of indirubin from Isatidis Folium based on Plackett-Burman design combined with central composite design-response surface methodology / 中草药

Objective: To investigate the significance of each influencing factor and optimize the process of extracting indirubin from Isatidis Folium by Plackett-Burman design combined with central composite design-response surface methodology (CCD-RSM). Methods: Plackett-Burman experimental design was used to screen the main influencing factors, and CCD-RSM was used to optimize the extraction process of indirubin. With the concentration of ethanol, the ratio of material to liquid, and the extraction time as independent variables and the extraction amount of indirubin as dependent variable, the optimum extraction process of indirubin from Isatidis Folium was predicted and analyzed by multiple linear regression and binomial fitting models with independent and dependent variables and the three-dimensional surface graph. Results; The optimal extraction process of indirubin was as follows: ethanol concentration 62%, solvent/sample ratio of 26, and extraction time 9 min. Under these conditions, the maximal extraction rate of indirubin was 4.37 mg/g which was consistent with model predictions. Conclusion: The optimal process is simple and convenient for extracting indirubin from Isatidis Folium with high precision, reproducibility, and predictability.

Optimization of Extraction Process for Active Ingredients in Seeds of Sophora alopecuroides by Plackett-Burman Design and Response Surface Analysis / 中国药学杂志

OBJECTIVE: To optimize extraction process for active ingredients in seeds of Sophora alopecuroides, to provide a reference for scale production. METHODS: Active ingredients from Sophora alopecuroides were extracted by ethanol, with average yield of oxysophocarpine and oxymatrine as index, some factors affecting index were firstly evaluated by Plackett-Burman design, then taking oxysophocarpine and oxymatrine as indexes respectively, extraction conditions were optimized by Box-Behnken design, experimental data was fitted by multiple linear regression and binomial formula fitting, extraction process was optimized by response surface method, and prediction was carried out through comparing the observed and predicted value. RESULTS: Extracting times, crushing degree and solvent times had significant effects on yields of oxysophocarpine and oxymatrine; binomial equation fitted well with good predictability. optimum extraction technology of Sophora alopecuroides was as following:crushed through 65 mesh sieve, extracted 4 times with 12-fold the amount of 60% ethanol for 2 h each time; yield of oxysophocarpine and oxymatrine was 92.3%, 78.6% respectively, both deviations were small by comparing with the predicted value. CONCLUSION: This extraction process is reasonable and feasible by Plackett-Burman design and response surface analysis with good predictability. This study can provide experimental basis for further scale production of Sophora alopecuroides.

Study on the Extraction Process of Herbal Pair Puerarin-Berberine by Plackett-Burman Design and Box-Behnken Response Surface Methodology / 中国药师

Objective:To optimize the alcohol extraction process of herbal pair puerarin-berberine.Methods:Based on the etha-nol reflux extraction,the extraction quantity of total flavonoids,total alkaloid,puerarin and berberine hydrochloride were used as the e-valuation index,and the independent variables included the drug particle,ratio of solid to liquid, ethanol concentration, reflux dura-tion and reflux times. Significance analysis was evaluated by Plackett-Burman design,and then the extraction process was optimized by Box-Behnken response surface methodology.Results:The optimal extraction conditions of drug pair puerarin-berberine were as follows:the drug particle was 80 mesh,the ratio of solid to liquid was 1:13,the ethanol concentration was 75%,the reflux time was 60 min, and the reflux times was 4. Under the above conditions, the extraction quantity of total flavonoids, alkaloid, puerarin and berberine hydrochloride was 120.34,56.99,109.63 and 39.26 mg ·ml-1, respectively.Conclusion: The extraction process of herbal pair puerarin-berberine is reasonable and feasible optimized by Plackett-Burman design and Box-Behnken response surface methodology.

Technology Optimization of Hydrolyzing Aralia Saponins from Alcohol Extracts of Aralia elata / 中国中医药信息杂志

Objective To optimize process of hydrolyzed oleanolic acid from Aralia elata medicinal alcohol extract. Methods Plackett-Burman combined with CCD response surface design method (method 1) and orthogonal experimental design method (method 2) were used to optimize process of hydrolyzed oleanolic acid from Aralia elata medicinal alcohol extract. In method 1, the content of oleanolic acid was the dependent variable, and the hydrolysis time, the hydrochloric acid concentration, and the ratio of material to liquid were set as the independent variables; In method 2, the content of oleanolic acid was the dependent variable, and the hydrolysis time, hydrolysis temperature, the hydrochloric acid concentration, and the ratio of material to liquid were set as the independent variables. Design expert 8.0.6 Trial software was used to analyze the data. Results Considering the actual production situation, method 1 determined the optimum process was to add 15–18 times the amount of 10% to 12% hydrochloric acid, reflux hydrolysis 45–70 min; method 2 determined the optimum process was 15 times the amount of 10% hydrochloric acid, reflux hydrolysis 60 min. Conclusion By comparing method 1 and method 2, the optimum process was selected, and the difference of the content of oleanolic acid is not great. However, the former is more intuitive and convenient, with high precision repeatability, and predictability.

Response Surface Methodology (RSM) for analysing culture conditions of Acidocella facilis strain USBA-GBX-505 and Partial Purification and Biochemical Characterization of Lipase 505 LIP / Metodología de Superficie de Respuesta (RSM) para el análisis de las condiciones de cultivo de Acidocella facilis cepa USBA-GBX-505 y la purificación parcial y caracterización bioquímica de la lipasa 505 LIP / Metodologia de Superfície de Resposta (MSR) para a análise das condições de cultivo de Acidocella facilis cepa USBA-GBX-505 e purificação parcial e caracterização bioquímica da lip ase 505 LIP

Abstract Using Response Surface Methodology (RSM) we evaluated the culture conditions (nitrogen source, carbon source, pH and agitation rate) that increase the biomass of Acidocellafaalis strain USBA-GBX-505 and therefore enhance the production of its lipolytic enzyme, 505 LIP. RSM results revealed that yeast extract and agitation were key culture factors that increased the growth-associated lipolytic activity by 4.5-fold (from 0.13 U.mg-1 to 0.6 U.mg-1). The 505 LIP lipase was partially purified using size-exclusion chromatography and ion-exchange chromatography. Its molecular weight was >77 kDa. The enzyme shows its optimum catalytic activity at 55 °C and pH 7.5. EDTA, PMSF, 1-butanol and DMSO inhibited enzymatic activity, whereas Tween 20, acetone, glycerol and methanol increased it. Metallic ions are not required for the activity of 505 LIP, and even have an inhibitory effect on the enzyme. This study shows the potential use of A. facilis strain USBA- GBX-505 for the production of a newly identified lipolytic enzyme, 505 LIP, which is stable at moderate temperatures and in the presence of organic solvents. These are important characteristics for the synthesis of many useful products.

Resumen Por medio de la Metodología de Respuesta de Superficie (RSM) evaluamos las condiciones de cultivo (fuente de N, fuente de C, pH y tasa de agitación) que incrementan la biomasa de Acidocella facilis cepa USBA-GBX-505 y, como consecuencia, la producción de su enzima lipolítica, llamada 505 LIP. Los resultados de la RSM revelaron que el extracto de levadura y la agitación fueron factores de cultivo claves, que incrementaron de 4 a 5 veces la actividad lipolítica asociada al crecimiento (de 0.13 U.mg-1 a 0.6 U.mg-1). La lipasa 505 LIP se purificó parcialmente usando cromatografía de exclusión por tamaño y cromatografía de intercambio iónico. Su peso molecular fue > 77 kDa. La enzima muestra su actividad catalítica óptima a 55 °C y pH 7.5. El EDTA, el PMSF, el 1-butanol y el DMSO inhibieron la actividad enzimática, mientras que el Tween 20, la acetona, el glicerol y el metanol la incrementaron. La enzima 505 LIP no requiere iones metálicos para su actividad, e incluso se inhibe en presencia de ellos. Este estudio muestra el uso potencial de A. facilis cepa USBA-GBX-505 para la producción de una nueva enzima lipolítica, 505 LIP, que es estable a temperaturas moderadas y en la presencia de solventes orgánicos. Estas son características importantes en la síntesis de muchos productos útiles.

Resumo Utilizando a Metodologia de Superfície de Resposta (MSR) avaliamos as condições de cultivo (fontes de nitrogénio e carbono, pH e taxa de agitação) que aumentam a biomassa de Acidocella facilis cepa USBA-GBX-505, e, portanto, elevam a produção de sua enzima lipolítica 505 LIP. Os resultados da MSR revelaram que o extrato de levedura e a agitação foram fatores de cultivo chave que permitiram aumentar 4 a 5 vezes a atividade lipolítica associada ao crescimento (de 0,13 U.mg-1 a 0,6 U.mg-1). A lipase 505 LIP foi parcialmente purificada utilizando cromatografia por exclusão de tamanho e cromatografia de intercambio iónico. Seu peso molecular foi > 77 kDa. A enzima mostra sua atividade catalítica ótima a 55 °C e pH 7,5. EDTA, PMSF, 1-butanol e DMSO inibiram a atividade enzimática, enquanto que Tween 20, acetona, glicerol e metanol aumentaram esta atividade. Íons metálicos não são necessários para a atividade da 505 LIP, apresentando inclusive efeito inibitório da enzima. Este estudo demonstra o potencial uso de A. facilis cepa USBA-GBX-505 para a produção de uma nova enzima lipolítica, 505 LIP, a qual é estável a moderadas temperaturas e na presença de solventes orgánicos. Estas características são importantes para a síntese de diversos produtos úteis.

Development method of fingerprint of Compound Danshen Dripping Pills based on analytical quality by design (AQbD) / 中草药

Objective To study the development of analytical quality by design (AQbD) method for the identification of Compound Danshen Dripping Pills (CDDP) by UPLC-UV. Methods The number of peaks and the target peak content (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1) were taken as the analytical target profile (ATP), Plackett-Burman design (PBD) method was used to select critical method parameters (CMPs) and critical method attributes (CMAs) on the basis of risk assessment, and then the response model between CMAs and CMPs was established by Box-Behnken design (BBD). The optimal conditions were obtained via multivariable regression analysis and verified. Results From the seven factors of risk assessment, the flow rate (A), sample weight (B), and C18 column weight (C) were selected as CMPs. And CMAs were the number of peaks (N), notoginsenoside R1 (CR1) and ginsenoside Re (CRe). The variation trends of each factor and model were analyzed by response surface analysis. The optimal conditions were as follows: flow rate of 0.28 mL/min, sample weight of 0.30 g, and C18 column weight of 1.10 g. Under the conditions, N was 12, CR1 was 1.676 5 mg/g, and CRe was 0.669 6 mg/g, with differences of 1.34%, 1.33%, and 2.94% respectively from the predictive values. The final method was as follows: chromatographic column was Acquity UPLC BEH C18 chromatographic column (50 mm × 2.1 mm, 1.7 μm), mobile phase was acetonitrile-water gradient elution, detection wavelength was 203 nm, column temperature was 30 ℃, flow rate was 0.28 mL/min, and injection volume was 2 μL. Conclusion The results prove that the method is reliable.

Purification technology of Yinju Jiedu Oral Liquid / 中草药

Objective To multi-objectively optimize the purification process parameters of Yinju Jiedu Oral Liquid (YJOL), mixed weight of AHP-CRITIC, Plackett-Burman design (PBD), and Box-Behnken design (BBD) were adopted on the basis of HPLC fingerprints. Methods HPLC was used to establish HPLC fingerprint of YJOL. Recovery rate of fingerprints of six components (chlorogenic acid, linarin, harpagoside, R,S-epigoitrin, psoralen, and isobalin) and HPLC fingerprint similarity was taken as the index to optimize the type of macroporous resin among 10 types. Weights of the recovery rates of six components and similarity were determined by mixed weight of AHP-CRITIC, in order to obtain the comprehensive index as the evaluation criterion. The significantly influencing factors were firstly evaluated by PBD, and purification conditions were optimized by BBD. Results HPD-400 type resin showed a high selectivity for six components. The optimized purification technology was as follows: the ratio of dia-height was 1:7, pH value was 3.5, sample concentration was 0.18 g/mL, ratio of sample to resin was 0.96 g/g, and 77% ethanol's dosage was 8 BV. Under the conditions, the recovery rates of six components were 78%-98%, and the similarity of fingerprint was higher than 0.99. These components could get balanced recovery. Moreover, the theoretical and actual comprehensive indexes were 94.28% and 93.69%, respectively, with a relative error of 0.59%. Conclusion Based on HPLC fingerprints, mixed weight of AHP-CRITIC, combined with PBD and BBD-RSM used to optimize the purification process for the YJOL in this study is scientific and feasible, this way can improve the purity of the active components and keep the uniformity of main components in the YJOL as well.

Optimization of Extraction Technology of Total Polyphenols and Total Flavonoids in Gardenia jasminoides by Response Surface Method / 中国药房

OBJECTIVE:To optimize the extraction technology of total polyphenol and total flavonoid in Gardenia jasminoi-des. METHODS:Plackett-burman(PB)design was used to select the ethanol volume fraction,liquid-solid ratio,particle size,ex-traction time and extraction temperature to determine the key factor affecting the extraction of total polyphenol and total flavonoids in G. jasminoides. Then central composite design (CCD) was combined with response surface method to optimize the extraction technology,and verification test was conducted. RESULTS:The optimal extraction conditions of total polyphenol were 40% etha-nol,particle size of 0.20 mm,extraction temperature of 60 ℃,liquid-solid ratio of 20,and extraction time of 20 min;the optimal extraction conditions of total flavonoids were 40% ethanol,particle size of 0.20 mm,extraction temperature of 30 ℃,liquid-solid ratio of 20,and extraction time of 20 min. In verification test,the contents of total polyphenol and total flavonoids in G. jasminoi-des were 1.70%(RSD=1.43%,n=3),3.23%(RSD=3.72%,n=3),with relative error of 1.80%,8.75% with predicted val-ues,respectively. CONCLUSIONS:The response surface method based on PB and CCD is simple,reasonable and feasible to opti-mize the extraction technology of total polyphenol and total flavonoids in G. jasminoides. The method can provide reference for its industrial extraction.

Optimization of Extraction Technology of Total Polyphenols and Total Flavonoids in Gardenia jasminoides by Response Surface Method / 中国药房

OBJECTIVE:To optimize the extraction technology of total polyphenol and total flavonoid in Gardenia jasminoi-des. METHODS:Plackett-burman(PB)design was used to select the ethanol volume fraction,liquid-solid ratio,particle size,ex-traction time and extraction temperature to determine the key factor affecting the extraction of total polyphenol and total flavonoids in G. jasminoides. Then central composite design (CCD) was combined with response surface method to optimize the extraction technology,and verification test was conducted. RESULTS:The optimal extraction conditions of total polyphenol were 40% etha-nol,particle size of 0.20 mm,extraction temperature of 60 ℃,liquid-solid ratio of 20,and extraction time of 20 min;the optimal extraction conditions of total flavonoids were 40% ethanol,particle size of 0.20 mm,extraction temperature of 30 ℃,liquid-solid ratio of 20,and extraction time of 20 min. In verification test,the contents of total polyphenol and total flavonoids in G. jasminoi-des were 1.70%(RSD=1.43%,n=3),3.23%(RSD=3.72%,n=3),with relative error of 1.80%,8.75% with predicted val-ues,respectively. CONCLUSIONS:The response surface method based on PB and CCD is simple,reasonable and feasible to opti-mize the extraction technology of total polyphenol and total flavonoids in G. jasminoides. The method can provide reference for its industrial extraction.

Preparation and Pharmaceutical Properties of Solid Lipid Nanoparticles Containing the Total Saponins of Paris Polyphylla / 中国药师

Objective:To design and develop a formula of solid lipid nanoparticles containing the total saponins of Paris Polyphylla using a quality by design ( QbD) method. Methods:The target product profile of solid lipid nanoparticles was determined according to the properties of dosage form and administration. The risk assessment was carried out according to the theoretical knowledge and experi-ence to define the critical variables influencing the properties of solid lipid nanoparticles. Firstly, Plackett-Burman test was used to screen out the key variables significantly affecting the pharmacological properties of solid lipid nanoparticles, and then the Box-Behnken effect surface method was use to further optimize the selected variables. The physicochemical properties of solid lipid nanoparticles con-taining the total saponins of Paris Polyphylla were studied, such as the particle size distribution, polydispersity index ( PdI) , zeta po-tential, morphology and in vitro drug release behavior. Results: The optimum formula and preparation process were as follows: the concentration of glycerol monostearate was 5. 5％, the concentration of soybean phospholipid was 8. 0％, the number of homogenization was 6 times, the concentration of drug was 5. 0％, the surfactant was Tween 80, the mass pressure was 600 bar and the homogeneous temperature was 65℃. The mean particle size, PdI and zeta potential of the optimized solid lipid nanoparticles was (116. 5 ± 32. 1) nm, (0. 198 ± 0. 018) and ( -23. 6. 5 ± 0. 9) mV, respectively. Transmission electron microscopy showed that the solid lipid nanop-articles were spherical. The results of in vitro release showed a sustained release property, and the cumulative release was 63. 5％ in 24 h. Conclusion:It is feasible to design and develop solid lipid nanoparticles containing the total saponins of Paris Polyphylla by using the QbD method, which can ensure the product quality to meet the requirements.