Otimização de método extrativo para raízes de Arctium lappa L / Extractive method optimization for Arctium lappa l. Roots / Optimización de un método de extracción de raíces de Arctium lappa L

Arctium lappa L. é indicada no Formulário de Fitoterápicos da Farmacopeia Brasileira para o tratamento de distúrbios urinários leves. Estudos já demonstraram o potencial antioxidante, anti-inflamatório e antidiabético deste extrato, onde foram identificados fenóis, lignanas, taninos e flavonoides. O objetivo deste trabalho foi otimizar o método extrativo de raízes de A. lappa. Realizou-se o preparo de extratos por diferentes métodos: Ultrassom, Soxhlet, maceração e turbo extração. A otimização foi realizada por turbo extração seguindo um planejamento fatorial 23, empregando como fatores: teor alcoólico, concentração da matéria prima e tempo de extração. Os extratos foram avaliados quanto ao resíduo seco, teores de fenóis e flavonoides, e atividade antioxidante. Com relação ao resíduo seco, e aos teores de fenóis e flavonoides, os métodos de ultrassom e turbo extração demonstraram melhor poder extrativo. Devido ao menor tempo e custo operacional, a otimização foi realizada por turbo extração, e o extrato otimizado foi obtido utilizando álcool 60%, em proporção matéria prima solvente 1:10 e tempo de extração de 15 minutos. Estas análises poderão nortear futuros testes de transposição de método para escala industrial, diminuindo mão de obra, tempo e custos, visando obter produtos fitoterápicos mais eficientes, com valor acessível à população.

Arctium lappa L. is indicated in the Brazilian Pharmacopeia Herbal Medicines Form for the treatment of mild urinary disorders. Studies have already demonstrated the antioxidant, anti-inflammatory and antidiabetic potential of this extract, where phenols, lignans, tannins and flavonoids were identified. The objective of this work was to optimize the extractive method of A. lappa roots. Extracts were prepared by different methods: Ultrasound, Soxhlet, maceration and vortical extraction. The optimization was performed by vortical extraction following a 23 full factorial design, using as factors: alcohol content, drug concentration and extraction time. The extracts were evaluated for dry residue, phenols and flavonoids contents, and antioxidant activity. Regarding the dry residue, and the phenols and flavonoids contents, the ultrasound and vortical extraction methods showed better extractive power. Due to the lower operating time and cost, the optimization was performed by vortical extraction, and the optimized extract was obtained using 60% alcohol, in a 1:10 drug solvent ratio and extraction time of 15 minutes. These assessments guide the future tests of transposition of the method to an industrial scale, reducing manpower, time and costs, aiming to obtain more efficient phytotherapic products, with affordable value for the population.

Arctium lappa L. está indicado en la Formulacao de Fitoterápicos da Farmacopeia Brasileira para el tratamiento de trastornos urinarios leves. Los estudios han demostrado el potencial antioxidante, antiinflamatorio y antidiabético de este extracto, donde se identificaron fenoles, lignanos, taninos y flavonoides. El objetivo de este trabajo fue optimizar el método extractivo de las raíces de A. lappa. Los extractos se prepararon por diferentes métodos: Ultrasonido, Soxhlet, maceración y turboextracción. La optimización se realizó mediante turboextracción siguiendo una planificación factorial de 23, empleando como factores: tenor alcohólico, concentración de materia prima y tiempo de extracción. Se evaluaron los extractos para determinar el residuo seco, el contenido de fenoles y flavonoides y la actividad antioxidante. En cuanto al contenido de residuo seco, fenoles y flavonoides, los métodos de extracción por ultrasonidos y turbo demostraron un mejor poder de extracción. Debido al menor tiempo y coste operativo, la optimización se realizó mediante turboextracción, y el extracto optimizado se obtuvo utilizando alcohol 60%, en proporción disolvente-materia 1:10 y tiempo de extracción de 15 minutos. Estos análisis podrán orientar futuros ensayos de transposición del método para escala industrial, reduciendo mano de obra, tiempo y costes, con el objetivo de obtener productos fitoterapéuticos más eficientes, con valor accesible para la población.

Extracción e identificación de lípidos polares de las microalgas Nannochloropsis oceanica y Desmodesmus asymmetricus / Extraction and identification of polar lipids from microalgae Nannochloropsis oceanica and Desmodesmus asymmetricus / Extração e identificação os lipídios polares das microalgas Nannochloropsis ocean ca e Desmodesmus asymmetricus

Resumen Los lípidos polares de las microalgas sor de gran interés debido a su aplicación como ingredientes naturales novedosos para las industrias cosmética, nutricional y farmacéutica. Por ello, el presente trabajo buscó determinar el efecto de los principales factores en la extracción e identificación de los lípidos polares de las microalgas Nannochloropsis oceanica y Desmodesmus asymmetricus, mediante el diseño de superficie de respuesta de Box-Behnken y el diseño factorial completo, respectivamente. Estas cepas del Banco de Germoplasma de Organismos Acuáticos (BGOA - IMARPE) fueron cultivadas en un invernadero, en biorreactores de 30 litros, centrifugadas y liofilizadas. Los lípidos fueron extraídos con cloroformo-metanol, fraccionados y analizados con un espectrómetro de masas Waters Xevo G2-XS QTOF. La maximización de la extracción de los lípidos totales determinó un valor óptimo de la relación masa-solvente de 25mg/3 mL, una proporción 1:1 de cloroformo-metanol, aproximadamente, y un tiempo del baño de ultrasonido entre 10 y 30 min. Los principales lípidos polares identificados para N. oceanica fueron lisofosfatidilcolina (LPC), diacilgliceril-N,N,N-trimetilhomoserina (DGTS), digalactosil diacilglicerol (DGDG) y monogalactosil diacilglicerol (MGDG) y para D. asymmetricus fueron sulfoquinovosil diacilglicerol (SQDG), LDGTS, DGTS, DGDG y MGDG.

Abstract The microalgae polar lipids are of great interest due to their application as novel natural ingredients for the cosmetic, nutritional, and pharmaceutical industry. For this reason, the present work sought to determine the effect of the main factors in the extraction and identification of polar lipids from the microalgae Nannochloropsis oceanica and Desmodesmus asymmetricus, using Box-Behnken response surface methodology design and full factorial design, respectively. These strains from the Germplasm Bank of Aquatic Organisms (BGOA - IMARPE) were grown in a greenhouse, in 30 L bioreactors, centrifuged and lyophilized. The lipids were extracted with chloroform-methanol, fractionated and analyzed with the Waters Xevo G2-XS QTOF mass spectrometer. The maximization of total lipid extraction determined an optimal value of the mass-solvent ratio of 25 mg / 3 mL, an approximate ratio of chloroform-methanol 1:1 and an ultrasound bath time between 10 and 30 min. The main polar lipids identified for the N. oceanica microalgae were lysophophatidylcholine (LPC), diacylglyceryl-N,N,N-trimethylhomoserine (DGTS), digalactosyldiacylglycerol (DGDG), and monogalactosyldiacylglycerol (MGDG) and for D. asymmetricus were sulfoquinovosyl diacylglycerol (SQDG), LDGTS, DGTS, DGDG, and MGDG.

Resumo Os lipídios polares das microalgas possuem grande interesse em sua aplicação como novos ingredientes naturais na indústria cosmética, nutricional e farmacêutica. A presente pesquisa procura determinar o efeito dos principais fatores na extração e identificação dos lipídios polares das microalgas Nannochloropsis oceanica e Desmodesmus asymmetricus, por meio do um experimento de Box-Behnken e um experimento fatorial completo, respectivamente. As cepas do Banco do Germoplasma do Organismos Aquáticos (BGOA - IMARPE), foram cultivadas em casa de vegetação, em biorreatores do 30 L, centrifugadas e liofilizadas. Os lipídios foram extraídos com clorofórmio-metanol, fracionados e analisados com o espectrómetro do massa Waters Xevo G2-XS QTOF. A maximização da extração lipídica determinou um valor ótimo da razão massa-solvente de 25 mg / 3 mL, uma proporção aproximadamente clorofórmio-metanol de 1:1 e no tempo do banho de ultrassom entre 10 e 30 minutos. Os principais lipídios polares identificados para das microalgas N. oceanica foram lisofosfatidilcolina (LPC), diacilgliceril-N,N,N-trimetil-homoserina (DGTS), digalactosil diacilglicerol (DGDG) e monogalactosil diacilglicerol (MGDG), e para D. asymmetricus foram sulfoquinovosil diacilglicerol (SQDG), LDGTS, DGTS, DGDG e MGDG.

Experimental design approach for development of novel microemulsion system and immediate release self microemulsifying tablet of nebivolol HCl

The objective of this study was to determine specific combination of pharmaceutical excipients that lead to formulation of efficient nebivolol hydrochloride SMEDDS and its subsequent formulation into IR-SET (Immediate release- Self emulsifying tablet) which will enhance its solubility and dissolution. Solubility and Pseudo-ternary phase studies were carried out to identify the excipients showing highest solubility and to identify the zone of microemulsion with selected ingredients. Liquid-SMEDDS (L-SMEDDS) were optimized for Concentration of oil(X1) and Smix(X2) and formulated using a combination of Kollisolv GTA as oil, Tween 80 as surfactant and propylene glycol as co-surfactant which gave smaller droplet size(Y1) 55.98nm , Emulsification time (Y2) 16±1.5 s,% transmittance (Y3) 99.94±0.47%. Neusilin US2 was used as solid carrier for solidification of L-SMEDDS in to Solid-SMEDDS (S-SMEDDS) by adsorption technique. IR-SET of nebivolol were formulated with S-SMEDDS and optimized for the concentration of binder (X1) (PVP K30) and superdisintegrant (X2) (KOLLIDON CL) which showed low Disintegration time (Y1) (92±0.5s) and low Friability(Y2)(0.424±0.03%). Also the DSC and XRD data revealed the molecular state of the drug in S-SMEDDS. The extent of in-vivo drug release and ex-vivo diffusion values from L-SMEDDS and IR-SET was much higher than pure drug and marketed tablet. In conclusion, the results showed potential of SMEDDS to improve solubility and thus the bioavailability.

Design, preparation and in vitro characterizations of fluconazole loaded nanostructured lipid carriers

Present study was aimed to prepare and characterize fluconazole loaded nanostructured lipid carriers (FLZ-NLCs) for the treatment of fungal infections. Fungal infections are tremendously widespread and are the often faced dermatological condition worldwide. FLZ-NLCs was prepared by ultrasonication emulsion technique using stearic acid (SA) as solid lipid, castor oil as liquid lipid and tween 20 as a surfactant. The mean diameter of optimized FLZ-NLCs were found to be 359.15 ± 9.83 nm. The drug content and entrapment efficiency of NLCs was found to be 102.97 ± 7.45% and 87 ± 0.59%, respectively. In vitro drug release studies of FLZ-NLCs showed 37.34 ± 2.08% drug release over a period of 72 h. The above studies confirmed the prepared FLZ-NLCs may be useful for the treatment of fungal infections.

Cytotoxic effect of transdermal invasomal anastrozole gel on MCF-7 breast cancer cell line

Introduction: Anastrozole is an anti-cancer drug, an effective aromatase inhibitor for the treatment of breast cancerin post-menopausal women. As it undergoes extensive first-pass metabolism and has many side effects related to oraluse, it has been envisaged to develop anastrozole invasomes in the form of transdermal gel.Objective: The objective of this work was to prepare, characterize, and evaluate invasomal gel of anastrozole.Materials and Methods: Invasomes were prepared by thin layer film hydration method using Phospholipon 80H,fenchone (terpene), and ethanol. The optimized invasomes were incorporated into sodium carboxy methyl cellulosegel. Prepared formulations were evaluated and cytotoxic study on Michigan cancer foundation (MCF)-7 cancer cellline was studied.Results and Discussion: The scanning electron microscope results of the optimized formulation showed sphericalshaped vesicles.The ex vivo permeation of invasomes and the skin deposition (73%) were studied on male Wistar ratskin. Cell line studies on MCF-7 cells showed cytotoxic effect of optimized formulation at 5 µl/ml.Conclusion: It was concluded that the developed anastrozole invasomes enhanced the transdermal flux and theresults obtained encouraged the use of the anastrozole invasomal gel for the potential treatment of breast cancer inpost-menopausal women.

Cytotoxic effect of transdermal invasomal anastrozole gel on MCF-7 breast cancer cell line

Introduction: Anastrozole is an anti-cancer drug, an effective aromatase inhibitor for the treatment of breast cancerin post-menopausal women. As it undergoes extensive first-pass metabolism and has many side effects related to oraluse, it has been envisaged to develop anastrozole invasomes in the form of transdermal gel.Objective: The objective of this work was to prepare, characterize, and evaluate invasomal gel of anastrozole.Materials and Methods: Invasomes were prepared by thin layer film hydration method using Phospholipon 80H,fenchone (terpene), and ethanol. The optimized invasomes were incorporated into sodium carboxy methyl cellulosegel. Prepared formulations were evaluated and cytotoxic study on Michigan cancer foundation (MCF)-7 cancer cellline was studied.Results and Discussion: The scanning electron microscope results of the optimized formulation showed sphericalshaped vesicles.The ex vivo permeation of invasomes and the skin deposition (73%) were studied on male Wistar ratskin. Cell line studies on MCF-7 cells showed cytotoxic effect of optimized formulation at 5 µl/ml.Conclusion: It was concluded that the developed anastrozole invasomes enhanced the transdermal flux and theresults obtained encouraged the use of the anastrozole invasomal gel for the potential treatment of breast cancer inpost-menopausal women

Optimization of ex vivo permeability characteristics of berberine in presence of quercetin using 32 full factorial design

The aim of the present work was to investigate permeability characteristics of an anticancer berberine chloride, inpresence and absence of bioenhancer quercetin on goat intestine using Franz diffusion cell. A 32 full factorial designapproach was employed to investigate the effect of independent variables such as the concentration of bioenhancer(X1) and pretreatment time (X2) on dependent variable % cumulative drug release (% CDR) (Y) using design expertsoftware. The effect of quercetin was examined at three different levels of pretreatment time (30, 45, and 60 minutes)and at three different concentrations (2, 6, and 10 mg) on goat intestine. The apparent permeability (Papp), flux (J), andenhancement ratio (ER) were determined. Further, in vitro anticancer activity of optimized batch was performed onvarious cancer cell lines K562, A459, and Hela. During pretreatment studies, it was observed that an increase in theconcentration of quercetin yielded a positive effect on % CDR while the increase in pretreatment time by quercetinhad a detrimental effect on % CDR. When goat intestine was pre-treated for 30 minutes with 10 mg of quercetin,90.91% ± 1.66% CDR was obtained while the minimum value of 17.45% ± 2.12% CDR was observed at 2 mgquercetin pre-treated for 60 minutes. In vitro anticancer activity of optimized batch demonstrated non-significanteffect as compared with parent drug. In conclusion, quercetin could be successfully utilized as bioenhancer to improveex vivo permeability of berberine chloride, which would be expected to improve its bioavailability and reduce the doseresulting in improved patient compliance.

Preparation, Characterization, In Vitro Release and Taste-masking Effect of Paroxetine Resinate / 中国药学杂志

OBJECTIVE: To prepare and characterize paroxetine resinate, and evaluate the in vitro drug release rate and taste-masking effect. METHODS: A full factorial design was first conceived and applied to screen some process and formulation parameters (reaction temperature, stirring speed, drug concentration in solution and the ratio of resin to drug) on the key responses of resinationprocess, such as drug utilization ratio, drug loading and complexation constant. The paroxetine resinate was then characterized and evaluated by scanning electronic microscope (SEM), differential scanning calorimetry (DSC), in vitro drug release test and panel test of taste-masking. RESULTS: The resin/drug ratio and reaction temperature were identified as the most important factors on paroxetine resinate preparation.The drug-resin complex was successfully formed via ion exchange mechanism rather than physical absorption with complete in vitro drug release (>96%) in acidic or salt solution and good taste-masking effect. CONCLUSION: Paroxetine resinate with good performance can be prepared via optimization of process and formulation parameters, which will facilitate the development of generic paroxetine suspension.

Optimized gastroretentive floating carvedilol tablets: an approach for prolonged gastric residence time and enhanced absorption.

Carvedilol (CVD) is an antihypertensive agent with short elimination half-life, pH-dependent solubility, and narrow absorption window. So, the present study aimed to prolong its gastric residence time that entailed a development of an optimized gastro retentive floating tablets (GRFTs) using 32full factorial design. The tablets were fabricated by direct compression using hydroxypropyl methylcellulose and carbopol 940 as release retarding polymers. The quality attributes of the tablets were evaluated. The buoyancy lag time, total floating time, swelling ability and in vitro release studies were also carried out in 0.1 N HCl (pH 1.2) at 37 ± 0.5 °C. Statistical data analysis revealed that the optimized formulation containing 21.91% HPMC and 15% carbopol 940 had acceptable hardness, optimum floating behavior and 24h controlled-release pattern. The design succeeded to develop CVD-GRFTs with floating ability and controlled release behavior that could improve its solubility, and improve its availability at the best absorptive site.

Formulation of fast disintegrating domperidone tablets using Plantago ovata mucilage by 32 full factorial design.

The present work was carried out to study the disintegrant property of plantago ovata mucilage. The objective of the work was to formulate fast disintegrating tablets of Domperidon with a view to enhance patient compliances and dissolution rate by direct compression method using 3² full factorial design. Plantago ovata mucilage (2-10% w/w) was used as natural superdisintegrant and microcrystalline cellulose (0-30% w/w) was used as diluent, along with directly compressible mannitol to enhance mouth feel. The tablets were evaluated for hardness, friability, thickness, drug content uniformity, in vitro dispersion time, wetting time and water absorption ratio. Based on in vitro dispersion time (approximately 10s); the formulation containing 10% w/w Plantago ovata mucilage and 30%w/w microcrystalline cellulose was found to be promising and tested for in vitro drug release pattern (in 0.1 N HCl), short-term stability (at 40º/75% RH for 3 month) and drug-excipient interaction. Surface response plots are presented to graphically represent the effect of independent variables (concentrations of Plantago ovata mucilage and microcrystalline cellulose) on the in vitro dispersion time. The validity of the generated mathematical model was tested by preparing two extra-design check point formulations. The optimized tablet formulation was compared with conventional commercial tablet formulation for drug release profiles. This formulation showed nearly four-fold faster drug release (t50% 2.85 min) compared to the conventional commercial tablet formulation (t50% 7.85 min). Short-term stability studies on the formulation indicated that there are no significant changes in drug content and in vitro dispersion time (p < 0.05).

Formulation and in vitro evaluation of berberine containing liposome optimized by 32 full factorial designs.

The present study demonstrates the application of 32 full factorial design for optimization of berberine loaded liposome for oral administration. Thin film hydration method was used to prepare liposome and optimization was done by 32 full factorial designs combined with desirability function. Nine formulations were prepared by using different drug : lipid and soyphosphatidylcholine : cholesterol (SPC:CHOL) ratios and evaluated for entrapment efficiency and vesicle size. The statistical validity of model was done by analysis of variance (ANOVA). Response surface graph and contour plots were used to understand the effect of variables on responses. The optimized formulation with 0.782 desirability value was prepared and evaluated for responses. The results of entrapment efficiency and vesicle size were found to be very close with the predicted values. In addition, an optimized formulation was also characterized for zeta potential, in vitro drug release and morphology. The formulation was found to be spherical shape with an average diameter of 0.823 nm and -1.93 mV zeta potential and also shows sustained release pattern. These results support the fact that 32 full factorial designs with desirability function could be effectively used in optimization of berberine loaded liposome.

Optimization of Polyphenols Extraction Method from Kola Nuts (Cola nitida Vent. Schott & Endl.) Using Experimental Design.

Aims: The aim of this study is to optimize extraction process of phenolics compounds from kola nuts by using experimental design. Study Design: Kola nuts were collected in October 2014-February 2015 in south of Côte d’Ivoire. Harvested kola nuts were transferred to the laboratory until used in the experiments Place and Duration of Study: This study was carried out during season 2014-2015 in the Laboratory of Biochemistry and Food Science, Félix Houphouët-Boigny University, Côte d’Ivoire. Methodology: Nuts were divided into two groups and subdivided to obtain four groups according to their variety (traditional or improved) and morphotype or cultivar (white and red). After drying and powder processing, the effect of six parameters (solvent type, solid-liquid ratio, extraction mode, variety, cultivar and extraction time) on polyphenol extraction from kola nuts were studied. Firstly, a Plackett-Burman design (8 experiments) was used to highlight the most important factors which influence the extraction process. Then, a full factorial design (2k, k=4) was used to optimize the extraction conditions. Results: Results showed that solvent, ratio (w/v), extraction mode and variety of nuts had significant effect on polyphenols extraction. The predicted optimal conditions for the highest polyphenol content from kola nuts were found with infusion of traditional variety at 1/100 (w/v) ratio with aqueous ethanol 50% (v/v). In the predicted optimal conditions, experimental values were 350 mg/L GAE, 1460 mg/L QE and 264.33 mg/L CE for total polyphenol, total flavonoid and condensed tannin, respectively. Experimental data were very close to the predicted values. Conclusion: The extractive capability of kola nuts polyphenol is considerably depended on the solvent type, the extraction mode, the solid-liquid ratio (w/v) and nuts variety. Thus, kola nuts can be considered as a natural source of phenolics compounds with good antioxidant capacity. This optimization of the extraction parameters of phenolics compounds from kola nuts is very original, it is the first on a current scale of research on kola nut.

Formulation of fast disintegrating domperidone tablets using Plantago ovata mucilage by 32 full factorial design.

The present work was carried out to study the disintegrant property of plantago ovata mucilage. The objective of the work was to formulate Fast disintegrating tablets of Domperidon with a view to enhance patient compliances and dissolution rate by direct compression method using 3² full factorial design. Plantago ovata mucilage (2-10% w/w) was used as natural superdisintegrant and microcrystalline cellulose (0-30% w/w) was used as diluent, along with directly compressible mannitol to enhance mouth feel. The tablets were evaluated for hardness, friability, thickness, drug content uniformity, in vitro dispersion time, wetting time and water absorption ratio. Based on in vitro dispersion time (approximately 10s); the formulation containing 10% w/w Plantago ovata mucilage and 30%w/w microcrystalline cellulose was found to be promising and tested for in vitro drug release pattern (in 0.1 N HCl), short-term stability (at 40º/75% RH for 3 month) and drug-excipient interaction. Surface response plots are presented to graphically represent the effect of independent variables (concentrations of Plantago ovata mucilage and microcrystalline cellulose) on the in vitro dispersion time. The validity of the generated mathematical model was tested by preparing two extra-design check point formula-tions. The optimized tablet formulation was compared with conventional commercial tablet formulation for drug release profiles. This formulation showed nearly four-fold faster drug release (t50% 2.85 min) compared to the conventional commercial tablet formulation (t50% 7.85 min). Short-term stability studies on the formulation indicated that there are no significant changes in drug content and in vitro dispersion time (p < 0.05).