Higher Oxidative Stability of Alpha-linolenic Acid Than Linoleic Acid in Nanoemulsions: a Comparison Between Bulk Flaxseed Oil and its O/W Nanoemulsions

In this study, the effects of different dispersed phase volume fractions (Φ 0.025 and 0.1) and storage temperature (4 and 25 °C) were determined on lipid oxidation, fatty acids profile, β-carotene degradation, and other physicochemical properties of flaxseed oil-in-water nanoemulsions. Nanoemulsions containing small anionic droplets (≈ 100 nm) were fabricated using high-pressure homogenization. Although an increase in the viscosity and physical stability of nanoemulsions was observed with increasing Φ, but mean droplet diameter and chemical stability decreased. β-carotene degradation, free fatty acids formation, as well as thiobarbituric acid-reactive substances production, were all faster in the more concentrated emulsions. As the storage temperature raised, physical and chemical stability both decreased. Interestingly, while the ratio of α-linolenic acid to linoleic acid in bulk oil decreased over time, an opposite trend was observed in the nanoemulsions. This effect was due to differences in the location of different unsaturated fatty acids inside the oil nanodroplets.

     

Physical properties of emulsion-based hydroxypropyl methylcellulose films: Effect of their microstructure

The initial characteristics of emulsions and the rearrangement of the oil droplets in the film matrix during film drying, which defines its microstructure, has an important role in the physical properties of the emulsion-based films. The objective of this work was to study the effect of the microstructure (two droplet size distributions) and stability (with or without surfactant) of HPMC oil-in-water emulsions over physical properties of HPMC emulsion-based edible films. HPMC was used to prepare sunflower oil-in-water emulsions containing 0.3 or 1.0% (w/w) of oil with or without SDS, as surfactant, using an ultrasonic homogenizer. Microstructure, rheological properties and stability of emulsions (creaming) were measured. In addition, microstructure, coalescence of oil droplets, surface free energy, optical and mechanical properties and water vapor transfer of HPMC films were evaluated. Image analysis did not show differences among droplet size distributions of emulsions prepared at different oil contents; however, by using SDS the droplet size distributions were shifted to lower values. Volume mean diameters were 3.79 and 3.77μm for emulsions containing 0.3 and 1.0% without surfactant, respectively, and 2.72 and 2.71μm for emulsions with SDS. Emulsions formulated with 1.0% of oil presented higher stability, with almost no change during 5 and 3 days of storage, for emulsions with and without SDS, respectively. Internal and surface microstructure of emulsion-based films was influenced by the degree of coalescence and creaming of the oil droplets. No effect of microstructure over the surface free energy of films was found. The incorporation of oil impaired the optical properties of films due to light scattering of light. Addition of oil and SDS decreased the stress at break of the emulsion-based films. The replace of HPMC by oil and SDS produce a lower "amount" of network structure in the films, leading to a weakening of their structure. The oil content and SDS addition had an effect over the microstructure and physical properties of HPMC-based emulsions which lead to different microstructures during film formation. The way that oil droplets were structured into the film had an enormous influence over the physical properties of HPMC films.     

Influence of Nanoemulsion Addition on the Stability of Conventional Emulsions

Interest in using nanoemulsions as delivery systems for lipophilic food ingredients is growing due to their high optical clarity, good physical stability, and ability to increase bioavailability. Nanoemulsion-based delivery systems may need to be incorporated into food matrices that also contain conventional emulsions. The aim of this work was to evaluate the effect of adding nanoemulsions (d?<?200 nm) to conventional emulsions (d?>?200 nm) on the creaming stability and microstructure of the mixed systems. Droplet flocculation and rapid creaming was observed when the nanoemulsion concentration exceeded a particular level: the critical flocculation concentration (CFC) was 3.75 % and 0.25 % (v/v) for conventional emulsions with average droplet diameters of 350 and 250 nm, respectively. Confocal microscopy indicated that there was appreciable droplet flocculation, and the fraction of individual droplets with diameters?<?100 nm decreased after 14 days storage, which was probably due to Ostwald ripening and/or coalescence. The results of the present study might have important implications for the incorporation of nanoemulsion-based delivery systems into food products containing larger fat droplets, such as dressings, sauces, or beverages.     

Crossflow microfiltration of recombinant Escherichia coli lysates after high pressure homogenization

Crossflow membrane filtration was used to process recombinant Escherichia coli cell lysates containing protein inclusion bodies after high pressure homogenization. The number of passes through the high pressure homogenizer changed the viscosities and average particle sizes of the cell lysates. The different cell lysates were processed with a hollow fiber unit containing microfiltration membranes and a plate and frame unit with either ultrafiltration or microfiltration membranes. There were differences in permeate flux and protein transmission for the various membranes with the best performing membranes giving permeate fluxes greater than 60 L m(-2) h(-1) and protein transmissions greater than 90%. For a given membrane, no differences were observed between the cell lysates following homogenization with one, two, and three passes at 83 MPa. The lack of a difference between the three lysates is due to their similarities with respect to the released macromolecules and the presence of small (<0.1 mum) cell debris. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 304-310, 1997.     

Investigating the Parameters Affecting the Stability of Superparamagnetic Iron Oxide-Loaded Nanoemulsion Using Artificial Neural Networks

Nanoemulsions are increasingly being investigated for their fascinating capability of loading both hydrophobic and hydrophilic molecules while their stability is still an issue, being affected by various factors. In this study, to evaluate the dominant factors affecting the stability of nanoemulsions, artificial neural networks (ANNs) were implemented. Nanoemulsions of almond oil in water containing oleic acid-coated superparamagnetic iron oxide nanoparticles were prepared using a mixture of Tween 80 and Span 80 as surfactant system and ethanol as a co-surfactant. The ratio of transparency of the samples at 30 min and 7 days after preparation was taken as an indication of the stability of samples. Four independent variables, namely, concentration of nanoparticle, surfactant, oil, and alcohol were investigated to find their relations with the dependent variable (i.e., transparency ratio). Using ANNs modeling, it was concluded that the stability is affected by all variables, with all variables showing reverse effect on the stability beyond an optimum amount.Key words: artificial neural networks, nanoemulsion, optimization, stability, superparamagnetic iron oxide     

In Vitro Characterization and Mosquito (Aedes aegypti) Repellent Activity of Essential-Oils-Loaded Nanoemulsions

The nanoemulsions composed of citronella oil, hairy basil oil, and vetiver oil with mean droplet sizes ranging from 150 to 220 nm were prepared and investigated both in vitro and in vivo. Larger emulsion droplets (195–220 nm) shifted toward a smaller size (150–160 nm) after high-pressure homogenization and resulted in higher release rate. We proposed that thin films obtained from the nanoemulsions with smaller droplet size would have higher integrity, thus increasing the vaporization of essential oils and subsequently prolonging the mosquito repellant activity. The release rates were fitted with Avrami’s equations and n values were in the same range of 0.6 to 1.0, implying that the release of encapsulated limonene was controlled by the diffusion mechanism from the emulsion droplet. By using high-pressure homogenization together with optimum concentrations of 5% (w/w) hairy basil oil, 5% (w/w) vetiver oil (5%), and 10% (w/w) citronella oil could improve physical stability and prolong mosquito protection time to 4.7 h due to the combination of these three essential oils as well as small droplet size of nanoemulsion.     

Disruption of Saccharomyces cerevisiae using enzymatic lysis combined with high-pressure homogenization

The disruption of commercially-available pressed Bakers' yeast (Saccharomyces cerevisiae) was studied using a relatively new high-pressure homogenizer (the Microfluidizer). Initial experiments using only mechanical disruption generally gave low disruption yields (i.e., less than 40% disruption in 5 passes). Consequently combinations of two disruption methods, namely enzymatic lysis and subsequent homogenization, were tested to identify achievable levels of disruption. The enzyme preparation employed was Zymolyase, which has been shown to effectively lyse the walls of viable yeast. Yeast cell suspensions ranging in concentration from 0.6 to 15 gDW/L were disrupted with and without enzymatic pre-treatment. Final total disruption obtained using the combined protocol approached 100% with 4 passes at a pressure of 95 MPa, as compared to only 32% disruption with 4 passes at 95 MPa using only homogenization. A model is presented to predict the fraction disrupted while employing this novel enzymatic pretreatment.Nomenclature a exponent of pressure (-) - b exponent of number of passes (-) - K disruption constant (MPa^-a) - N number of passes (-) - P pressure (MPa) - R total fraction of cells disrupted (-) - Ro fraction of cells disrupted after enzymatic pre-treatment (-) - X cell concentration (dry weight) (gDW/L) abbreviation DW dry weight     

Influence of Disperse Phase Transfer on Properties of Nanoemulsions Containing Oil Droplets with Different Compositions and Physical States

Food Biophysics - The impact of the initial oil droplet composition and physical state on molecular exchange processes in mixed oil-in-water nanoemulsions was investigated. Nanoemulsions consisting...     

Neem oil nanoemulsions: characterisation and antioxidant activity

The aim of the present work is to develop nanoemulsions (NEs), nanosized emulsions, manufactured for improving the delivery of active pharmaceutical ingredients. In particular, nanoemulsions composed of Neem seed oil, contain rich bioactive components, and Tween 20 as nonionic surfactant were prepared. A mean droplet size ranging from 10 to 100?nm was obtained by modulating the oil/surfactant ratio. Physicochemical characterisation was carried out evaluating size, ζ-potential, microviscosity, polarity and turbidity of the external shell and morphology, along with stability in simulated cerebrospinal fluid (CSF), activity of Neem oil alone and in NEs, HEp-2 cell interaction and cytotoxicity studies. This study confirms the formation of NEs by Tween 20 and Neem oil at different weight ratios with small and homogenous dimensions. The antioxidant activity of Neem oil alone and in NEs was comparable, whereas its cytotoxicity was strongly reduced when loaded in NEs after interaction with HEp-2 cells.     

Critical analysis of quantitative indicators of cell disruption applied to Saccharomyces cerevisiae processed with an industrial high pressure homogenizer

A comparison of quantification techniques was performed on suspensions of Saccharomyces cerevisiae which had been disrupted with a high pressure homogenizer. The quantification techniques included cell counting, monitoring protein release, UV absorbance, turbidity, sample mass loss analysis, variations in viscosity and measuring the particle size distribution of the homogenate. It was found that all quantification techniques resulted in similar relationships between the measured extent of disruption and number of passes through the homogenizer. The data from all techniques (except particle sizing) could be fitted to simple exponential decay models at various homogenization pressures. Turbidity, particle sizing and UV absorbance generally gave more conservative estimates of the extent of cell disruption compared to protein release and cell counting. Measuring both the turbidity and monitoring the release of cellular metabolites using UV absorbance gave simple, reliable and reproducible measures of disruption and were identified as being the most applicable to on-line disruption monitoring.     

Simulation of particle size distribution changes occurring during high-pressure disruption of bakers' yeast

Measurements of size distributions are provided for the breakage of commercial packed bakers' yeast cells as a function of operating pressure and number of passes through a Manton Gaulin high-pressure homogenizer. A two parameter model was developed, based upon the use of a Boltzmann function, to simulate the changes in size distribution that accompany the cell breakage process. The effects of operating pressure and number of passes are incorporated in the model and the result is used to simulate the particle size distribution of the cell homogenate. The results show that there is little breakage below a threshold pressure of 115 bar and above which breakage is critically dependent upon the pressure and number of passes through the homogenizer. The analysis provides a means of studying the efficiency of centrifugation that may follow cell disruption and provides the basis for further studies of size distribution changes accompanying cell disruption. (c) 1996 John Wiley & Sons, Inc.     

A novel fiber-optic photometer for in situ stability assessment of concentrated oil-in-water emulsions

The purpose of this research was to evaluate a novel fiberoptic photometer for its ability to monitor physical instabilities occurring in concentrated emulsions during storage. For this, the fiber-optic photometer was used to measure transmission of oil-in-water emulsions stabilized with hypromellose (HPMC) as a function of oil volume fraction and droplet size distribution (DSD). To detect physical instabilities like creaming and coalescence, the transmissivity of the samples was studied at 2 different hight levels over a certain period of time. The corresponding droplet size distributions were determined by laser diffraction with PIDS. Transmissivity was found to depend on the number of dispersed droplets and thus is sensitive to both the variation of phase volume fraction as well as the emulsions droplet size distribution. At constant DSD, light transmission decreased linearly with increasing oil content within a large interval of phase volume fractions from 0.01 to 0.3. At constant phase volume fraction, an increase in droplet size increased light transmission. Investigation of creaming on emulsions with different droplet size distributions showed changes in the initial delay times and creaming velocities. In contrast to creaming phenomenon coalescence can be identified by height independent changes of the transmissivity. In conclusion, transmissivity of oil-in-water emulsions observed by the novel fiber-optic photometer is sensitive to phase volume fraction, droplet size distribution, and thus can be used as a tool for stability studies on concentrated emulsions. Published: August 31, 2007     

Oxidative kinetics of salmon oil in bulk and in nanoemulsion stabilized by marine lecithin

Salmon oil (Salmo salar) rich in long-chain polyunsaturated fatty acids was submitted to various oxidative stresses (temperature, atmosphere, presence of natural antioxidant or pro-oxidant). Oxidative kinetics of oil formulations and nanoemulsions prepared by high-pressure homogenization (1700 bars) were carried out by using conjugated dienes and infrared spectroscopy measurements. Droplet size nanoemulsion and polyene index were calculated from the fatty acid composition of oils, and the lipid fraction of emulsions were followed up over a period of 40 days.The particle size of different nanoemulsions was about 160–207 nm. The oxidative stability of salmon lecithin was higher than that of other oil samples in spite of its high LC-PUFA concentration. Crude salmon oil was better protected by its natural antioxidants (tocopherols and astaxanthin). In this study, a high concentration of α-tocopherol (0.2%) was used for its pro-oxidant role. The addition of quercetin improves slightly the oxidative stability of oils. However, no effect of this antioxidant was observed in emulsion, due to its insolubility in water and lipid phases.     

Cumulative sedimentation analysis of Escherichia coli debris size

A new method to measure Escherichia coli cell debris size after homogenization is presented. It is based on cumulative sedimentation analysis under centrifugal force, coupled with Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis of sedimented proteins. The effects that fermentation and homogenization conditions have on the resulting debris distributions were investigated using this method. Median debris size decreased significantly from approximately 0.5 mum to 0.3 mum as the number of homogenization passes increased from 2 to 10. Under identical homogenization conditions, uninduced host cells in stationary phase had a larger debris size than exponential cells after 5 homogenizer passes. This difference was not evident after 2 or 10 passes, possibly because of confounding intact cells and the existence of a minimum debris size for the conditions investigated. Recombinant cells containing protein inclusion bodies had the smallest debris size following homogenization. The method was also used to measure the size distribution of inclusion bodies. This result compared extremely well with an independent determination using centrifugal disc photosedimentation (CDS), thus validating the method. This is the first method that provides accurate size distributions of E. coli debris without the need for sample pretreatment, theoretical approximations (e.g. extinction coefficients), or the separation of debris and inclusion bodies prior to analysis. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioang 55: 556-564, 1997.     

Antioxidant Effect of Nanoemulsions Containing Extract of <Emphasis Type="Italic">Achyrocline satureioides</Emphasis> (Lam) D.C.—Asteraceae

Ethanolic extracts of Achyrocline satureioides have pronounced antioxidant activity mainly due to the presence of the flavonoid quercetin. However, direct topical application of the extract is not possible due to the presence of high amounts of ethanol. In this sense, nanoemulsions arise as an alternative for topical formulation associating molecules with limited aqueous solubility. This article describes the development of topical nanoemulsions containing either A. satureioides extract or one of its most abundant flavonoid, quercetin. Nanoemulsions composed of octyldodecanol, egg lecithin, water and extract (NEE), or quercetin (NEQ) were prepared by spontaneous emulsification. This process led to monodisperse nanoemulsions presenting a mean droplet size of approximately 200–300 nm, negative zeta potential, and high association efficiency. A study of quercetin skin retention using porcine skin which was performed using a Franz diffusion cell revealed a higher accumulation of quercetin in skin for NEE when compared to NEQ. Finally, the antioxidant activity of formulations was measured by thiobarbituric acid-reactive species and the APPH model. A lower lipoperoxidation for the extract in respect to quercetin solution was observed. However, no difference between NEQ and NEE lipoperoxidation could be seen. The protection against lipoperoxidation by the formulations was also measured in the skin, where lower formation of reactive species was observed after treatment with NEE. In conclusion, this study shows the formulation effect on the physicochemical properties of nanoemulsions as well as on the skin retention and antioxidant activity of quercetin.     

Formulation development and optimization using nanoemulsion technique: A technical note

Summary and Conclusion  Ramipril nanoemulsion formulations were successfully prepared by the spontaneous emulsification method (titration method). Sefsol 218 was selected as the oil phase for the development of the formulation on the basis of the solubility studies. The differences in the droplet size between the formulations selected from the phase diagram was not statistically significant, although the polydispersity was at a minimum for the formulation containing 20% oil, 27% Smix, and 53% vol/vol aqueous phase. The droplet size was found to be 34.5 nm. Therefore, nanoemulsion, a multipurpose technology, can be exploited in drug delivery for poorly soluble drugs. Nanoemulsions have a higher solubilization capacity than simple micellar solutions, and their thermodynamic stability offers advantages over unstable dispersions, such as emulsions and suspensions, because they can be manufactured with little energy input (heat or mixing) and have a long shelf life. This technical note explains the basis for calculation and construction of pseudoternary phase diagrams and, most important, explains selection of the formulations from the phase diagrams to avoid metastable formulations having minimum surfactant concentration in the least possible time. Published: April 6, 2007     

Etoposide-loaded nanoparticles made from glyceride lipids: Formulation,characterization, in vitro drug release,and stability evaluation

The aim of the study was to prepare etoposide-loaded nanoparticles with glyceride lipids and then characterize and evaluate the in vitro steric stability and drug release characteristics and stability. The nanoparticles were prepared by melt emulsification and homogenization followed by spray drying of nanodispersion. Spray drying created powder nanoparticles with excellent redispersibility and a minimal increase in particle size (20–40 nm). Experimental variables, such as homogenization pressure, number of homogenization cycles, and surfactant concentration, showed a profound influence on the particle size and distribution. Spray drying of Poloxamer 407-stabilized nanodispersion lead to the formation of matrix-like structures surrounding the nanoparticles, resulting in particle growth. The in vitro steric stability test revealed that the lipid nanoparticles stabilized by sodium tauroglycocholate exhibit excellent steric stability compared with Poloxamer 407. All 3 glyceride nanoparticle formulations exhibited sustained release characteristics, and the release pattern followed the Higuchi equation. The spray-dried lipid nanoparticles stored in black polypropylene containers exhibited excellent long-term stability at 25°C and room light conditions. Such stable lipid nanoparticles with in vitro steric stability can be a beneficial delivery system for intravenous administration as long circulating carriers for controlled and targeted drug delivery. Published: September 30, 2005     

Factors Affecting Foamed Emulsions Prepared with an Extract from <Emphasis Type="Italic">Quillaja saponaria</Emphasis> Molina: Oil Droplet Size,pH and Presence of Beta-Lactoglobulin

Oil is well-known to act as antifoam and to destabilize foam lamellae by bridging between two adjacent foam bubbles. It was hypothesized that an optimal oil droplet size exists with respect to the stability of a foamed emulsions, where the oil droplets are sufficiently small to postpone bridging and the amount of free surfactant is sufficient to stabilize the oil/water-interface and the air/water-interface. Emulsions with 0.3% Quillaja saponin and a median oil drop-let size between 0.2 and 2.0 μm were prepared under varying homogenization conditions and characterized in a dynamic foam analyzer. Results confirmed the above mentioned hypothesis. Stability of the foamed emulsions considerably increased with increasing pH, which was attributed to electrostatic repulsion between oil droplets and the effect on the balance between disjoining pressure and capillary pressure. In a binary system containing proteins and saponins, stability of foamed emulsions can be further increased when emulsifiers are added sequentially. When the emulsion is stabilized by β-LG and QS is added after emulsification stability of the foamed emulsion is distinctly higher compared to systems, where QS and β-LG are added prior to emulsification. Future studies should deepen our understanding of these complex dispersed systems by investigating the molecular interactions including other proteins and additional food constituents.     

Monitoring the Stability of Perfluorocarbon Nanoemulsions by Cryo-TEM Image Analysis and Dynamic Light Scattering

Perfluorocarbon nanoemulsions (PFC-NE) are disperse systems consisting of nanoscale liquid perfluorocarbon droplets stabilized by an emulsifier, usually phospholipids. Perfluorocarbons are chemically inert and non-toxic substances that are exhaled after in vivo administration. The manufacture of PFC-NE can be done in large scales by means of high pressure homogenization or microfluidization. Originally investigated as oxygen carriers for cases of severe blood loss, their application nowadays is more focused on using them as marker agents in ¹⁹F Magnetic Resonance Imaging (¹⁹F MRI). ¹⁹F is scarce in organisms and thus PFC-NE are a promising tool for highly specific and non-invasive imaging of inflammation via ¹⁹F MRI. Neutrophils, monocytes and macrophages phagocytize PFC-NE and subsequently migrate to inflamed tissues. This technique has proven feasibility in numerous disease models in mice, rabbits and mini pigs. The translation to clinical trials in human needs the development of a stable nanoemulsion whose droplet size is well characterized over a long storage time. Usually dynamic light scattering (DLS) is applied as the standard method for determining particle sizes in the nanometer range. Our study uses a second method, analysis of transmission electron microscopy images of cryo-fixed samples (Cryo-TEM), to evaluate stability of PFC-NE in comparison to DLS. Four nanoemulsions of different composition are observed for one year. The results indicate that DLS alone cannot reveal the changes in particle size, but can even mislead to a positive estimation of stability. The combination with Cryo-TEM images gives more insight in the particulate evolution, both techniques supporting one another. The study is one further step in the development of analytical tools for the evaluation of a clinically applicable perfluorooctylbromide nanoemulsion.     

Physical-Mechanical and Antifungal Properties of Pectin Nanocomposites / Neem Oil Nanoemulsion for Seed Coating

Biodegradable polymers, when reinforced with nanostructures, are considered good sustainable coatings and viable alternatives to replace conventional coatings. In addition, biopesticides are also considered safe, biodegradable and environmentally friendly; therefore there is a growing interest in nanoemulsions based on phytochemical mixtures. In this context, the aim of this study is to aggregate Neem oil nanoemulsions and pectin matrices to produce nanocomposite films, as well as evaluate the nanoemulsions effect on the film properties for coating soybean seeds. Nanoemulsions were characterized assessing their average diameter and stability, while the nanocomposite antifungal, morphology, mechanical and barrier properties were analyzed. In general, the nanoemulsions had an average diameter close to 59 ± 0.61 nm, showed good stability and its addition improved film mechanical properties: reduced stiffness, resistance, and water vapor permeability, and increased extensibility. In addition, Neem oil provided antifungal properties against Aspergillus Flavus and Penicillium Citrinum. The seed coatings promoted a positive effect on the germination process of soybean seeds. Thus, antifungal nanocomposite films from renewable sources were successfully produced. The fungicidal inhibition of Neem oil as a nanoemulsion makes these new materials promising for the production of seed coatings.