Hanna Salminen

Formation of solid shell nanoparticles with liquid ω-3 fatty acid core

A major challenge for food and pharmaceutical industries is the engineering of nanostructures that can efficiently encapsulate bioactive compounds with enhanced physical and chemical stability, and high load. The influence of surfactant properties on the physical and chemical stability of (i) nanostructured lipid carriers (NLC) containing tristearin and ω-3 fish oil, (ii) tristearin solid lipid nanoparticles (SLN), and (iii) ω-3 fish oil-in-water emulsions was investigated. As surfactants we used low (LM)- and high-melting (HM) lecithins. Results indicated that the presence of fish oil reduced the crystallisation temperature, melting temperature, and melting enthalpy of tristearin. NLC stabilized with HM-lecithin inhibited the oxidation of ω-3 fatty acids ≥90% compared to those stabilized with LM-lecithin. This was attributed to the solidified surfactant layer of HM-lecithin inducing crystallisation of the shell by interfacial heterogeneous nucleation. The results showed that the saturated HM-lecithin was the key in controlling the crystallisation behaviour, and thereby enabled the formation of oxidatively and physically stable lipid nanoparticles.

Influence of surfactant composition on physical and oxidative stability of Quillaja saponin-stabilized lipid particles with encapsulated ω-3 fish oil.

The purpose of this study was to investigate the potential of a saponin-rich extract of Quillaja saponaria to replace bile salts in the surfactant formulations for stabilization of nanostructured lipid carriers (NLC). The influence of Quillaja extract and/or high-melting lecithin at different concentrations on physical and oxidative stability was evaluated in (i) NLC containing tristearin and ω-3 fish oil, (ii) ω-3 fish oil-in-water emulsion, and (iii) solid lipid nanoparticles (SLN) containing tristearin. Best physical, polymorphic and oxidative stability of NLC were achieved with a surfactant combination of 2.4% (w/w) Quillaja extract and 0.6% (w/w) high-melting lecithin. The results showed that encapsulation of ω-3 fish oil into NLC inhibited the formation of lipid hydroperoxides, propanal and hexanal by 72, 53 and 57%, respectively, compared to the fish oil-in-water emulsion prepared with the same surfactants. This indicated that the low oxidation observed in NLC cannot be due to potential antioxidative effects of the surfactant combination itself. Evidence is accumulating that tristearin is able to form a protective shell around the ω-3 fish oil, when crystallization is induced via high-melting phospholipids in the solidified interfacial layer.

Influence of encapsulated functional lipids on crystal structure and chemical stability in solid lipid nanoparticles: Towards bioactive-based design of delivery systems

We investigated the influence of physicochemical properties of encapsulated functional lipids--vitamin A, β-carotene and ω-3 fish oil--on the structural arrangement of solid lipid nanoparticles (SLN). The relationship between the crystal structure and chemical stability of the incorporated bioactive lipids was evaluated with different emulsifier compositions of a saponin-rich, food-grade Quillaja extract alone or combined with high-melting or low-melting lecithins. The major factors influencing the structural arrangement and chemical stability of functional lipids in solid lipid dispersions were their solubility in the aqueous phase and their crystallization temperature in relation to that of the carrier lipid. The results showed that the stabilization of the α-subcell crystals in the lattice of the carrier lipid is a key parameter for forming stable solid lipid dispersions. This study contributes to a better understanding of SLN as a function of the bioactive lipid.

Oil-in-water emulsions as a delivery system for n-3 fatty acids in meat products.

The oxidative and physical stabilities of oil-in-water emulsions containing n-3 fatty acids (25 wt.% oil, 2.5 wt.% whey protein, pH 3.0 or pH 6.0), and their subsequent incorporation into meat products were investigated. The physical stability of fish oil emulsions was excellent and neither coalescence nor aggregation occurred during storage. Oxidative stability was better at pH 6.0 compared to pH 3.0 likely due to antioxidative continuous phase proteins. Incorporation of fish oil emulsions into pork sausages led to an increase in oxidation compared to sausages without the added fish oil emulsion. Confocal microscopy of pork sausages with fish oil emulsions revealed that droplets had coalesced in the meat matrix over time which may have contributed to the decreased oxidative stability. Results demonstrate that although interfacial engineering of n-3 fatty acids containing oil-in-water emulsions provides physical and oxidative stability of the base-emulsion, their incorporation into complex meat matrices is a non-trivial undertaking and products may incur changes in quality over time.

Influence of co-surfactants on crystallization and stability of solid lipid nanoparticles.

HYPOTHESIS The purpose of this study was to find a suitable co-surfactant to replace non-food grade bile salts in solid lipid nanoparticle (SLN) formulations. The hypothesis was that the molecular structure and physical properties of co-surfactant modulate the stabilization of SLNs upon polymorphic transition. EXPERIMENTS Tristearin SLNs were prepared by using two main surfactants: saturated high-melting lecithin, and unsaturated low-melting lecithin. As co-surfactants we used sodium taurodeoxycholate (i.e. bile salt), Pluronic F68, Tween 60 and 80, and amino acids tyrosine, tryptophan, and phenylalanine. The influence of co-surfactants on crystallization behavior and physical stability of SLNs was investigated by differential scanning calorimetry and static light scattering, respectively. FINDINGS The results showed that the aromatic amino acids had optimal structures and properties to act as effective co-surfactants in SLNs. Our study suggests that ideal co-surfactants are amphiphilic with pronounced hydrophobic areas, but highly water soluble so that they can have a reservoir of molecules readily available for interfacial stabilization. They adsorb fast to the interfaces, but without inducing polymorphic transition. This work demonstrates how the right structure can facilitate the desired function.

Formation of transparent solid lipid nanoparticles by microfluidization: influence of lipid physical state on appearance.

HYPOTHESIS This study investigated the influence of liquid-solid transition and particle size on the optical properties of nanoemulsions. The hypothesis was that the crystallization of lipid droplets influences the nanoemulsion appearance. EXPERIMENTS Liquid and solid nanoemulsions (10 wt% octadecane, 1-5 wt% sodium dodecylsulfate) were formed by high-pressure microfluidization (5000-28,500 psi) at 45 °C. Solid lipid nanoparticles were formed by cooling the nanoemulsions to 5 °C and then heating to ambient temperature, whereas liquid nanoemulsions were formed by maintaining them at 25 °C. FINDINGS Results indicated that lipid nanoparticles ranging from 136 nm down to 36 nm were generated, and were stable to particle aggregation. The melting and onset temperatures of the nanoparticles decreased with decreasing particle diameter. Upon crystallization of the lipid, the absorbance increased by about 140% for nanoemulsions with 136 nm particle diameter, but only 5% for nanoemulsions with 36 nm particle diameter. These results were explained in terms of changes in refractive index upon droplet solidification that alter their scattering behavior. These results show that solidification of nanoemulsions results in a shift of the transparent-to-turbid transition regime. The practical consequences for emulsion manufacturers are that solid nanoemulsions must be smaller than liquid nanoemulsions to remain transparent.

Miscibility of Quillaja Saponins with other Co-surfactants under Different pH Values

The miscibility behavior of mixed surfactant systems and the influence of extrinsic parameters are crucial for their application as emulsifiers. Therefore, the objective of this study was to evaluate the miscibility behavior of mixed systems composed of commercial Quillaja saponin and a co-surfactant, namely sodium caseinate, pea protein, rapeseed lecithin, or egg lecithin. These mixtures were evaluated macro- and microscopically at different concentration ratios (maximum concentration 5% w/v) at pH 3, 5, and 7 at 25 °C. The individual ingredients were also assessed for their charge properties and surface hydrophobicity. The results showed that Quillaja saponin-caseinate mixtures were miscible only at pH 7, and showed aggregation and precipitation at lower pH due to increasing electrostatic attraction forces. Rheological measurements showed that Quillaja saponin-pea protein mixtures formed gelled structures at all tested pH values mainly via association of hydrophobic patches. Quillaja saponins mixed with rapeseed lecithin were miscible at all tested pH values due to electrostatic repulsion. Quillaja saponin-egg lecithin mixtures aggregated independent of pH and concentration ratio. The microscopic analysis revealed that the lower the pH and the higher the Quillaja saponin ratio, the denser were the formed Quillaja saponin-egg lecithin aggregates. The results are summarized in ternary phase diagrams that provide a useful tool in selecting a surfactant system for food applications.

Influence of heat on miscibility of Quillaja saponins in mixtures with a co-surfactant

Thermal treatment of mixed surfactant systems can have a major impact on their phase behavior through modified interactions between the surfactants. In this study, we investigated the miscibility behavior of aqueous binary surfactant systems composed of Quillaja saponin extract and sodium caseinate, pea protein, rapeseed lecithin, or egg lecithin at different concentration ratios (0-5% w/v) at pH3, 5, and 7 upon heat treatment (25-75°C). The results revealed that the heat-treated Quillaja saponin-sodium caseinate mixtures at pH7 remained miscible when the ratio of Quillaja saponins was equal or higher to the ratio of caseinate, otherwise the mixtures flocculated due to increased hydrophobic interactions. At pH3, the aggregation of Quillaja saponin-sodium caseinate structures was intensified by heating mainly through self-association of casein molecules. In Quillaja saponin-pea protein mixtures as well as in pure pea protein samples heating led to weakening of the gel structures at all tested pH values. In contrast, heating did not affect Quillaja saponin-rapeseed lecithin mixtures, which stayed miscible independent of pH due to electrostatic repulsive forces. Furthermore, the flocculated (pH5, 7) or aggregated (pH3) Quillaja saponin-egg lecithin mixtures were only slightly affected by heating. These results are important for understanding the interactions of binary surfactant systems when subjected to heating, which is a common processing step in many food applications.

Isothermal titration calorimetric analysis on solubilization of an octane oil-in-water emulsion in surfactant micelles and surfactant-anionic polymer complexes.

Polymers may alter the ability of surfactant micelles to solubilize hydrophobic molecules depending on surfactant-polymer interactions. In this study, isothermal titration calorimetry (ITC) was used to investigate the solubilization thermodynamics of an octane oil-in-water emulsion in anionic sodium dodecylsulphate (SDS), nonionic polyoxyethylene sorbitan monooleate (Tween 80), cationic cetyltrimethylammonium bromide (CTAB) surfactant micelles and respective complexes formed by these micelles and an anionic polymer (carboxymethyl cellulose). Results indicated that the oil solubilization in single ionic micelles was endothermic, while in nonionic micelles or mixed ionic/nonionic micelles it was exothermic. The addition of carboxymethyl cellulose did not influence the solubilization behavior in these micelles, but affected the solubilization capacities of these systems. The solubilization capacity of cationic micelles or mixed cationic/nonionic micelles was enhanced while that of nonionic or anionic micelles was decreased. Based on the phase separation model, a molecular pathway mechanism driven by enthalpy was proposed for octane solubilization in surfactant micelles and surfactant-polymer complexes.

Solubilization of octane in electrostatically-formed surfactant–polymer complexes

Polymers can be used to modulate the stability and functionality of surfactant micelles. The purpose of this study was to investigate the solubilization of an octane oil-in-water emulsion in mixtures of an anionic polymer (carboxymethyl cellulose) and anionic sodium dodecylsulphate (SDS), nonionic polyoxyethylene sorbitan monooleate (Tween 80) and cationic cetyltrimethylammonium bromide (CTAB) surfactant micelles using dynamic light scattering, microelectrophoresis and turbidity measurements. The results showed that the addition of anionic carboxymethyl cellulose accelerated octane solubilization in cationic CTAB and CTAB-Tween 80 micelles, but did not affect the solubilization behaviors of micelles that were nonionic and anionic. The surfactant-polymer interactions were also studied using isothermal titration calorimetry (ITC) to characterize different physiochemical interaction regions depending on surfactant concentration in surfactant-polymer systems. Upon octane solubilization in CTAB-carboxymethyl cellulose mixtures, shape transitions of polymer-micelle complexes may have taken place that altered light scattering behavior. Based on these results, we suggest a mechanism for oil solubilization in electrostatically-formed surfactant-polymer complexes.