Thrandur Helgason

Effect of surfactant surface coverage on formation of solid lipid nanoparticles (SLN)

The effect of surfactant surface coverage on formation and stability of Tween 20 stabilized tripalmitin solid lipid nanoparticles (SLN) was investigated. A lipid phase (10% w/w tripalmitin) and an aqueous phase (2% w/w Tween 20, 10 mM phosphate buffer, pH 7) were heated to 75 degrees C and then homogenized using a microfluidizer. The resulting oil-in-water emulsion was kept at a temperature (37 degrees C) above the crystallization temperature of the tripalmitin to prevent solidification of emulsion droplets, and additional surfactant at various concentrations (0-5% w/w Tween 20) was added. Droplets were then cooled to 5 degrees C to initiate crystallization and stored at 20 degrees C for 24 h. Particle size and/or aggregation were examined visually and by light scattering, and crystallization behavior was examined by differential scanning calorimetry (DSC). Excess Tween 20 concentration remaining in the aqueous phase was measured by surface tensiometry. Emulsion droplets after homogenization had a mean particle diameter of 134.1+/-2.0 nm and a polydispersity index of 0.08+/-0.01. After cooling to 5 degrees C at low Tween 20 concentrations, SLN dispersions rapidly gelled due to aggregation of particles driven by hydrophobic attraction between insufficiently covered lipid crystal surfaces. Upon addition of 1-5% w/w Tween 20, SLN dispersions became increasingly stable. At low added Tween 20 concentration (<1% w/w) the SLN formed gels but only increased slightly at higher surfactant concentrations (>1% w/w). The Tween 20 concentration in the aqueous phase decreased after tripalmitin crystallization suggesting additional surfactant adsorption onto solid surfaces. At higher Tween 20 concentrations, SLN had increasingly complex crystal structures as evidenced by the appearance of additional thermal transition peaks in the DSC. The results suggest that surfactant coverage at the interface may influence crystal structure and stability of solid lipid nanoparticles via surface-mediated crystal growth.

Impact of Surfactant Properties on Oxidative Stability of β-Carotene Encapsulated within Solid Lipid Nanoparticles

The impact of surfactant type on the physical and chemical stability of solid lipid nanoparticle (SLN) suspensions containing encapsulated beta-carotene was investigated. Oil-in-water emulsions were formed by homogenizing 10% w/w lipid phase (1 mg/g beta-carotene in carrier lipid) and 90% w/w aqueous phase (surfactant + cosurfactant) at pH 7 and 75 degrees C and then cooling to 20 degrees C. The impact of surfactant type was investigated using aqueous phases containing different water-soluble surfactants [2.4% w/w high-melting (HM) lecithin, 2.4% w/w low-melting (LM) lecithin, and 1.4% w/w Tween 60 or 1.4% w/w Tween 80] and a cosurfactant (0.6% taurodeoxycholate). The impact of the physical state of the carrier lipid was investigated by using either a high melting point lipid (tripalmitin) to form solid particles or a low melting point lipid (medium chain triglycerides, MCT) to form liquid droplets. A higher fraction of alpha-crystals was detected in solid particles prepared with high-melting surfactants (HM-lecithin and Tween 60) than with low-melting surfactants (LM-lecithin and Tween 80). With the exception of the HM-lecithin-coated solid particles, the suspensions were stable to particle aggregation during 21 days of storage. beta-Carotene degradation after 21 days of storage was 11, 97, 100, and 91% in the solid particles (tripalmitin) and 16, 21, 95, and 90% in the liquid droplets (MCT) for HM-lecithin, LM-lecithin, Tween 80, and Tween 60, respectively. These results suggest that beta-carotene may be stabilized by (1) LM- or HM-lecithin when liquid carrier lipids are used and (2) HM-lecithin when solid carrier lipids are used. The origin of this latter effect is attributed to the impact of the surfactant tails on the generation of a crystal structure better suited to maintain the chemical stability of the encapsulated bioactive.

Influence of lipid physical state on the in vitro digestibility of emulsified lipids.

The objective of this study was to investigate the influence of the physical state of emulsified lipids on their in vitro digestibility by pancreatic lipase. A 10 wt % tripalmitin oil-in-water emulsion stabilized by sodium dodecyl sulfate (0.9 wt % SDS) was prepared at a temperature (>70 degrees C) above the melting point of the lipid phase (T(m) approximately 60 degrees C). A portion of this emulsion was cooled to a temperature (0 degrees C for 15 min) well below the crystallization temperature of the emulsified lipid (T(c) approximately 22 degrees C) and then warmed to 37 degrees C so as to have completely solid lipid particles. Another portion of the emulsion was directly cooled from 70 to 37 degrees C (which is above the T(c)) to have completely liquid (supercooled) lipid particles. Pancreatic lipase (8 mg/mL) and bile extract (5.0 mg/mL) were then added to each emulsion at 37 degrees C, and the evolution of the particle charge, particle size, appearance, and free fatty acid release were measured over a period of 2 h. It was found that the rate and extent of lipid digestion were higher in the emulsion containing liquid particles but that appreciable lipid digestion still occurred in the emulsion containing solid particles (i.e., >35% lipid digestion after 2 h). These results may have important consequences for controlling the digestion rate of lipids or for developing solid lipid particle delivery systems for lipophilic functional components.

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 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.

Examination of the Interaction of Chitosan and Oil-in-Water Emulsions Under Conditions Simulating the Digestive System Using Confocal Microscopy

ABSTRACT Chitosan is a cationic biopolymer that has been used extensively in dietary supplements to reduce fat absorption in the fight against obesity. The mechanism of fat binding of chitosan is still not fully understood and has been the subject of controversy. This study was designed to improve the understanding of the underlying mechanism by investigating the interaction of chitosan with oil-in-water emulsion droplets. Our results indicated that (0.1% w/w) chitosan adsorbed to a 20% w/w phosphatidylcholine-stabilized anionic primary emulsion to form a secondary cationic emulsion by electrostatic attraction forces under conditions resembling the stomach (pH 2). Bile salts (6 mM) were added to simulate secretion in the small intestine and pH increase. Bile salts adsorbed to the chitosan secondary emulsion, which resulted in aggregation of oil droplets followed by coalescence due to close packing of droplets. Increased viscosity (267–2531 cp) and increased degree of deacetylation (40–92% DDA) of chitosan enhanced emulsion breakdown. Increasing the pH to 7.5 without addition of bile salts yielded little aggregation. Pronounced aggregation is thought to decrease the accessibility of lipase to the oil resulting in lower bioavailability and reduced caloric intake. Understanding how chitosan interacts with oil droplets in the digestive tract is vital to developing a comprehensive model of the influence of chitosan on the bioavailability of dietary lipids. The information gained in this study may be useful for the interpretation and experimental design of animal and human feeding studies and for the rational design of chitosan-based functional foods for fat reduction.

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.

Temperature scanning ultrasonic velocity study of complex thermal transformations in solid lipid nanoparticles

The purpose of this study was to determine whether temperature scanning ultrasonic velocity measurements could be used to monitor the complex thermal transitions that occur during the crystallization and melting of triglyceride solid lipid nanoparticles (SLNs). Ultrasonic velocity ( u) measurements were compared with differential scanning calorimetry (DSC) measurements on tripalmitin emulsions that were cooled (from 75 to 5 degrees C) and then heated (from 5 to 75 degrees C) at 0.3 degrees C min (-1). There was an excellent correspondence between the thermal transitions observed in deltaDelta u/delta T versus temperature curves determined by ultrasound and heat flow versus temperature curves determined by DSC. In particular, both techniques were sensitive to the complex melting behavior of the solidified tripalmitin, which was attributed to the dependence of the melting point of the SLNs on particle size. These studies suggest that temperature scanning ultrasonic velocity measurements may prove to be a useful alternative to conventional DSC techniques for monitoring phase transitions in colloidal systems.

Tuning of shell thickness of solid lipid particles impacts the chemical stability of encapsulated ω-3 fish oil

HYPOTHESIS This study demonstrates that tuning the shell thickness of lipid particles can modulate their oxidative stability. We hypothesized that a thick crystallized shell around the incorporated fish oil would improve the oxidative stability due to the reduced diffusion of prooxidants and oxygen. EXPERIMENTS We prepared solid lipid nanoparticles (5%w/w lipid phase, 1.5%w/w surfactant, pH 7) by using different ratios of tristearin as carrier lipid and ω-3 fish oil as incorporated liquid lipid stabilized by high- or low-melting lecithin. The physical, polymorphic and oxidative stability of the lipid particles was assessed. FINDINGS The high-melting lecithin was the key in inducing the formation of a solidified tristearin shell around the lipid particles by interfacial heterogeneous nucleation. Lipid particles containing a higher ratio of tristearin showed a better oxidative stability. The results revealed that a crystallized tristearin layer above 10nm was required to inhibit oxidation of the incorporated fish oil. This cut-off was shown for lipid particles containing 50-60% fish oil. This research gives important insights into understanding the relation between the thickness of the crystallized shell around the lipid particles and their chemical stability.