Food Biophysics (2008) 3:146154 DOI 10.

1007/s11483-008-9065-8

SPECIAL ISSUE ARTICLE

Solid Lipid Nanoparticles as Delivery Systems for Bioactive Food Components

Jochen Weiss & Eric A. Decker & D. Julian McClements & Kristberg Kristbergsson & Thrandur Helgason & Tarek Awad

Received: 30 November 2007 / Accepted: 4 February 2008 / Published online: 1 March 2008 # Springer Science + Business Media, LLC 2008

Abstract The inclusion of bioactive compounds, such as carotenoids, omega-3 fatty acids, or phytosterols, is an essential requisite for the production of functional foods designed to improve the long-term health and well-being of consumers worldwide. To incorporate these functional components successfully in a food system, structurally sophisticated encapsulation matrices have to be engineered, which provide maximal physical stability, protect ingredients against chemical degradation, and allow for precise control over the release of encapsulated components during mastication and digestion to maximize adsorption. A novel encapsulation system initially developed in the pharmaceutical industries to deliver lipophilic bioactive compounds is solid lipid nanoparticles (SLN). SLN consist of crystallized nanoemulsions with the dispersed phase being composed of a solid carrier lipidbioactive ingredient mixture. Contrary to larger colloidal solid lipid particles, specific crystal structures can be dialed-in in SLN by using specific surfactant mixtures and ensuring that mean particle sizes are below 100200 nm. Moreover, in SLN, microphase separations of the bioactive compound from the solidifying lipid matrix can be prevented resulting in an even dispersion of the encapsulated compound in the solid matrix thereby improving chemical and physical stability of the bioactive. In this

review article, we will briefly introduce the structure, properties, stability, and manufacturing of solid lipid particles and discuss their emerging use in food science. Keywords Bioactive compounds . Nanoemulsions . Nutraceuticals . Functional foods . Delivery systems . Lipid crystallization

Introduction Increases in dietary-intake-related illnesses such as obesity, cardiovascular disease, hypertension, and cancer have led to the food industry making the development of health-andwellness-promoting foods using bioactive compounds a priority.1 Clinical studies have demonstrated tangible health benefits that may be derived from intake of bioactive compounds as part of consumers daily diet.2 However, many bioactive compounds tightly bind to the food matrix and/or are highly lipophilic resulting in poor absorption and limited bioavailability.3 More importantly, many of these compounds once extracted from the animal- or plant-based source tissue are chemically unstable. This applies to a wide variety of classes of bioactive compounds with demonstrated efficacies such as, for example, carotenoids (e.g., -carotene, lutein, and lycopene), strongly lipophilic compounds that possess virtually no water solubility. In the food industry, it has become apparent that current difficulty associated with inclusion of lipophilic bioactives in food matrices is one of the major problems that manufacturers struggle with when developing health-and-wellness-promoting foods. This problem is not unlike the one that the pharmaceutical industry faced in the past, where oral administration of lipophilic drugs resulted in low efficacies. The pharmaceutical industry solved this problem by developing a wide variety of

J. Weiss (*) : E. A. Decker : D. J. McClements : T. Awad Department of Food Science, University of Massachusetts, 100 Holdsworth Way, Amherst, MA 01003, USA e-mail: jweiss1@foodsci.umass.edu K. Kristbergsson : T. Helgason Department of Food Science, University of Iceland, Hjardarhagi 2-6, Reykjavik, 107, Iceland

Food Biophysics (2008) 3:146154

147

delivery systems that not only resulted in significant improvements in efficacies of the administered drug but also allowed for better control of release rate and targeting of the encapsulated compounds, improved the physicochemical stability by decreasing the reactivity of the encapsulated material in relation to the outside environment, and promoted easier handling of the compounds by achieving uniform dispersions.4 With a suitable carrier system, the in vivo fate of the active compound is no longer only determined by its properties but by those of the carrier system.

Microcapcules Versus Nanocapsules In pharmaceutical systems, it has been well established that colloidal delivery systems with sizes in the nanometer range such as nanoemulsions, microemulsions, and liposomes exhibit superior functionalities compared to conventional microencapsulation systems resulting in a dramatic decline in the use of microcapsules and a rapid rise in the use of nanoscalar colloidal carrier systems. Microencapsulation techniques include production of liquidliquid dispersions, e.g., emulsions, liposomes, or manufacturing of solid microcapsules from biopolymers (proteins of carbohydrates) including gel microcapsules, spray-dried and freeze-dried emulsions, and coated microparticles.5,6 Microcapsules however have proven to be less than ideal carrier systems. The particles are typically relatively large (>1 m), they are polydisperse, i.e., their size is distributed, and they are often thermodynamically unstable; that is, they tend to break down over time. The large size in particular slows down absorption. Physical and chemical instability processes can render the carrier system and the bioactive compound nonfunctional before consumption of foods. Over the past 5 years, research in our and other laboratories has shown that encapsulation of bioactives in nanoscalar colloidal carrier systems is feasible and can overcome issues associated with slow and low uptake and thermodynamic instability.7 Nanoparticles with less that 100 nm have been investigated as carrier systems for bioactives. Examples of colloidal carrier systems include nanoemulsions, microemulsions, and biopolymeric nanoparticles.5,8,9 These systems are either monodisperse or quasi-monodisperse, are thermodynamically stable, or have shown to exhibit long-term kinetic stability. Their small and uniform size ensures rapid release of encapsulated compounds, making them far superior to microparticles. At sizes below 100 nm, the particles appear to readily adsorb at the mucus where direct absorption of encapsulated bioactives may take place.10 With the exception of biopolymeric nanoparticles, all these systems are liquid and considerably less stable and more difficult to handle than solid particles.

Production of solid biopolymeric nanoparticles on the other hand has proved difficult because of the low availability of suitable biopolymers and the need to use organic solvents that may not be approved for food systems.11 Cytotoxicity and lack of large-scale production methods have further decreased the interest of the food industry in biopolymeric nanoparticles as a delivery system. Therefore, interest in microemulsions and nanoemulsions has increased. Despite the similarity in name, the two systems are fundamentally different.5 Nanoemulsions are extremely fine-dispersed emulsion droplets but are emulsions nevertheless. They are thermodynamically unstable and eventually phase separate albeit, because of their small size, over a considerably longer time than conventional emulsions. Microemulsions on the other hand are self-assembled association colloids that are thermodynamically stable. They are small enough to not scatter light, and microemulsion dispersions are thus transparent. However, they are prone to disintegration upon dilution. Interestingly, nanoemulsions and microemulsions could theoretically be composed of the same components, of course at different concentration ratios, but differ in their physicochemical properties.

Solid Lipid Nanoparticles Solid lipid nanoparticles (SLN) have gained increased attention in the pharmaceutical and food industries because of their ability to overcome deficiencies of both microcapsules and the previously mentioned nanoscalar colloidal carrier systems. They are the latest generation of nanoscalar encapsulation systems7,1158 and combine the advantages of the parent liquid nanoemulsions or microemulsions of high dissolution velocities associated with high permeability of the active compound through the gut wall, while simultaneously solving existing problems associated with physical and chemical stability of the encapsulated compound and ease of handling. Structure and Properties of Solid Lipid Nanoparticles SLN consist of a core of solid lipid with the bioactives being a part of the lipid matrix5962 (Figure 1). The particle is stabilized by a surfactant layer, which may consist of a single surfactant, but typically is composed of a mixture of surfactants.39,45 SLN can be manufactured from either nanoemulsions- or microemulsions-yielding particles with distinctly different properties (see below). In general, the use of crystallized lipids instead of liquid lipids has been shown to increase control over release and stability of incorporated bioactives. This is because mobility of bioactives can be controlled by controlling the physical state of the lipid matrix6367 (Figure 1).

148 Fig. 1 Structure of liquid nanoemulsions (left) and solid lipid nanoparticles (right) stabilized by a surfactant layer carrying a lipophilic bioactive compound

Food Biophysics (2008) 3:146154

Lipid Composition The composition of the parent carrier is one of the key parameters in controlling the properties and structure of SLN.19,24,25,32,33,6877 Firstly, the choice of the parent carrier determines the conditions under which the SLN has to be manufactured, e.g., homogenization temperature and cooling rates, as the lipid-bioactive mixture is first melt-homogenized and then solidified by cooling. Secondly, the choice of the carrier lipid will influence the loading capacity for the selected lipophilic bioactive. The generated crystal structure has a limited capacity to carry a second lipophilic compound. If maximum loading capacities are exceeded, the excess bioactive may be expelled from the generated crystal, which may lead to physical and chemical breakdown of the system. Thirdly, the crystallization temperature of the bioactive lipid in relation to the crystallization temperature of the carrier lipid will influence the generated structure of the SLN. For example, if the bioactive lipid has crystallization temperature that is significantly below that required to crystallize the carrier and below the temperature at which the SLNs are being used or stored at, the bioactive will remain liquid at all times. In this case, SLN with coreshell structures may emerge where either the shell or the core may be liquid. Conversely, if the bioactive has a crystallization temperature significantly above that of the melting temperature of the carrier, a mixed crystal matrix may be generated. Most importantly, the lipid composition will determine the type of crystal that is generated upon cooling, thereby influencing the stability of SLN and the release characteristics of the encapsulated bioactive from the SLN. Triacylglycerides (TAGs) are commonly used as carrier lipids. TAGs exhibit polymorphism upon cooling; that is, the individual chains of the lipid molecules may assume a variety of possible association configurations giving rise to longitudinal stacking of TAG molecules in lamellae that lead to the formation of , , and crystals with hexagonal, orthorhombic, and triclinic unit structures, respectively.78 These crystals differ in their

lattice spacing from 4.15 for the thermodynamically least stable and lowest melting -form to 4.6 for the thermodynamically most stable and highest melting -form. Pure homogenous lipids tend to form predominantly perfect crystals with the typical plated-like pattern of -modification (Figures 2 and 3). This can lead to aggregation and destabilization in the cooling process and higher oxidative

Fig. 2 Polymorphism of triacylcglycerides. The figure denotes the structural arrangement of fatty acid chains in TAGs leading to hexagonal, cubic, and orthogonal unit cells with distinctively different x-ray scattering patterns corresponding to , , and -crystal modifications

Food Biophysics (2008) 3:146154 Fig. 3 An x-ray scattering pattern of a triacylglyceride in bulk or in the form of a nanodispersion during cooling (adapted from Westensen, Bunjes and Koch 59)

149

instability due to increased surface aria and must therefore be avoided if possible. Using heterogeneous lipids will favor formation of spherical particles due to higher presence of crystals in the lipid phase.79 In summary, a solid understanding of the polymorphic features of the carrier lipid in combination with the bioactive lipid is required to form SLN that are physically and chemically stable. Surfactant Type One of the most important decisions that manufacturers of SLN have to make is to choose an appropriate surfactant or surfactant combination. In conventional emulsions, the emulsifier predominantly influences the final particle size that can be achieved during the homogenization and influences the stability of the dispersions after manufacturing by providing sufficient repulsive interaction forces that prevent droplets from coming into close contact leading to flocculation and or coalescence. However, in SLN, the surfactant plays an additional very important role in controlling the crystallization process. Because of the small size of the parent nanoemulsions, the number of lipid molecules interacting with the hydrophobic emulsifier tail groups is large enough to modulate the crystallization process. Moreover, the surfactant can subsequently improve the kinetic stability of the generated crystal structure even if that crystal structure is thermodynamically less stable than that of a corresponding alternative polymorphic form and thereby prevent re-crystallization during storage that may lead to destabilization of the dispersion during storage.80 To fulfill both the demand of particle stabilization and crystal modulation, mixtures of nonionic surfactants providing for the modulation and ionic surfactants providing for the repulsive interactions are commonly used.81

Surfactant Concentration Surfactant concentration influences the final size that can be achieved during homogenization and therefore the physical properties of the system. High concentration of surfactants will decrease the surface tension and stabilize newly formed surfaces during homogenization leading to smaller droplets.82 In addition, in SLN, insufficient amount of surfactant can lead to increased instability if recrystallization processes.83 For example, the thermodynamically more unfavorable crystal modification may transition into the -form. This transition is associated with morphological changes, i.e., the -crystals may undergo directional growth leading to formation of needle-like structures with increased surface areas. If these surfaces are not stabilized by surfactant molecules, hydrophobic interactions may lead to flocculation and destabilization of the SLN suspension. Droplet Size As mentioned earlier, the unique properties and functionalities of SLN arise due to their small size that allows for a specific crystal structure associated with a specific melting profile to be engineered.84 A reduction in particle size leads firstly to increased surface curvatures and, secondly, to an increase in the number of molecules per particle that actively interact with molecules in the interface (e.g., surfactants). With increasing surface curvature, lipid molecules are restricted in their ability to assume the shape of a perfect crystal. Consequently, smaller particles have increasing concentrations of - and -crystals, while bulk fat tend to crystallize predominantly in the form of -crystals (Figure 3).85,86 Furthermore, as -crystals tend to crystallize at lower temperature because of their decreased thermodynamic

150

Food Biophysics (2008) 3:146154

stability, increasingly more supercooling is required to transform the liquid lipid into a solid matrix. Conversely, if the particle size increases, for example, because of partial coalescence, the oil starts to behave more like a bulk fat and crystallizes at higher temperatures.8688 Cooling Conditions One of the most important parameters to control crystal structure is the cooling rate, which is very often ignored, leading to uncontrolled crystal formation, sometimes resulting in formation of plated-shaped particles and destabilization. Figure 4 shows results of a simple melting experiment after cooling a nanoemulsion composed of tripalmitin and stabilized by Tween 20 at 0.2, 2 and 20 C/min.83 The differential scanning calorimetry thermogram reveals that as the cooling speed is increased from 0.2 to 20 C/min, the concentration of -crystals as visible by the transition peak at 43 C increases while the concentration of -crystals that are formed at ~61 C in the SLN suspension remains virtually unchanged. The effect of cooling speed on crystal formation has also been reported by Jenning and coauthors62 who found that cooling at 10 C/min resulted in formation of -crystals but that, at 2 C/min, -crystals were formed predominantly. Stability of Solid Lipid Nanoparticles The physical and chemical stability of SLN is governed by two distinctly different processes, that is, the ability of the SLN suspension to remain homogeneous (suspension stability) and, secondly, the ability of the crystal matrix to resist recrystallization (matrix stability). Instability processes leading to the breakdown of the suspension include those typically encountered in many other dispersions such as flocculation and sedimentation or creaming and coalescence. In addition to the processes leading to suspension instability, recrystallization, due to polymorphic transformations, can

lead to network formation of crystallized particles resulting in a gel-like behavior.83 Suspension Stability Flocculation is the process whereby two or more droplets associate with each other but maintain their individual integrity. Flocculation can cause a pronounced increase in the viscosity of liquidliquid and liquidsolid dispersions and can sometimes lead to gel formation.82 Flocculation is typically caused by insufficient repulsive interaction forces between particles. A reduction in repulsive forces can be caused by changes in the ionic strength or pH of the system, which influence the charge of ionic surfactants adsorbed at the surface of the particles.89 Moreover, flocculation may be increased by stirring or by increasing the temperature, both of which may increase the number of droplet collisions per unit time and area.90 Finally, flocculation may occur in the presence of biopolymers due to depletion or bridging.82,91 Flocculation typically leads to an acceleration of gravitationdriven separation, e.g., sedimentation or creaming. Gravitational separation is driven by the density difference between the continuous and the dispersed phase, the viscosity of the continuous phase, and the particle size as described by the well-known Stokes equation. In stable SLN, gravitational separation is not a major concern, as particles have sizes at which the process occurs over a period of sometimes years. Coalescence is the process where two or more liquid droplets merge together to form a single larger droplet.82 Coalescence requires a liquid lipid matrix and is therefore not usually a problem in SLN. However, coalescence may occur if, during a manufacturing or production process, the previously formed SLN are heated above their melting temperature, e.g., during sterilization, pasteurization, evaporation, or drying.82,92 Above the melting temperatures, the melted SLN (nanoemulsions) may coalesce during the heating cycle. Upon cooling, significantly different matrix structures will be obtained, as the particle size has increased

Fig. 4 Heat flow between reference and sample measured by differential scanning calorimetry for tripalmitin solid lipid nanoparticles with mean diameters of ~120 nm stabilized by Tween 20 upon heating from 35 to 70 C after cooling at rates of 0.2, 2, and 20 C/min

Food Biophysics (2008) 3:146154

151

leading to the above-mentioned changes in crystallization behavior. The other area where coalescence can be a problem in production of SLN is in the homogenization where elevated temperatures reduce the interfacial tension leading to insufficient repulsive forces between particles and also where the lipid matrix is melted, such as during heat processing, sterilization, and drying. Matrix Stability Westesen and Siekmann93 postulated that recrystallization of the matrix lipids destabilized the suspension via transformation of the initially spherical particles to needle-shaped particles, a phenomenon that they observed with cryo-transmission electron microscopy. They suggested that particles became needle shaped because the lipid matrix adopted the thermodynamically more stable, highly ordered -structure.86,9395 Moreover, the transformation from spherical to needle-shaped particles resulted in increases in surface area, and authors suggested that surfactants needed to cover the newly formed surfaces were unable to diffuse rapidly enough to the newly formed surfaces and therefore resulted in flocculation due to hydrophobic attraction between the two lipophilic patches. Recent research in our laboratory has shown that, in Tween-20-stabilized tripalmitin SLN, a strong relationship between onset of gelation and a reduction in the -crystal content in SLN exists. The recent study confirmed that polymorphic transformation from -form to the more

stable -form may pose a significant problem to the overall stability of the system.83,96 Manufacturing of Solid Lipid Nanoparticles The production of SLN can be achieved with little capital investment because few processing equipment is required. Three principal manufacturing techniques are available,13,38,63,97100 and selection of a proper production technique depends predominantly on the selection of surfactants and lipid components. Melt Homogenization Figure 5 shows the principal steps involved in the production of SLN via melt homogenization. Melt homogenization can be carried out by high-pressure homogenization or highintensity ultrasonication; however, the possibility of metal contamination due to probe degradation when using highintensity ultrasound should be considered unless a coated probe is used. The presence of metal fragments could lead to increased oxidative degradation of the bioactive. In melt homogenization, the carrier lipid is first mixed with the bioactive ingredient using a simple stirrer at a temperature above the melting temperature of the carrier lipid. The mixture is then dispersed in a hot surfactant solution to form a coarse hot emulsion premix. The premix is fed into a highpressure homogenizer equipped with a thermostatted ho-

Fig. 5 Left Flow chart of the melt homogenization process denoting the individual steps leading to formation of solid lipid nanoparticles. Right Schematics of continuous or batch process to manufacture solid lipid nanoparticles by melt homogenization

152

Food Biophysics (2008) 3:146154

Fig. 6 Left Schematic diagram depicting the structures formed during the production of solid lipid nanoparticles using the melt microemulsification process. Right Flow chart of the melt microemulsifica-

tion process denoting the individual steps leading to formation of solid lipid nanoparticles

mogenization chamber to form the fine-disperse nanoemulsion. The hot nanoemulsion is then cooled under controlled conditions, e.g., by pumping the solution through a heat exchanger. Typical lipid content of SLN produced by melt homogenization range from 5 to 10%, although successful production of up SLN suspensions with a lipid content of up to 40% has been reported. Typical conditions for melt highpressure homogenization include a minimum of three to five passes through the homogenizer at homogenization pressures of up to 1,500 bar. Melt microemulsification A preparation technique for SLN using microemulsion was first described by Gasco and co-workers (Figure 6).101 As previously mentioned, microemulsions arise from the spontaneous self-assembly of the hydrophobic or hydrophilic parts of surfactant molecules6 in a solvent giving rise to micelles, wormlike micelles, bilamellar phases, and reverse micelles. The structures are capable of solubilizing significant amounts of one or more lipophilic components. Their sizes range from several nanometers to several tens of nanometers. They are thermodynamically stable, and because they are spontaneously formed virtually, no mechanical energy is required to initiate their formation. SLN can be made from microemulsions by first melting the to-bedispersed lipid and mixing it with hot micellar surfactant solution to form an optically transparent microemulsion. Typically, a number of different surfactants such as lecithin,

polysorbate 20, polysorbate 60, and co-emulsifier, such as bile salts and butanol, are used to form the hot microemulsion. The hot solution is then diluted into cold water (23 C) under stirring thereby solidifying the lipid and/or the surfactant in the micelle. The dilution process is of critical importance, and dilution factors typically range between 1:25 and 1:50.6 Cold Homogenization If the bioactive lipids that are to be delivered in the SLN are temperature-sensitive, degradation during the manufacturing process using melt homogenization or melt microemulsification can occur. In this case, cold homogenization may be used. In cold homogenization, the lipid is briefly heated, and the bioactive compound dispersed followed by rapid cooling of the bulk lipid mixture using liquid nitrogen. The bulk lipid mixture is then milled to form lipid microparticles using, for example, a ball mill. In this step, it is important to make sure that the temperatures during the milling processes do not exceed those of the melting temperatures of either of the components in the lipid matrix. The lipid microparticles are then dispersed in a cold surfactant solution and further milled to produce fine disperse solid lipid particles. While the rapid cooling can lead to formation of the desired crystal structure, the particles are typically larger than those obtained with the other two techniques and therefore do not have the same functionalities in terms of dissolution velocity during mastication and digestion.

Food Biophysics (2008) 3:146154

153 8. H. Chen, J. Weiss, F. Shahidi, Food Technol. 03(06), 3036 (2006) 9. T. M. Taylor, P. M. Davidson, B. D. Bruce, J. Weiss, Crit. Rev. Food Sci. Technol. 45, 587605 (2005) 10. M. P. Desai, V. Labhasetwar, G. L. Amidon, R. J. Levy, Pharm. Res. 13(12), 18381845 (1996) 11. R. H. Muller, K. Mader, S. Gohla, Eur. J. Pharm. Sci. 50(1), 161177 (2000) 12. J. Liu, W. Hu, H. Chen, Q. Ni, H. Xu, X. Yang, Int. J. Pharm. 382(2), 191195 (2007) 13. Y. Luo, D. Chen, L. Ren, X. Zhao, J. Qin, J. Control. Release 114(1), 5359 (2006) 14. R. H. Muller, S. Runge, V. Ravelli, W. Mehnert, A. F. Thunemann, E. B. Souto, Int. J. Pharm. 317(1), 8289 (2006) 15. B. Lu, S.-B. Xiong, H. Yang, X.-D. Yin, R.-B. Chao, Eur. J. Pharm. Sci 28(12), 8695 (2006) 16. N. Pedersen, S. Hansen, A. V. Heydenreich, H. G. Kristensen, H. S. Poulsen, Eur. J. Pharm. Biopharm. 62(2), 155162 (2006) 17. M. A. Schubert, M. Harms, C. C. Muller-Goymann, Eur. J. Pharm. Sci. 27(23), 226236 (2006) 18. H. Chen, X. Chang, D. Du et al., J. Control. Release 110(2), 296306 (2006) 19. S. Lombardi Borgia, M. Regehly, R. Sivaramakrishnan et al., J. Control. Release 110(1), 151163 (2005) 20. I. Friedrich, S. Reichl, C. C. Muller-Goymann, Int. J. Pharm. 305 (12), 167175 (2005) 21. C. Charcosset, A. El-Harati, H. Fessi, J. Control. Release 108(1), 112120 (2005) 22. R. Pandey, S. Sharma, G. K. Khuller, Tuberculosis 85(56), 415420 (2005) 23. A. Lippacher, R. H. Muller, K. Mader, Eur. J. Pharm. Biopharm. 58(3), 561567 (2004) 24. R. H. Muller, C. M. Keck, J. Biotechnol. 113(13), 151170 (2004) 25. S. A. Wissing, O. Kayser, R. H. Muller, Adv. Drug Deliv. Rev. 56(9), 12571272 (2004) 26. F. Q. Hu, Y. Hong, H. Yuan, Int. J. Pharm. 273(12), 2935 (2004) 27. V. Venkateswarlu, K. Manjunath, J. Control. Release 95(3), 627 638 (2004) 28. Z. Mei, H. Chen, T. Weng, Y. Yang, X. Yang, Eur. J. Pharm. Biopharm. 56(2), 189196 (2003) 29. D. Hou, C. Xie, K. Huang, C. Zhu, Biomaterials 24(10), 1781 1785 (2003) 30. J. Kristl, B. Volk, P. Ahlin, K. Gombac, M. Sentjurc, Int. J. Pharm. 256(12), 133140 (2003) 31. S. A. Wissing, R. H. Muller, Int. J. Pharm. 254(1), 6568 (2003) 32. R. H. Muller, M. Radtke, S. A. Wissing, Adv. Drug Deliv. Rev. 54(Suppl 1), S131S155 (2002) 33. R. H. Muller, M. Radtke, S. A. Wissing, Int. J. Pharm. 242(12), 121128 (2002) 34. E. Ugazio, R. Cavalli, M. R. Gasco, Int. J. Pharm. 241(2), 341 344 (2002) 35. S. A. Wissing, R. H. Muller, J. Control. Release. 81(3), 225233 (2002) 36. F. Q. Hu, H. Yuan, H. H. Zhang, M. Fang, Int. J. Pharm. 239(1 2), 121128 (2002) 37. R. Cavalli, M. R. Gasco, P. Chetoni, S. Burgalassi, M. F. Saettone, Int. J. Pharm. 238(12), 241245 (2002) 38. W. Mehnert, K. Mader, Adv. Drug Deliv. Rev. 47(23), 165196 (2001) 39. V. Jenning, K. Mader, S. H. Gohla, Int. J. Pharm. 205(12), 15 21 (2000) 40. V. Jenning, S. Gohla, Int. J. Pharm. 196(2), 219222 (2000) 41. V. Jenning, A. Gysler, M. Schafer-Korting, S. H. Gohla, Eur. J. Pharm. Biopharm. 49(3), 211218 (2000)

Conclusions SLN are an exciting new delivery system with potential for multiple applications in the food and agricultural industries. They are of particular interest to manufacturers of functional foods that are looking for novel ways to include lipophilic but chemically sensitive bioactive compounds. It is important to keep in mind though that SLN cannot be simply regarded as nanoemulsions with a solid core. SLN are structurally much more complex systems, and because of this, they have clear advantages and disadvantages when compared with other colloidal carrier system. For example, based on the chosen manufacturing methodology, it is possible to have simultaneously other colloidal systems such as micelles, mixed micelles, nanocrystals, and even liposomes present. Moreover, due to the complexity involved in the crystallization process in SLN, transformation between the modifications may occur, and if the process is not carried out properly, even supercooled melts may be obtained instead of solid particles. On the other hand, large-scale production is possible, and no organic solvents are required, making this an ideal system for use in the food industry. More importantly, high concentrations of functional compounds in each particle can be achieved without incurring phase separation or destabilization of the particles. SLN also offer a number of advantages in terms of manufacturing and handling procedures as particles can, for example, be lyophilized or spray-dried if carrier lipids with melting temperatures below 70 C are used to obtain shelf-stable powders.

Acknowledgments This material is based upon the work supported by the Cooperative State Research, Extension, Education Service, United State Department of Agriculture, Massachusetts Agricultural Experiment Station (Project No. 911 and 836) and United States Department of Agriculture, CREES, NRI Grants (Award Number 2004-35201 and 2005-35503).

References
1. CDC National Center for Chronic Disease Prevention (Office of Communication) (2005), http://www.cdc.gov/od/oc/media/pressrel/ fs051026.htm. Accessed 21 November 2007 2. B. H. Stewart, P. Kleihues, World Cancer ReportInternational Agency for Research on Cancer. (World Health Organization, New York, 2003) 3. IFIC (2006), http://www.ific.org/nutrition/functional/index.cfm. Accessed 20 November 2007 4. D. A. Groneberg, M. Giersig, T. Welte, U. Pison, Current Drug Targets 7(6), 643648 (2006) 5. J. Weiss, P. Takhistov, D. J. McClements, J. Food Sci. 71(9), 107116 (2006) 6. J. Flanagan, H. Singh, Crit. Rev. Food Sci. Nutr. 46(3), 221237 (2006) 7. A. zur Muhlen, C. Schwarz, W. Mehnert, Eur. J. Pharm. Biopharm. 45(2), 149155 (1998)

154 42. C. S. Maia, W. Mehnert, M. Schafer-Korting, Int. J. Pharm. 196 (2), 165167 (2000) 43. G. Lukowski, J. Kasbohm, P. Pflegel, A. Illing, H. Wulff, Int. J. Pharm. 196(2), 201205 (2000) 44. E. Zimmermann, R. H. Muller, K. Mader, Int. J. Pharm. 196(2), 211213 (2000) 45. V. Jenning, K. Mader, S. H. Gohla, Int. J. Pharm. 205(12), 15 21 (2000) 46. N. Scholer, E. Zimmermann, U. Katzfey, H. Hahn, R. H. Muller, O. Liesenfeld, Int. J. Pharm. 196(2), 235239 (2000) 47. R. Cavalli, E. Peira, O. Caputo, M. R. Gasco, Int. J. Pharm. 182 (1), 5969 (1999) 48. H. Heiati, R. Tawashi, N. C. Phillips, Int. J. Pharm. 174(12), 7180 (1998) 49. C. Freitas, R. H. Muller, Eur. J. Pharm. Biopharm. 46(2), 145 151 (1998) 50. C. Schwarz, W. Mehnert, Int. J. Pharm. 157(2), 171179 (1997) 51. R. H. Muller, S. Maassen, C. Schwarz, W. Mehnert, J. Control. Release 47(3), 261269 (1997) 52. A. J. Almeida, S. Runge, R. H. Muller, Int. J. Pharm. 149(2), 255265 (1997) 53. R. Cavalli, O. Caputo, M. E. Carlotti, M. Trotta, C. Scarnecchia, M. R. Gasco, Int. J. Pharm. 148(1), 4754 (1997) 54. H. Heiati, R. Tawashi, R. R. Shivers, N. C. Phillips, Int. J. Pharm. 146(1), 123131 (1997) 55. R. H. Muller, D. Ruhl, S. A. Runge, Int. J. Pharm. 144(1), 115 121 (1996) 56. R. H. Muller, C. Freitas, A. zur Muhlen, W. Mehnert, Eur. J. Pharm. Sci. 4(Supp 1), S75S75 (1996) 57. A. Dingler, S. Runge, R. H. Muller, Eur. J. Pharm. Sci. 4(Suppl 1), S132S132 (1996) 58. S. Runge, W. Mechnert, R. H. Muller, Eur. J. Pharm. Sci. 4 (Suppl 1), S132S132 (1996) 59. K. Westesen, H. Bunjes, M. H. J. Koch, J. Control. Release 48 (23), 223236 (1997) 60. A. Radomska-Soukharev, R. H. Muller, Pharmazie 61(5), 425 430 (2006) 61. M. A. Schubert, C. C. Muller-Goymann, Eur. J. Pharm. Biopharm. 61(12), 7786 (2005) 62. V. Jenning, A. F. Thunemann, S. H. Gohla, Int. J. Pharm. 199(2), 167177 (2000) 63. Y. Yang, J. F. Feng, H. Zhang, J. Y. Luo, Zhongguo Zhong Yao Za Zhi 31(8), 650653 (2006) 64. L. K. Zhang, S. X. Hou, S. J. Mao, D. P. Wei, X. R. Song, Y. Lu, Int. J. Pharm. 287(12), 155162 (2004) 65. R. Sivaramakrishnan, C. Nakamura, W. Mehnert, H. C. Korting, K. D. Kramer, M. Schafer-Korting, J. Control. Release 97(3), 493502 (2004) 66. M. A. Videira, M. F. Botelho, A. C. Santos, L. F. Gouveia, J. J. de Lima, A. J. Almeida, J. Drug Target. 10(8), 607613 (2002) 67. Y. Wang, W. Wu, Drug Deliv. 13(3), 189192 (2006) 68. A. Saupe, K. C. Gordon, T. Rades, Int. J. Pharm. 314(1), 5662 (2006) 69. F.-Q. Hu, S.-P. Jiang, Y.-Z. Du, H. Yuan, Y.-Q. Ye, S. Zeng, Colloids Surf B Biointerfaces 45(34), 167173 (2005)

Food Biophysics (2008) 3:146154 70. F. Castelli, C. Puglia, M. G. Sarpietro, L. Rizza, F. Bonina, Int. J. Pharm. 304(12), 231238 (2005) 71. E. B. Souto, S. A. Wissing, C. M. Barbosa, R. H. Muller, Int. J. Pharm. 278(1), 7177 (2004) 72. E. B. Souto, S. A. Wissing, C. M. Barbosa, R. H. Muller, Eur. J. Pharm. Biopharm. 58(1), 8390 (2004) 73. K. Jores, W. Mehnert, M. Drechsler, H. Bunjes, C. Johann, K. Mader, J. Control. Release 95(2), 217227 (2004) 74. E. B. Souto, R. H. Muller, Pharmazie 61(5), 431437 (2006) 75. M. Uner, Pharmazie 61(5), 375386 (2006) 76. E. B. Souto, R. H. Muller, J. Microencapsul. 23(4), 377388 (2006) 77. E. B. Souto, R. H. Muller, J. Microencapsul. 22(5), 501510 (2005) 78. K. Sato, N. Garti, Crystallization and polymorphism of fats and fatty acids (Marcel Dekker, New York, 1988) 79. W. Mehnert, K. Mader, Adv. Drug Deliv. Rev. 47(23), 165196 (2001) 80. H. Bunjes, M. H. J. Koch, K. Westesen, J. Pharm. Sci. 92(7), 15091520 (2003) 81. G. Worle, B. Siekmann, M. H. Koch, H. Bunjes, Eur. J. Pharm. Sci. 27(1), 4453 (2006) 82. D. J. McClements, Food emulsions: principles, practice, and techniques, 2nd edn. (CRC, Boca Raton, 1999) 83. T. Helgason, T. Awad, E. Decker, K. Kristbergsson, D. J. McClements, J. Weiss, J. Am. Oil Chem. Soc. In press (2008) 84. T. Unruh, H. Bunjes, K. Westesen, M. H. J. Koch, J. Phys. Chem. B 103(47), 1037310377 (1999) 85. K. Westesen, B. Siekmann, M. H. J. Koch, Int. J. Pharm. 93(1 3), 189199 (1993) 86. V. Jenning, M. Schafer-Korting, S. Gohla, J. Control. Release 66 (23), 115126 (2000) 87. B. Siekmann, K. Westesen, Colloids Surf. 3, 159175 (1994) 88. J. Palanuwech, J. N. Coupland, Colloids Surf. A Physicochem. Eng. Asp. 223(13), 251262 (2003) 89. C. Freitas, R. H. Muller, J. Microencapsul. 16(1), 5971 (1999) 90. C. Freitas, R. H. Muller, Int. J. Pharm. 168(2), 221229 (1998) 91. D. J. McClements, Food Hydrocoll. 14(2), 173177 (2000) 92. E. Dickinson, D. J. McClements, in Advances in food colloids, ed. by E. Dickinson, D. J. McClements (Blackie Academic and Professional, Glasgow, 1995), pp. 211244 93. K. Westesen, B. Siekmann, Int. J. Pharm. 151(1), 3545 (1997) 94. H. Bunjes, F. Steiniger, W. Richter, Langmuir. 23(7), 40054011 (2007) 95. S. Hatziantoniou, G. Deli, Y. Nikas, C. Demetzos, G. T. Papaioannou, Micron. 38(8), 819823 (2007) 96. T. Awad, T. Helgason, K. Kristbergsson E. A. Decker, J. Weiss, D. J. McClements, Food Biophys. In press (2008) DOI 10.1007/ s11483-008-9057-8 97. A. Dingler, S. Gohla, J. Microencapsul. 19(1), 1116 (2002) 98. S. H. Gohla, A. Dingler, Pharmazie. 56(1), 6163 (2001) 99. S. R. Mao, P. Wang, D. Z. Bi, Pharmazie. 60(4), 273277 (2005) 100. M. A. Casadei, F. Cerreto, S. Cesa, et al., Int. J. Pharm. 325(1), 140160 (2006) 101. M. R. Gasco, Patent No. 5,250,236 (1993)