Innovative Food Science and Emerging Technologies 19 (2013) 29–43

Contents lists available at SciVerse ScienceDirect

Innovative Food Science and Emerging Technologies

journal homepage: www.elsevier.com/locate/ifset

Nanostructured lipid carriers (NLC): A potential delivery

system for bioactive food molecules
Fardin Tamjidi a,⁎, Mohammad Shahedi a, 1, Jaleh Varshosaz b, 1, Ali Nasirpour a
a
Department of Food Science and Technology-Center of Excellence for Safety and Quality of Foods, College of Agriculture, Isfahan University of Technology, Isfahan 84156-83111, Iran
b
Department of Pharmaceutics, Faculty of Pharmacy and Novel Drug Delivery Systems Research Center, Isfahan University of Medical Sciences, Isfahan 81746-73461, Iran

a r t i c l e i n f o a b s t r a c t

Article history: Particle size and physical state of the lipid phase are major factors influencing the permanence of lipid disper-
Received 11 November 2012 sions. Nanostructured lipid carriers (NLC) are a delivery system in which partial-crystallized lipid particles
Accepted 17 March 2013 with mean radii ≤100 nm are dispersed in an aqueous phase containing emulsifier(s), as a potential delivery
system may have some advantages in certain circumstances when compared with other colloidal carriers. NLC
Editor Proof Receive Date 17 April 2013
are a useful nutraceutical delivery system with high drug loading, encapsulation efficiency and stability. They
Keywords:
may increase, bioavailability and stability of bioactive compounds, and shelf-life, consumer acceptability, func-
Lipid nanocarriers tionality, nutritional value and safety of food systems, and provide controlled release of encapsulated materials.
Lipid nanoparticles In this review, beneficial aspects of NLC are presented and valuable information about ingredients, production
Lipid crystallization methods, structure and characteristics of them provided. Moreover, potential applications and disadvantages
Bioactive compounds of NLC as emerging delivery system in food science are introduced.
Nutraceuticals Industrial relevance: With the increasing public perception of a strong correlation between food and disease pre-
Functional foods vention, producers are trying to enrich staple foods and beverages with nutraceuticals and produce functional
foods. Nonetheless, fortification of aqueous-based food with many of nutraceuticals is greatly limited owing to
their poor water-solubility, chemical instability, and low bioavailability. NLC are a novel nanocarrier that may
dispel these limitations, combine the advantages of other lipid nanocarriers and avoid some of their disadvan-
tages. They may be suitable for application within foods and transparent/opaque beverages.
© 2013 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
2. NLC versus liposomes, nanoemulsions, microemulsions and SLN . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
2.1. Liposomes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
2.2. Nanoemulsions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
2.3. Microemulsions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
2.4. SLN . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
2.5. NLC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
3. NLC ingredients . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.1. Lipids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.1.1. Liquid lipids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.1.2. Solid lipids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
3.2. Emulsifiers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
4. Production of NLC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
4.1. Hot homogenization method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
4.2. Cold homogenization method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
4.3. Solvent emulsification–evaporation method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
5. Structure of NLC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
6. Characterization of NLC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

⁎ Corresponding author. Tel.: + 98 311 3913384; fax: + 98 311 3912254.

E-mail addresses: f.tamjidi@ag.iut.ac.ir (F. Tamjidi), shahedim@cc.iut.ac.ir (M. Shahedi), varshosaz@pharm.mui.ac.ir (J. Varshosaz), ali.nasirpour@cc.iut.ac.ir (A. Nasirpour).
1
Both authors contributed equally to this work.

1466-8564/$ – see front matter © 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.ifset.2013.03.002
30 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43

6.1. Particle size and zeta (ζ)-potential . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

6.2. Particle morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
6.3. Chemical stability of bioactive compounds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
6.4. Encapsulation efficiency (EE) and drug loading (DL) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
6.5. Crystallinity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
7. Food applications of NLC — Perspective and challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
8. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

1. Introduction
According to National Nanotechnology Initiative, nanotechnology is
generally defined as “the understanding and control of matter at the
nanoscale, at dimensions between roughly 1 and 100 nm, where unique
phenomena enable novel applications”. A nanomaterial is also defined as
“a discrete entity that has one or more dimensions of the order of
100 nm or less” elsewhere (SCENIHR, 2007). The physicochemical prop-
erties (such as color, solubility, viscosity, diffusivity, material strength
and toxicity) and biological properties of structures and systems at nano-
scale are substantially different than the macro-scale counterparts owing
to the interactions of individual atoms and molecules thereby offering
unique and novel functional applications (Neethirajan & Jayas, 2011).
The potential benefits for application of nanotechnologies in food include
many aspects, such as packaging materials, efficient nutrient delivery
systems, formulations with improved bioavailability and new tools for
molecular and cellular biology (Chaudhry et al., 2008; Chen, Weiss, &
Shahidi, 2006; Das, Saxena, & Dwivedi, 2009; Maynard et al., 2006;
Moraru, Huang, Takhistov, Dogan, & Kokini, 2009; Weiss, Takhistov, &
McClements, 2006). Delivery system is defined as the entrapment of a
bioactive material in a carrier to control its release rate (Fathi, Mozafari,
& Mohebbi, 2012). In comparison to micro-delivery systems, the nano-
ones have some advantages. Nanocarriers are quasi-monodisperse, are
thermodynamically stable or show long-term kinetic stability (Weiss et
al., 2008). These systems provide more surface area and have the poten-
tial to enhance the solubility and bioavailability of the bioactive materials;
finally the nano-delivery systems improve controlled release and
targeting of the encapsulated food ingredients (Mozafari, 2006; Weiss,
Gaysinsky, Davidson, & McClements, 2009). It should be pointed that larg-
er size ranges mostly up to 600 nm have been reported for nanocarriers in
the literature. This skew from the “nano-scale definition” presumably
arises from these reasons: these systems contain a portion of particles
that are 1–100 nm in size; the particles with the sizes up to 600 nm
have same effects in the human gastrointestinal (GI) tract as ones with
1–100 nm as; the particles are assumed to be spherical and the dimen-
sion may be considered either the radius or the diameter.
Nano-delivery systems are valuable prospects for nanotechnology in
pharmaceutics, cosmetics as well as food industries. These systems can
generally be divided into two groups: polymer- and lipid-based systems
(Sawant & Dodiya, 2008); The number of products based on the former
in the market is limited because of the potential toxicity of polymers,
the lack of suitable large-scale production methods (Chen, Tsai, Huang,
& Fang, 2010; Weiss et al., 2008), the low availability of suitable biopoly-
mers and the need to use organic solvents (Müller, Mäder, & Gohla,
2000). In addition, most of the bioactive compounds (e.g. fatty acids,
carotenoids, tocopherols, flavonoids, polyphenols, phytosterols, oil
soluble vitamins and nutraceuticals), flavors, and preservatives have
hydrophobic nature. For encapsulating these lipophilic compounds,
colloidal dispersions suitable in aqueous environments (i.e., oil in
water type) are often required. Finally, the presence of digestible lipid
often facilitates the absorption of bioactive compounds in the small
intestine because it increases the amount of mixed micelles available
to solubilize and transport hydrophobic compounds (McClements,
2012a; Porter, Trevaskis, & Charman, 2007; Pouton, 2006). Thus, a
great interest has focused on the major lipid-based nanocarriers includ-
ing nanoemulsions, microemulsions, liposomes, solid lipid nanoparticles
(SLN), and nanostructured lipid carriers (NLC). It seems that NLC have
some advantages over the other lipidic nanocarriers in certain condi-
tions. Therefore this paper particularly concentrates on the presentation
including composition, manufacturing, structure and features of NLC.
Moreover, we will briefly discuss their emerging and potential uses in
food science as a novel delivery system. So this article does not consider
nanoemulsions, microemulsions, liposomes and SLN, which have been
reviewed elsewhere (Flanagan & Singh, 2006; Garti, 2003; Keller, 2001;
McClements, 2012b; McClements & Rao, 2011; Mehnert & Mäder,
2012; Mozafari, Johnson, Hatziantoniou, & Demetzos, 2008; Taylor,
Weiss, Davidson, & Bruce, 2005; Weiss et al., 2008).
2. NLC versus liposomes, nanoemulsions, microemulsions and SLN
Before discussing NLC, it is useful to begin with a brief overview of
the other major lipid nanocarriers in general, focusing on the defects
of these nanocarriers in comparison to NLC.
2.1. Liposomes
Liposomes are spherical vesicles that form with the hydration of
surfactants such as phospholipids when mixed with water under low
shear force. Phospholipids arrange themselves in sheets; by joining tails
and tails of them, these sheets form a bilayer membrane that contains
some water inside a phospholipid vesicle (Jin et al., 2008). Liposome is a
flexible carrier, can contain both water-soluble and water-insoluble com-
pounds simultaneously and provides targetability (Mozafari et al., 2008),
but currently, high cost, low payload, drug leakage and fast release are
main disadvantages of this delivery system. Employing this nanocarrier
might incur stability problems during storage (Mu & Zhong, 2006).
2.2. Nanoemulsions
A nano- or mini-emulsion can be considered to be an emulsion
that contains very small droplets with mean radii between about
10 to 100 nm (Mason, Wilking, Meleson, Chang, & Graves, 2006;
Tadros, Izquierdo, Esquena, & Solans, 2004). The droplet is typically
stabilized by a layer that consists of a mixture of surfactant and
co-surfactant. Similar to microemulsions, nanoemulsions are optically
transparent or only slightly turbid. Both nano- and micro-emulsions
have much better stability to gravitational separation and aggregation
than conventional-emulsions (i.e., emulsions contain droplets with
mean radii larger than 100 nm) (McClements & Rao, 2011; Tadros et
al., 2004; Wooster, Golding, & Sanguansri, 2008). However unlike
microemulsions, nanoemulsions (and also liposomes) are thermodynam-
ically unstable, are long-term kinetically stable. This is the major distinc-
tion between a nanoemulsion and a microemulsion (McClements,
2012b). Nanoemulsions have toxicological safety, high content of
the lipid phase and the possibility of large-scale production (Fathi,
Mozafari, et al., 2012). However, they still undergo instabilities
such as Ostwald ripening (Klang, Matsko, Valenta, & Hofer, 2012;
Mason et al., 2006; McClements & Rao, 2011), flocculation,
 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
membrane permeability and drug release (Fig. 1), creaming, gelling,
and particle aggregation during storage (Varshosaz, Eskandari, &
Tabbakhian, 2012). Moreover nanoemulsions are unstable in acidic
conditions and controlled drug release from nanoemulsions
is very improbable due to their small size and the liquid state of the
carrier (Fathi, Mozafari, et al., 2012). Lastly, the chemical stability of
nanoemulsions is questionable.
2.3. Microemulsions
Microemulsion also known as swollen micelle, typically contains
droplets with radii between the range of 2 to 100 nm (McClements &
Rao, 2011). Fabricating nanoemulsions and microemulsions usually
requires fairly similar ingredients: an oil phase, an aqueous phase, a
surface active agent, and probably a co-surfactant (McClements,
2012b). Microemulsions have complicated nature and require special
oils and synthetic surfactants, very high concentration of surfactant
and precise production approach. Moreover they are prone to disinte-
gration upon the changes of environmental conditions (e.g. composi-
tion and temperature) (McClements & Rao, 2011). Finally, it is often
difficult to incorporate large lipophilic molecules within the particle
structure while maintaining the optimum curvature of the surfactant
monolayer (Coupland, Weiss, Lovy, & McClements, 1996). Currently,
these difficulties limit use of microemulsion as delivery system for
many of food applications.
2.4. SLN
Functional properties of oil-in-water (O/W) nano-/micro-emulsions
can often be improved by structurally designing and fabricating emulsion
systems using appropriate ingredients and processing operations. It is
possible to produce lipidic nanoparticles from nano-/micro-emulsions
by using a lipid phase that is able to fully or partially crystallize at the
final temperature of application (body/room temperature) (Müller,
Radtke, & Wissing, 2002a). SLN contain lipid droplets that are fully crys-
tallized and have a highly-ordered crystalline structure with the bioactive
components being a part of the lipid matrix (Radomska-Soukharev &
Müller, 2006; Uner, 2006; Weiss et al., 2008), were introduced at the
beginning of the 1990s. SLN were designed to combine the advantages
of polymeric particles, liposomes and emulsions (Gramdorf et al., 2008)
and to avoid some of their disadvantages. The SLN are produced from
one solid lipid or a blend of solid lipids (Müller et al., 2002a). The mobility
Fig. 1. Schematic presentation of defects of nanoemulsion [in comparison to SLN; Upper: Modified from Ref.(Weiss, Decker, McClements, Kristbergsson, Helgason, & Awad, 2008)]
and SLN (Lower). After preparation of SLN by cooling a hot nanoemulsion, the droplets crystallize, at least partially, in higher energy modifications α and β′. During storage, these
modifications transform to the low energy, more ordered β modification. A burst release is observed for nanoemulsion and for SLN during polymorphism.
31
of bioactive component can be controlled by controlling the physical state
of the lipid matrix of SLN (Wang & Wu, 2006; Yang, Feng, Zhang, &
Luo, 2006). Bioactive substance has much lower diffusion rate,
therefore prolonged release can be achieved by SLN (Fig. 1). Overall,
proposed advantages for SLN include: possibility of controlled drug
release and drug targeting, having high encapsulation efficiency,
avoidance of organic solvents, feasibility for incorporation of lipo-
philic and hydrophilic drugs, possibility of large-scale production
and sterilization, no bio-toxicity of the carrier, increased drug stability
(Mäder & Mehnert, 2005; Mehnert & Mäder, 2001), having high physi-
cal stability and excellent tolerability (Singhal, Patel, Prajapati, & Patel,
2011). However, common deficiencies of SLN include low drug loading
capacity and drug expulsion (burst release) after polymorphic transi-
tion (i.e., α-crystal → β -crystal → β-crystal) during storage (Fig. 1)
(Mehnert & Mäder, 2001; Müller, Radtke, & Wissing, 2002b; Müller et
al., 2002a; Yuan et al., 2007), particularly if the lipid matrix consists of
similar molecules (Almeida & Souto, 2007); the increase in perfection
of the crystal leaves less space to accommodate drug molecules. In addi-
tion to polymorphic transition, the formation of drug-enriched shell
leads to burst release (Müller, Petersen, Hommoss, & Pardeike, 2007).
Moreover SLN have high water content (70–99.9%) (Mehnert &
Mäder, 2001; Müller et al., 2002a,b; Radtke & Müller, 2001). These
disadvantages make SLN improper for some of food applications as
delivery system.
2.5. NLC
The process of full-crystallization or recrystallization of fat reduces
the drug solubility, leads to drug expulsion from the lipid nanoparticles
especially when the drug concentration in the formulation is too high.
Majority of drugs have higher solubility in a liquid lipid rather than a
solid lipid. NLC are a modified SLN in which the lipidic phase contain
both solid (fat) and liquid (oil) lipids at room temperature (Müller et
al., 2002a,b). The NLC or oil-loaded SLN contain lipid droplets that are
partially crystallized and have a less-ordered crystalline structure or an
amorphous solid structure, which were developed for overcoming the
limitations of SLN by Müller et al. at the end of the 1990s. The purpose
of NLC-formulation is to produce particles in which the oil is incorporat-
ed into the core of a solid lipid; this leads to a higher loading capacity and
controlled drug release as the drug dissolves in the oil and simultaneous-
ly encapsulates in the solid lipid (Varshosaz, Eskandari, & Tabakhian,
2010). The lipid blend in NLC has slower polymorphic transition and
32 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43

Table 1
Commonly used lipids and emulsifiers for preparation of NLC.

Ingredient Reference

Liquid lipid

Soybean oil Chinsriwongkul et al. (2011); Zhang, Chen, Zhang, Shen, & Pan (2011)
Medium chain triglycerides (MCT)/caprylic- and Cirri, Bragagni, Mennini, & Mura (2012), Esposito, Mariani, Ravani, Contado, Voltado, Bido et al. (2012), Fathi, Varshosaz,
capric-triglycerides Mohebbi, & Shahidi (2012), Guo, Yanga, Li, Tan, Xi, Liud et al. (2012), Haag, Chen, Peters, Keck, Taskoparan, Fahr et al. (2011),
Han, Li, Yin, Liu, & Xu (2008), Hu, Jiang, Du, Yuan, Ye, & Zeng (2006), Jia, Zhang, Li, Duan, Wang, Feng et al. (2010), Jores,
Mehnert, Drechsler, Bunjes, Johan, & Mäder (2004), Li, Weng, Wang, He, Yang, & Tang (2010), Li, Lin, Yu, Li, Geng, & Zheng
(2009), Lin, Li, Zheng, Yu, Zhang, & Liu (2007), Luo, Zhao, Zhang, & Pan (2011), Mitri, Shegokar, Gohla, Anselmi, & Müller
(2011), Nayak, Tiyaboonchai, Patankar, Madhusudhan, & Souto (2010), Obeidat, Schwabe, Müller, & Keck (2010), Puglia,
Frasca, Musumecim, Rizza, Puglisi, Bonina et al. (2012), Sanad, AbdelMalak, elBayoomy, & Badawi (2010), Saupe, Gordon, &
Rades (2006), Saupe, Wissing, Lenk, Schmidt, & Müller (2005), Teeranachaideekul, Souto, Junyaprasert, & Müller (2007),
Varshosaz, Hassanzadeh, Sadeghi, & Khadem (2012), Xia, Saupe, Müller, & Souto (2007), Zhang, Liu, Qiao, Liu, Ni, Zhang et al.
(2010), Zhuang, Li, Wang, Zhang, Pan, Peng et al. (2010)
Oleic acid Chinsriwongkul et al. (2011), Fathi, Varshosaz, Mohebbi, & Shahidi (2012), Hu, Jiang, Du, Yuan, Ye, & Zeng (2005), Kuo &
Chung (2011), Li, Weng, Wang, He, Yang, & Tang (2010), Liu, Liu, Wang, Zhang, & Zhang (2011), Pardeike et al. (2011), Sanad,
AbdelMalak, elBayoomy, & Badawi (2010), Tiwari & Pathak (2011), Varshosaz, Hassanzadeh, Sadeghi, & Andalib (2012),
Varshosaz, Hassanzadeh, Sadeghi, & Khadem (2012), Yuan, Wang, Du, You, Hu, & Zeng (2007)
α-tocopherol/vitamin E Souto & Müller (2005), Tsai et al. (2012)
Corn oil Liu & Wu (2010), Mitri, Shegokar, Gohla, Anselmi, & Müller (2011), Nguyen, Hwang, Park, & Park (2012)
Squalene Araújo, Nikolic, Egea, Souto, & Garcia (2011), Chen, Tsai, Huang, & Fang (2010), Fang, Fang, Liu, & Su (2008)

Solid lipid

Stearic acid Fathi, Varshosaz, Mohebbi, & Shahidi (2012), Guo et al. (2012), Hu, Jiang, Du, Yuan, Ye, & Zeng (2005), Kuo & Chung (2011), Liu, Liu,
Wang, Zhang, & Zhang (2011), Yuan, Wang, Du, You, Hu, & Zeng (2007)
Carnauba wax (Cera carnauba) Mitri, Shegokar. Gohla, Anselmi, & Müller (2011), Xia, Saupe, Müller & Souto (2007)
Glyceryl tristearate/tristearin Ali, El-Sayed, Sylvester, & Nazzal (2010), Esposito et al. (2012), Haag et al. (2011), Nayak, Tiyaboonchai, Patankar,
Madhusudhan, & Souto (2010)
Glyceryl monostearate Fathi, Varshosaz, Mohebbi, & Shahidi (2012), Guo et al. (2012), Han, Li, Yin, Liu, & Xu (2008), Hu, Jiang, Du, Yuan, Ye, & Zeng
(2006), Jia et al. (2010), Lacatusu, Badea, Ovidiu, Bojin, & Meghea (2012), Li, Weng, He, Yang, & Tang (2010), Liu, Liu, Wang,
Zhang, & Zhang (2011) Müller, Radtke, & Wissing (2002b), Nayak, Tiyaboonchai, Patankar, Madhusudhan, & Souto (2010),
Sanad, AbdelMalak, elBayoomy, & Badawi (2010), Teeranachaideekul, Müller, & Junyaprasert (2007), Tiwari & Pathak (2011),
Varshosaz, Hassanzadeh, Sadeghi, & Khadem (2012), Yuan, Wang, Du, You, Hu, & Zheng (2007), Zhang, Chen, Zhang, Shen, &
Pan (2011), Zhang et al. (2010), Zheng et al. (2013), Zhuang et al. (2010)
Cetyl palmitate Ali, El-Sayed, Sylvester, & Nazzal (2010), Chinsriwongkul et al. (2011), Eskandari, Varshosaz, Minaiyan, & Tabbakhian (2011),
Müller, Radtke, & Wissing (2002b), Obeidat, Schwabe, Müller, & Keck (2010), Saupe, Gordon, & Rades (2006), Saupe, Wissing,
Lenk, Schmidt, & Müller (2005), Teeranachaideekul, Souto, Junyaprasert, & Müller (2007), Varshosaz, Eskandari, & Tabakhian
(2010), Varshosaz, Eskandari, & Tabbakhian (2012), Xia, Saupe, Müller, & Souto (2007)
Glyceryl monocaprate Li, Lin, Yu, Li, Geng, & Zheng (2009), Lin, Li, Zheng, Yu, Zhang, & Liu (2007)
Glyceryl palmitostearate Ali, El-Sayed. Sylvester, & Nazzal (2010) Araújo, Nikolic, Egea, Souto, & Garcia (2011), Chen, Tsai, Huang, & Fang (2010),
Doktorovová, Araújo, Garcia, Rakovsky, & Souto (2010), Fang, Fang, Liu, & Su (2008), Kasongo, Pardeike, Müller, & Walker
(2011), Li & Ge (2012), Liu & Wu (2010), Pardeike et al. (2011), Puglia et al. (2012)
Glyceryl behenate Ali, El-Sayed, Sylvester, & Nazzal (2010), Cirri, Bragagni, Mennini, & Mura (2012), Fathi, Varshosaz, Mohebbi, & Shadidi (2012) Jores,
Mehnert, Drechsler, Bunjes, Johann & Mäder (2004), Kuo & Chung (2011), Luo, Zhao, Zhang, & Pan (2011), Müller, Radtke, & Wissing
(2002b), Souto & Müller (2005), Varshosaz, Eskandari, & Tabakhian (2010), Xia, Saupe, Müller, & Souto (2007), Zhuang et al. (2010)
Propylene glycol monostearate Gramdorf et al. (2008), Hentschel, Gramdorf, Müller, & Kurz (2008)

Emulsifier

Tween 80 Chinsriwongkul et al. (2011), Doktorovová, Araújo, Garcia, Rakovsky, & Souto (2010), Fang, Fang, Liu, & Su (2008), Fathi, Varshosaz,
Mohebbi, & Shahidi (2012), Gramdorf et al. (2008), Han, Li, Yin, Liu, & Xu (2008), Hentschel, Gramdorf, Müller, & Kurz (2008), Hung,
Basri, Tejo, Ismail, Nang, Hassan et al. (2011), Kuo & Chung (2011), Lacatusu, Badea, Ovidiu, Bojin, & Meghea (2012), Li & Ge (2012), Li,
Lin, Yu, Li, Geng, & Zheng (2009), Lin, Li, Zheng, Yu, Zhang & Liu (2007), Luo, Zhao, Zhang, & Pan (2011), Nayak, Tiyaboonchai,
Patankar, Madhusudhan, & Souto (2010), Nguyen, Hwang, Park, & Park (2012), Puglia et al. (2012), Teeranachaideekul, Müller, &
Junyaprasert (2007), Varshosaz, Eskandari, & Tabakhian (2010), Varshosaz, Hassanzadeh, Sadeghi, & Andalib (2012), Varshosaz,
Hassanzadeh, Sadeghi, & Khadem (2012), Zhang et al. (2010)
Lecithin Chen, Tsai, Huang, & Fang (2010), Chinsriwongkul et al. (2011), Doktorovová, Araújo, Garcia, Rakovsky, & Souto (2010), Eskandari,
Varshosaz, Minaiyan, & Tabbakhian (2011), Fang, Fang, Liu, & Su (2008), Guo et al. (2012), Han, Li, Yin, Liu, & Xu (2008), Jia et al. (2010),
Kuo & Chung (2011), Lacatusu, Badea, Ovidiu, Bojin, & Meghea (2012), Li & Ge (2012), Liu, Liu, Wang, Zhang, & Zhang (2011), Liu, Wang,
& Xia (2012), Nguyen, Hwang, Park, & Park (2012), Tsai et al. (2012), Varshosaz, Eskandari, & Tabbakhian (2010), Varshosaz, Eskandari,
& Tabbakhian (2012), Varshosaz, Hassanzadeh, Sadeghi, & Andalib (2012), Varshosaz, Hassanzadeh, Sadeghi, & Khadem (2012), Zhang,
Chen, Zhang, Shen, & Pan (2011), Zheng et al. (2013), Zhuang et al. (2010)
Poloxamer 188 Ali, El-Sayed. Sylvester, & Nazzal (2010), Araújo, Nikolic, Egea, Souto, & Garcia (2011), Chen, Tsai, Huang, & Fang (2010), Cirri,
Bragagni, Mennini, & Mura (2012), Eskandari, Varshosaz, Minaiyan, & Tabbakhian (2011), Esposito et al. (2012), Fang, Fang, Liu, &
Su (2008), Han, Li, Yin, Liu & Xu (2008), Hu, Jiang, Du, Yuan, Ye, & Zeng (2006), Jia et al. (2010), Jores, Mehnert, Drechsler, Bunjes,
Johann, & Mäder (2004), Li, Weng, Wang, He, Yang, & Tang (2010), Liu & Wu (2010), Liu, Liu, Wang, Zhang (2011), Nayak,
Tiyaboonchai, Patankar, Madhusudhan, & Souto (2010), Puglia et al. (2012), Souto & Müller (2005), Tsai et al. (2012), Varshosaz,
Eskandari, & Tabbakhian (2010), Varshosaz, Eskandari, & Tabbakhian (2012), Zhang et al. (2010), Zhuang et al. (2010)
Myverol 18-04K Chen, Tsai, Huang, & Fang (2010). Fang, Fang, Liu, & Su (2008), Liu & Wu (2010), Nayak, Tiyaboonchai, Patankar,
Madhusudhan, & Souto (2010)
Polyglyceryl-3-methylglucose distearate Obeidat, Schwabe, Müller, & Keck (2010), Saupe, Gordon, & Rades (2006), Saupe, Wissing, Lenk, Schmidt, & Müller (2005),
Teeranachaideekul, Müller, & Junyaprasert (2007), Teeranachaideekul, Souto, Junyaprasert, & Müller (2007)
Sodium dodecyl sulfate (SDS) Hu, Jiang, Du, Yuan, & Zeng (2005, 2006)
Sodium deoxycholate (SDC) Han, Li, Yin, Liu, & Xu (2008), Li, Weng, Wang, He, Yang, & Tang (2010), Souto & Müller (2005)
Tween 20 Lacatusu, Badea, Ovidiu, Bojin, & Meghea (2012), Varshosaz, Hassanzadeh, Sadeghi, & Andalib (2012), Yuan, Wang, Du, You,
Hu, & Zeng (2007), Zheng et al. (2013)
 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
low crystallinity index. Along with existing oil in the NLC-core, the spher-
ical shape of the particle may account for these improving properties
(Jores et al., 2004; Müller et al., 2002a). It therefore seems that NLC dispel
the SLN drawbacks and possess the advantages of SLN and thereupon
other lipid nanocarriers. Thus, NLC may increase encapsulation efficien-
cy, drug loading and physical stability and it seems to be a valuable
option for improving the chemical stability, bioavailability and controlled
release of functional lipophilic compounds in food. NLC immobilize the
drug in the solid particle matrix and protect the incorporated bioactive
compound from degradation (Pardeike, Hommoss, & Müller, 2009;
Teeranachaideekul, Müller, & Junyaprasert, 2007); the lipid matrix as a
physical barrier may protect encapsulated sensitive active compound
from damaging factors in aqueous phase. NLC as a new delivery system
are in entering stage of food domain.
3. NLC ingredients
NLC are a nano-particulate carrier system derived from O/W
nanoemulsions. Like nano- and micro-emulsions, the major ingredients
of NLC are lipid, surface active agent and water. The most used lipids
and surfactants for preparation of active material-loaded NLC are listed
in Table 1. In NLC, a major portion of the oil constituent of the O/W
emulsion is replaced by a solid lipid leading to a solid particle matrix
of this carrier system at body temperature (Jenning, Mäder, & Gohla,
2000; Jores et al., 2004). To obtain the blends for the particles matrix,
solid lipids are mixed with liquid lipids (oils), preferably in a ratio of
70:30 up to a ratio of 99.9:0.1. SLN are composed of 0.1% to 30% (w/w)
solid lipid dispersed in an aqueous medium and if necessary stabilized
with preferably 0.5% (w/w) to 5% (w/w) surfactant (Pardeike et al.,
2009) while the overall solid content of NLC can be up to 95% (w/w)
(Müller, Mäder, Lippacher, & Jenning, 1999). The higher the solid
content, the higher the drug payload; this is a significant advantage
both in terms of cost and impact on food. Less NLC is required to deliver
similar drug content in comparison to SLN. Moreover, to produce
nanoparticles powder, water removal from NLC is more cost-effective.
3.1. Lipids
Selection of an appropriate lipidic blend is crucial for successful pro-
duction of NLC with suitable physical and chemical characteristics. It
was reported that the lipid blend has a significant impact on chemical
stability of sensitive bioactive materials (Mitri et al., 2011; Müller et al.,
2002a). The requirements that should be considered for choice of a suit-
able lipid blend, are listed below:
1. The solubility of the active compound in lipid matrices is essential,
prior to attempting to incorporate the molecule into lipid nanocarriers
because this invariably influences the drug loading capacity, encapsu-
lation efficiency, and subsequent usefulness of NLC (Jaspart, Piel,
Delattre, & Evrard, 2005; Kasongo et al., 2011; Müller et al., 1995,
2000). One of the most important factors that determines the loading
capacity of the drug in the lipid phase is the solubility of drug in the
liquid lipid. The solubility of drugs in various liquid lipids, surfactants,
co-surfactants, solubilizers or a mixture of these can briefly be
determined by adding an excess amount of drug to them, stirring
and/or sonicating the mixture for a sufficient time to achieve equilibri-
um, removing the undissolved drug by centrifugation and/or filtration
(e.g. with a membrane filter 0.2 μm or 0.45 μm) and finally quantifying
the dissolved drug using UV–Vis spectroscopy or chromatography
apparatuses (Joshi, Pathak, Sharma, & Patravale, 2008; Joshi &
Patravale, 2008; Liu et al., 2012). This action can be performed in
interval times for determination of dissolution rate. Likewise,
the equilibrium solubility can be determined by measurement of
the amount of liquid excipient required to dissolve a fixed amount
of drug. The second approach can also be used for determination of
solubility of drugs in melted solid lipids (Joshi & Patravale, 2008). For
33
bioactive compounds which are sensitive to deterioration inhibitive
plans should be applied; such as flushing with N2 and light shielding.
2. The molecules of liquid and solid lipids should be spatially incompat-
ible together as much as possible; this means that the oil molecules
shouldn't be participated in the solid crystalline matrix of solid lipid
and the crystals of solid lipid shouldn't be dissolved in the liquid
lipid. Moreover, it is a prerequisite that the solid and liquid lipids
used to form NLC are miscible at the specific concentrations to be
used (Kasongo et al., 2011; Müller et al., 2002b; Radtke & Müller,
2001; Radtke, Souto, & Müller, 2005); this means that macroscopic
phase separation shouldn't be occurred at a temperature below melt-
ing point of fat; i.e., the blend of bulk lipids should be homogenous
and the liquid lipid should be situated as nano-capillaries/-holes with-
in the fat matrix. The lipid mixtures creating one single phase only
should be used (Doktorovová et al., 2010). Kasongo et al. used a differ-
ential scanning calorimetry (DSC) analysis and a hydrophilic filter
paper to determine the miscibility between solid and liquid lipids.
They smeared a cooled sample of the lipids mixture onto filter
paper, followed by visual observation to determine the presence of
any liquid oil droplets on the filter paper. A binary mixture exhibiting
a melting point above 40 °C and which did not reveal the presence of
oil droplets on the filter paper was selected for use in the NLC produc-
tion (Kasongo et al., 2011).
3. The lipid phase should be more stable to chemical degradation such
as oxidation and lipolysis.
4. The lipids should be biodegradable and be capable to produce parti-
cles in the nanometric scale. It is usually easier to produce very fine
O/W emulsion when the oil phase has a low viscosity and/or interfa-
cial tension than when it has a high viscosity and/or interfacial tension
(Walstra, 2003).
5. The lipids should have an acceptable toxicological profile and should
not lead to production of any toxic residues during NLC preparation
(Müller et al., 2000).
Usually, the lipids used to prepare NLC are mono-, di- and
tri-acylglycerols, fatty acids and waxes.
3.1.1. Liquid lipids
Medium chain triglycerides (MCT) and oleic acid are most used
liquid lipids for production of NLC (Table 1).
MCT are a unique class of saturated lipids composed mainly of caprylic
(C8:0; 50–80%) and capric (C10:0; 20–50%) fatty acids with a minor level
of caproic (C6:0; ≤ 2%), lauric (C12:0; ≤ 3%) and myristic (C14:0; ≤ 1%)
fatty acids; they solidify at ~0 °C, have low viscosity (28–32 mPa.s at
20 °C) and are also considered as emulsifier and suspender rather than
solver (Rowe, Sheskey, & Quinn, 2009). MCT are food-grade and usually
produced by modification (hydrolysis/esterification) and fractionation
of natural oils, such as coconut or palm kernel oils. MCT are known to
be faster digestible than long chain triglycerides, e.g. corn oil (Porter et
al., 2007) and have high stability against oxidation (Hippalgaonkar,
Majumdar, & Kansara, 2010). They are odorless, however, if they hydro-
lyze, the released free fatty acids may significantly influence the aroma
of NLC-incorporated foods (hydrolytic rancidity). MCT have been ap-
proved as generally recognized as safe (GRAS) by the US Food and Drug
Administration (FDA), when added directly to many food types (includ-
ing beverages) for specific functional uses (e.g. as vehicle, solvent, release
agent or emulsifier). MCT with different molecular and physicochemical
properties are commercially available.
Oleic acid [cis-9-octadecenoic acid; C18:1 (9)] is a constituent of many
natural edible lipids. It is odorless and usually obtained by the hydrolysis
of various animal and vegetable lipids (e.g. olive oil) followed by separa-
tion of the liquid fatty acids. Based on the documented specifications by
European pharmacopeia (PhEur), its assay is 65–88% and has varying
amounts of saturated and other unsaturated fatty acids. It has low viscos-
ity (~26 mPa.s at 25 °C), solidifies at ~4 °C and as an emulsifying agent is
used in food and pharmaceutical formulations (Rowe et al., 2009). Its
34 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
health promoting value is also well documented (Lopez-Huertas, 2010).
Oleic acid is somewhat susceptible to oxidation. The relative rates of
oxidation for stearic acid, oleic acid and linoleic acid are 1, 100 and
1200, respectively (Belitz, Grosch, & Schieberle, 2009). Moreover, free
fatty acids are more surface-active than triacylglycerols and therefore
accumulate preferentially at oil–water interface, which increases their
susceptibility to oxidation (Coupland et al., 1996). Again, free fatty acids
are more susceptible to oxidation than are triglycerides. The free radicals
from oxidation process may damage the encapsulated sensitive bioactive
compound within NLC. Fatty acids manufactured from fats and oils
derived from edible sources (e.g. capric, caprylic, lauric, myristic, oleic,
palmitic, and stearic) are included on FDA list of food additives permitted
for direct addition to food (21CFR172.860). Oleic acid with several grades
is commercially available.
Natural edible oils such as corn oil, soybean oil (Table 1) and sunflow-
er oil (Gramdorf et al., 2008) can also be used as liquid lipid for NLC pro-
duction. These liquid lipids are more “label-friendly” and some of these
oils may contain natural antioxidants leading to effective protection of
drugs from oxidation deterioration; e.g. γ-tocopherol in corn oil (Valls,
Goicoechea, Muniz, Saez, & Cabo, 2003). Moreover, for food manufac-
turers, these oils are cheaper alternatives to MCT and oleic acid. Howev-
er, these oils are not efficient surface active, have high unsaturation
degree and high viscosity and may be poor-solver for some of bioactive
compounds. Thus, utilizing these oils for NLC production may lead to ox-
idative instability (e.g. for soybean oil), inconvenience of droplet
disruption during size reduction and low encapsulation efficiency.
Other naturally occurring substances such as vitamin E and squalene
have also been used in NLC formulation as liquid lipid (Table 1). Squalene
is a triterpene found in plants, animals and human. Vitamin E and squa-
lene have health-promoting effects in human but as NLC ingredients
they are susceptible to oxidation and relatively expensive. Moreover,
consuming high amounts of them may be controversial. These oils are
commercially available. Tocopherols as direct food additives are affirmed
as GRAS.
3.1.2. Solid lipids
The most commonly used solid lipids for NLC preparation are glyceryl
behenate, glyceryl palmitostearate, glyceryl monostearate/monostearin,
cetyl palmitate and stearic acid (Table 1). These fats are commercially
available and excluding cetyl palmitate, have been approved as GRAS
for direct addition to food. Many of these solid lipids are also particularly
surface active rather than vehicle and lattice former. Saturated lipids
undergo oxidation more slowly than unsaturated lipids (Nawar, 1996).
Glyceryl behenate mainly consists of diacylglycerols of behenic acid
(C22:0) together with variable quantities of mono- and tri-acylglycerols,
is used in cosmetics, foods and oral pharmaceutical formulations, and
accepted for use as a food additive in Europe. Glyceryl behenate obtains
nanoparticles with high encapsulation efficiency and stability (Castro,
Oréfice, Vilela, Andrade, & Ferreira, 2007). The high encapsulation
efficiency is attributed to many imperfections in the crystalline lattice,
which lead to higher entrapment of drug (Chawla & Saraf, 2011).
Glyceryl palmitostearate is a mixture of mono-, di-, and tri-
acylglycerols of palmitic (C16:0) and stearic (C18:0) fatty acids.
Glyceryl palmitostearate is a good solid lipid for sustained-release
applications (Vivek, Reddy, & Murthy, 2007).
Glyceryl monostearate may be a mixture of mono- and di-
acylglycerols. Different pharmacopeial specifications have been
reported for glyceryl monostearate in PhEur and United States pharma-
copeia (USP). Glyceryl monostearate contains at least 40% mono-
stearoylglycerol based on PhEur. Whiles, the USP describes glyceryl
monostearate as consisting of not less than 90% of monoacylglycerols
of saturated fatty acids, chiefly monostearoylglycerol and glyceryl
monopalmitate. Glyceryl monostearate is widely used in cosmetics,
foods, and pharmaceutical formulations and is generally regarded as a
nontoxic and nonirritant material. It is used as a nonionic emulsifier,
stabilizer, emollient, and plasticizer in a variety of foods and pharmaceu-
tical and cosmetic applications (Rowe et al., 2009).
The USP describes cetyl esters waxes as mixtures consisting primarily
of esters of saturated fatty alcohols (C14–C18) and saturated fatty acids
(C14–C18). Cetyl esters waxes (e.g. cetyl palmitate) are not permitted
for application in foods. However, naturally occurring waxes (from:
Carnauba, Candelilla and Bees) are approved as GRAS for direct addition
to foods and may therefore be used as solid lipid in NLC formulation for
food applications.
Stearic acid is an endogenous long-chain saturated fatty acid and a
primary component of lipids in animal and plant sources which has bet-
ter biocompatibility and lower toxicity than the synthesized counter-
parts (Fundaro et al., 2000). Moreover, it is a principal constituent of
most commercially hydrogenated fats. Stearic acid has a melting point
(69.6 °C) higher than the body temperature, it is biocompatible with
human tissues, and neutral with respect to physiological fluids
(Severino, Pinho, Souto, & Santana, 2011). Stearic acid is accepted as a
food additive in Europe.
Tristearin, propylene glycol monostearate and glyceryl monocaprate
can also be used as solid lipid in NLC formulation (Table 1). Tristearin
and propylene glycol monostearate are included on the FDA list of direct
food additives (21CFR172.811 & 21CFR172.856), while glyceryl
monocaprate is included on the FDA list of indirect food additives
(21CFR176.180).
3.2. Emulsifiers
In the food industry, the main types of emulsifiers are small molecule
surfactants, phospholipids, proteins, and polysaccharides. Proteins and
polysaccharides have the advantage of being natural ingredients that
are often considered more “label-friendly”, thus, there is a great interest
in producing lipid nanocarriers with them. Nevertheless to our best
knowledge, all of the NLC formulations were already stabilized only by
surfactants or rarely by a combination of surfactants and biopolymers
(Liu, Wang, & Xia, 2012; Zheng et al., 2013) rather than by biopolymers
individually. This is mainly because of the ability of surfactants to sponta-
neously form nanoemulsions by low-energy methods, and because of
their ability to rapidly adsorb to droplet surfaces and reduce the interfacial
tension in high-energy methods (such as high-pressure homogenization)
(McClements & Rao, 2011). In addition, the functional properties of many
biopolymers may significantly change during NLC production by the most
used method for production of lipid nanoparticles i.e. hot homogenization
technique (see Section 4). However, the use of biopolymers as emulsifier
should be further investigated for NLC production in food area, since they
have successfully been used for stabilization of nanoemulsions and
nanosuspensions.
Usually, a blend of (oil-soluble and water-soluble) surfactants is used
for preparation of NLC rather than a single surfactant, since improved
functional properties and good physical stability can often be achieved.
A complete list of food surfactants and their Legal Status is published
elsewhere (Hasenhuettl, 2008; McClements, 2005; Whitehurst, 2004).
The most used surfactants in the literature for preparation of NLC are
polyoxyethylene sorbitan monooleate (trade names: Polysorbate® 80,
Tween® 80), lecithin and Poloxamer 188 (Table 1).
Tween 80 is an oleate ester of sorbitol and its anhydrides
copolymerized with approximately 20 mol of ethylene oxide for each
mole of sorbitol and sorbitol anhydrides (Qiu et al., 2013). Tween 80 is
a water-soluble and non-ionic synthetic surfactant with hydrophile–
lipophile balance (HLB) number ~15 (Whitehurst, 2004) and is therefore
suitable for stabilization of NLC. One can usually assume that the steric re-
pulsion is the major colloidal interaction among NLC nanoparticles when
only stabilized with non-ionic surfactants. The particles stabilized by ste-
ric repulsion are fairly insensitive to both electrolyte concentration and
pH and have good freeze–thaw stability; however, they may undergo
weak flocculation (easily reversible), as well as, large amounts of emulsi-
fier is needed to cover particles surface compared to when they are
 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
stabilized by electrostatic repulsion (Hunter, 1986). Tween 80, as a direct
food additive is allowed for certain specific foods by FDA (21CFR172.840).
Its acceptable daily intake (ADI) value is 0–25 mg/kg body weight/day.
Non-ionic surfactants have low toxicity or irritation potential as com-
pared with ionic ones (McClements & Rao, 2011).
Lecithins are naturally occurring surface-active components that can
be extracted from a variety of sources, including soybean, rapeseed and
egg (Faergemand & Krog, 2003). Soy lecithin is the most widely used sur-
factant in the food industry because it can be extracted during the pro-
cessing of crude soybean oil, economically (Stauffer, 1999). Natural
lecithins are a complex mixture of different phospholipids and other
lipids in which phosphatidylcholin, phosphotidyletanolamine and phos-
phatidylinositol are the most common phospholipids (Faergemand &
Krog, 2003). Natural lecithins are essentially hydrophobic molecules
with intermediate HLB numbers (~8) and are not therefore suitable for
stabilizing NLC individually, but are widely used in combination with
other surfactants for stabilizing NLC. One of the two fatty acids of lecithin
can be chemically or enzymatically removed to produce more hydrophilic
lecithin known as lysolecithin that is capable of stabilizing O/W dispersion.
Lecithins are zwitterionic surfactants, and they can have a net negative,
neutral or positive charge depending on pH and electrolyte concentration.
Lecithins (especially soybean lecithin) may contain high amount of
unsaturated fatty acids which are sensitive to oxidation. Lecithins have
GRAS status and with various degrees of purity and specificity are com-
mercially available. They are widely used in the food and pharmaceutical
industries.
Poloxamer 188 (Lutrol® F68; Pluronic® F68) is a nonionic surfactant
with HLB number ~29 and is included in the FDA Inactive Ingredients
Database (Rowe et al., 2009). Poloxamer 188 is a copolymer based on
poly (ethylene oxide)-block-poly(propylene oxide)-block- poly(ethyl-
ene oxide) structure. Poloxamers exhibit minimum toxicities may en-
able controlled release and targeted delivery applications and are
stable at high temperatures; they are commonly used in industrial,
pharmaceutical, natural health product and cosmetic applications, but
they are not food-grade (Trujillo & Wright, 2010) and cannot be used
as direct additive in food.
Monoglycerides of hydrogenated palm oil (Myverol 18-04K) can also
be used for stabilization of NLC (Table 1). Myverol 18-04K is frequently
used in margarine and ice cream as an emulsifier (Martini & Herrera,
2008).
Sodium dodecyl sulfates (SDS), polyoxyethylene sorbitan
monolaurate (Polysorbate® 20/Tween® 20), sodium deoxycholate
(SDC) and polyglyceryl-3-methylglucose distearate have also been
used as surfactant in NLC formulation (Table 1). SDS or sodium lauryl
sulfate (SLS) is a highly hydrophilic anionic surfactant (HLB ~ 40)
which approved as a direct food additive by FDA (21 CFR 172.822).
Tween 20 is a hydrophilic non-ionic surfactant (HLB ~ 16.7) which
included on the FDA list of indirect food additives (21CFR178.3400).
SDC (HLB ~ 16) is the sodium salt of deoxycholic acid which along
with polyglyceryl-3-methylglucose distearate (HLB ~ 12) are not com-
mon emulsifiers for food applications.
Gelucires are multifunctional lipid excipients composed of mono-,
di-, and tri-acylglycerols and mono- and di-fatty acid esters of polyeth-
ylene glycol (PEG) (Siepmann et al., 2006).
Gelucires have interesting properties and can be used as surfactants,
co-surfactants, and lipid matrices in drug delivery systems (Shimpi,
Mahadik, & Paradkar, 2009). Moreover, they produce unique composi-
tions with surfactants, co-surfactants and lipid phases and have inter-
esting properties such as emulsifying agent, drug solubility enhancer
and granule former (Chambin & Jannin, 2005). Their incorporation in
lipid nanocarriers may be helpful in increasing drug loading of lipophilic
compounds (Tsai et al., 2012). However, fatty acid esters of PEG are
included on the FDA list of indirect food additives (21CFR176.170).
Other ingredients such as preservatives (e.g. antioxidant, antimicro-
bial agent), co-surfactants, co-solvents (e.g. glycerol) and solubilizers
(e.g. Transcutol, PEG 400) may also be used in NLC formulation.
35
4. Production of NLC
Production techniques of NLC are similar to that of SLN. Many different
methods have been described in the literature for production of lipid
nanoparticles, especially SLN. These methods are hot/cold homogeniza-
tion, microemulsion technique (Gasco, 1993; Joshi & Patravale, 2008;
Priano et al., 2007), solvent emulsification–evaporation, emulsification-
solvent diffusion (Hu, Yuan, Zhang, & Fang, 2002), solvent injection
(or solvent displacement) (Schubert & Müller-Goymann, 2003), phase
inversion technique (Heurtault, Saulnier, Pech, Proust, & Benoit, 2002),
multiple emulsion method (Garcy-Fuentes, Torres, & Alonso, 2002) and
membrane contractor technique (Charcosset, El-Harati, & Fessi, 2005).
Hot homogenization is the most used method for fabrication of both
SLN and NLC. This method has many advantages (such as easy scale up,
lack of organic solvents and short production time) compared to the
other methods (Pardeike et al., 2009). Since cold homogenization and
solvent emulsification–evaporation methods have potential to produce
NLC, are discussed along with hot homogenization there.
4.1. Hot homogenization method
In this approach, the drug is initially dissolved or dispersed in the
melted lipid mixture (Ca. 5–10 °C above the temperature of the lipid
with highest melting point). Then, the lipid melt is dispersed in aqueous
emulsifier solution at the same temperature by high speed stirring/
shearing. The obtained hot emulsion may further be homogenized at
the same temperature, by instruments such as high pressure homoge-
nizer (HPH), high intensity ultrasonic probe/jet/bath or microfluidizer,
to produce a hot nanoemulsion. Subsequently, NLC are produced by
cooling the hot nanoemulsion in cold water, room temperature or by
a heat exchanger to crystalize lipid droplet and precipitate the lipid
nanoparticles. Usually, HPH yields smaller particles with lower polydis-
persity index usually below 0.2 (Severino, Santana, & Souto, 2012).
The disadvantages of hot homogenization technique are that: a. The
high heating temperature promotes degradation of the labile active
compounds. b. Most surfactants have low cloud points; therefore, high
temperatures may reduce the emulsifying capability of them and
induce the instability of NLC. c. During homogenization, the hydrophilic
drug partitions to the aqueous phase resulting in low entrapment
efficiency (Uner, 2006); this may also lead to increase in solubility of
the lipophilic compound in aqueous phase at hot state and thereupon
the crystallization of it during cooling. Because the solid matrix of
lipid doesn't permit to return of the bioactive within itself.
4.2. Cold homogenization method
Cold homogenization is high-pressure milling of a suspension. To
our best knowledge rather than SLN none of the NLC formulations
were already produced by this method presumably because lower tem-
peratures are required during the manufacture. In this method, after
dissolving/dispersing bioactive compound in the melt lipid mixture, the
bulk lipid is rapidly cooled (e.g. by means of liquid nitrogen). Subse-
quently the bulk lipid matrix is milled to form lipid microparticles e.g.
by a ball mill. It is important to make sure that during the milling process
the temperature does not exceed the temperature of lipid with lowest
melting point. The microparticles are then dispersed in a cold emulsifier
solution and further homogenized to produce fine lipid nanoparticles.
Typically, the particles have larger particle size and wider size distribu-
tion than those obtained with the other techniques (Mehnert & Mäder,
2001) and therefore do not have the same functionalities in terms of dis-
solution velocity during mastication and digestion (Weiss et al., 2008).
Cold homogenization reduces thermal degradation of the bioactive com-
pound. Moreover, drug entrapment is improved and the high cooling
rate favors a uniform distribution of the drug within the lipid matrix.
Finally, crystallization process is controllable and the rapid cooling can
lead to formation of the desired crystal structure.
36 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
Fig. 2. Three structure types of NLC [Modified from Ref. (Radtke & Müller, 2001)]. NLC retard polymorphic transition to more thermodynamically stable forms and reduce the overall
crystallinity of lipid nanoparticles.
4.3. Solvent emulsification–evaporation method
In this method, lipids and bioactive compound are dissolved in a water
immiscible organic solvent with low boiling point (e.g. methylene chlo-
ride). The solution is then emulsified in the aqueous emulsifier solution.
Upon evaporation of the solvent under reduced pressure, the organic
solvent will be removed leaving the lipids and bioactive compound,
nanoparticles form. The benefit of this method is the minimization of
thermal exposure of the sample hence is suitable for heat sensitive
compounds. The particles have narrow distribution and small mean size
(approx. 30–100 nm) depending on the lipid load, emulsifier type and
production conditions. Nevertheless, disadvantages of this method are
the residues solvent in the final product and the low concentration of
final NLC (Wissing, Kayser, & Müller, 2004). Moreover, to reduce the
droplet radius to half, a portion of 7/8 of droplet volume should be
evaporated (Remembrance: Vsphere = (4πr3)/3). Finally, high amount of
emulsifier is used, which all of it is not required for stabilization of the
final NLC. Residues solvent in NLC may create toxicological problems
that can fairly be dispelled by using GRAS solvents like ethyl acetate.
The low concentration of final NLC is due to the limited solubility of the
lipids in the organic solvents used or the dilute organic solvent used in
the procedure.
5. Structure of NLC
Three types of structure have been proposed for NLC; which one of
these structures can be obtained, depends on the formulation composi-
tion and the production parameters.
(I) Imperfect type NLC (imperfectly structured solid matrix; Fig. 2a):
Distances between fatty acid chains of the glycerides and general
imperfections in the crystal, which are a prerequisite for good
drug accommodation, can be increased by using glycerides
being comprised of very different fatty acids (e.g. in length of C
chain, mixture of saturated and unsaturated acids). In order to
achieve ‘highest incompatibility’ rather than using a solid lipid
as for SLN, NLC is produced by mixing solid lipids with chemically
very different liquid lipids (oils). These imperfections/holes ac-
commodate the drug in molecular form and amorphous clusters;
increase the drug payload (Müller et al., 2002b). Imperfect type
NLC is therefore obtained by mixing spatially different lipids.
(II) Amorphous type NLC (structureless solid amorphous ma-
trix; Fig. 2b): As mentioned, crystallization process itself causes
expulsion of the drug that led to the concept of producing NLC
which are solid but not crystalline (Müller et al., 2002b); i.e.,
the lipidic core congeals in an amorphous rather than crystalline
state. This kind of NLC can be produced by mixing solid lipids
with special lipids such as hydroxyoctacosanylhydroxystearate,
isopropylmyristate or MCT (Radtke & Müller, 2001). Reduction
in mobility of lipids (i.e., the solid state) and the absence of a
melting process can be verified by nuclear magnetic resonance
(NMR) and DSC measurements, respectively (Jenning, 1999;
Müller et al., 2002b).
(III) Multiple type NLC (multiple oil-in-fat-in-water (O/F/W) carri-
er; Fig. 2c): Usually, the solubility of lipophilic bioactive com-
pounds is higher in liquid lipids (oils) compared to solid lipids.
Based on this, the multiple type NLC was developed, in which,
the solid matrix of the lipid nanoparticle contains tiny liquid
nano-compartments of oil. In these oil nano-compartments
the drug solubility is higher, thus increasing the total drug
loading capacity. The nano-compartments are surrounded by
solid lipid matrix, thus allowing controlled drug release. This
solid matrix prevents drug leakage (Jenning, Thunemann, &
Gohla, 2000). Multiple type NLC is produced by mixing a
solid lipid with a higher amount of oil. At low concentrations
of oil, the oil molecules are dispersed within the fat matrix
(no oily nano-compartments are formed); when oil concentra-
tion exceeds the solubility of crystallized lipid, phase separation
occurs and oily nano-compartments are formed; this occurs dur-
ing the cooling process after production of the nano-droplets by
the hot homogenization method (Müller et al., 2002a).
Therefore, the nano-compartments within a nanoparticle cannot be
generated by mechanical tools; they are formed by a phase separation
process during particle production. For example, a mixture of glycer-
yl behenate and MCT has been used for NLC production by hot homog-
enization method. Phase separation during the cooling process has been
led to formation of small MCT droplets. The presence of free liquid MCT
could be discerned by the crystallization peaks in DSC (Jenning, 1999).
Moreover, NMR and DSC measurements have been confirmed that
there was no presence of separate solid lipid particles and oil droplets;
i.e., the oil compartments are associated with the solid lipid matrix
(Zimmermann, Müller, & Mäder, 2005). Because, difference in the com-
position of NLC nanoparticles is thermodynamically unfavorable owing
to entropy of mixing; it is more favorable to have the two lipids distribut-
ed equally throughout all of the nanoparticles, rather than be located in
special nanoparticles. It was reported that the oil nano-compartments
localize on the surface of particles instead of inside the particles (Jores
et al., 2004; Saupe et al., 2006); this is challengeable because drug can
be damaged by environmental agents. Thus the formulation of type III
NLC requires further research.
6. Characterization of NLC
The major characteristics which are needed for description of NLC are
size and size distribution, particle charge (usually as zeta potential),
particle morphology, chemical stability during processing and storing
(alone or as a food additive), encapsulation efficiency, drug loading,
crystalline status, release manner and finally the biological fate of
 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
nanoparticles. Here, a brief description of some of these properties is
provided.
6.1. Particle size and zeta (ζ)-potential
The mean particle size and the particle size distribution (usually
as polydispersity index) are the most important characteristics for
nanodispersions which govern the physical stability, solubility, biologi-
cal performance (Lakshmi & Kumar, 2010), release rate, turbidity and
chemical stability. Based on pharmaceutical literature, the usual parti-
cles diameter of NLC is in the range of approximately 10–1000 nm
(Zhang et al., 2011). During NLC production, the particles size of final
NLC is influenced by a series of factors, depending on utilized method.
In the most used methods for production of NLC (i.e. hot homogeniza-
tion and solvent emulsification–evaporation) the oil/organic phase is
dispersed in an aqueous phase containing emulsifier and can therefore
be regarded as the homogenization of an emulsion. For a sufficient
residence time the achievable particle size in emulsions depends on
adsorption kinetic of the emulsifier, interfacial tension between dis-
persed and continuous phase, viscosities of dispersed and continuous
phases, volume fraction of dispersed phase and mechanical energy
input to deform and break up the droplets (Gramdorf et al., 2008).
Moreover, in the solvent emulsification–evaporation method, the ratio
of organic solvent to lipid mixture is another variable. Determining par-
ticle size changes of NLC is the best method for monitoring their physi-
cal stability. The stability of oil dispersions depends on the balance of
emulsifiers at the oil–water interface and the nature of the oil phase
(Martini & Herrera, 2008).
Analytical instruments based on dynamic light scattering (DLS) are
suitable to determine particle size of dispersions in submicron scale
with an accurate, reliable and repeatable estimation. DLS techniques
are based on measurements of the translational diffusion coefficient of
particles determined by analyzing the interaction between a laser
beam and a dispersion (Horne, 1995). In a dilute suspension, where
there are no particle–particle interactions, the size of the particles can
be calculated by the Stokes–Einstein equation (McClements, 2005).
Instruments based on conventional DLS techniques, i.e. photon correla-
tion spectroscopy (PCS) and Doppler shift spectroscopy (DSS) are
primarily designed to analyze dilute dispersions where the laser beam
only undergoes single scattering before reaching the detector (Horne,
1995). But, those based on diffusing wave spectroscopy (DWS) are
designed to analyze concentrated dispersions where the laser beam
undergoes extensive multiple scattering and propagates through the
system by diffusion before reaching the detector (Pine, Weitz, Chaikin,
& Herbolzheimer, 1988). PCS (and also DWS) measures the variations
of the intensity of the scattered laser beam while DSS measures the
shift in frequency of the scattered laser beam known as a Doppler
shift (Horne, 1995). DLS techniques are ordinarily capable of analyzing
particles with diameters between a few nanometers to about 3 microns
(Mehnert & Mäder, 2012) which have noticeable Brownian motion and
are not able to detect larger particles. The presence of larger particles
(3 μm b) in the NLC formulation can be excluded by static light
scattering also known as laser diffraction (LD). LD is based on the
dependence of the diffraction angle on the particle size. Smaller par-
ticles cause more intense scattering at wide-angles compared to the
larger ones. Particle size distributions can be determined by analyz-
ing the intensity of the scattered light as a function of angle between
the incident and scattered beams using a suitable light scattering
theory (McClements, 2005).
Majority of researchers have analyzed the particle size of NLC by
PCS. Before particle size analysis by PCS (and also LD), the dispersion
must properly be diluted to a concentration where multiple scattering
effects are negligible. PCS yields the particle size as intensity-weighted
mean of the hydrodynamic diameter and the polydispersity index (PI;
0 ≤ PI ≤ 1) as a measure of the width of the particle size distribution.
The PI has an important effect on physical stability of nanosuspensions
37
and as much as possible should be low for the long‐term stability of
nanosuspensions. PI values of 0.1–0.25 show a fairly narrow size distri-
bution where a PI value greater than 0.5 indicates a very broad distribu-
tion (Lakshmi & Kumar, 2010).
The ζ-potential is the electrical potential at the shear plane, which is
defined as the distance away from the particle surface below which the
counter-ions remain strongly attached to the particle when it moves in
an electrical field (McClements & Rao, 2011). It is an indirect measure
of physical stability of NLC and even influences the release kinetic and
the biological fate of nanoparticles. To achieve a nanosuspension with
good stability, for an electrostatically stabilized nanosuspension a mini-
mum ζ-potential of ± 30 mV is required whereas in the case of a com-
bined electrostatic and steric stabilization, a minimum ζ-potential of
± 20 mV is desirable (Lakshmi & Kumar, 2010; Mitri et al., 2011). If all
particles in the suspension have a high negative or positive ζ-potential
then they will repel each other and there will be no tendency for the
particles to come together (Wissing & Müller, 2002). The positively-
charged particles will tend to stick to negatively-charged (biological)
surfaces or components, and vice versa. The ζ-potential can be conve-
niently determined by analytical instruments based on electrophoretic/
electroacoustic mobility (McClements, 2005). In many of the commercial
instruments a ζ-potential unit is integrated to DLS equipment so that both
the particle charge and the particle size can be determined using the same
instrument.
6.2. Particle morphology
Usually, lipids prefer to crystallize in the non-spherical platelet form
(Sato, 1988). The non-spherical lipid nanoparticles have high surface
area, short diffusion pathways and low lipid layers as compared with
spherical ones (Mehnert & Mäder, 2012). Therefore, many characteristics
of NLC formulation such as physical and chemical stabilities, encapsula-
tion efficiency, drug loading, drug location within lipid nanoparticles
and drug release rate can also be influenced by the shape of lipid
nanoparticles significantly. Moreover, higher amounts of surfactants are
needed for stabilization of non-spherical lipid particles. Nevertheless,
analytical instruments based on PCS and LD estimate the particle size
while assuming that the particles have spherical shape. Likewise, the
aggregation (flocculation and partial coalescence) of NLC particles if
occurred cannot be detected by these techniques. These underline the
need for additional methods of analysis to provide direct information
about lipid nanoparticles.
Although, optical light microscopy can be used for monitoring of the
presence of larger particles and undissolved drug crystals in colloidal
carrier systems, but this technique is futile for the exact visualization
of nano-sized particles below 500 nm (Klang et al., 2012). Thus,
advanced microscopic techniques such as scanning electron microscopy
(SEM), transmission electron microscopy (TEM) and atomic force mi-
croscopy (AFM) are frequently used to provide critical information
with respect to size, size distribution, morphology, surface topography
and even internal structure of lipid nanoparticles. It should be kept in
mind that possible artifacts may be caused by the sample preparation
(e.g. fixation, dehydration). Moreover, the use of high energy electron
beams and vacuum condition by electron microscopy may cause
changes in the structure of unstable specimens.
SEM gives three-dimensional morphological and surface infor-
mation of the specimen. The sample is bombarded with a focused
electron beam and surface characteristics are obtained from the
secondary electrons emitted from the specimen surface (Domingo
& Saurina, 2012). NLC need to be converted first into a dry powder
and then they are gold/platinum coated before measurements in vacuum
conditions. A major benefit of SEM is the large depth of field, which means
that images of relatively large structures are all in-focus (Hunter, 1993)
but its resolving power (~3–4 nm) is lower than TEM's (McClements,
2005) and doesn't provide internal details (Reimer, 1993). Environmental
SEM (ESEM) instruments are also available that are compatible with
38 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
hydrated specimens; they work at pressures not as lower as SEM and
coating before measurement is not required (Domingo & Saurina, 2012)
but their resolution is not sufficient to obtain detailed information regard-
ing the surface properties and architecture of nanoscale structures (Klang
et al., 2012).
TEM usually provides two-dimensional images when a beam of
electrons is transmitted through a specimen. The fraction of transmit-
ted electrons depends on electron density of the specimen: the lower
the electron density, the greater the fraction of electrons transmitted
and the darker is the image. Components with different electron
densities thus appear as regions of different intensity on the image.
The density contrast between components is usually enhanced by
selectively staining the specimen with various heavy metal salts
(McClements, 2005). The NLC should already be stained, dried and
fixed. The thickness of TEM specimens is required to be in the order
of magnitude of 100 nm and thus the sample preparation results in
a complex procedure as compared to SEM (Domingo & Saurina,
2012). TEM can provide the information about internal structure of
lipid nanoparticles (Klang et al., 2012) and its resolution power is
around 0.4 nm (Hunter, 1993).
AFM instrument consists of a cantilever with a tiny probe at its end
and a laser detection system. The probe is held extremely close to the
specimen surface and three-dimensional images are obtained by mea-
suring the interaction forces between the tip and the sample surface. A
spatial resolution of up to 0.01 nm can be obtained by AFM (Mehnert
& Mäder, 2012). In general, surface profiles of non-conductive samples
without any coating under atmospheric conditions and liquid/wet envi-
ronment can be analyzed by AFM (Domingo & Saurina, 2012; Mehnert &
Mäder, 2012). However, submicron particles are moving rapidly because
of Brownian motion and fixation/dehydration of them is required to
assess their shape by the very tiny AFM tip (Mehnert & Mäder, 2012).
Thus, it may be needed to dry NLC onto an extremely flat plate surface
such as mica.
Other microscopic techniques such as cryogenic TEM (cryo-TEM),
cryogenic SEM (cryo-SEM) and freeze fracture TEM have been success-
fully employed for visualization of lipid nanoparticles (Das, Ng, & Tan,
2012; Dong, Ng, Shen, Kim, & Tan, 2012; Gramdorf et al., 2008; Jores et
al., 2004; Saupe et al., 2006) that allow the sample to be visualized
close to its natural state. Further details about advanced microscopic
techniques can be found in the literature (Klang et al., 2012).
6.3. Chemical stability of bioactive compounds
Chemical stability of the bioactive compound is the initial character-
istic required for biological performance of NLC and health-promoting
of NLC-added foods. Moreover, this is essential to calculate encapsula-
tion efficiency and drug loading. Chemical stability of lipid nanocarriers
can be measured according to two different approaches: destructive or
non-destructive assays.
Destructive assays usually rely on the destruction of the carrier struc-
ture to completely release the drug (encapsulated and unencapsulated) in
a proper medium and subsequently drug quantification. To attain this
purpose, organic solvents such as methanol, methylene chloride, acetoni-
trile, acetone or tetrahydrofuran are often used as destructive agent or
drug solver. Many researchers have applied this approach (Hentschel et
al., 2008; Jia et al., 2012; Liu et al., 2011; Qian, Decker, Xiao, &
McClements, 2012; Tsai et al., 2012). In destructive assay, the solvent
extraction itself may significantly impact the chemical stability of drug.
The drug content in the solution recovered from destructive assays is
often quantified by UV–Vis spectroscopy or HPLC. UV–Vis techniques are
simple but they are, in general, inefficient to detect the occurrence of
interfering components (Domingo & Saurina, 2012). In contrast, chro-
matographic techniques are appropriate to monitor side processes such
as drug degradation or release of unwanted components and impurities
from the carrier (Argemí, Vega, Subra-Paternault, & Saurina, 2009).
Non-destructive assays may also be used for determination of chem-
ical stability of some bioactive substances in lipid nanocarriers which rely
on the changes of a measurable physical characteristic such as NLC color
or NLC absorbance at the λmax of bioactive material (Helgason et al.,
2009; Mitri et al., 2011; Qian et al., 2012). Non-destructive assays are
simple and eliminate the need for solvent extraction but require an
accurate calibration curve, thus an accurate evaluation of the drug
percentage may be hindered, specifically if the drug crystallized within
aqueous or lipid phases.
6.4. Encapsulation efficiency (EE) and drug loading (DL)
The encapsulation efficiency (EE) is defined as the ratio of encapsu-
lated drug in nanoparticles to whole drug first incorporated into the
lipid phase of NLC multiplied by 100 (Das et al., 2012; Kumbhar &
Pokharkar, 2013; Liu & Wu, 2010; Yuan et al., 2007). But, it should be
kept in mind that the damaged portion of bioactive compound during
production and storage of NLC must be subtracted. The EE can be calcu-
lated after quantification of free portion or encapsulated portion of drug.
The unencapsulated free drug may crystallize and/or dissolve in the
aqueous phase. The crystallized portion usually has micron size and
precipitates in the NLC system and can conveniently be removed by
microfiltration (Das et al., 2012; Nguyen et al., 2012) or mild centrifuga-
tion (Agrawal, Petkar, & Sawant, 2010) and then quantified. The contin-
uous phase can be separated from the nanoparticles by techniques such
as ultrafiltration (Araújo et al., 2011; Das et al., 2012), (ultra-)centrifuga-
tion (Liu et al., 2011; Yuan et al., 2007) or size exclusion chromatography
(Chen et al., 2010; Huang et al., 2013) and the amount of dissolved or
encapsulated drug can be quantified. Before drug quantification, the
separated nanoparticles may be washed with water or aqueous phase
in order to remove the free drug adsorbed on their surface (Yuan et al.,
2007). Moreover, the percentage of dissolved drug in aqueous phase
may be determined by extraction with organic solvents (Liu & Wu,
2010).
A number of researchers reported the EE as the percentage of
non-crystallized drug within NLC system rather than as the per-
centage of drug entrapped within lipid nanoparticles (Agrawal et
al., 2010; Nguyen et al., 2012) presumably because of poor aqueous
phase-solubility of drug.
The EE influences the release characteristics of NLC and depends on
the NLC ingredients (which influence factors such as the solubility of
drug in the lipid phase, nanoparticle crystallinity index, viscosity of
continuous phase and thereby diffusion coefficient of drug), production
method and conditions used. In an O/W emulsion, nano-droplets in
comparison to micro-ones increase the water-solubility of the encapsu-
lated lipophilic bioactive compounds; thus it seems that micro-droplets
have higher EE. Nonetheless, the solid state of lipidic nanoparticles in
NLC immobilizes the incorporated drug and leads to higher EE in
comparison to nanoemulsion. If the drug which should be encapsulated,
has poor water-solubility and be sensitive to degradation under physi-
cochemical stresses during processing and storage of food and GI stress-
es (e.g. carotenoids), having a high EE is appropriate. But if the drug be
more stable to physicochemical stresses, and only has low water
solubility and thereupon low bioavailability, having a low EE is appro-
priate. In the later case, the nanoemulsion is preferred rather than
NLC provided that the drug doesn't crystallize in aqueous phase and
nanoemulsion has acceptable physicochemical stability. The increase
of water-solubility of bioactive compound by nanozation of dispersed
phase in the O/W dispersion is one of the reasons for the achievement
of higher bioavailability by nanoparticles (McClements & Rao, 2011).
The drug loading (DL) of NLC is defined as the ratio of encapsulated
drug to amount of lipid phase or lipid nanoparticles in NLC formulation
multiplied by 100 (Das et al., 2012; Nguyen et al., 2012; Yuan et al.,
2007). For food applications the ratio of whole intact drug (i.e. encapsu-
lated and water-dissolved) to NLC amount may be a better characteris-
tic rather than DL. DL is predominantly influenced by the polymorphic
 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
state of the lipid, solubility of the drug in the melted form of lipid, types
of lipids, and chemical and physical structure of the solid lipid matrix
(Müller et al., 2000). A higher DL is a significant advantage both in
terms of cost and impact on food; less NLC is required to deliver a
predetermined drug level into food.
6.5. Crystallinity
The physical status of the bioactive material in both aqueous and lipid
phases governs its bioavailability and efficacy of the NLC formulation as a
controlled release system. Moreover, the polymorphic changes and the
melting behavior of the dispersed phase in NLC govern the form of
crystals, particle morphology, release rate of bioactive molecule, drug
loading and crystallinity index of lipid nanoparticles and the maximum
temperature in which nanoparticles remain solid. In addition to correct
choice of the emulsifier, knowing the melting point of NLC nanoparticles
is a key temperature during heat treatment of NLC-added foods. Usually,
the lipid blend of NLC particles shows a melting point depression
compared to the original solid lipid (Müller et al., 2002a), but the
obtained blend is also solid at body temperature.
X‐ray spectroscopy and DSC are widely used to investigate the
status of drug and the polymorphic changes and melting behavior
of lipid nanoparticles in NLC (Jores, Mehnert, & Mäder, 2003;
Kumbhar & Pokharkar, 2013; Liu & Wu, 2010; Pardeike et al., 2011;
Teeranachaideekul, Souto, Junyaprasert, & Müller, 2007). The structural
organization of the oil fraction associated with the nanoparticles at least
at higher oil loads can also be investigated by NMR analysis (Garcia-
Fuentes, Alonso, & Torres, 2005; Jores et al., 2003).
7. Food applications of NLC — Perspective and challenges
With the increasing public perception of a strong correlation between
food and disease prevention, new technologies are being introduced to
enrich staple foods and beverages with “health promoting substances”
and produce functional foods (de Vos, Faas, Spasojevic, & Sikkema,
2010). As a result, enormous studies have been performed on identifying
and isolating bioactive compounds. Bioactive food components are
divided into bioactive molecules and bioactive living cells (probiotics).
The most important types of bioactive lipids that need to be delivered
within foods are fatty acids, carotenoids, oil-soluble antioxidants (tocoph-
erols and polyphenols), phytosterols, oil-soluble vitamins (A and D) and
other nutraceuticals (e.g. Co-enzyme Q10) (McClements & Li, 2010).
Lipid-soluble bioactive compounds pose a particularly difficult
challenge, especially in fat-free and low-fat foods, which are in growing
demand, and enrichment of aqueous-based food with these compounds
has been greatly restricted.
Omega-3 fatty acids, the major essential fatty acids, are susceptible to
oxidative deterioration. Moreover, the oxidation products of omega-3
fatty acids have unattractive effect on sensory acceptance of enriched
foodstuffs. Thus, these hydrophobic compounds require stabilization in
an aqueous medium and protection against deteriorating factors
(Zimet, Rosenberg, & Livney, 2011). It has been shown that encapsula-
tion of omega-3 fatty acids decreased its oxidation (in enriched foods)
significantly (Garg, Wood, Singh, & Moughan, 2006; Tamjidi, Nasirpour,
& Shahedi, 2012).
Oil-soluble vitamins are also susceptible to oxidation and may create
drug taste in fortified foods; for these reasons encapsulation of them is
often essential before addition to foods.
The current challenges to application of carotenoids as nutraceutical
compounds are poor water-solubility, high melting point, chemical
instability, and low bioavailability (Qian et al., 2012) and delivery
systems should therefore be designed to fortification of foods with carot-
enoids, efficiently.
Enrichment of foods with phytosterols is problematic because of
their high melting point and tendency to form insoluble crystals. There-
fore, for incorporation into aqueous-based foods, phytosterols should
39
be either suspended or emulsified (McClements, Decker, & Weiss,
2007). Oxidation of phytosterols is another challenge in enriched food
products which can be prevented using encapsulation techniques.
Flavonoids (e.g. quercetin) are a large group of natural polyphenols;
many of these molecules have low bioavailability (due to poor water-
solubility), low chemical stability and undesirable taste and suitable encap-
sulation system should therefore be designed to deliver them into foods.
NLC are particularly suitable for encapsulating and delivering of
lipophilic bioactive molecules and other lipophilic compounds such as
flavors, antimicrobials and drugs into aqueous-based foods. The possi-
bility of NLC as delivery system has been explored for many compounds
in pharmaceutical area. It has been shown that NLC was able to
encapsulate lipophilic drugs and to improve the stability and the
bioavailability of them. It was reported that hydrophilic drugs can also
be incorporated into lipid nanoparticles (Kasongo et al., 2011); but,
loading hydrophilic bioactive materials in lipid nanoparticles is difficult
because of the tendency of partitioning the encapsulated molecules in
the water during the production process of nanoparticles (Singh,
Dobhal, Jain, Pandit, & Chakraborty, 2010). Like nanoemulsions, NLC
are optically transparent or only slightly turbid and are therefore suit-
able for fortification of transparent beverages.
NLC can be produced on industrial scale but GRAS or food-grade
ingredients are required. Since many substances that are permitted
for application in pharmaceutical industries are not allowed in food
in large quantity, further studies is required for selection of appropriate
materials for food applications. The ingredients should not unpleasantly
affect the sensory properties of NLC-incorporated food (Du, 2011).
Moreover, the stability of NLC must be examined during processing
and storing of food against mechanical and physicochemical stresses
such as pressure, mixing, heat treatment, light, drying, freezing and
changes of ionic strength, pH and environmental storage condition.
As mentioned earlier, the NLC should remain solid at room and GI
tract temperatures. The solid lipid, like liquid lipid, can be digested in
body by lipase but digestion rate for solid lipid is lower than liquid
state (McClements & Li, 2010). Moreover, if the bioactive compound
immobilizes within lipid nanoparticle as crystalline form, its bioavail-
ability may decrease comparing to non-crystalline form.
When the lipid particles are partly crystalline a crystal from one par-
ticle can penetrate into the liquid oil portion of another particle during a
contact or collision which causes the particles to stick together; this
phenomenon is known as partial coalescence and as a potential disad-
vantage, may lead to a decrease in NLC stability to aggregation or gela-
tion. The various factors that influence partial coalescence have been
discussed elsewhere (Dickinson & McClements, 1995; McClements,
2005; Walstra, 2003). The types of lipids and emulsifiers, the lipids
ratio and the particle size influence the partial coalescence, significantly.
We would not expect to see any partial coalescence in NLC (if occurred)
when the nanoparticles have amorphous structure due to the absence
of any crystal. Although a good physical stability have been frequently
reported for NLC formulations in the literature (Das et al., 2012; Liu,
Wang, & Xia, 2012; Pardeike et al., 2011; Tan, Billa, Roberts, & Burley,
2010; Xia & Wang, 2011), further studies are needed to seek this insta-
bility mechanism, especially with food-grade and GRAS ingredients.
Lastly, it should be pointed out that the solid lipid must be careful-
ly selected and optimized for NLC production, whereas there is an in-
creasing tendency toward removing or reducing the amounts of food
constituents that have been associated with human health concerns
(e.g. saturated lipid and cholesterol).
8. Conclusions
Nanostructured lipid carriers (NLC) are a carrier system with which
the requirements of food science and technology can be fulfilled. They
may combine the advantages of different colloidal drug delivery systems
and avoid some of their disadvantages. NLC are a promising delivery
system for food application of the poor water-soluble bioactive
40 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
components, may increase their stability and bioavailability. NLC may
dispel the disadvantages of other lipid nanocarriers such as low encapsu-
lation efficiency, low drug loading and physicochemical instability.
Moreover, control of release is feasible by NLC system, is applicable for
both sustained and delayed releases which are time- or
site-dependent. Burst release can also be achieved by controlling the
physical state of core lipid; this is valuable for increasing flavor release
in fast foods and ready-meals during heating and consumption. NLC
have high loading capacity and their ingredients can be selected from
low-cost food-grade and GRAS materials. NLC have compatibility for in-
corporation into aqueous-based foods. High pressure homogenizer as in-
dustrially the most achievable instrument for the production of NLC, is
widely used in food industry; no regulatory problems exist for usage
this production device. The NLC produced by hot homogenization meth-
od may be directly added to pasteurized transparent, translucent and
opaque products such as beer, fruit juice and milk. NLC can also be
added to food systems before pasteurization. Depending on type and
concentration of ingredients and chemical stability of encapsulated
active molecule, NLC may be sterilized by autoclaving, gamma irradiation
or filtration (if the particle diameter is well below 200 nm). Moreover,
they can also be spray-dried or lyophilized. Thus, NLC may be an efficient
drug delivery system and may present all the necessary characteristics
for use as efficient vehicle or drug nanocarrier, for food industries. Food-
stuffs containing bioactive-loaded NLC can be regarded as possible
health-promoting nutraceutical ones. This review opens a new path for
specific applications and the development of innovative delivery systems
and/or functional food products.
Acknowledgment
This paper is an outcome of a research project financially supported
by Isfahan University of Technology.
References
Agrawal, Y., Petkar, K. C., & Sawant, K. K. (2010). Development, evaluation and clinical
studies of Acitretin loaded nanostructured lipid carriers for topical treatment of
psoriasis. International Journal of Pharmaceutics, 401(1–2), 93–102.
Ali, H., El-Sayed, K., Sylvester, P. W., & Nazzal, S. (2010). Molecular interaction and localiza-
tion of tocotrienol-rich fraction (TRF) within the matrices of lipid nanoparticles: Evi-
dence studies by Differential Scanning Calorimetry (DSC) and Proton Nuclear
Magnetic Resonance spectroscopy (1H NMR). Colloids and Surfaces. B, Biointerfaces,
77(2), 286–297.
Almeida, A. J., & Souto, E. (2007). Solid lipid nanoparticles as a drug delivery system for
peptides and proteins. Advanced Drug Delivery Reviews, 59(6), 478–490.
Araújo, J., Nikolic, S., Egea, M. A., Souto, E. B., & Garcia, M. L. (2011). Nanostructured
lipid carriers for triamcinolone acetonide delivery to the posterior segment of
the eye. Colloids and Surfaces. B, Biointerfaces, 88(1), 150–157.
Argemí, A., Vega, A., Subra-Paternault, P., & Saurina, J. (2009). Characterization of
azacytidine/poly(L-lactic) acid particles prepared by supercritical antisolvent pre-
cipitation. Journal of Pharmaceutical and Biomedical Analysis, 50(5), 847–852.
Belitz, H. -D., Grosch, W., & Schieberle, P. (2009). Food chemistry (4th ed.). Berlin: Springer.
Castro, G. A., Oréfice, R. L., Vilela, J. M., Andrade, M. S., & Ferreira, L. A. (2007). Develop-
ment of a new solid lipid nanoparticle formulation containing retinoic acid for top-
ical treatment of acne. Journal of Microencapsulation, 24(5), 395–407.
Chambin, O., & Jannin, V. (2005). Interest of multifunctional lipid excipients: Case of
Gelucire® 44/14. Drug Development and Industrial Pharmacy, 31(6), 527–534.
Charcosset, C., El-Harati, A., & Fessi, H. (2005). Preparation of solid lipid nanoparticles
using a membrane contactor. Journal of Controlled Release, 108(1), 112–120.
Chaudhry, Q., Scotter, M., Blackburn, J., Ross, B., Boxall, A., Castle, L., et al. (2008). Appli-
cations and implications of nanotechnologies for the food sector. Food Additives
and Contaminants, 25(3), 241–258.
Chawla, V., & Saraf, S. A. (2011). Glyceryl behenate and its suitability for production of
aceclofenac solid lipid nanoparticles. Journal of the American Oil Chemists' Society,
88(1), 119–126.
Chen, C. -C., Tsai, T. -H., Huang, Z. -R., & Fang, J. -Y. (2010). Effects of lipophilic emulsi-
fiers on the oral administration of lovastatin from nanostructured lipid carriers:
Physicochemical characterization and pharmacokinetics. European Journal of
Pharmaceutics and Biopharmaceutics, 74(3), 474–482.
Chen, H. D., Weiss, J. C., & Shahidi, F. (2006). Nanotechnology in nutraceuticals and
functional foods. Food Technology, 60(3), 30–36.
Chinsriwongkul, A., Chareanputtakhun, P., Ngawhirunpat, T., Rojanarata, T., Sila-on, W.,
Ruktanonchai, U., et al. (2011). Nanostructured Lipid Carriers (NLC) for parenteral
delivery of an anticancer drug. AAPS Pharmaceutical Science and Technology, 13(1),
150–158.
Cirri, M., Bragagni, M., Mennini, N., & Mura, P. (2012). Development of a new delivery sys-
tem consisting in “drug – in cyclodextrin – in nanostructured lipid carriers” for
ketoprofen topical delivery. European Journal of Pharmaceutics and Biopharmaceutics,
80(1), 46–53.
Coupland, J. N., Weiss, J., Lovy, A., & McClements, D. J. (1996). Solubilization kinetics of
triacyl glycerol and hydrocarbon emulsion droplets in a micellar solution. Journal
of Food Science, 61(6), 1114–1117.
Coupland, J. N., Zhu, Z., Wan, H., McClements, D. J., Nawar, W. W., & Chinachotti, P.
(1996). Droplet composition affects the rate of oxidation of emulsified ethyl lino-
leate. Journal of the American Oil Chemists' Society, 73(6), 795–801.
Das, S., Ng, W. K., & Tan, R. B. H. (2012). Are nanostructured lipid carriers (NLCs) better
than solid lipid nanoparticles (SLNs): Development, characterizations and compara-
tive evaluations of clotrimazole-loaded SLNs and NLCs? European Journal of Pharma-
ceutical Sciences, 47(1), 139–151.
Das, M., Saxena, N., & Dwivedi, P. D. (2009). Emerging trends of nanoparticles applica-
tion in food technology: Safety paradigms. Nanotoxicology, 3(1), 10–18.
de Vos, P., Faas, M. M., Spasojevic, M., & Sikkema, J. (2010). Encapsulation for preservation
of functionality and targeted delivery of bioactive food components. International
Dairy Journal, 20(4), 292–302.
Dickinson, E., & McClements, D. J. (1995). Advances in food colloids. London: Chapman &
Hall.
Doktorovová, S., Araújo, J., Garcia, M. L., Rakovsky, E., & Souto, E. B. (2010). Formulating
fluticasone propionate in novel PEG-containing nanostructured lipid carriers
(PEG-NLC). Colloids and Surfaces. B, Biointerfaces, 75(2), 538–542.
Domingo, C., & Saurina, J. (2012). An overview of the analytical characterization of
nanostructured drug delivery systems: towards green and sustainable pharmaceu-
ticals: A review. Analytica Chimica Acta, 744, 8–22.
Dong, Y., Ng, W. K., Shen, S., Kim, S., & Tan, R. B. H. (2012). Solid lipid nanoparticles:
Continuous and potential large-scale nanoprecipitation production in static
mixers. Colloids and Surfaces. B, Biointerfaces, 94, 68–72.
Du, Y. (2011). Fabrication and characterization of low crystalline curcumin loaded lipid
nanoparticles. MSc Thesis, Graduate School-New Brunswick Rutgers, The State University
of New Jersey, New Jersey.
Eskandari, S., Varshosaz, J., Minaiyan, M., & Tabbakhian, M. (2011). Brain delivery of
valproic acid via intranasal administration of nanostructured lipid carriers: in
vivo pharmacodynamic studies using rat electroshock model. International Journal
of Nanomedicine, 6, 363–371.
Esposito, E., Mariani, P., Ravani, L., Contado, C., Volta, M., Bido, S., et al. (2012).
Nanoparticulate lipid dispersions for bromocriptine delivery: Characterization and in
vivo study. European Journal of Pharmaceutics and Biopharmaceutics, 80(2), 306–314.
Faergemand, M., & Krog, N. (2003). Using emulsifiers to improve food texture. In B. M.
McKenna (Ed.), Texture in foods. Semi-solid foods, 1. (pp. 216–274). Florida: CRC Press.
Fang, J. -Y., Fang, C. -L., Liu, C. -H., & Su, Y. -H. (2008). Lipid nanoparticles as vehicles for topical
psoralen delivery: Solid lipid nanoparticles (SLN) versus nanostructured lipid carriers
(NLC). European Journal of Pharmaceutics and Biopharmaceutics, 70(2), 633–640.
Fathi, M., Mozafari, M. R., & Mohebbi, M. (2012). Nanoencapsulation of food ingredients
using lipid based delivery systems. Trends in Food Science & Technology, 23(1), 13–27.
Fathi, M., Varshosaz, J., Mohebbi, M., & Shahidi, F. (2012). Hesperetin-loaded solid
lipid nanoparticles and nanostructure lipid carriers for food fortification:
Preparation, characterization, and modeling. Food and Bioprocess Technology.
http://dx.doi.org/10.1007/s11947-012-0845-2.
Flanagan, J., & Singh, H. (2006). Microemulsions: A potential delivery system for bioac-
tives in food. Critical Reviews in Food Science and Nutrition, 46(3), 221–237.
Fundaro, A., Cavalli, R., Bargoni, A., Vighetto, D., Zara, G. P., & Gasco, M. R. (2000).
Non-stealth and stealth solid lipid nanoparticles (SLN) carrying doxorubicin: Phar-
macokinetics and tissue distribution after i.v. administration to rats. Pharmacolog-
ical Research, 42(4), 337–343.
Garcia-Fuentes, M., Alonso, M. J., & Torres, D. (2005). Design and characterization of a
new drug nanocarrier made from solid–liquid lipid mixtures. Journal of Colloid
and Interface Science, 285(2), 590–598.
Garcy-Fuentes, M., Torres, D., & Alonso, M. (2002). Design of lipid nanoparticles for the
oral delivery of hydrophilic macromolecules. Colloids and Surfaces. B, Biointerfaces,
27(2–3), 159–168.
Garg, M. L., Wood, L. G., Singh, H., & Moughan, P. J. (2006). Means of delivering
recommended levels of long chain n-3 polyunsaturated fatty acids in human
diets. Journal of Food Science, 71(5), R66–R71.
Garti, N. (2003). Microemulsions as microreactors for food applications. Current Opin-
ion in Colloid and Interface Science, 8(2), 197–211.
Gasco, M. R. (1993). Method for producing solid lipid microspheres having a norrow
size distribution. USA Patent, 5 250 236.
Gramdorf, S., Hermann, S., Hentschel, A., Schrader, K., Müller, R. H., Kumpugdee-Vollrath,
M., et al. (2008). Crystallized miniemulsions: Influence of operating parameters dur-
ing high-pressure homogenization on size and shape of particles. Colloids and Sur-
faces A: Physicochemical and Engineering Aspects, 331(1–2), 108–113.
Guo, C., Yanga, C., Li, Q., Tan, Q., Xi, Y., Liud, W., et al. (2012). Development of a
Quercetin-loaded nanostructured lipid carrier formulation for topical delivery. In-
ternational Journal of Pharmaceutics, 430(1–2), 292–298.
Haag, S. F., Chen, M., Peters, D., Keck, C. M., Taskoparan, B., Fahr, A., et al. (2011). Nanostruc-
tured lipid carriers as nitroxide depot system measured by electron paramagnetic res-
onance spectroscopy. International Journal of Pharmaceutics, 421(2), 364–369.
Han, F., Li, S., Yin, R., Liu, H., & Xu, L. (2008). Effect of surfactants on the formation and char-
acterization of a new type of colloidal drug delivery system: Nanostructured lipid car-
riers. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 315(1–3),
210–216.
Hasenhuettl, G. L. (2008). Overview of food emulsifiers. In G. L. Hasenhuettl, & R. W.
Hartel (Eds.), Food emulsifiers and their applications (pp. 1–10). New York: Springer.
 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
Helgason, T., Awad, T. S., Kristbergsson, K., Decker, E. A., Mcclements, D. J., & Weiss, J.
(2009). Impact of surfactant properties on oxidative stability of β-carotene encap-
sulated within solid lipid nanoparticles. Journal of Agricultural and Food Chemistry,
57(17), 8033–8040.
Hentschel, A., Gramdorf, S., Müller, R. H., & Kurz, T. (2008). β-carotene-loaded nano-
structured lipid carriers. Journal of Food Science, 73(2), N1–N6.
Heurtault, B., Saulnier, P., Pech, B., Proust, J. E., & Benoit, J. P. (2002). A novel phase
inversion-based process for the preparation of lipid nanocarriers. Pharmaceutical Re-
search, 19(6), 875–880.
Hippalgaonkar, K., Majumdar, S., & Kansara, V. (2010). Injectable lipid emulsions-
advancements, opportunities and challenges. AAPS Pharmaceutical Science and Technolo-
gy, 11(4), 1526–1540.
Horne, D. S. (1995). Light scattering studies of colloid stability and gelation. In E.
Dickinson (Ed.), New physicochemical techniques for the characterization of complex
food systems (pp. 240–267). London: Blackie Academic & Professional.
Hu, F. -Q., Jiang, S. -P., Du, Y. -Z., Yuan, H., Ye, Y. -Q., & Zeng, S. (2005). Preparation and
characterization of stearic acid nanostructured lipid carriers by solvent diffusion
method in an aqueous system. Colloids and Surfaces. B, Biointerfaces, 45(3–4),
167–173.
Hu, F. -Q., Jiang, S. -P., Du, Y. -Z., Yuan, H., Ye, Y. -Q., & Zeng, S. (2006). Preparation and
characteristics of monostearin nanostructured lipid carriers. International Journal of
Pharmaceutics, 314(1), 83–89.
Hu, F. Q., Yuan, H., Zhang, H. H., & Fang, M. (2002). Preparation of solid lipid
nanoparticles with clobetasol propionate by a novel solvent diffusion method in
aqueous system and physicochemical characterization. International Journal of
Pharmaceutics, 239(1–2), 121–128.
Huang, X., Chen, Y. -J., Peng, D. -Y., Li, Q. -L., Wang, X. -S., Wang, D. -L., et al. (2013).
Solid lipid nanoparticles as delivery systems for Gambogenic acid. Colloids and Surfaces.
B, Biointerfaces, 102, 391–397.
Hung, L. C., Basri, M., Tejo, B. A., Ismail, R., Nang, H. L. L., Hassan, H. A., et al. (2011). An
improved method for the preparations of nanostructured lipid carriers containing
heat-sensitive bioactives. Colloids and Surfaces. B, Biointerfaces, 87(1), 180–186.
Hunter, R. J. (1986). Foundations of colloid science, Vol. 1, Oxford: Oxford University
Press.
Hunter, R. J. (1993). Introduction to modern colloid science. Oxford: Oxford University Press.
Jaspart, S., Piel, G., Delattre, L., & Evrard, B. (2005). Solid lipid microparticles: Formula-
tion, preparation, characterization, drug release and applications. Expert Opinion on
Drug Delivery, 2(1), 75–87.
Jenning, V. (1999). Feste Lipid-Nanopartikel (SLN) als Tragersystem fur die dermale
Applilkation von Retinol. PhD thesis, Department of Pharmaceutics, Biopharmaceutics
and Biotechnology, The Free University of Berlin, Berlin.
Jenning, V., Mäder, K., & Gohla, S. H. (2000). Solid lipid nanoparticles (SLN) based on
binary mixtures of liquid and solid lipids: A H-NMR study. International Journal of
Pharmaceutics, 205(1–2), 15–21.
Jenning, V., Thunemann, A. F., & Gohla, S. H. (2000). Characterization of a novel solid
lipid nanoparticle carrier system based on binary mixtures of liquid and solid
lipids. International Journal of Pharmaceutics, 199(2), 167–177.
Jia, L., Shen, J., Zhang, D., Duan, C., Liu, G., Zheng, D., et al. (2012). In vitro and in vivo eval-
uation of oridonin-loaded long circulating nanostructured lipid carriers. International
Journal of Biological Macromolecules, 50(3), 523–529.
Jia, L., Zhang, D., Li, Z., Duan, C., Wang, Y., Feng, F., et al. (2010). Nanostructured lipid
carriers for parenteral delivery of silybin: Biodistribution and pharmacokinetic
studies. Colloids and Surfaces. B, Biointerfaces, 80(2), 213–218.
Jin, Y., Perrie, C., Zhang, W., Diepen, C. V., Curtis, J., & Barrow, C. J. (2008). Microencap-
sulation of marine lipids as a vehicle for functional food delivery. In C. J. Barrow, &
F. Shahidi (Eds.), Marine nutraceuticals and functional foods (pp. 115–154). London:
CRC Press.
Jores, K., Mehnert, W., Drechsler, M., Bunjes, H., Johann, C., & Mäder, K. (2004). Inves-
tigations on the structure of solid lipid nanoparticles (SLN) an oil-loaded solid
lipid nanoparticles by photon correlation spectroscopy, field-flow fractionation
and transmission electron microscopy. Journal of Controlled Release, 95(2),
217–227.
Jores, K., Mehnert, W., & Mäder, K. (2003). Physicochemical investigations on solid
lipid nanoparticles and on oil-loaded solid lipid nanoparticles: A nuclear magnetic
resonance and electron spin resonance study. Pharmaceutical Research, 20(8),
1274–1283.
Joshi, M., Pathak, S., Sharma, S., & Patravale, V. (2008). Design and in vivo pharmacodynamic
evaluation of nanostructured lipid carriers for parenteral delivery of artemether:
Nanoject. International Journal of Pharmaceutics, 364(1), 119–126.
Joshi, M., & Patravale, V. (2008). Nanostructured lipid carrier (NLC) based gel of
celecoxib. International Journal of Pharmaceutics, 346(1–2), 124–132.
Kasongo, K. W., Pardeike, J., Müller, R. H., & Walker, R. B. (2011). Selection and character-
ization of suitable lipid excipients for use in the manufacture of Didanosine-loaded
solid lipid nanoparticles and nanostructured lipid carriers. Journal of Pharmaceutical
Sciences, 100(12), 5185–5196.
Keller, B. C. (2001). Liposomes in nutrition. Trends in Food Science & Technology, 12(1),
25–31.
Klang, V., Matsko, N. B., Valenta, C., & Hofer, F. (2012). Electron microscopy of
nanoemulsions: An essential tool for characterisation and stability assessment. Mi-
cron, 43(2–3), 85–103.
Kumbhar, D. D., & Pokharkar, V. B. (2013). Engineering of a nanostructured lipid carrier
for the poorly water-soluble drug, Bicalutamide: Physicochemical investigations.
Colloids and Surfaces A: Physicochemical and Engineering Aspects, 416, 32–42.
Kuo, Y. -C., & Chung, J. -F. (2011). Physicochemical properties of nevirapine-loaded
solid lipid nanoparticles and nanostructured lipid carriers. Colloids and Surfaces.
B, Biointerfaces, 83(2), 299–306.
41
Lacatusu, I., Badea, N., Ovidiu, O., Bojin, D., & Meghea, A. (2012). Highly antioxidant
carotene-lipid nanocarriers: Synthesis and antibacterial activity. Journal of Nano-
particle Research, 14(6), 1–16.
Lakshmi, P., & Kumar, G. A. (2010). Nanosuspension technology: A review. International
Journal of Pharmacy and Pharmaceutical Sciences, 2(4), 35–40.
Li, B., & Ge, Z. -Q. (2012). Nanostructured lipid carriers improve skin permeation and
chemical stability of Idebenone. AAPS Pharmaceutical Science and Technology,
13(1), 276–283.
Li, Z., Lin, X., Yu, L., Li, X., Geng, F., & Zheng, L. (2009). Effects of chloramphenicol on the
characterization of solid lipid nanoparticles and nanostructured lipid carriers. Journal
of Dispersion Science and Technology, 30(7), 1008–1014.
Li, F., Weng, Y., Wang, L., He, H., Yang, J., & Tang, X. (2010). The efficacy and safety
of bufadienolides-loaded nanostructured lipid carriers. International Journal of
Pharmaceutics, 393(1–2), 203–211.
Lin, X., Li, X., Zheng, L., Yu, L., Zhang, Q., & Liu, W. (2007). Preparation and characterization
of monocaprate nanostructured lipid carriers. Colloids and Surfaces A: Physicochemical
and Engineering Aspects, 311(1–3), 106–111.
Liu, D., Liu, Z., Wang, L., Zhang, C., & Zhang, N. (2011). Nanostructured lipid carriers as novel
carrier for parenteral delivery of docetaxel. Colloids and Surfaces. B, Biointerfaces, 85(2),
262–269.
Liu, W., Tian, R., Hu, W., Jia, Y., Jiang, H., Zhang, J., et al. (2012). Preparation and evaluation of
self-microemulsifying drug delivery system of baicalein. Fitoterapia, 83(8), 1532–1539.
Liu, G. Y., Wang, J. M., & Xia, Q. (2012). Application of nanostructured lipid carrier in food for
the improved bioavailability. European Food Research and Technology, 234(3), 391–398.
Liu, C. -H., & Wu, C. -T. (2010). Optimization of nanostructured lipid carriers for lutein
delivery. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 353(2–3),
149–156.
Lopez-Huertas, E. (2010). Health effects of oleic acid and long chain omega-3 fatty acids
(EPA and DHA) enriched milks. A review of intervention studies. Pharmacological
Research, 61(3), 200–207.
Luo, Q., Zhao, J., Zhang, X., & Pan, W. (2011). Nanostructured lipid carrier (NLC) coated
with chitosan oligosaccharides and its potential use in ocular drug delivery system.
International Journal of Pharmaceutics, 403(1–2), 185–191.
Mäder, K., & Mehnert, W. (2005). Solid lipid nanoparticles-concepts, procedures, and
physicochemical aspects. In C. Nastruzzi (Ed.), Lipospheres in drug targets and delivery:
Approaches, methods, and applications (pp. 1–22). New York: CRC Press.
Martini, S., & Herrera, M. L. (2008). Physical properties of shortenings with low-trans
fatty acids as affected by emulsifiers and storage conditions. European Journal of
Lipid Science and Technology, 110(2), 172–182.
Mason, T. G., Wilking, J. N., Meleson, K., Chang, C. B., & Graves, S. M. (2006).
Nanoemulsions: Formation, structure, and physical properties. Journal of Physics.
Condensed Matter, 18(41), R635–R666.
Maynard, A. D., Aitken, R. J., Butz, T., Colvin, V., Donaldson, K., Oberdörster, G., et al.
(2006). Safe handling of nanotechnology. Nature, 444(7117), 267–269.
McClements, D. J. (2005). Food emulsions: principles, practice, and techniques (2th ed.).
London: CRC Press.
McClements, D. J. (2012a). Crystals and crystallization in oil-in-water emulsions: Implica-
tions for emulsion-based delivery systems. Advances in Colloid and Interface Science,
174, 1–30.
McClements, D. J. (2012b). Nanoemulsions versus microemulsions: Terminology, differ-
ences, and similarities. Soft Matter, 8(6), 1719–1729.
McClements, D. J., Decker, E. A., & Weiss, J. (2007). Emulsion-based delivery systems for
lipophilic bioactive components. Journal of Food Science, 72(8), R109–R124.
McClements, D. J., & Li, Y. (2010). Structured emulsion-based delivery systems: Controlling
the digestion and release of lipophilic food components. Advances in Colloid and Interface
Science, 159(2), 213–228.
McClements, D. J., & Rao, J. (2011). Food-grade nanoemulsions: Formulation, fabrication,
properties, performance, biological fate, and potential toxicity. Critical Reviews in
Food Science and Nutrition, 51(4), 285–330.
Mehnert, W., & Mäder, K. (2001). Solid lipid nanoparticles: Production, characterization
and applications. Advanced Drug Delivery Reviews, 47(2–3), 165–196.
Mehnert, W., & Mäder, K. (2012). Solid lipid nanoparticles: Production, characterization
and applications. Advanced Drug Delivery Reviews, 64, 83–101.
Mitri, K., Shegokar, R., Gohla, S., Anselmi, C., & Müller, R. H. (2011). Lipid nanocarriers for
dermal delivery of lutein: Preparation, characterization, stability and performance.
International Journal of Pharmaceutics, 414(1–2), 267–275.
Moraru, C., Huang, Q., Takhistov, P., Dogan, H., & Kokini, J. (2009). Food nanotechnology:
current developments and future prospects. In G. Barbosa-Canovas, A. Mortimer, D.
Lineback, W. Spiess, K. Buckle, & P. Colonna (Eds.), Global issues in food science and
technology (pp. 369–399). Burlington: Academic Press.
Mozafari, M. R. (2006). Bioactive entrapment and targeting using nanocarrier technologies:
An introduction. In M. R. Mozafari (Ed.), Nanocarrier technologies: frontiers of
nanotherapy (pp. 1–16). Amsterdam: Springer.
Mozafari, M. R., Johnson, C., Hatziantoniou, S., & Demetzos, C. (2008). Nanoliposomes and
their applications in food nanotechnology. Journal of Liposome Research, 18(4),
309–327.
Mu, X., & Zhong, Z. (2006). Preparation and properties of poly(vinyl alcohol)-stabilized
liposomes. International Journal of Pharmaceutics, 318(1–2), 55–61.
Müller, R. H., Mäder, K., & Gohla, S. (2000). Solid lipid nanoparticles (SLN) for controlled
drug delivery — a review of the state of the art. European Journal of Pharmaceutics and
Biopharmaceutics, 50(1), 161–177.
Müller, R. H., Mäder, K., Lippacher, A., & Jenning, V. (1999). Fest-flüssig (halbfeste)
Lipidpartikel (Nano-Compartiment-Carrier-NCC) und Verfahren zur Herstellung
hochkonzentrierter Lipidpartikel. Germany Patent, DE19945203A1.
Müller, R. H., Mehnert, W., Lucks, J. S., Chwarz, C. S., zur Muhlen, A., Wyhers, H., et al.
(1995). Solid lipid nanoparticles (SLN): An alternative colloidal carrier system for
42 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
controlled drug delivery. European Journal of Pharmaceutics and Biopharmaceutics,
41(1), 62–69.
Müller, R. H., Petersen, R. D., Hommoss, A., & Pardeike, J. (2007). Nanostructured lipid
carriers (NLC) in cosmetic dermal products. Advanced Drug Delivery Reviews, 59(6),
522–530.
Müller, R. H., Radtke, M., & Wissing, S. A. (2002a). Nanostructured lipid matrices for
improved microencapsulation of drugs. International Journal of Pharmaceutics,
242(1–2), 121–128.
Müller, R. H., Radtke, M., & Wissing, S. A. (2002b). Solid lipid nanoparticles (SLN) and
nanostructured lipid carriers (NLC) in cosmetic and dermatological preparations.
Advanced Drug Delivery Reviews, 54(Supplement 1), S131–S155.
Nawar, W. W. (1996). Lipids. In O. R. Fennema (Ed.), Food chemistry (pp. 225–319).
New York: Marcel Dekker.
Nayak, A. P., Tiyaboonchai, W., Patankar, S., Madhusudhan, B., & Souto, E. B. (2010).
Curcuminoids-loaded lipid nanoparticles: Novel approach towards malaria treat-
ment. Colloids and Surfaces. B, Biointerfaces, 81(1), 263–273.
Neethirajan, S., & Jayas, D. S. (2011). Nanotechnology for the food and bioprocessing in-
dustries. Food and Bioprocess Technology, 4(1), 39–47.
Nguyen, H. M., Hwang, I. C., Park, J. W., & Park, H. J. (2012). Enhanced payload and
photo-protection for pesticides using nanostructured lipid carriers with corn oil
as liquid lipid. Journal of Microencapsulation, 29(6), 596–604.
Obeidat, W. M., Schwabe, K., Müller, R. H., & Keck, C. M. (2010). Preservation of
nanostructured lipid carriers (NLC). European Journal of Pharmaceutics and
Biopharmaceutics, 76(1), 56–67.
Pardeike, J., Hommoss, A., & Müller, R. H. (2009). Lipid nanoparticles (SLN, NLC) in cosmetic
and pharmaceutical dermal products. International Journal of Pharmaceutics, 366(1–2),
170–184.
Pardeike, J., Weber, S., Haber, T., Wagner, J., Zarfl, H. P., Plankb, H., et al. (2011). Development
of an Itraconazole-loaded nanostructured lipid carrier (NLC) formulation for pulmonary
application. International Journal of Pharmaceutics, 419(1–2), 329–338.
Pine, D. J., Weitz, D. A., Chaikin, P. M., & Herbolzheimer, E. (1988). Diffusion wave spectros-
copy. Physical Review Letters, 60(12), 1134–1137.
Porter, C. J. H., Trevaskis, N. L., & Charman, W. N. (2007). Lipids and lipid-based formulations:
Optimizing the oral delivery of lipophilic drugs. Nature Reviews. Drug Discovery, 6(3),
231–248.
Pouton, C. W. (2006). Formulation of poorly water-soluble drugs for oral administration:
Physicochemical and physiological issues and the lipid formulation classification sys-
tem. European Journal of Pharmaceutical Sciences, 29(3–4), 278–287.
Priano, Esposti, D., Esposti, R., Castagna, G., DeMedici, C., Fraschini, F., et al. (2007). Solid
lipid nanoparticles incorporating melatonin as new model for sustained oral and trans-
dermal delivery systems. Journal of Nanoscience and Nanotechnology, 7(10), 3596–3601.
Puglia, C., Frasca, G., Musumeci, T., Rizza, L., Puglisi, G., Bonina, F., et al. (2012). Curcumin
loaded NLC induces histone hypoacetylation in the CNS after intraperitoneal admin-
istration in mice. European Journal of Pharmaceutics and Biopharmaceutics, 81(2),
288–293.
Qian, C., Decker, E. A., Xiao, H., & McClements, D. J. (2012). Physical and chemical sta-
bility of β-carotene-enriched nanoemulsions: Influence of pH, ionic strength, tem-
perature, and emulsifier type. Food Chemistry, 132(3), 1221–1229.
Qiu, S., Liu, Z., Hou, L., Li, Y., Wang, J., Wang, H., et al. (2013). Complement activation
associated with polysorbate 80 in beagle dogs. International Immunopharmacology,
15(1), 144–149.
Radomska-Soukharev, A., & Müller, R. H. (2006). Chemical stability of lipid excipients
in SLN-production of test formulations, characterisation and short-term stability.
Pharmazie, 61(5), 425–430.
Radtke, M., & Müller, R. H. (2001). NLC™ nanostructured lipid carriers: The new gener-
ation of lipid drug carriers. New Drugs, 2, 48–52.
Radtke, M., Souto, E. B., & Müller, R. H. (2005). Nanostructured lipid carriers: A novel gener-
ation of solid lipid drug carriers. Pharmaceutical Technology Europe, 17(4), 45–50.
Reimer, L. (1993). Image formation in low-voltage scanning electron microscopy. Bellingham:
SPIE Optical Engineering Press.
Rowe, R. C., Sheskey, P. J., & Quinn, M. E. (2009). Handbook of pharmaceutical excipients
(6th ed.). London: Pharmaceutical Press.
Sanad, R. A., AbdelMalak, N. S., elBayoomy, T. S., & Badawi, A. A. (2010). Formulation of
a novel oxybenzone-loaded Nanostructured Lipid Carriers (NLCs). AAPS Pharma-
ceutical Science and Technology, 11(4), 1684–1694.
Sato, K. (1988). Crystallization of fats and fatty acids. In N. Garti, & K. Sato (Eds.), Crystalliza-
tion and polymorphism of fats and fatty acids (pp. 227–266). New York: Marcel Dekker
Inc.
Saupe, A., Gordon, K. C., & Rades, T. (2006). Structural investigations on nanoemulsions,
solid lipid nanoparticles and nanostructured lipid carriers by cryo-field emission
scanning electron microscopy and Raman spectroscopy. International Journal of
Pharmaceutics, 314(1), 56–62.
Saupe, A., Wissing, S. A., Lenk, A., Schmidt, C., & Müller, R. H. (2005). Solid Lipid
Nanoparticles (SLN) and Nanostructured Lipid Carriers (NLC) — Structural investi-
gations on two different carrier systems. Bio-Medical Materials and Engineering,
15(5), 393–402.
Sawant, K. K., & Dodiya, S. S. (2008). Recent advances and patents on solid lipid
nanoparticles. Recent Patents on Drug Delivery & Formulation, 2(2), 120–135.
SCENIHR (Scientific Committee on Emerging and Newly-Identified Health Risks)
(2007). The existing and proposed definitions relating to products of nanotechnol-
ogies. : Health & Consumer Protection Directorate-General, European Commission.
Schubert, M. A., & Müller-Goymann, C. C. (2003). Solvent injection as a new approach for
manufacturing lipid nanoparticles-evaluation of the method and process parameters.
European Journal of Pharmaceutics and Biopharmaceutics, 55(1), 125–131.
Severino, P., Pinho, S. C., Souto, E. B., & Santana, M. H. A. (2011). Polymorphism, crystallin-
ity and hydrophilic–lipophilic balance of stearic acid and stearic acid-capric/caprylic
triglyceride matrices for production of stable nanoparticles. Colloids and Surfaces. B,
Biointerfaces, 86(1), 125–130.
Severino, P., Santana, M. H. A., & Souto, E. B. (2012). Optimizing SLN and NLC by 22 full
factorial design: Effect of homogenization technique. Materials Science and Engineering:
C, 32(6), 1375–1379.
Shimpi, S. L., Mahadik, K. R., & Paradkar, A. R. (2009). Study on mechanism for amor-
phous drug stabilization using Gelucire 50/13. Chemical & Pharmaceutical Bulletin,
57(9), 937–942.
Siepmann, F., Muschert, S., Flament, M. P., Leterme, P., Gayot, A., & Siepmann, J. (2006).
Controlled drug release from Gelucire-based matrix pellets: Experiment and theo-
ry. International Journal of Pharmaceutics, 317(2), 136–143.
Singh, S., Dobhal, A. K., Jain, A., Pandit, J. K., & Chakraborty, S. (2010). Formulation and
evaluation of solid lipid nanoparticles of a water soluble drug: Zidovudine. Chemical
and Pharmaceutical Bulletin, 58(5), 650–655.
Singhal, G. B., Patel, R. P., Prajapati, B. G., & Patel, N. A. (2011). Solid lipid nanoparticles
and nano lipid carriers: As novel solid lipid based drug carrier. International Research
Journal of Pharmacy, 2(2), 40–52.
Souto, E. B., & Müller, R. H. (2005). SLN and NLC for topical delivery of ketoconazole.
Journal of Microencapsulation, 22(5), 501–510.
Stauffer, C. E. (1999). Emulsifiers. St. Paul, MN: Eagen Press.
Tadros, T., Izquierdo, R., Esquena, J., & Solans, C. (2004). Formation and stability of
nano-emulsions. Advances in Colloid and Interface Science, 108–109, 303–318.
Tamjidi, F., Nasirpour, A., & Shahedi, M. (2012). Physicochemical and sensory properties of
yogurt enriched with microencapsulated fish oil. Food Science and Technology Interna-
tional, 18(4), 381–390.
Tan, S. W., Billa, N., Roberts, C. R., & Burley, J. C. (2010). Surfactant effects on the phys-
ical characteristics of Amphotericin B-containing nanostructured lipid carriers.
Colloids and Surfaces A: Physicochemical and Engineering Aspects, 372(1–3), 73–79.
Taylor, T. M., Weiss, J., Davidson, P. M., & Bruce, B. D. (2005). Liposomal nanocapsules in food
science and agriculture. Critical Reviews in Food Science and Nutrition, 45(7–8), 587–605.
Teeranachaideekul, V., Müller, R. H., & Junyaprasert, V. B. (2007). Encapsulation of ascorbyl
palmitate in nanostructured lipid carriers (NLC)-effects of formulation parameters on
physicochemical stability. International Journal of Pharmaceutics, 340(1–2), 198–206.
Teeranachaideekul, V., Souto, E. B., Junyaprasert, V. B., & Müller, R. H. (2007). Cetyl
palmitate-based NLC for topical delivery of Coenzyme Q10 — Development, phys-
icochemical characterization and in vitro release studies. European Journal of
Pharmaceutics and Biopharmaceutics, 67(1), 141–148.
Tiwari, R., & Pathak, K. (2011). Nanostructured lipid carrier versus solid lipid
nanoparticles of simvastatin: Comparative analysis of characteristics, pharmacoki-
netics and tissue uptake. International Journal of Pharmaceutics, 415(1–2), 232–243.
Trujillo, C. C., & Wright, A. J. (2010). Properties and stability of solid lipid particle dis-
persions based on canola stearin and Poloxamer 188. Journal of the American Oil
Chemists' Society, 87(7), 715–730.
Tsai, M. -J., Wu, P. -C., Huang, Y. -B., Chang, J. -S., Lin, C. -L., Tsai, Y. -H., et al. (2012).
Baicalein loaded in tocol nanostructured lipid carriers (tocol NLCs) for enhanced sta-
bility and brain targeting. International Journal of Pharmaceutics, 423(2), 461–470.
Uner, M. (2006). Preparation, characterization and physico-chemical properties of
Solid Lipid Nanoparticles (SLN) and Nanostructured Lipid Carriers (NLC): Their
benefits as colloidal drug carrier systems. Pharmazie, 61(5), 375–386.
Valls, V., Goicoechea, M., Muniz, P., Saez, G. T., & Cabo, J. R. (2003). Effect of corn oil and
vitamin E on the oxidative status of adipose tissues and liver in rat. Food Chemistry,
81(2), 281–286.
Varshosaz, J., Eskandari, S., & Tabakhian, M. (2010). Production and optimization of
valproic acid nanostructured lipid carriers by the Taguchi design. Pharmaceutical
Development and Technology, 15(1), 89–96.
Varshosaz, J., Eskandari, S., & Tabbakhian, M. (2012). Freeze-drying of nanostructure
lipid carriers by different carbohydrate polymers used as cryoprotectants. Carbohy-
drate Polymers, 88(4), 1157–1163.
Varshosaz, J., Hassanzadeh, F., Sadeghi, H., & Andalib, S. (2012). Synthesis of
octadecylamine-retinoic acid conjugate for enhanced cytotoxic effects of 5-FU
using LDL targeted nanostructured lipid carriers. European Journal of Medicinal
Chemistry, 54, 429–438.
Varshosaz, J., Hassanzadeh, F., Sadeghi, H., & Khadem, M. (2012). Galactosylated nano-
structured lipid carriers for delivery of 5-FU to hepatocellular carcinoma. Journal of
Liposome Research, 22(3), 224–236.
Vivek, K., Reddy, H., & Murthy, R. S. R. (2007). Investigations of the effect of the lipid matrix
on drug entrapment, in vitro release, and physical stability of olanzapine-loaded solid
lipid nanoparticles. AAPS Pharmaceutical Science and Technology, 8(4), 16–24.
Walstra, P. (2003). Physical chemistry of foods. New York: Marcel Decker.
Wang, Y., & Wu, W. (2006). In situ evading of phagocytic uptake of stealth solid lipid
nanoparticles by mouse peritoneal macrophages. Drug Delivery, 13(3), 189–192.
Weiss, J., Decker, E. A., McClements, D. J., Kristbergsson, K., Helgason, T., & Awad, T.
(2008). Solid lipid nanoparticles as delivery systems for bioactive food compo-
nents. Food Biophysics, 3(2), 146–154.
Weiss, J., Gaysinsky, S., Davidson, M., & McClements, D. J. (2009). Nanostructured en-
capsulation systems: Food antimicrobials. In G. Barbosa-Canovas, A. Mortimer, D.
Lineback, W. Spiess, K. Buckle, & P. Colonna (Eds.), Global issues in food science
and technology (pp. 425–479). Burlington: Academic Press.
Weiss, J., Takhistov, P., & McClements, D. J. (2006). Functional materials in food nanotechnol-
ogy. Journal of Food Science, 71(9), R107–R116.
Whitehurst, R. J. (2004). Emulsifiers in food technology. Oxford: Blackwell Publishing Ltd.
Wissing, S. A., Kayser, O., & Müller, R. H. (2004). Solid lipid nanoparticles for parenteral
drug delivery. Advanced Drug Delivery Reviews, 56(9), 1257–1272.
Wissing, S. A., & Müller, R. H. (2002). Solid lipid nanoparticles as carrier for sunscreens:
in vitro release and in vivo skin penetration. Journal of Controlled Release, 81(3),
225–233.
 F. Tamjidi et al. / Innovative Food Science and Emerging Technologies 19 (2013) 29–43
Wooster, T., Golding, M., & Sanguansri, P. (2008). Impact of oil type on nanoemulsion
formation and Ostwald ripening stability. Langmuir, 24(22), 12758–12765.
Xia, Q., Saupe, A., Müller, R. H., & Souto, E. B. (2007). Nanostructured lipid carriers as
novel carrier for sunscreen formulations. International Journal of Cosmetic Science,
29(6), 473–482.
Xia, Q., & Wang, H. (2011). Study on stability of coenzyme Q10-loaded nanostructured
lipid carriers. Integrated Ferroelectrics, 129(1), 208–214.
Yang, Y., Feng, J. F., Zhang, H., & Luo, J. Y. (2006). Optimization preparation of
chansu-loaded solid lipid nanoparticles by central composite design and response
surface method. Zhongguo Zhong Yao Za Zhi, 31(8), 650–653.
Yuan, H., Wang, L. -L., Du, Y. -Z., You, J., Hu, F. -Q., & Zeng, S. (2007). Preparation and
characteristics of nanostructured lipid carriers for control-releasing progester-
one by melt-emulsification. Colloids and Surfaces. B, Biointerfaces, 60(2), 174–179.
Zhang, T., Chen, J., Zhang, Y., Shen, Q., & Pan, W. (2011). Characterization and evaluation
of nanostructured lipid carrier as a vehicle for oral delivery of etoposide. European
Journal of Pharmaceutical Sciences, 43(3), 174–179.
43
Zhang, X., Liu, J., Qiao, H., Liu, H., Ni, J., Zhang, W., et al. (2010). Formulation optimization
of dihydroartemisinin nanostructured lipid carrier using response surface methodol-
ogy. Powder Technology, 197(1–2), 120–128.
Zheng, K., Zou, A., Yang, X., Liu, F., Xia, Q., Ye, R., et al. (2013). The effect of
polymeresurfactant emulsifying agent on the formation and stability of a-lipoic acid
loaded nanostructured lipid carriers (NLC). Food Hydrocolloids, 32(1), 72–78.
Zhuang, C. -Y., Li, N., Wang, M., Zhang, X. -N., Pan, W. -S., Peng, J. -J., et al. (2010). Preparation
and characterization of vinpocetine loaded nanostructured lipid carriers (NLC) for im-
proved oral bioavailability. International Journal of Pharmaceutics, 394(1–2), 179–185.
Zimet, P., Rosenberg, D., & Livney, Y. D. (2011). Re-assembled casein micelles and
casein nanoparticles as nano-vehicles for ω-3 polyunsaturated fatty acids. Food
Hydrocolloids, 25(5), 1270–1276.
Zimmermann, E., Müller, R. H., & Mäder, K. (2005). Physicochemical investigations on the
structure of drug-free and drug-loaded solid lipid nanoparticles (SLN) by means of
DSC and 1H-NMR. European Journal of Pharmaceutics and Biopharmaceutics, 60(7),
508–513.