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Abstract
Over the past few decades, nanoparticle (NP) formulation has been the subject of extensive research. The
choice of a suitable NP formulation technique is dependent on the physicochemical properties of the drug,
such as solubility and chemical stability. Different NP manufacturing methods enable modification of the
physicochemical characteristics such as size, structure, morphology and surface texture, but also affect the
drug loading, drug entrapment efficiency and release kinetics. This review covers an update on the state of
art of the manufacturing of polymeric NPs from preformed polymers. Both, conventional methods for NP
preparation, such as spontaneous formulation and emulsification-based methods, and new approaches in
NP technology are presented. A comparative analysis is given for polymer, drug and solvent nature, toxicity,
purification, drug stability and scalability of the method. The information obtained allows establishing
criteria for selecting a method for preparation of NPs according to its advantages and limitations.
Keywords: nanoparticles, polymer, formulation, preparation methods, drug delivery
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Introduction
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NPs composed of polymeric materials have been extensively investigated for their use in delivery and controlled
release of low molecular weight drugs, peptides and nucleotides via oral, topical and parenteral routes (Table 1).
Different polymers, both synthetic and natural, have been
developed to improve site-specific delivery, controlled
release and drug-loading capabilities. The most common
types of polymer used for the production of NP carrier
systems are Food and Drug Administration approved
hydrophobic materials such as poly-lactic acid (PLA),
poly-(lactic-co-glycolic acid) (PLGA), polycaprolactone
(PCL) and hydrophilic polymers such as chitosan, albumin,
gelatine and alginate. Advanced polymer chemistry and
engineering are leading to the production of increasingly
intricate polymer structures, including multivalent polymers, branched polymers, graft polymers, dendrimers,
dendronized polymers, block copolymers, star-like
polymers and hybrid glyco- and peptide polymeric derivatives. Their advantages include better defined chemical
composition, tailored surface multivalency and defined
three-dimensional architecture. Synthetic hydrophobic
polymers have the advantage of sustaining the release of
Address for correspondence: Pegi Ahlin Grabnar, Faculty of Pharmacy, University of Ljubljana, Askerceva 7, 1000 Ljubljana, Slovenia. Tel: 386 1 4769 621.
Fax: 386 1 4258 031. E-mail: ahlinp@ffa.uni-lj.si
(Received 8 Nov 2010; accepted 28 Feb 2011)
http://www.informahealthcare.com/mnc
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phase separation
nanoprecipitation
EMULSIFICATION TECHNIQUES
ionic gelation
coacervation
solvent evaporation
W/O emulsification
solvent displacement
solvent diffusion
salting-out
AQUEOUS PHASE
(water + stabilizer)
ORGANIC PHASE
(solvent + polymer + drug)
Precipitation
Nanoparticles
polymer/solvent
Water
NP
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AQUEOUS PHASE
(water + albumin)
Turbidimetry
control
ORGANIC PHASE
(oil)
Crosslinking
Aqueous solution
of protein
Nanoparticles
Emulsification
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W/O emulsion
Albumin nanospheres
Figure 4. W/O emulsification technique.
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ORGANIC PHASE
(solvent + polymer + drug)
Salt (only in
salting out
method)
protein or a peptide drug is the rapid diffusion of the molecule into the outer aqueous phase during the emulsification. This can result in poor entrapment efficiency, i.e. drug
loading. Therefore, it is critical to ensure immediate formation of a polymer membrane during the first water-in-oil
emulsification (Zambaux et al., 1998).
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Emulsification
O/W emulsion
Water
addition
3
SOLVENT
EVAPORATION
SOLVENT
DIFFUSION
SALTING
OUT
Nanoparticles
Figure 5. Emulsification-solvent evaporation (1), emulsification-solvent
diffusion (2) and salting-out method (3).
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emulsified in an aqueous stabilizer solution (PVA, poloxamer). Emulsification is carried out by sonication or under
high-energy homogenization to reduce the size of the
emulsion droplets and an O/W emulsion is formed. The
organic solvent is then removed by evaporation at room
temperature under stirring or under reduced pressure,
which leads to polymer precipitation and NP formation.
Two factors contribute to rapid solvent loss high volatility
of the solvent and the increased tendency of the polymer to
precipitate as it becomes more and more exposed to water.
As the precipitation process takes control over diffusion,
the solvent is expelled from the particles. The physicochemical properties of a drug are decisive in influencing
its loading and distribution between particles and aqueous
medium (Ahlin et al., 2000a, 2000b, 2003). The method
provides NPs with high drug entrapment efficacy, although
it is limited to lipophilic drugs that are soluble in the same
solvent as the polymer. Hydrophilic drugs are distributed
into the external water phase and not retained in the polymer phase. Limitations are imposed by the scale up of the
high energy requirements in homogenization and the use
of toxic chlorinated solvents.
The double emulsion solvent evaporation technique is a
modification of the emulsification-solvent evaporation
method and can be employed to entrap small hydrophilic
drugs and proteins. An aqueous drug solution is emulsified
in an organic polymer solution, usually by sonication. This
pre-emulsion is then added to an aqueous phase containing a stabilizer. This results in W/O/W emulsion formation
which is followed by evaporation of the organic solvent.
The polymer precipitate and NPs are formed. The main
problem with encapsulating a hydrophilic molecule like a
Emulsification-solvent diffusion
The emulsification-solvent diffusion method (Leroux et al.,
1995) involves the use of an organic solvent that is partially
water-soluble, for example benzyl alcohol, propylene carbonate, ethyl acetate and others. Polymer and drug are
dissolved in this organic solvent, which is then emulsified
in an aqueous phase containing stabilizer to form an O/W
emulsion (Figure 5). A large amount of water is then added
to the system in order to overcome the miscibility ratio of
organic solvent. This causes the solvent to diffuse into the
external phase. The polymer precipitates as a result of
the diffusion of organic solvent into the water, leading to
the formation of NPs. The solvent can be eliminated by
cross-flow filtration. The method is suitable for hydrophobic drugs, such as estrogen (Kwon et al., 2001), enalaprilat
(Ahlin et al., 2002) and ibuprofen (Galindo-Rodriguez et al.,
2005). The advantages of the method are the use of low
toxicity solvents (benzyl alcohol) and high batch-to-batch
reproducibility. High entrapment efficiency (generally
470%) has been reported. The disadvantage of the
method is the large volume of water that has to be eliminated from the suspension.
The double emulsion solvent diffusion method combines
the advantages of two well-established techniques, double
emulsion solvent evaporation and emulsification solvent
diffusion, and it is suitable for incorporation of hydrophilic
drugs (Kocbek et al., 2007; Cohen-Sela et al., 2009). The
double emulsion system minimizes escape of the hydrophilic drug to the aqueous medium, and ethyl acetate is
more acceptable pharmaceutically and considered to be
less toxic than dichlormethane. The preparation technique
results in improved formulation characteristics over those
of the classical method, including smaller size, lower size
distribution, higher drug entrapment efficiency and more
biocompatible ingredients (Tween and Pluronic rather
than PVA). The authors explained these findings by the
different mechanisms of NP formation. The dichloromethane used in the double emulsion method is relatively
poorly soluble in water and the solvent does not diffuse
extensively through the aqueous phase. NPs are formed
during evaporation once the critical concentration for polymer precipitation is reached, with a single particle formed
from each emulsion droplet. On the other hand, ethyl acetate, used in the double emulsion solvent diffusion method,
is partially water-miscible, and the solvent diffuses freely
via the aqueous phase, creating regions of local supersaturation near the interface (Cohen-Sela et al., 2009).
A double emulsification-solvent diffusion technique has
been demonstrated to maintain protein function (Cegnar
et al., 2004). Maintenance of protein activity during the
fabrication process involves a delicate balance between
energy input (mechanical stirring, homogenization and
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Salting-out
The emulsification-solvent diffusion method can be
improved by salting-out effects. In the salting-out technique a water-miscible solvent, like acetone, ethanol or
N-methyl-2-pyrrolidone, is used instead of chlorinated solvents. It was first proposed by Bindschaedler et al. (Ibrahim
et al., 1992; Kristl et al., 1996; Bindschaedler et al., 1988).
The mixing of organic and aqueous phases is prevented by
saturating the aqueous phase with electrolytes such as
magnesium acetate, magnesium chloride or calcium chloride. An organic solution of polymer and drug is emulsified
into an aqueous phase, which contains salt and colloidal
stabilizer, to form an O/W emulsion (Figure 5). The emulsion is then diluted with sufficient water to enhance the
diffusion of acetone into the aqueous phase, thus inducing
polymer precipitation. Both the solvent and the salting-out
agent are then eliminated by cross-flow filtration. The
method does not require an increase of temperature and
avoids the use of organic chlorinated solvents and large
amounts of stabilizer during formulation. High drug loadings can be achieved, depending on the solubility of the
drug in acetone and on the nature of the salting-out agent.
The disadvantages are exclusive application to lipophilic
drugs and the extensive NP washing steps. The use of acetone and large amounts of salts may raise some concern
about recycling of the salts and about compatibility with
active compounds.
The application of emulsification techniques is greatly
limited by disadvantages such as working with toxic solvents (dichloromethane, chloroform) and the requirement
of high energy apparatus (ultrasound probe or homogenizer). In contrast, methods for spontaneous formation of
NPs do not require a source of external energy or organic
solvents, except in nanoprecipitation where less toxic
organic solvents such as acetone (Class 3 according to
the ICH specifications) are used (Table 2).
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SCF technology
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Hydrophobic
Hydrophilic
Hydrophilic
Hydrophilic
Hydrophobic
Hydrophobic
Hydrophobic
Hydrophobic
Hydrophobic
Spontaneous formation
Nanoprecipitation
Coacervation
Ionic gelation
Emulsification-based methods
W/O emulsification technique
Hydrophobic
Hydrophilic
Hydrophobic
Hydrophilic
Hydrophobic
Hydrophilic
Lipophilic
Hydrophilic
Hydrophilic
Drug nature
Class 3/Class 2
Class 3/Class 2
Class 3
Class 3
Class 3
Class 3/Class 2
Class 3
No organic solvent
No organic solvent
Solvent nature
Residual solvents
Hardening agents (glutaraldehyde)
Low toxicity
Toxicity
Low
Low
Moderate
Moderate
High
High
Low
Moderate
Low
Purificationa
Moderate
Moderate*
High
Moderate*
High
Moderate
High
Moderate
High
Drug stablityb
Yes
Yes
Yes
Yes
Yes
NR
Yes
NR
NR
Scalability
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Polymer nature
Method
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High
pressure
vessle
Rapid expansion of
SCF after leaving
the nozzle and
polymer
precipitation
NP in
collection
chamber
High
pressure
collection
chamber
Expanded
solution
Supercritical
anti-solvent
GAS/SASS
RESS
Figure 6. SCF technologies: RESS and gas/supercritical anti-solvent technique (GAS/SAS) (adapted from Ginty et al. (2005) with permission from
Elsevier).
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Syringe pump
(solvent + polymer + drug)
High voltage
power supplies
Particle
collector
Air inlet
4
Heater
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Membrane reactor
The membrane reactor (contactor) controls the addition of
one reactant (the organic phase) to another reactant (the
aqueous phase; Figure 8). This process can be compared to
the membrane emulsification process, where the oil (or the
water phase) permeates through the membrane pores to
form droplets in water (or oil phase) for the preparation of
O/W or W/O emulsions (Charcosset and Fessi, 2005b). The
organic phase is pressed through the membrane pores,
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of the aqueous
phase
Nanoparticles
Organic phase
(solvent + polymer + drug)
Figure 8. Membrane reactor method (adapted from Charcosset and Fessi
(2006) with permission from Elsevier).
Coarse
emulsion
Porous glass
membrane
Nanoemulsion
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Aerosol flow reactors are similar to spray dryers. They operate by atomizing a solution of polymer and drug in a
common solvent or solvent mixture (water, ethanol, etc.)
via a collision-type air jet atomizer into an inert carrier gas
which is then passed through a heated tubular chamber
(Figure 10; Peppas et al., 2007). The solvent is evaporated
in a controlled manner using a heated tubular laminar flow
reactor, and the particles produced are collected as a dry
powder using a low-pressure impactor (Eerikainen et al.,
2004). Compared to conventional spray drying, the aerosol
flow reactor method provides better control of the thermal
history and residence time of each droplet and product
particle, due to the laminar flow in the heated zone of
the reactor where droplet drying and particle formation
take place (Eerikainen et al., 2003). In spray drying, hot
gas is used as a source of heat to evaporate the solvent
and the spray-drying chamber is only used as a place for
heat transfer to occur and is not heated by itself.
The temperature of the gas changes across the chamber
as heat transfer occurs between the cold feed and the
hot gas. On the contrary, in the aerosol flow reactor
method, the droplets, which are generated by a jet
nebulizer, ultrasonic nebulizer or electrospray, are already
suspended in the carrier gas before they are fed into
the tubular flow reactor that is located in an oven and
maintained at a constant temperature. The carrier gas
flows evenly in the tubular reactor at a constant rate,
controlled temperature field and non-circulating flow
(Peppas et al., 2007).
The method is a simple and efficient one-step
process that can produce particles directly within a
desirable particle size range with consistent and controlled
properties (Raula et al., 2007). It produces dry particles
directly without the need for further purification, and
no additives are required (Eerikainen et al., 2004;
Raula et al., 2004).
Eerikainen et al. reported that corticosteroid-loaded
Eudragit NPs with mean diameter of 90 nm can be produced by an aerosol flow reactor method using ethanol.
The composition of the NPs and drying temperature did
not affect the particle size distribution, particle morphology
or structure. The problems of drug leaking and uneven
drug distribution, which are commonly encountered in
emulsion processes, are avoided using this novel method
(Eerikainen and Kauppinen, 2003).
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COLLECTION
DILLUTION
VIBRATION
SYSTEM
PULSATION CHAMBER
Heating zone
NOZZLE
PARTICLE
FORMATION
ELECTRODE
Carrier gas
mixed with
droplets
DROPLET
GENERATION
STROBOSCOPE
solvent + drug
+ polymer
AQUEOUS PHASE
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Declaration of interest
The authors report no conflicts of interest.
References
Ahlin P, Kristl J, Kristl A, Vrecer F. Investigation of polymeric nanoparticles
as carriers of enalaprilat for oral administration. Int J Pharm,
2002;239:11320.
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