Cavitation technology A greener processing technique

for the generation of pharmaceutical nanoemulsions

Manickam Sivakumar
a,
, Siah Ying Tang
b
, Khang Wei Tan
c
a
Manufacturing and Industrial Processes Research Division, Faculty of Engineering, University of Nottingham Malaysia Campus, 43500 Semenyih, Selangor, Malaysia
b
Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 46150 Bandar Sunway, Selangor, Malaysia
c
Department of Chemical Engineering, UCSI University, Kuala Lumpur, Malaysia
a r t i c l e i n f o
Article history:
Received 30 November 2013
Received in revised form 24 March 2014
Accepted 24 March 2014
Available online 3 April 2014
Keywords:
Nano
Emulsion
Ultrasound
Cavitation
Pharmaceutical
Hydrodynamic
a b s t r a c t
Novel nanoemulsion-based drug delivery systems (DDS) have been proposed as alternative and effective
approach for the delivery of various types of poorly water-soluble drugs in the last decade. This nanofor-
mulation strategy signicantly improves the cell uptake and bioavailability of numerous hydrophobic
drugs by increasing their solubility and dissolution rate, maintaining drug concentration within the ther-
apeutic range by controlling the drug release rate, and reducing systemic side effects by targeting to spe-
cic disease site, thus offering a better patient compliance. To date, cavitation technology has emerged to
be an energy-efcient and promising technique to generate such nanoscale emulsions encapsulating a
variety of highly potent pharmaceutical agents that are water-insoluble. The micro-turbulent implosions
of cavitation bubbles tear-off primary giant oily emulsion droplets to nano-scale, spontaneously leading
to the formation of highly uniform drug contained nanodroplets. A substantial body of recent literatures
in the eld of nanoemulsions suggests that cavitation is a facile, cost-reducing yet safer generation tool,
remarkably highlighting its industrial commercial viability in the development of designing novel nano-
carriers or enhancing the properties of existing pharmaceutical products. In this review, the fundamen-
tals of nanoemulsion and the principles involved in their formation are presented. The underlying
mechanisms in the generation of pharmaceutical nanoemulsion under acoustic eld as well as the advan-
tages of using cavitation compared to the conventional techniques are also highlighted. This review
focuses on recent nanoemulsion-based DDS development and how cavitation through ultrasound and
hydrodynamic means is useful to generate the pharmaceutical grade nanoemulsions including the com-
plex double or submicron multiple emulsions.
2014 Elsevier B.V. All rights reserved.
1. What is nanoemulsion?
Nanoemulsions are isotropic colloidal systems composed of oil
droplets dispersed in continuous aqueous media and stabilized
by surfactant molecules. They are also frequently known as ne-
dispersed emulsions, miniemulsions, ultrane emulsions and sub-
micron emulsions [1]. The nanoemulsions can be in the form of
simple emulsion of oil-in-water, O/W or water-in-oil, W/O types.
Typically the particle size or the mean droplet diameter covers a
size range of 20500 nm [2]. They are a class of transparent, trans-
lucent or sometimes milky emulsion systems which cannot be
formed spontaneously as being non-equilibrium systems. Unlike
microemulsions, which are also transparent and thermodynami-
cally stable, nanoemulsions have a higher solubilization capacity
for lipophilic drugs and better resistance toward droplet collisions
induced by Brownian motion, thus giving rise to an excellent
kinetic colloidal stability. Although they are kinetically stable,
nanoemulsions are thermodynamically unstable dispersions as
their free energy of formation is greater than that of their phase
separated state. However, due to their nanometer-sized droplets,
the long-term physical stability of nanoemulsions makes them
unique without any apparent occulation or coalescence during
storage. Therefore these tiny emulsions of nano-dimension are
sometimes referred to as approaching thermodynamic stability
[3]. In the eld of pharmaceuticals, nanoemulsion is usually a
homogeneous mixture consisting of oil droplets encapsulating bio-
active ingredients that are well-dispersed in an aqueous medium
in the presence of a surfactant or emulsier. In pharmaceutical
nanoemulsions, the oil droplets serve as the reservoir for hydro-
phobic drugs. The most widely used oils include ax-seed oil, olive
oil, castor oil, soybean oil, and other vegetable oils that are rich in
http://dx.doi.org/10.1016/j.ultsonch.2014.03.025
1350-4177/ 2014 Elsevier B.V. All rights reserved.

Corresponding author. Tel.: +60 3 8924 8156; fax: +60 3 8924 8017.
E-mail address: Sivakumar.Manickam@nottingham.edu.my (M. Sivakumar).
Ultrasonics Sonochemistry 21 (2014) 20692083
Contents lists available at ScienceDirect
Ultrasonics Sonochemistry
j our nal homepage: www. el sevi er . com/ l ocat e/ ul t son
Omega-3 and Omega-6-fatty acids and contain useful vitamins and
minerals. Non-ionic surfactants are preferred since they are of low
toxicity and known to be less affected by pH and ionic strength,
and they are generally regarded as safe (GRAS) and are biocompat-
ible. Combination of various surfactants has also been used to con-
trol droplet size and improve the stability of nanoemulsions. These
pharmaceutical nanoemulsions generally appear to be clear or
transparent with the mean droplet diameter no larger than
200 nm while some appear optically milky with the droplet size
up to 500 nm [4].
2. Why are pharmaceutical nanoemulsions?
Nanoemulsions are useful means of encapsulating, protecting
and delivering the poorly water-soluble bioactive components for
both functional food and pharmaceutical applications [5,6].
Although nanoemulsions have applications in wider technological
areas (Fig. 1), their vital importance could be felt more in the phar-
maceutical eld. Admittedly, innovations in material chemistry
and nanotechnology have synergistically fuelled the development
of novel drug delivery systems and nanocarriers that are biode-
gradable, biocompatible, targeting and stimulus-responsive [7].
Increasing attention has been devoted to these systems due to
their unique structure and properties, such as extremely smaller
droplet size with larger surface area to volume ratio, increasing
drug solubility and dissolution rate, protecting the bioactive com-
pounds against degradation, improving diffusion across intestinal
membrane and enhancing mucosal permeability and bioavailabil-
ity [813]. Nanoemulsion formulations offer appealing alternative
for the easy administration of poorly water-soluble drugs (hydro-
phobic bioactive agents) with reduced dosing frequency and drug
originated systemic toxic effects and thus they could be considered
as a promising drug delivery technology. The outstanding advanta-
ges of nanoemulsions over microemulsions in the pharmaceutical
area could be mentioned in terms of enhanced drug properties
and enhanced dosing requirements as shown in the Table 1. Over-
all nanoemulsions represent an effective, convenient, exible for-
mulation strategy and in fact, they are more patient compliant as
compared to other existing dosage forms which make them a
promising candidate for interesting pharmaceutical applications.
Approximately 40% of the newly discovered drugs or bioactive
agents intended for transdermal, parenteral, intravenous route or
oral ingestion are highly hydrophobic compounds with low
water solubility which reduce their dissolution-rate limited cell
absorption. The in vivo bioavailability of these drugs remains prob-
lematic due to their extremely poor solubility in the body uids,
thus greatly hinders their clinical translations. Several strategies
and formulations have been employed to overcome this limitation.
For these reasons, many of the hydrophobic drugs are frequently
formulated in the conventional dosage forms such as tablets, cap-
sules, pellets and injectable suspensions. Despite their established
manufacturing techniques and good reproducibility, these phar-
maceutical dosage forms suffer from the disadvantages of low drug
absorption rate across gastrointestinal (GI) tract due to the low
surface area, uncontrolled drug release prole due to the fast dis-
solution rate, non cell/tissue-specic drug delivery due to the
absence of targeting ligands, higher doses and frequent dosing
due to rapid metabolism rate of drugs in gastric uid. The gastric
uid is essentially a colorless digestive uid which is produced
from the mucous membrane of the stomach, consisting of hydro-
chloric acid, pepsin, rennin and mucin whereas for GI tract, it is a
9-meter long, hollow digestive tube inside the human body, start-
ing from mouth where food enters, to the rectum and anus where
food is expelled, which is also known as alimentary canal. Nano-
emulsions, on contrary, offer many crucial benets over the exist-
ing solid dosage forms. By virtue of their ne droplet diameter,
nanoemulsions provide ultra low interfacial tension and extremely
large surface area that could signicantly enhance the drug absorp-
tion in GI tract [14]. Therefore, nanoemulsion technology could
improve the therapeutic index of drugs by enhancing their efcacy
and increasing their tolerability in the human body. Nanoemulsion
could also carry large payloads (e.g. drugs, protein, peptides,
nucleic acids), protect the therapeutic agents from physiological
barriers, as well as enable the development and synthesis of novel
classes of polysaccharide nanoparticles [1517].
The efciency of drug delivery via different routes of adminis-
tration is of vital importance in medicine and healthcare. Paren-
teral route is generally regarded as one of the major routes of
drug administration due to its quickest onset of action caused by
rapid access to blood circulation. However, it has been proven to
be a challenging task to deliver hydrophobic drugs via parenteral
route. For a majority of the existing hydrophobic drugs, solvents
such as ethanol and propylene glycol are often used to solubilize
water-insoluble drugs prior to parenteral administration [18]. Nev-
ertheless, drug precipitation caused by co-solvent [19], severe pain
during injection and also hemolysis at injection site often occur
which strongly limits its application practically. One strategy
to tackle these problems is through intravenous fat emulsion
therapy. Commercially available intravenous fat emulsions such
Fig. 1. Applications of nanoemulsions.
2070 M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083
as Intralipid

are oil-in-water emulsion consisting of one or more

medium chain triglyceride, a phospholipid emulsier and glycerin.
The potential shortcoming of this therapy is that fat emboli may, if
coalescing of emulsion droplets occurs, be produced leading to
pulmonary embolism and potentially fatal consequences. It had
been reported that the droplets greater than 5 lm are large enough
to occlude small blood capillaries and risk of pulmonary emboli
[20]. Nanoemulsions seem to be ideal liquid vehicles for various
routes of drug delivery since they provide all the possible require-
ments of a therapeutic liquid system including easy formation, low
viscosity with Newtonian behavior, high drug loading and very
small droplet size [21]. For oral administration, dilution or degra-
dation of the drug vehicle in the body uids is not desirable before
a particular site of action is reached. The delay of vehicle degrada-
tion and drug release could be achieved by using nanoemulsions as
drug vehicle. In nanoemulsion, the drug moieties are encapsulated
in the oil droplets to preserve drug activity and particle integrity,
as well as to prevent partitioning of drug from inner oil phase to
external aqueous phase during the delivery through GI tract.
Lipophilic antineoplastic agents such as paclitaxel, curcumin,
tamoxifen, and dacarbazine have been encapsulated in nanoemul-
sions to increase cytotoxicity and to overcome multi-drug resis-
tance [9,12,22,23].
Nanoemulsions, on account of their higher ratio of surface
area to volume, with the intestine provide high concentration
gradient and improved pharmacokinetics and biodistribution of
therapeutic agents and thus minimize toxicity by their preferen-
tial accumulation at the target site. They improve the solubility of
hydrophobic compounds and render them suitable for oral,
parenteral, ocular and transdermal administration. Furthermore,
they increase the stability of a variety of therapeutic agents, like
peptides, and oligonucleotides. In particular, the bioavailability of
compounds encapsulated into emulsions is enhanced when
emulsion droplets are of nanometric size [24]. Pharmaceutical
scientists have shown that reducing the droplet size to values
below cell size (500 nm) produces higher absorption of the active
ingredient and higher particle uptake, by enhancing the mecha-
nisms of passive transport through the intestinal walls [25].
Due to their extremely smaller droplet size, injectable liquid
nanoemulsions meet no difculty to pass through the biological
barrier in the blood vessel, eliminating the risk of emboli. It has
been reported that droplets smaller than 20 nm will be cleared
from circulation via reticuloendothelial system (RES) within few
hours when injected intravenously, larger ones trapped in the
liver and the spleen within minutes [26]. Many studies have
shown that positively charged nanoemulsions possess improved
transdermal permeation of anti-inammatory drugs over the con-
ventional emulsions and gels due to their enhanced interaction
with negatively charged skin epithelia [27,28]. Briey, in improv-
ing therapeutic efcacy, nanotechnology based DDS (nanoemul-
sions/liposomes/nanoparticles) has beneted tens of millions of
patients by means of relieving the suffering and prolonging life
[7]. They have also changed the economics of drug development.
Making an existing drug into nanoemulsion-based formulations
may not only improve its performance but also extend its patent
life as a new pharmaceutical product. Not surprisingly the world
market for advanced DDS utilizing nanomaterials is expected to
reach $2.6 trillion by 2015 [29].
3 Principles of nanoemulsion formation
The dispersion of oil into water can be explained with the help
of Gibbs free energy, DG, based on thermodynamic theory as
expressed in the following Eq.(1) [1]:
DG l
2
l
1
1
where l
1
and l
2
are the chemical potentials of state 1 and 2. It sug-
gests that, when DG > 0, the phase transformation is thermodynam-
ically impossible. Whereas, when DG = 0, oil and water mixture is in
thermodynamic equilibrium. But, when DG < 0, the transformation
from heterogeneous multi-phase oil and water mixture to single
phase emulsion is possible, which is spontaneous and reversible.
When DG < 0, one bulk phase is dispersed into another bulk phase.
The congurational entropy change, DS
conf
can be described as
below Eq. (2) [30]:
DS
conf
nk
B
ln/ f1 /=/gln1 / 2
where n is the number of droplets of dispersed phase, k
B
is the
Boltzmann constant, and / is the dispersed phase volume fraction.
By combining the above two equations, Gibbs free energy can be
expressed as the sum of free energy (to form new interfacial area)
(DA
c12
) and congurational entropy as follows Eq. (3):
DG DA
c12
TDS
conf
3
From the above equation, DA
c12
has to be much lower than
TDS
conf
in order to full DG < 0. Owing to this, surfactants are
always required to achieve an ultra-low interfacial tension,
DA
c12
. However, the formation of nanoemulsion is not spontane-
ous and requires an intensive external energy. As a result, high
energy emulsication devices such as high pressure homogenizer
or Microuidizer are always in order. The high amount of external
energy needed for the formation can be described by using the fol-
lowing Eq. (4),
p c
1
R
1

1
R
2

4
where p indicates the Laplace pressure, R
1
and R
2
are the principal
radii of curvature of the droplets and c is the interfacial tension.
Theoretically, this happens only when the subjected shear rate is
much higher than the Laplace pressure of tiny droplets. As a result,
this partly supports the existence of a surfactant, as its employment
can signicantly reduces the interfacial tension, c. Therefore, the
applied stress can be reduced and the deformation of tiny droplets
can be avoided. A parameter that has been widely used to describe
the droplet deformation is Weber number (We) which can be
expressed as below Eq. (5):
We
G
g
g
c
R
2c
5
Table 1
Advantages of pharmaceutical nanoemulsions.
M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083 2071
where G
g
is the velocity gradient, g
c
is the viscosity of continuous
phase, and R is radius of curvature of the droplet. It suggests that
an increase in Weber number could increase the possibility for
the deformation of droplets and when it reaches a specic value
called as critical Weber number, the breakdown of droplets
occurs.
4. Synthesis and basic processes for the fabrication of
nanoemulsions
To synthesize nanoemulsion, it requires, in general, four compo-
nents: (a) oil phase (b) water phase (c) surfactant and (d) energy.
Fig. 2 shows the components involved and their role in the gener-
ation of nanoemulsions. Nanoemulsions cannot be formed sponta-
neously and consequently higher energy input is required.
Although the importance and promise of this nanoscale thera-
peutic system have been recognized in the past decade, the prepa-
ration of extremely ne emulsions could be a key challenge, as the
pharmacological properties of nanoemulsions can be largely inu-
enced by the physicochemical characteristics of the droplets i.e.
droplet size, size distribution, morphology, viscosity etc. Conven-
tionally, different homogenizing/dispersing technologies such as
simple agitation, colloidal mills, static mixers and high shear mix-
ers have been employed and are useful to generate emulsions. As
intense shear should be applied in order to overcome the Laplace
pressure so that larger droplets are broken into smaller droplets
of nanoregime, the formation of nanoemulsion is not possible with
these conventional mixing systems. The preparation of nanoemul-
sions with the uniformdroplet size as well as the choice of its prep-
aration method is of crucial importance in the current
pharmaceutical industries. This is because both the physicochem-
ical and organoleptic properties of nanoemulsions such as stability,
color, appearance, texture, and rheology are greatly affected by the
droplet size of nanoemulsions. More often than not, smaller drop-
let size leads to a creamier mouth feel and greater emulsion stabil-
ity [31]. In general, the basic processes used to fabricate the
nanoemulsions are classied under (a) low-energy emulsication
methods that include phase inversion temperature, solvent-diffu-
sion and spontaneous emulsication and (b) high-energy emulsi-
cation methods that require larger mechanical energy generated
by high-pressure homogenizer, ultrasonicator and Microuidizer.
Of the emulsifying methods used to design and formulate pharma-
ceutical naonoemulsions, the most common formulation technique
is focused on low-energy emulsication method i.e. phase inver-
sion temperature (PIT) technique. PIT method generates chemical
potential through the use of surfactants. However, several limita-
tions have been coupled with PIT method such as, the use of a large
quantity of surfactant, careful selection of both surfactant-cosur-
factant, high polydispersity index (PDI) induced instability after
long-term storage and relatively heavy and tedious workload to
identify the system inversion temperature. In comparison, nano-
emulsions prepared by ultrasonic cavitation offers a greater
improvement in terms of stability with lower Ostwald ripening
rate than the systems prepared using PIT method. The high energy
input originating from micro-turbulent implosions of cavitation
bubbles provides enormous forces that are able to deform and
break-off droplets into nanometer scale, provided the Laplace pres-
sure is overcome. However, in order to obtain smaller droplet sizes,
higher energy input is required for further droplet break-off [32]. It
was found that the droplet sizes of ultrasonically-induced nano-
emulsions are much smaller with superior physical stability than
the dispersion systems prepared by homogenizer [33]. Although
nanoemulsions with much smaller droplet sizes can be produced
by applying Microuidizer, a comparative study revealed that
microuidization is not convenient as compared to using an
ultrasound probe sonicator. The latter uses a probe and is much
easier to clean and is free of line blockage whereas for Microuidiz-
er, the contamination of materials still remains a main issue [34].
Furthermore, they are also less viable with respect to production
and maintenance costs. Besides, most of the conventionally
employed technologies are less energy intensive as only a small
part of the applied energy is effective for the droplet break-off
[35]. All these facts continue to make ultrasonic cavitation the
most effective and promising method in the preparation of
pharmaceutical nanoemulsions.
5. Why ultrasonic cavitation in the generation of
nanoemulsions?
Introducing a large amount of mechanical energy is essential to
produce ne emulsion droplets. In general, the mechanical mixing
could be achieved by using inline rotor/stator mixers, colloid mills
and high pressure homogenizers, etc. However, most of the tradi-
tional methods usually fail to provide a satisfactory control over
the particle size distribution and often result in poor dispersion
stability. Compared to mechanical agitation, high intensity ultra-
sound offers a better alternative yet energy efcient technique in
the generation of nanoemulsions. Ultrasound cavitation is dened
as a combined phenomenon of bubble formation, growth and
implosive collapse of the bubbles in a liquid medium. A transient
collapse of bubbles initiated by compressed air generates localized
hot spots with the temperatures up to 5000 K, pressure of approx-
imately 1000 bar and heating/cooling rates above 10
10
K/s [36].
Ultrasound emulsication was rst reported in 1927 by Wood
and Loomis [37], yet the rst patent was only granted in 1944
[38]. Li and Fogler [39,40] proposed two-stage ultrasound emulsi-
cation mechanism (Fig. 3). First stage is the generation of primary
droplets, where the acoustic eld produces the interfacial waves,
the instability of which causes eruption of oil phase into water in
the form of droplets. Second stage involves the break-up of pri-
mary droplets where the acoustic cavitation causes localized
intense turbulence and shear forces that produces violently and
asymmetrically imploding bubbles and causes microjets which
effectively further breaking the primary droplets of dispersed oil
generated in the rst stage into droplets of nanometric-sized. The
rst stage of break-up of droplets is primarily controlled by the
type and amount of shear applied to droplets. Resistance of drop-
lets to deformation is determined by the surfactant. In the second
stage (coalescence of droplets) where the rate at which the surfac-
tant is adsorbed into the newly formed droplets is controlled by
the surfactant surface activity and its concentration. Thus, selec-
tion of appropriate surfactant is of paramount importance in pro-
ducing nanoemulsions with high stablity. HLB (Hydrophilic-
Lipophilic balance) serves as an useful parameter in the selection
of surfactant. HLB is an empirical expression to indicate which sur-
factant is hydrophilic or hydrophobic. The lower the HLB value the
more lipophilic or oil soluble the surfactant is. The higher the HLB
value, the more hydrophilic or water soluble the surfactant is. In
general, surfactants with a HLB range of 27 are used for the for-
mation of water-in-oil emulsion systems, whereas in producing
oil-in-water emulsions, surfactants with a HLB range of 816 is
preferred.
Many studies have revealed that ultrasonic emulsication could
serve as a feasible and energy efcient tool in the generation of
pharmaceutical grade nanoemulsions. For instance, parenteral
nanoemulsions had been successfully produced by Wulff-Prez
et al. via low frequency ultrasound (20 kHz) using three different
types of natural oils: olive, sesame and soybean oils [41]. The
resulting mean droplet diameters for olive, sesame and soybean
oil nanoemulsions were found to be 379 nm, 368 nm and 380 nm
2072 M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083
respectively. Stability of droplets against coalescence was con-
rmed using Turbiscan MA 2000 even with a relatively small
amount of surfactant (7.8 10
4
3.1 10
3
mol/L) and with vol-
umes of oil phase as high as 25% (v/v). In a comparative study with
a Microuidizer, Kentish et al. proved that nanoemulsions with the
mean droplet size as low as 135 nm could be achieved by using a
ow-through ultrasonic cell (2024 kHz) [35]. The obtained drop-
let sizes are comparable to those for nanoemulsions prepared
using a Microuidizer operated at 100 MPa. With ultrasonic cavita-
tion, Abismail et al. successfully prepared nanoemulsion in a
water/kerosene/Tween 60 model system having droplets with
the average size no larger than 300 nm [33]. They suggested that
ultimate droplet size of nanoemulsion was greatly inuenced by
the time of emulsication, concentration of emulsier, ultrasonic
power and volume of oil fraction when low frequency ultrasound
(20 kHz, 130 W) was applied. Their results showed that the desir-
able nanoemulsion could be obtained with 30 s of emulsication
time, with 6% oil fraction and 10 g/L of Tween 60. Another study
conducted by Abismail et al. demonstrated that ultrasound was
more efcient and superior than rotorstator systems in terms of
producing ner emulsion droplets [42]. Similarly, Behrend et al.
applied ultrasonic waves to an emulsion system consisting of
water/vegetable oil/sodium dodecyl sulfate and successfully pro-
duced nanodroplets with minimum sizes of below 200 nm in the
presence of glycerine or polyethylene glycol as stabilizers [43].
Recently, novel dextrin nanoparticles had been successfully devel-
oped by Manchun et al. from nanoemulsion templates utilizing a
high power ultrasonicator [15]. Where, nanoemulsions with drop-
let sizes of about 200 nm were achieved using mixed surfactants
(Span 80/Tween 80) at HLB 6 after subjecting to sonication for
30 min. The nal droplet size of nanoemulsion templates was
found to decrease with increasing emulsication time, surfactant
concentration, and water content. On the other hand, a topical
nanoemulsion encapsulating eucalyptus oil has also been designed
and reported recently [44]. It has been observed that the transpar-
ency and stability of nanoemulsion were enhanced with sonication
time as compared to the emulsion generated using hand blender.
The most stable nanoemulsion with the mean droplet diameter
of 3.8 nm was obtained after 30 min of ultrasonication.
Many authors have reported that in the emulsication process
ultrasound has emerged as an excellent yet powerful emulsifying
tool as compared to that of other mechanical alternatives in terms
of obtaining a smaller droplet size and high energy efciency
[3,33,35]. Additionally, under the same conditions, in case of
producing a nanoemulsion with desired diameter, the required
amount of surfactant was signicantly reduced; the energy con-
sumption was considerably lowered than other classical mechani-
cal devices. It would then appear that ultrasound emulsication
may be used in place of high-pressure homogenization and micro-
uidization as it is capable of achieving similar local power densi-
ties with lower operating costs, if a suitable treatment chamber is
used. However, it is worth mentioned here that the nal droplet
size distribution of a nanoemulsion is mainly controlled by the rate
between two opposing processes; droplet break-up and droplet re-
coalescence [45]. Studies of the effect of ultrasonic operating
parameters and the nanoemulsion composition on the droplet
break-up and coalescence are particularly important in obtaining
smaller nano-sized emulsion droplets with narrower distribution.
The balance between these two dominant processes strongly
depends on the energy input during emulsication and the geom-
etry of design of the ultrasound emulsication device. For this rea-
son, several ultrasonic parameters such as irradiation power,
irradiation time, volume ratio of phases, surfactant concentration,
viscosity, liquid purity, gas content, position of the ultrasonic
source with respect to the liquidliquid interface, tip diameter
and vessel geometry must be properly controlled and optimized,
so that the effectiveness of ultrasound-engineered nanoemulsions
and their physiochemical properties could be guaranteed [46].
6. Examples of pharmaceutical nanoemulsions generated using
ultrasonic cavitation
In general, especially in the pharmaceutical and cosmetic indus-
tries, emulsions with mean droplet size of less than 1 lm are
essential in order to impart an enhanced bioavailability, as well
as high physical stability of the emulsion [47]. However, an ef-
cient approach to produce such nanocarriers still remains ambigu-
ous in the last two decades. To date, self-nanoemulsifying drug
delivery systems (SNEDDS) have been emerged to be one of the
most promising approaches to provide the transient formation of
hydrophobic drug encapsulated nanoemulsions. Several successful
cases had been reported such as Cefpodoxime proxetil [48], Ibu-
profen [49], Ramipril [50], CoQ10 [51], Oridonin [52], b-lactamase
[53], etc. However, a higher concentration of surfactant was always
required (normally in the range of 2050%), and this may lead to an
increased incidence of unwanted side-effects in the gastrointesti-
nal (GI) tract. A transient formation of nanoemulsion without
Fig. 2. Components and their role in the formation of nanoemulsions.
M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083 2073
introducing surplus stabilizer to the drug capsule could be easily
achieved using ultrasonic transducer [35,54]. In fact, the develop-
ment of such energy-intensive techniques for the preparation of
pharmaceutical grade nanoemulsions is a much more recent phe-
nomenon [2,14,5558].
Li et al. had successfully produced two types of Bufadienolides-
loaded nanoemulsions which were separately prepared by ultra-
sonication at an intensity of 40 W at 40 C for 60 min via a high
intensity ultrasonic processor [58]. The mean diameter, PDI and
zeta potential of both the Bufadienolides-loaded nanoemulsions
were 43.5 nm versus 161.4 nm, 0.100 versus 0.143 and 19.7 to
26.2 mV versus 29.4 to 35.3 mV, respectively. The study
showed that both the lyophilized nanoemulsions were stable over
a period of up to 3 months with no change in the appearance,
reconstitution ability, particle size, drug concentration and entrap-
ment efciency when maltose (20%) and trehalose (20%) were used
as cryoprotectants. Using combined technique of solvent diffusion
with ultrasonication, Araujo et al. successfully achieved a novel
cationic nanoemulsion formulation containing 2-(butylamino)-1-
phenyl-1-ethane thiosulfuric acid (BphEA), a schistosomicidal
agent [59]. The cationic nanoemulsion was formulated with a com-
bination of surfactants where the emulsion was generated by
slowly adding the organic phase to the aqueous phase under mag-
netic stirring at 150 rpm for 30 min at 25 C. The formation of
coarse oil-in-water emulsion occurred via diffusion of the organic
solvent into external phase and the solvent was then evaporated
under reduced pressure at the end of the process. The nal step
was accomplished by the size reduction using an ultrasonic proces-
sor with a tapered microtip probe of 1/8 (3 mm) diameter. This
combined approach had effectively rendered the emulsion to be
homogenous with a smaller droplet size in the nanometric range
around 225 nm with the PDI of 0.294. In vitro schistosomiasis study
showed that Schistosoma mansoni worms incubated with nanofor-
mulation (3 h) could gradually reduce their activity, and all female
worms were died after 48 h of incubation with the formulation.
Table 2 shows the details of representative nanoemulsions of
water-insoluble drugs that have been generated by ultrasonication.
By employing ultrasound probe sonicator alone, Doh et al. had
successfully developed a novel oral lipid nanoemulsion system
containing Granisetron, an antiemetic drug and the mean diame-
ters of ultrasonically-induced nanoemulsion were around 50 nm
with the PDI less than 0.2 [14]. The resulting nanoemulsion exhib-
ited a higher drug dissolution prole and in vitro drug permeation
rate compared to Granisetron powder. For topical applications,
Kong et al. had successfully fabricated a new hyaluronic acid
(HA) based nanoemulsions with higher degree of substitution
using combined solvent evaporation and ultrasonic cavitation
approach [2]. The acoustically-formed HA nanoemulsions showed
smaller droplet sizes of approximately 50 nm. The size distribution
and morphology qualities of obtained nanoemulsions were found
to be signicantly improved to be more uniform and the droplet
occulation was also alleviated with the addition of Ca
2+
as a cross-
linker. In addition, the highest encapsulation efciency of about
94% was achieved by using an ultrasonic probe. It is worth men-
tioned here that without using any chemical enhancer, ultra-
sound-engineered HA nanoemulsion showed desirable skin
permeability with no irritation in dermis compared with the con-
trol group, suggesting that it could be a potential colloidal trans-
dermal carrier for the targeted delivery of active lipophilic drugs.
An O/W nanoemulsion had also been formulated and characterized
by Sandig et al. for the percutaneous delivery of Imipiramine and
Doxepin [27]. The mean droplet size of Imipramine loaded nano-
emulsion was 17.8 nm, and for Doxepine loaded nanoemulsion it
was 19.9 nm. It was found that the average droplet size was
slightly but not signicantly decreased after loading the drug.
PDI values were observed to be 0.216 for Imipramine nanoemul-
sion and 0.133 for Doxepine nanoemulsion. The stability test
showed that both the nanoemulsions had an excellent physical sta-
bility with no phase separation, creaming, cracking or precipitation
being observed upon centrifugation. More importantly, the in vivo
analgesic and antiallodynic active ex vivo studies suggested that
topically applied Imipramine and Doxepin loaded nanoemulsions
were safe and an alternative analgesic therapy. Sood et al. sug-
gested that an anionic surfactant could provide an electrostatic
barrier on the droplets surface against coagulation, and also retards
molecular diffusion of monomer from the particle surface, thereby
lowering down the effect of Ostwald ripening [45]. Wulff-Perez
et al. reported that the concentration of surfactant in the formula-
tion could be further reduced at the expense of an increased ultra-
sonic power [41]. Interestingly, a reversible destabilization was
observed when higher amount of surfactant was employed. In
some of the cases, phase diagram has been used as a useful sche-
matic guideline as it provides the most straightforward solution
to identify the minimum surfactant needed to form a nanoemul-
sion with desirable droplet sizes [60,61].
Besides, a novel nanoemulsion encapsulating the natural avo-
noid has been recently reported by Ragelle et al. [57]. They have
successfully formulated a nanoemulsion of Fisetin with high load-
ing capacity (5 mg/ml with the use of combined surfactants, Tween
80 and Labrasol). Where, after subjecting to ultrasound and subse-
quent ltration, the nanoemulsion possessed an oil droplet diame-
ter of 153 nm, a negative zeta potential of 28.4 mV and a PDI of
0.129. The dispersion was stable at 4 C for 30 days. Interestingly,
when the Fisetin nanoemulsion was administered intraperitone-
ally, a 24-fold increase in Fisetin relative bioavailability was noted
as compared to free Fisetin. Additionally, the antitumour activity of
Fisetin nanoemulsion in Lewis lung carcinoma bearing mice
occurred at lower doses (36.6 mg/kg) compared to free Fisetin
(223 mg/kg). A similar nding has been reported by Gao et al.
[56] who have performed a modied emulsication solvent
Fig. 3. Two-stage ultrasound emulsication: droplet formation and break-up.
2074 M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083
evaporation to produce a Candesartan cilexetil-loaded nanoemul-
sion to improve the oral bioavailability of this inactive racemic pro-
drug. The ultrasound prepared ne emulsion was nanometer-sized
(35.5 nm) with the negative zeta potential. It was found that the
absorption of nanoemulsion containing Candesartan cilexetil was
signicantly improved as compared to free dug solution. In partic-
ular, the area under concentrationtime curve (AUC
0 ? t
) of Cande-
sartan was improved over 10-fold with the oral administration of
formulated nanoemulsion. Exciting advances in nanoemulsion-
based technology have been witnessed by Shiraishi et al., where
they have successfully designed a novel nano-sized emulsion con-
taining Peruoropentane (PFC5) in a diameter range of 200 nm by
using a bath-type sonicator [55]. Additionally, the PFC5 nanoemul-
sions were prepared in high yield at 40 C, which is higher than the
boiling temperature of PFC5. This ultrasonically prepared nano-
emulsion was delivered into tumor tissue which was transformed
into a micron-sized bubble upon ultrasound irradiation through
the phase transition of PFC5. This very facile preparation method
using ultrasound offers immediate and realistic potential for a
large-scale production of this pharmaceutically valuable nano-
emulsion for efcient theranostics of solid tumors.
7. Nanoemulsions from our sonochemical engineering
laboratory
The aim of a good nanoemulsion should be towards high loaded
drug capacity stabilized with a very low concentration of surfac-
tant with high homogeneity and excellent stability over long-term
storage. In our laboratory, we have successfully generated a variety
of nanoemulsions encapsulated with pharmaceutically active
ingredients such as ganoderic acid, curcumin, and aspirin using
cavitation.
7.1. Nanoemulsions encapsulated with ganoderic acid
Ganoderic acid (GA), a major bioactive ingredient isolated from
oriental medicinal mushroom ganoderma tsugae, has been found to
have signicant benecial effects either in the prevention or in the
treatment of a variety of diseases such as hepatitis B, hypertension,
AIDS, and cancers. However, the insolubility of GA in water poses
difculty in the formulation and in oral administration. The limita-
tions of exisiting approach via reverse microemulsion process
often involves the addition of a toxic solvent (e.g. hexane or chlo-
roform), and results in a lower product yield. To overcome this, we
have successfully encapsulated crude ganoderic acids by means of
nanoemulsication using ultrasound cavitation [62,63]. By taking
advantage of the surfactants chemical potential at oilwater inter-
face, the free energy required to full the entropy of micelles for-
mation (S) had been minimized, which subsequently led to
shorter emulsication time needed to generate the nanometric-
sized emulsion. The resultant formulations consisting of water,
surfactants and ganoderic acid solution were subjected to 5, 10,
20, 30 and 60 min of ultrasonication in an ultrasonic bath at room
temperature. In our studies, only 5 min of sonication time was
required to form tiny droplets and the generation of minimum
droplet sizes had been conrmed though DLS particle size analysis.
The resultant nanodispersions containing ganoderic acid in the
hydrophobic oil core were found to have a mean droplet no larger
than 200 nm. An increase in the time of ultrasound did not have a
signicant inuence on the size reduction of droplets. This may
represent the limiting value for the minimum droplet size for these
nanoemulsions. However, a slightly narrower particle size distri-
bution was observed, as the PDI of droplets decreased from 0.187
to 0.158 after 5 min of ultrasonication. In general, under a given
set of emulsication conditions at xed emulsion composition,
there exists a minimum emulsion droplet size which cannot be
reduced with repeated or prolonged ultrasound treatment time,
and therefore any longer application of ultrasound for the emulsi-
cation did not lead to any further reduction in the droplet size. It
should however be noted that the ultimate or minimum droplet
diameter is also the function of the physical and interfacial proper-
ties of the oil phase and the emulsier present in the emulsion sys-
tem. Ganta et al. observed a similar trend and reported that after
reaching a critical droplet size, extending the ultrasonication fur-
ther did not reduce the size of emulsion droplets [64].
7.2. Nanoemulsions encapsulated with curcumin
Curcumin, a yellowpigment extracted fromcurcuma longa dried
rhizome, has been reported to have signicant pharmaceutical
activities such as anti-cancer, anti-carcinogenic, anti-inammatory
and anti-oxidant properties [65]. However, the bioavailability of
curcumin after administration is intrinsically low due to its extre-
mely low water solubility. In order to improve the absorption rates
of curcumin in vivo, attempts to incorporate curcumin into the
nanoemulsion systems have been made. The formation of curcu-
min-loaded nanoemulsions has been achieved using an ultrasonic
bath (38 kHz) [63]. Colloidal stability of the resulting formulations
has been measured in terms of particle size distribution and PDI.
The mean droplet diameters and PDI of the nanoformulations were
approximately 120 nm and 0.35 respectively. MTS assay test shows
encouraging inhibition activity levels on MCF-7 human breast ade-
nocarcinoma cell lines after administration of 20 ll of curcumin
nanoformulation. Approximately 68% inhibition has been recorded
in the cultured cell line after an incubation period of 3 days with
the drug formulation.
Fig. 4 shows the SEM images of unhomogenized curcumin oily
droplets in water prior to ultrasound treatment. Figs. 5 and 6 illus-
trate the SEM images of generated curcumin nanoemulsions after
subjected to ultrasonic cavitation. The microscopic results show
that with merely 5 min of ultrasonication, curcumin-loaded nano-
emulsions were produced. Effect of ultrasonication time i.e. 5, 10,
20 and 30 min on the droplet size of nanoemulsion has been inves-
tigated. The study suggests that the mean droplet diameter of
nanoemulsions readily decreased with an increase in the duration
of sonication, and a more uniform particle size distribution could
be obtained. It is also interesting to note that the rate of size reduc-
tion was not affected by the weight percentage of isopropyl palmi-
tate which acts as an organic oil phase.
7.3. Nanoemulsions of O/W and multiple W/O/W encapsulated with
aspirin
Aspirin (acetylsalicylic acid) is a well-known non-steroidal anti-
inammatory drug (NSAID) widely used for its anti-inammatory,
anti-pyretic, analgesic, and platelet anti-aggregation properties.
Although adequate inammation and pain relief could be achieved
with the currently available aspirin dosage forms such as tablets or
capsules, some of their serious side effects in the GI, kidney and
platelets are major limitations to their routine use in therapy.
Due to the small droplet size, nanoemulsions are expected to afford
an efcient delivery of therapeutic agents to target sites in the
body and increase the oral bioavailability. On the other hand, the
development of complex multiple nanoemulsion system is aimed
to facilitate a sustained release and transport of entrapped active
aspirin drug, thereby reducing transient drug overload and severity
of drug-related gastrointestinal adverse effects. Hence, incorporat-
ing aspirin in the core nanoemulsion and multiple nanoemulsion
formulations could be useful in the treatment of inammation
and pain. Most importantly, both nanoformulations would be ef-
cient, convenient, exible and more patient compliant approaches
M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083 2075
in comparison to the conventional drug tablets and capsules. Gen-
eration of novel nanoemulsions and multiple nanoemulsions con-
taining aspirin using ultrasonic cavitation approach has been
presented [46]. For the former, response surface methodology
(RSM) has been adopted to produce an optimal cremophore-EL
based oil-in-water (O/W) nanoemulsion with the desired droplet
size and PDI [66,67]. As to the generation of new water-in-oil-in-
water (W/O/W) multiple nanoemulsion, a modied two-step cavi-
tational emulsication technique has been employed [68]. Besides,
the impact of gelling and osmotic pressure in the stabilization of
highly stable single core water-in-oil-in-water (W/O/W) multiple
nanoemulsions of aspirin through this two-stage ultrasonic assis-
tance has also been studied [69].
Fig. 7 shows the microscopic observations of aspirin nanoemul-
sions and reveals that the droplets were almost spherical in shape
by the employed three different processes. More interestingly, it
shows that the nanoemulsion generated by ultrasonic probe soni-
cator exhibited superior homogeneity with its nanosized oil drop-
lets, as compared to other emulsions prepared by both the
magnetic stirrer and high-speed homogenizer (Ultra-Turrax T18,
IKA, Labortechnik, Germany). In addition to this, the oil droplets
of ultrasonic-treated emulsion were all nely dispersed in the
water medium in a more uniform manner relative to the emulsion
prepared by homogenizer. The result is in line with dynamic light
scattering (DLS) particle size analysis, showing that the aspirin
emulsion droplets are present in the nanometer scale (ranging
around 200 nm) (Table 3).
Fig. 8(AD) images reveal that aspirin-containing nanoemulsion
droplets were almost spherical in shape with homogenous nano-
metric size distribution. The STEM measured droplet size corrobo-
rates with the results obtained from the droplet size analysis using
Zetasizer, showing that emulsion droplets are present in the nano-
meter range, varying from about 200 to 300 nm. Furthermore,
there is no signicant difference in terms of droplet size between
aspirin-containing nanoemulsion and the free formulation without
aspirin (control) (Fig. 8C). No change in the droplet size due to oc-
culation, coalescence, and drug degradation was observed over a
storage period of 15 days (Fig. 8D). It clearly shows that the incor-
poration of drug into oily core phase has no signicant inuence on
the droplet size and the stability of formulated nanoemulsion.
The performance of industrial scale high intensity ultrasound in
the generation of nanoemulsion has been compared with a bench-
top Microuidizer. In this study, for both of these techniques, the
importance of pre-homogenization on the resultant characteristics
of nanoemulsions has been studied [70]. With the pre-homoge-
nized emulsion feed, the morphology from the images revealed
that all the nanoemulsions containing aspirin produced by ultra-
sonic processor and Microuidizer were comprised of spherical
droplets and almost of uniform size (Fig. 9C and D). In comparison
with the emulsion generated by ultrasound or Microuidizer in the
absence of pre-homogenization (Fig. 9A and B), it could be seen
that there is a variation in the size distribution which was consid-
erably reduced with pre-homogenized emulsion feed. The droplet
size measurements by SEM indicated that the oil droplets in the
aspirin-loaded nanoemulsions were primarily between 200 and
250 nm in diameter, which is in agreement with the DLS particle
size analysis. In contrast, much larger droplets were observed in
both the non pre-homogenized/coarse emulsions treated by high
power ultrasound and Microuidizer where the measured droplet
diameters ranging from 300 to 450 nm (Fig. 9A and B). However, in
non pre-homogenized nanoemulsions, they appeared to be
clumped together in clusters (Fig. 9B and C), with few groups of
densely populated oil droplets. It has been observed that to achieve
a similar droplet size of 160 nm, ultrasound (60% amplitude, 12.5 J/
cm
3
) is 18 times more energy efcient than a Microuidizer
(200 bar, 226 J/cm
3
). STEM images of the ultrasonically-induced
and microuidized nanoemulsions using standard negative-stain-
ing procedure have been shown in Fig. 9AD.
Besides, the anti-inammatory activities of acoustically formed
nanoemulsion were determined using the k-carrageenan-induced
paw edema model. The analgesic activities of both the nanoformu-
lations were determined using acetic acid induced writhing
response and hot plate assay. For comparison, the effect of
pretreatment with blank nanoemulsion and reference aspirin
Table 2
Representative nanoemulsions of water-insoluble drugs that are generated by ultrasonication.
Drug and route of
administration
Dosage Oil Surfactant Co-surfactant/ solvent Indication Mean
droplet
diameter
(nm)
Stability References
Saquinavir (SQV) (oral) 400 lg/
ml
Flaxseed oil Lipoid E80 Deoxycholic acid HIV/AIDS 218.0 13.9 2 months (stored at
4 C in the dark
place)
[13]
Bufadienolides (mBU)
(i.v.)
0.28 mg/
ml
Soybean oil Lipoid E80,
pluronic F68,
Tween 80
Sodium oleate, glycerine Antitumor
analgesic
43.5 13.8 3 months (stored in
a dessicator with
silica-gel at 25 C)
[58]
Paclitaxel (oral) Pine nut oil Egg lecithin Stearylamine,
deoxycholic acid
Anticancer 90120 [23]
Granisetron (GRN)(oral) 0.4 mg/
ml
Isopropyl
myristate,
lauroglycol
90
Lipoid E80 Pluronic F68,
hydroxypropyl-b-
cyclodextrin,
polyvinylpyrrolidone K25
Antiemetic 50 3 months (stored at
4 C in the dark
place)
[14]
2-(Butylamino)-1 -
phenyl-1-
ethanethiosulfuric
acid (BphEA) (i.n.)
10 mg Medium
chain
triglyceride
Tween 80,
Span 80,
stearylamine
Ethanol, glycerol Schistosomiasis 225.8 32.1 3 months (stored at
4 C in the dark
place)
[59]
Imipramine /doxepin
(topical)
3% w/w D-limonene Labrasol,
plurol
oleique
Propylene glycol,
transcutol, oleic acid,
isostearyl isostearate
Analgesic anti-
allodynic
1820 3 months (stored at
room temperature)
[27]
Fisetin (i.p) 36.6 mg/
kg
Miglyol 812 Tween 80,
labrasol
Lipoid E80 Antitumor 153 2 1 month (stored at
4 C in the dark
place)
[57]
2076 M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083
suspension were also studied for their anti-inammatory and
antinociceptive activities. The results showed that oral administra-
tion of nanoemulsion containing aspirin (60 mg/kg) signicantly
reduced paw edema induced by k-carrageenan injection [71].
Nanoformulations decreased the number of abdominal constric-
tion in acetic acid induced writhing model. Pretreatment with
nanoformulations led to a signicant increase in reaction time in
the hot plate assay. Nanoemulsion demonstrated an enhanced
anti-inammatory and analgesic effects compared to reference
suspension. These experimental studies suggest that nanoemul-
sion produced a pronounced anti-inammatory and analgesic
effect in rats and may be candidates as new nanocarriers for phar-
macological NSAIDs in the treatment of inammatory disorders
and alleviating pains. Briey, the aspirin-loaded nanoemulsion
exhibits enhanced anti-inammatory and analgesic properties
while multiple nanoemulsion demonstrated mild or sustained
Fig. 4. Unhomogenized curcumin emulsion droplets prior to ultrasonication.
Fig. 5. (Left) Curcumin nanodroplets in ethanolic solution (micellar solution) prepared by ultrsonication with surfactant concentration of 0.2 wt%. (Right) Curcumin coarse
emulsion droplets using isopropyl palmitate (IPP) as an organic oil phase.
Fig. 6. Scanning transmission electron microscopy (STEM) images of curcumin nanoemulsions obtained after 5 min of ultrasonication.
M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083 2077
anti-inammatory and analgesic activities. Fig. 10 shows the over-
all anti-inammatory and analgesic activity rank among the aspi-
rin formulations.
7.4. Submicron multiple emulsion encapsulated with ferrous fumarate
Ferrous fumarate, [C
4
H
2
FeO
4
] is widely used in the effective
treatment and prevention of iron deciency anemia (IDA), but its
administration has been oftentimes linked with quite a few side
effects than ferric products. For this reason, multiple or double
emulsion of water-in-oil-in-water (W/O/W) type formulation had
been proposed as an alternative drug delivery system to reduce
the transient drug overload and to overcome drug-related side
effects caused by daily iron supplementation as well as masking
the unpleasant taste of the iron supplements. However, the
production of multiple emulsions with controlled droplet sizes
Fig. 7. Microscopic views (200) of emulsions obtained through three different methods (after 2-weeks of storage): (A) magnetic stirring (7 h); (B) high-speed
homogenization (5 min at 13,500 rpm); (C) ultrasonic emulsication (1 min using 25% amplitude).
Table 3
Comparison of droplet size of emulsions of aspirin produced by three different methods.
Emulsion preparation method Magnetic stirring High-speed homogenization Ultrasonic emulsication
Droplet size (d.nm) 1160 359 232
Polydispersity index (PDI) 0.971 0.379 0.309
Fig. 8. Scanning transmission electron microscopy (STEM) images of aspirin-containing nanoemulsion (A) 5000 (B) 10,000 (C) blank and (D) aspirin-containing
nanoemulsions prepared via ultrasonication after 15-days of storage at room temperature (30,000).
2078 M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083
Fig. 9. Scanning transmission electron microscopy (STEM) images of formulation of aspirin-containing nanoemulsions prepared by (A) ultrasound, and (B) Microuidizer,
with coarse emulsion feed (20,000) (C) ultrasound and (D) Microuidizer, with pre-homogenized emulsion feed (50,000).
Nanoemulsion Drug Suspension
Multiple
Nanoemulsion
Blank
Nanoemulsion
Overall Anti-inflammatory and Analgesic Activity Rank
Decreasing Activity Order
Fig. 10. Overall anti-inammatory and analgesic activity rank of aspirin formulations.
High
manufacturing &
maintenance cost
High
manufacturing &
maintenance cost
Less homogenous
Less Stable
Large amount of
surfactant
Broad size
Less stable
Phase
Invesion
Two-step
Micro-
channel
Membrane
Fig. 11. Preparative methods of multiple emulsions with their characteristics.
M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083 2079
and internal structures could be a major challenge and there have
been intensive studies of emulsication techniques for producing
uniform and stable multiple emulsion droplets in the past decade
[7275]. Recently, a major breakthrough that has been achieved
is the controlled preparation of ne and monodispersed multiple
emulsions by various microstructured processing techniques, such
as microchannel and membrane emulsication but only on the
smaller scale of product throughput. Fig. 11 shows the conven-
tional preparative methods that are employed in the generation
of multiple emulsions with their characteristics.
We have employed a novel yet alternative strategy of hydrody-
namic cavitation reactor of liquid whistle type, called Sonolator, for
the generation of stable and highly monodispersed submicron
multiple emulsions (600 nm) with the droplet size distribution
(PDI) in the range of 0.350.40 containing pharmaceutical grade
ferrous fumarate via a two-stage method of droplet formation
[76]. The inuence of variation in the operating conditions of such
unique liquid whistle hydrodynamic cavitation based reactor
(LWHCR) on the generation of submicron multiple emulsions has
been investigated. An increase in the number of passes has been
proved to be capable of producing smaller and stable W/O/W sub-
micron multiple emulsions with the narrow droplet size distribu-
tion. Although an increase in the inlet pressure and number of
emulsication passes led to enhanced stability of the W/O/W mul-
tiple emulsion, but, extreme pressure was not recommended for
preparing W/O/W submicron multiple emulsions in the second
stage since the intense cavitational bubble collapse could promote
the rupture of internal aqueous droplets, which subsequently led
to the loss of inner water phase encapsulating the active pharma-
ceutical or nutraceutical ingredients.
Unlike primary W/O emulsion, an excess or intense cavitational
forces caused by high pressure and uid acceleration can lead to
the eventual rupture of the oil lm separating the internal droplets
and the outer continuous phase in the multiple emulsion prepara-
tion. For this reason during the secondary emulsication, the
LWHCR was operated at three very low inlet pressures: 100, 160
and 200 psi. The resultant primary W/O coarse emulsion and sec-
ondary W/O/W multiple submicron emulsion containing ferrous
fumarate have been shown in Fig. 12.
The microscopic results demonstrated that variation in the mul-
tiple globule size distribution is greatly reduced when the
operating pressures were increased from 100 to 200 psi, indicating
an improved homogeneity of the ne dispersion. Selection of an
optimumset of operating parameters including operating pressure,
number of treatment passes, temperature of the continuous phase
and the distance between the blade and orice is essential to
achieve maximum benets in the emulsication process. Thus,
LWHCR is an important technological key for large-scale produc-
tion of these medicinally valuable multiple emulsions.
7.5. Nanoemulsion encapsulated with curcumin using LWHCR
Besides ferrous fumarate, the encapsulation of curcumin in a
palm oil-based nanoemulsion has been achieved by employing
LWHCR [77]. At rst, the effect of premixing on the size of
generated droplets was also investigated using a high-speed
homogenizer. Different orice plate-blade distances i.e. 0.5 cm,
0.6 cm and 0.8 cm for each of the inlet operating pressures i.e.
400, 600 and 800 psi have been considered. It has been reported
that for the orice plate-blade distance of 0.5 cm at 800 psi, the
lowest droplet size of 415 nm with a PDI of 0.5 was observed.
Effect of co-surfactant, Span 80 on the generation of nanoemul-
sions was also examined and it has been observed that its
addition did not have any inuence on the nal droplet size.
Finally, the nanoencapsulation of curcumin and its efciency
were studied using the optimized conditions of nanoemulsion
generation. The nanoencapsulation efciency of curcumin was
found to be 88%, indicating the superiority of LWHCR. The stabil-
ity studies conrmed that the particle size was found to be
decreasing from 740 nm to 560 nm for Span 80 nanoemulsions
and from 708 nm to 532 nm for curcumin based nanoemulsions
over a storage period of 30 days.
8. Future works
Following our investigations as described above, three main
potential research areas need to be explored for future works in
the formulation development and evaluation of the newnano/mul-
tiple emulsion-based drug delivery systems using cavitation
approach. (1) Kinetic modelling of in vitro drug release proles of
O/W nanoemulsion and W/O/W multiple nanoemulsions gener-
ated using ultrasonic cavitation: aspirin nanoemulsion and its
multiple nanoemulsion intended for oral administration have been
Fig. 12. Primary W/O coarse emulsion (A) and secondary W/O/W submicron multiple emulsion (B) containing ferrous fumarate produced by LWHCR technology.
2080 M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083
developed using ultrasonic cavitation approach. Besides, the anti-
inammatory and analgesic activities of both the nanoformula-
tions have been evaluated and compared to the control. However,
the release mechanisms and drug release kinetics from both nano-
emulsion and multiple nanoemulsion have not yet been explored.
Thus, a proper study and verication of the drug release models for
both nanoformulations should be considered. Besides, the thera-
peutic efcacy of W/O/W multiple nanoemulsion is reported to
be different from the simple O/W nanoemulsion. Hence, further
investigation on the drug release mechanisms and kinetics of both
the ultrasonically-formed nanoformulations should be carried out.
(2) Comparative study of the continuous emulsication process by
ultrasonic cavitation and microuidization in the generation of
pharmaceutical nanoemulsions: A comparison between an indus-
trial scale high power ultrasound horn and a bench-top air-driven
microuidizer in the batch production of aspirin nanoemulsions
has been investigated. However, a detailed comparative work of
both emulsifying techniques in the generation of pharmaceutical
grade nanoemulsion in continuous mode has not been reported
in the open literature. This study is of highly relevance and in fact
it merits further investigation as it would then allow pharmaceuti-
cal scientists in selecting an appropriate production method to be
applied to develop and manufacture of a new nanoemulsion based
drug delivery system in a continuous manner. (3) Optimization of
process parameters in the generation of highly stable pharmaceu-
tical W/O/W submicron multiple emulsions produced via novel
LWHCR technology: Application of a very interesting yet novel
Sonolator technology in the emulsication process where a liquid
whistle hydrodynamic cavitation based reactor (LWHCR) was
employed to produce W/O/W submicron multiple emulsions con-
taining ferrous fumarate via two-stage droplet formation in a cav-
itation chamber. The effect of operating pressure and number of
emulsication passes on the mean droplet size and stability of
W/O/W multiple emulsions had been investigated and analyzed
thoroughly. Apart from the operating pressure and number of
emulsication passes, other important parameters such as
orice diameter, distance between blade and orice, processing
temperature, and viscosity of emulsion should be explored in the
future work to explain how different processing conditions of
LWHCR affect the physicochemical properties of the resultant mul-
tiple emulsions. In this aspect, a standard RSM could be used as an
optimization tool in conjunction with the design and manufacture
of optimum submicron multiple emulsion formulation with
desired properties. Furthermore, this should allow a better inter-
pretation of spectra of the variations of W/O/W multiple emulsion
properties caused by different processing parameters used in
LWHCR system.
9. Conclusions
This review has outlined the principles relating to the cavita-
tional emulsication and its potential use in producing the phar-
maceutical nanoemulsions in the current drug delivery system.
In the generation of nanoemulsion, cavitation technology is proved
to be a powerful and promising approach for the efcient produc-
tion of droplets with smaller size compared to other mechanical
alternatives. It is competitive or even superior in terms of energy
efciency and thus more economical compared to classical homog-
enizing systems. Most importantly, by using cavitation technique,
the requirement of surfactant is signicantly reduced and equip-
ment contamination is less and thus aseptic processing is feasible
and appropriate for pharmaceutical processing. Additionally, the
acoustically formed nanoemulsions were all found to be stable
excellently and more homogeneous as compared to those disper-
sions produced by conventional mechanical methods.
However, in most of the pharmaceutical technology operations,
the achievement of size reduction by ultrasonic cavitation tech-
nique still requires proper study. This is due to the fact that it is
usually difcult, in pharmaceutical industry, to alter one process-
ing parameter without adversely affecting another. Besides, the
quality of nanoemulsion for pharmaceutical use or application
must be monitored. Monitoring the critical parameters of ultrason-
ication process will ensure that the product specications are met.
A clear understanding of the ultrasonic processing parameters, the
choice of recipients (including oils and surfactant), and compatibil-
ity of formulations and the overall efcacy of formulated nano-
emulsion is therefore necessary to carry out a successful
optimization endeavor in the design and manufacture of pharma-
ceutical nanoemulsions. To fully realize the potential of nanoemul-
sion-based drug delivery system with controlled size and shape,
nanofabrication and nanomanufacturing by means of cavitation
approach will play a prominent role in the future. Overall, ultra-
sonic cavitation is simple, energy efcient and cost-effective in
the generation of ne pharmaceutical nanoemulsions and multiple
nanoemulsions with improved efcacy. Whereas, hydrodynamic
cavitation is facile, highly exible and scalable in the production
of monodispersed multiple submicron emuslion for the pharma-
ceutical industries. Conclusively, cavitation technology provides
innovating and exciting perspectives for modern pharmaceutical
industries which are looking for signicant and competitive pro-
cess improvements with sonochemistry.
Acknowledgements
Our sincere gratitude goes to the Malaysian Ministry of Science,
Technology & Innovation (MOSTI) for their nancial support
through eScience (M0058.54.01).
References
[1] N. Anton, J.-P. Benoit, S. Patrick, Design and production of nanoparticles
formulated from nano-emulsion templates A review, J. Control. Release 128
(3) (2008) 185199.
[2] M. Kong, X.G. Chen, D.K. Kweon, H.J. Park, Investigations on skin permeation of
hyaluronic acid based nanoemulsion as transdermal carrier, Carbohydr. Polym.
86 (2) (2011) 837843.
[3] T.F. Tadros, P. Izquierdo, J. Esquena, C. Solans, Formation and stability of nano-
emulsions, Adv. Colloid Interface Sci. 108109 (2004) 303318.
[4] W.Y. Zhou, M. Wang, W.L. Cheung, B.C. Guo, D.M. Jia, Synthesis of carbonated
hydroxyapatite nanospheres through nanoemulsion, J. Mater. Sci. Mater. Med.
19 (1) (2008) 103110.
[5] D.J. McClements, Emulsion design to improve the delivery of functional
lipophilic components, Annu. Rev. Food Sci. Technol. 1 (1) (2010) 241269.
[6] H. Chen, C. Khemtong, X. Yang, X. Chang, J. Gao, Nanonization strategies for
poorly water-soluble drugs, Drug Discov. Today 16 (78) (2011) 354360.
[7] Y. Zhang, H.F. Chan, K.W. Leong, Advanced materials and processing for drug
delivery: the past and the future, Adv. Drug Deliv. Rev. 65 (1) (2013) 104120.
[8] V. Bali, M. Ali, J. Ali, Study of surfactant combinations and development of a
novel nanoemulsion for minimising variations in bioavailability of ezetimibe,
Colloids Surf. B Biointerfaces 76 (2) (2010) 410420.
[9] J.-B. Tagne, S. Kakumanu, D. Ortiz, T. Shea, R.J. Nicolosi, A nanoemulsion
formulation of tamoxifen increases its efcacy in a breast cancer cell line, Mol.
Pharm. 5 (2) (2008) 280286.
[10] F. Kuo, B. Subramanian, T. Kotyla, T.A. Wilson, S. Yoganathan, R.J. Nicolosi,
Nanoemulsions of an anti-oxidant synergy formulation containing gamma
tocopherol have enhanced bioavailability and anti-inammatory properties,
Int. J. Pharm. 363 (12) (2008) 206213.
[11] K.K. Singh, S. Vingkar, Formulation, antimalarial activity and biodistribution of
oral lipid nanoemulsion of primaquine, Int. J. Pharm. 347 (2008) 136143.
[12] S. Ganta, M. Amiji, Coadministration of paclitaxel and curcumin in
nanoemulsion formulation to overcome multidrug resistance in tumor cells,
Mol. Pharm. 6 (3) (2009) 928939.
[13] T.K. Vyas, A. Shahiwala, M.M. Amiji, Improved oral bioavailability and brain
transport of Saquinavir upon administration in novel nanoemulsion
formulations, Int. J. Pharm. 347 (2008) 93101.
[14] H.-J. Doh, Y. Jung, P. Balakrishnan, H.-J. Cho, D.-D. Kim, A novel lipid
nanoemulsion system for improved permeation of granisetron, Colloids Surf.
B Biointerfaces 101 (2013) 475480.
M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083 2081
[15] S. Manchun, C.R. Dass, P. Sriamornsak, Designing nanoemulsion templates for
fabrication of dextrin nanoparticles via emulsion cross-linking technique,
Carbohydr. Polym. 101 (2014) 650655.
[16] K. Burapapadh, H. Takeuchi, P. Sriamornsak, Novel pectin-based nanoparticles
prepared from nanoemulsion templates for improving in vitro dissolution and
in vivo absorption of poorly water-soluble drug, Eur. J. Pharm. Biopharm. 82
(2) (2012) 250261.
[17] A. Ethirajan, K. Schoeller, A. Musyanovych, U. Ziener, K. Landfester, Synthesis
and optimization of gelatin nanoparticles using the miniemulsion process,
Biomacromolecules 9 (9) (2008) 23832389.
[18] M.J. Akers, Excipientdrug interactions in parenteral formulations, J. Pharm.
Sci. 91 (11) (2002) 22832300.
[19] D.M. Vyas, Paclitaxel (Taxol

) formulation and prodrugs, Pharmacochem. Libr.

22 (1995) 103130.
[20] C. Cocchio, A.J. Geib, K.O. Rynn, Physiochemical stability of intravenous fat
emulsion in combination with medications used for resuscitation, SOJ Pharm.
Pharm. Sci. 1 (1) (2014) 3.
[21] A. Kogan, N. Garti, Microemulsions as transdermal drug delivery vehicles, Adv.
Colloid Interface Sci. 123126 (2006) 369385.
[22] S. Kakumanu, J.B. Tagne, T.A. Wilson, R.J. Nicolosi, A nanoemulsion formulation
of dacarbazine reduces tumor size in a xenograft mouse epidermoid carcinoma
model compared to dacarbazine suspension, Nanomed 7 (3) (2011) 277283.
[23] S.B. Tiwari, M.M. Amiji, Improved oral delivery of paclitaxel following
administration in nanoemulsion formulations, J. Nanosci. Nanotechnol. 6
(2006) 32153221.
[24] E. Acosta, Bioavailability of nanoparticles in nutrient and nutraceutical
delivery, Curr. Opin. Colloids Interface Sci. 14 (1) (2009) 315.
[25] Y. Luo, D. Chen, L. Ren, X. Zhao, J. Qin, Solid lipid nanoparticles for enhancing
vinpocetines oral bioavailability, J. Control. Release 114 (1) (2006) 5359.
[26] H. Carrstensen, R.H. Mller, B.W. Mller, Particle size, surface hydrophobicity
and interaction with serum of parenteral fat emulsions and model drug
carriers as parameters related to RES uptake, Clin. Nutr. 11 (5) (1992) 289
297.
[27] A.G. Sandig, A.C.C. Campmany, F.F. Campos, M.J.M. Villena, B.C. Naveros,
Transdermal delivery of imipramine and doxepin from newly oil-in-water
nanoemulsions for an analgesic and anti-allodynic activity: development,
characterization and in vivo evaluation, Colloids Surf. B Biointerfaces 103
(2013) 558565.
[28] F. Shakeel, S. Baboota, A. Ahuja, J. Ali, A. Mohammed, S. Shaq, Nanoemulsions
as vehicles for transdermal delivery of aceclofenac, AAPS Pharm. Sci. 9 (4)
(2007) 191199.
[29] S. Raj, S. Jose, U.S. Sumod, M. Sabitha, Nanotechnology in cosmetics:
opportunities and challenge, J. Pharm. Bioallied Sci. 4 (3) (2012) 186193.
[30] J.Th.G. Overbeek, The rst rideal lecture. Microemulsions, a eld at the border
between lyophobic and lyophilic colloids, Faraday Discuss. Chem. Soc. 65
(1978) 719.
[31] D.J. McClements, Food Emulsions: Principles, Practices, and Techniques,
second ed., CRC Press, Boca Raton, Florida, 2004.
[32] A.-M. Shi, D. Li, L.-J. Wang, B.-Z. Li, B. Adhikari, Preparation of starch-based
nanoparticles through high-pressure homogenization and miniemulsion
cross-linking: Inuence of various process parameters on particle size and
stability, Carbohydr. Polym. 83 (4) (2011) 16041610.
[33] B. Abismail, J.P. Canselier, A.M. Wilhelm, H. Delmas, C. Gourdon, Emulsication
by ultrasound: droplet size distribution and stability, Ultrason. Sonochem. 6
(12) (1999) 7583.
[34] S.M. Jafari, A. Elham, Y. He, B. Bhandari, Re-coalescence of emulsion droplets
during high-energy emulsication, Food Hydrocolloids 22 (7) (2008) 1191
1202.
[35] S. Kentish, T.J. Wooster, M. Ashokkumar, S. Balachandra, R. Mawson, L. Simons,
The use of ultrasonics for nanoemulsion preparation, Innovative Food Sci.
Emerg. Technol. 9 (2) (2008) 170175.
[36] K.S. Suslick, Sonoluminescence and sonochemistry, in: R.A. Meyers (Ed.),
Encyclopedia of Physical Science and Technology, Academic Press, San Diego,
2001.
[37] R.W. Wood, A.L. Loomis, Physical and biological effects of high-frequency
sound waves, Philos. Mag. 4 (1927) 417436.
[38] Swiss Patent, 394.390, 1944.
[39] M.K. Li, H.S. Fogler, Acoustic emulsication. Part 1. The instability of the oil-
water interface to form the initial droplets, J. Fluid Mech. 88 (3) (1978) 499
511.
[40] M.K. Li, H.S. Fogler, Acoustic emulsication. Part 2. Break-up of the
larger primary oil droplets in a water medium, J. Fluid Mech. 88 (3) (1978)
513528.
[41] M. Wulff-Prez, A. Torcello-Gmez, M.J. Glvez-Ruz, A. Martn-Rodrguez,
Stability of emulsions for parenteral feeding: preparation and characterization
of o/w nanoemulsions with natural oils and Pluronic f68 as surfactant, Food
Hydrocolloids 23 (4) (2009) 10961102.
[42] B. Abismail, J.P. Canselier, A.M. Wilhelm, H. Delmas, C. Gourdon, Emulsication
processes: on-line study by multiple light scattering measurements, Ultrason.
Sonochem. 7 (2000) 187192.
[43] O. Behrend, K. Ax, H. Schubert, Inuence of continuous phase viscosity on
emulsication by ultrasound, Ultrason. Sonochem. 7 (2000) 7785.
[44] S. Sugumar, V. Ghosh, M.J. Nirmala, A. Mukherjee, N. Chandrasekaran,
Ultrasonic emulsication of eucalyptus oil nanoemulsion: antibacterial
activity against Staphylococcus aureus and wound healing activity in Wistar
rats, Ultrason. Sonochem. 21 (3) (2014) 10441049.
[45] A. Sood, Coagulative stability of miniemulsion droplets, J. Appl. Polym. Sci. 109
(2) (2008) 12621270.
[46] S.Y. Tang, Formulation development and evaluation of pharmaceutical
nanoemulsions and multiple nanoemulsions using ultrasonic cavitation
technique (Ph.D. thesis), University of Nottingham Malaysia Campus, 2012.
[47] H. Schubert, Hochschulkurs Emulgiertechnik am Institut fur
Lebensmittelverfahrenstechnik, Karlsruhe University (1998).
[48] A.A. Date, M.S. Nagarsenker, Design and evaluation of self-nanoemulsifying
drug delivery systems (SNEDDS) for cefpodoxime proxetil, Int. J. Pharm. 329
(12) (2007) 166172.
[49] L. Wang, J. Dong, J. Chen, J. Eastoe, X. Li, Design and optimization of a new self-
nanoemulsifying drug delivery system, J. Colloid Interface Sci. 330 (2009) 443
448.
[50] S. Shaq, S. Faiyaz, T. Sushma, F.J. Ahmad, R.K. Khar, A. Mushir, Development
and bioavailability assessment of ramipril nanoemulsion formulation, Eur. J.
Pharm. Biopharm. 66 (2007) 227243.
[51] P. Balakrishnan, B.-J. Lee, D.H. Oh, J.O. Kim, Y.-I. Lee, D.-D. Kim, J.-P. Jee, Y.-B.
Lee, J.S. Woo, C.S. Yong, H.-G. Choi, Enhanced oral bioavailability of coenzyme
Q10 by self-emulsifying drug delivery systems, Int. J. Pharm. 374 (12) (2009)
6672.
[52] P. Zhang, Y. Liu, N. Feng, J. Xu, Preparation and evaluation of self-
microemulsifying drug delivery system of oridonin, Int. J. Pharm. 355 (12)
(2008) 269276.
[53] S.V.R. Rao, J. Shao, Self-nanoemulsifying drug delivery systems (SNEDDS) for
oral delivery of protein drugs. I. Formulation development, Int. J. Pharm. 362
(12) (2008) 29.
[54] T.S.H. Leong, T.J. Wooster, S.E. Kentish, M. Ashokumar, Minimizing oil droplet
size using ultrasonic emulsication, Ultrason. Sonochem. 16 (2009) 721727.
[55] K. Shiraishi, R. Endoh, H. Furuhata, M. Nishihara, R. Suzuki, K. Maruyama, Y.
Oda, J.-I. Jo, Y. Tabata, J. Yamamoto, M. Yokoyama, A facile preparation method
of a PFC-containing nano-sized emulsion for theranostics of solid tumors, Int. J.
Pharm. 421 (2) (2011) 379387.
[56] F. Gao, Z. Zhang, H. Bu, Y. Huang, Z. Gao, J. Shen, C. Zhao, Y. Li, Nanoemulsion
improves the oral absorption of candesartan cilexetil in rats: performance and
mechanism, J. Control. Release 149 (2) (2011) 168174.
[57] H. Ragelle, S. Crauste-Manciet, J. Seguin, D. Brossard, D. Scherman, P. Arnaud,
G.G. Chabot, Nanoemulsion formulation of setin improves bioavailability and
antitumour activity in mice, Int. J. Pharm. 427 (2) (2012) 452459.
[58] F. Li, T. Wang, H. He, X. Tang, The properties of bufadienolides-loaded nano-
emulsion and submicro-emulsion during lyophilisation, Int. J. Pharm. 349
(2008) 291299.
[59] S.C.A. Araujo, A.C.A. Mattos, H.F. Teixeira, P.M.Z.C. Coelho, D.L. Nelson, M.C.
Oliveira, Improvement of in vitro efcacy of a novel schistosomicidal drug by
incorporation into nano-emulsions, Int. J. Pharm. 337 (2007) 307315.
[60] F. Zhong, M. Yu, C. Luo, C.F. Shoemaker, Y. Li, S. Xia, J. Ma, Formation and
characterisation of mint oil/S and CS/water microemulsions, Food Chem. 115
(2) (2009) 539544.
[61] V. Domnguez-Villegas, B. Clares-Naveros, M.L. Garca-Lpez, A.C. Calpena-
Campmany, P. Bustos-Zagal, M.L. Garduo-Ramrez, Development and
characterization of two nano-structured systems for topical application of
avanones isolated from Eysenhardtia platycarpa, Colloids Surf. B. Biointerfaces
116 (2014) 183192.
[62] K.W. Tan, M. Sivakumar, Synthesis and characterization of ganoderic acid
nanoemulsion or reverse nanoemulsion as colloidal carrier, in: International
Conference on Nanotechnology Research and Commercialization (ICONT),
2009, Langkawi: SIRIM.
[63] K.W. Tan, Process development and optimization of nano colloidal carriers for
the delivery of chemopreventive phytochemicals using ultrasonic cavitation
(Ph.D. thesis), University of Nottingham Malaysia Campus, 2011.
[64] S. Ganta, J.W. Paxton, B.C. Baguley, S. Garg, Pharmacokinetics and
pharmacodynamics of chlorambucil delivered in parenteral emulsion, Int. J.
Pharm. 360 (12) (2008) 115121.
[65] C.-L. Lin, J.-K. Lin, Curcumin: a potential cancer chemopreventive agent
through suppressing NF-jB signaling, J. Cancer Mol. 4 (1) (2008) 1116.
[66] S.Y. Tang, M. Sivakumar, A novel formulation and optimization of aspirin
nano-emulsion prepared by cavitation induced by ultrasonic waves, J. Ind.
Technol. 19 (1) (2010).
[67] S.Y. Tang, M. Sivakumar, K.H. Tan, B. Nashiru, Formulation development and
optimization of a novel cremophore EL-based nanoemulsion using ultrasound
cavitation, Ultrason. Sonochem. 19 (2) (2011) 330345.
[68] S.Y. Tang, M. Sivakumar, Design and evaluation of aspirin-loaded water-in-oil-
in-water submicron multiple emulsions generated using two-stage ultrasonic
cavitational emulsication technique, Asia-Pac. J. Chem. Eng. 7 (2012) S145
S156.
[69] S.Y. Tang, M. Sivakumar, B. Nashiru, Impact of osmotic pressure and gelling in
the generation of highly stable single core water-in-oil-in-water (W/O/W)
nano multiple emulsions of aspirin assisted by two-stage ultrasonic
cavitational emulsication, Colloids Surf. B. Biointerfaces 102 (2013) 653658.
[70] S.Y. Tang, P. Shridharan, M. Sivakumar, Impact of process parameters in the
generation of novel aspirin nanoemulsions-comparative studies between
ultrasound cavitation and microuidizer, Ultrason. Sonochem. 20 (1) (2013)
485497.
[71] S.Y. Tang, M. Sivakumar, N. Min-Hwei, P. Shridharan, Anti-inammatory and
analgesic activity of novel oral aspirin-loaded nanoemulsion and nano
multiple emulsion formulations generated using ultrasound cavitation, Int. J.
Pharm. 430 (12) (2012) 299306.
2082 M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083
[72] S.T. Dogru, S. alis, F. ner, Oral multiple w/o/w emulsion formulation of a
peptide salmon calcitonin: in vitroin vivo evaluation, J. Clin. Pharm. Ther. 25
(6) (2000) 435443.
[73] G.T. Vladisavljevic, M. Shimizu, T. Nakashima, Preparation of monodisperse
multiple emulsions at high production rates by multi-stage premix membrane
emulsication, J. Membr. Sci. 244 (12) (2004) 97106.
[74] H. Wei, F. Zhong, J. Ma, Z. Wang, Formula optimization of emulsiers for
preparation of multiple emulsions based on articial neural networks,
J. Dispersion Sci. Technol. 29 (2008) 319326.
[75] J. Weiss, I. Scherze, G. Muschiolik, Polysaccharide gel with multiple emulsion,
Food Hydrocolloids 19 (2005) 605615.
[76] S.Y. Tang, M. Sivakumar, A novel and facile liquid whistle hydrodynamic
cavitation reactor to produce submicron multiple emulsions, AIChE J. 59 (1)
(2013) 155167.
[77] S. Parthasarathy, T. Siah Ying, S. Manickam, Generation and optimization of
palm oil-based oil-in-water (O/W) submicron-emulsions and encapsulation of
curcumin using a liquid whistle hydrodynamic cavitation reactor (LWHCR),
Ind. Eng. Chem. Res. 52 (34) (2013) 1182911837.
M. Sivakumar et al. / Ultrasonics Sonochemistry 21 (2014) 20692083 2083