Industrial Crops and Products 67 (2015) 18–24

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Industrial Crops and Products

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

Use of various vegetable oils in designing photoprotective

nanostructured formulations for UV protection and antioxidant
activity
Gabriela Badea, Ioana Lăcătuşu ∗ , Nicoleta Badea, Cristina Ott, Aurelia Meghea
University POLITEHNICA of Bucharest, Faculty of Applied Chemistry and Material Science, 1 Polizu Street, 011061 Bucharest, Romania

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

Article history: Vegetable oils are well-known for their beneficial health effects, mainly due to their antioxidant activity,
Received 30 September 2014 which is maintained or enhanced when they are encapsulated at nanoscale. This study aims to design new
Received in revised form nanostructured lipid carriers (NLCs) containing various vegetable oils and their combinations in order
19 December 2014
to obtain efficient formulations with UV protection performance and antioxidant activity. Pomegranate
Accepted 22 December 2014
seed oil, wheat germ oil, blackcurrant seed oil, sesame seed oil, carrot root oil, raspberry seed oil and rice
bran oil were used for obtaining NLCs, as carriers for a photoprotective agent that absorbs the UVA radia-
Keywords:
tion: diethylamino hydroxybenzoyl hexyl benzoate. The NLCs were characterized by mean particle size,
Vegetable oils
Nanostructured lipid carriers
physical stability over time, and entrapment efficiency of the UVA filter. The UV protection performance
Antioxidant was assessed using both in vitro sun protection factor (SPF) and the UVA protection factor (UVAPF). The
UV protection antioxidant activity was determined by chemiluminescence analysis. All vegetable oils used led to the
development of appropriate NLCs, having mean particle sizes ranging between 100 nm and 145 nm and
good physical stability with zeta potential values less negative than −35 mV. The UV protection factors
were evaluated on the individual vegetable oils and on cream formulations based on NLCs. Out of the
seven cream formulations, the best UV protection was assured by the pomegranate seed oil based cream
resulting in a SPF of 4.1 and an UVAPF of 7.8. Knowing that a synergistic interaction can occur among the
active compounds of different oils, new NLCs were developed by using a mixture of pomegranate seed
oil and one of the other oils. The NLCs based on pomegranate seed oil combined with wheat germ oil
have shown the best entrapment efficiency (70%) and photoprotection (SPF of 5.1 and a UVAPF of 9.5). All
vegetable oils showed a good antioxidant activity, which was improved by their incorporation into NLCs.
The results have shown that vegetable oils and especially their combinations can be used as renewable
raw materials in designing effective and eco-friendly photoprotective nanostructured formulations.
© 2014 Elsevier B.V. All rights reserved.

1. Introduction (tocopherols and tocotrienols), triterpene alcohols, hydrocarbons

In the last years, there is a growing demand for natural, non-
animal, industrial products in all sectors of life. In the cosmetic
industry, vegetable oils, which are abundant renewable resources
and readily available being mainly derived from seed waste, exhibit
a great interest as raw materials in the development of natural and
eco-friendly cosmetics (Balboa et al., 2014; Lubbe and Verpoortea,
2011). Oils are composed of triglycerides of different fatty acids,
accompanied by mono- and di-glycerides and free fatty acids. In
small amounts they may contain phospholipids, free sterols, tocols
∗ Corresponding author. Tel.: +40 213154 193; fax: +40 213154193.
E-mail address: ioana laca@yahoo.com (I. Lăcătuşu).
and fat soluble vitamins (Karak, 2012). By their natural antioxidants
such as tocols, triterpene, carotenoids, flavonoids and polyphe-
nols, the oils can provide skin protection against reactive oxygen
species (ROS) (Tehranifar et al., 2011; Dhavamani et al., 2014;
Nichols and Katiyar, 2010), but also present anti-carcinogenic and
anti-inflammatory activity (Beutner, 2001; Saha and Ghosh, 2011;
Cicerale et al., 2012). In cosmetic formulations, oils are used as
moisturizers and emollients by increasing the hydration of the skin
(Stamatas et al., 2008; Saraf et al., 2010). Thus, besides their main
role as natural constituent of the lipid fraction, vegetable oils also
have multiple skin benefits and a therapeutic activity by counter-
acting the oxidative stress.
ROS are the main natural byproducts of the oxidative metabolic
process within cells and contribute to cell oxidative damage lead-
ing to the process of aging. ROS act by initiating autocatalytic

http://dx.doi.org/10.1016/j.indcrop.2014.12.049
0926-6690/© 2014 Elsevier B.V. All rights reserved.
 G. Badea et al. / Industrial Crops and Products 67 (2015) 18–24 19

reactions and the molecules they react to are converted into free NLCs as carriers for a synthetic sunscreen that absorbs in the
radicals to propagate the chain of damage (Carocho and Ferreira, UVA wavelength: diethylamino hydroxybenzoyl hexyl benzoate
2013; Chen et al., 2012). Additionally, the UV radiation, X-rays, (DHHB). Pomegranate seed oil, blackcurrant seed oil, sesame seed
gamma rays and atmospheric pollutants generate ROS, contribut- oil, raspberry seed oil, carrot root oil, wheat germ oil and rice bran
ing to an even greater extent to skin damage and the aging process oil were the seven vegetable oils used. Also, vegetable oil-NLCs were
(Junkins-Hopkins, 2010; Wang et al., 2011). The UVA radiation formulated using a mixture of pomegranate seed oil and one of the
(320–400 nm) has the most important contribution to ROS genera- other oils in order to obtain enhanced antioxidant and photopro-
tion, skin chromophores transitioning into an excited and unstable tective actions. The hot high pressure homogenization, a solvent
state by absorbing most of the energy from the UVA wavelengths free method, was used for preparing the NLCs. The vegetable oil-
(Chen et al., 2012). In addition, the UVA radiation has the ability NLCs were characterized in terms of particle size, zeta potential,
to penetrate significantly deeper into the skin, reaching the der- entrapment efficiency and antioxidant activity. The UV protection
mis, and it is by 20 times more abundant in sunlight than UVB performance of the cosmetic formulations developed with veg-
(Kullavanijaya and Lim, 2005). etable oils-NLCs was evaluated based on the in vitro sun protection
Photoprotective cosmetic products use synthetic sunscreens or factor (SPF) and UVA protection factor (UVAPF).
UV filters, which absorb, reflect or scatter the UVA and UVB radia-
tion to some extent, in order to protect the skin against the harmful
2. Materials and methods
effects of the sun’s rays (González et al., 2008). Adding antioxidants
to photoprotective formulations can also be beneficial to the skin
2.1. Materials
by neutralizing the ROS generated from the residual UV rays that
were not filtered by the sunscreen and that reached the skin (Wang
Sodium cholate and Synperonic PE/F68 (block copolymer
et al., 2011; Mishra et al., 2011; Chiari et al., 2014). For example,
of polyethylene and polypropylene glycol) were supplied by
gamma-oryzanol from rice bran oil was found to have a protec-
Sigma–Aldrich Chemie GmbH (Germany). Polyoxyethylenesor-
tive role in the UV-light induced lipid peroxidation (Patel and Naik,
bitan monolaurate (Tween 20) was purchased from Merck
2004), sesamol from sesame oil offers potent photoprotection to
(Germany). Glycerol monostearate (GM) was obtained from Cog-
lymphocytes against UVB induced lipid peroxidation (Prasad et al.,
nis GmbH (Germany) and Emulgade SE/PF (Em) was provided
2005), catechin and ellagic acid from pomegranate seed oil show to
by Elmiplant-Sarantis S.A. (Romania). The UVA filter, diethy-
inhibit markers for UVB-induced skin damage in cultured human
lamino hydroxybenzoyl hexyl benzoate (DHHB), was purchased
skin fibroblasts (Melo et al., 2014), and tocopherol found in wheat
from Sigma–Aldrich Chemie GmbH (Germany). The cream base
germ and raspberry seed oil protects the cell membranes from
(containing stearats, glycerine, fatty alcohols, emulsifier, emol-
light-induced damage (Korać and Khambholja, 2011; Oomah et al.,
lients and an antioxidant–butylhydroxyanisole) was provided
2000). Also, in vitro studies have demonstrated the ability of the
by Elmiplant-Sarantis S.A. Tris[hydroxymethyl] aminomethane,
carotenoids, which can be found in carrot root oil, to protect fibrob-
5-amino-2,3-dihydro-1,4-phthalazinedione (Luminol) were pur-
lasts against UV irradiation-induced mitochondrial DNA mutations
chased from Sigma–Aldrich Chemie GmbH and hydrogen peroxide
(Zussman et al., 2010).
was supplied by Merck (Germany). The vegetable oils purchased
Furthermore, some vegetable oils found in literature are consid-
from Elemental S.R.L. (Romania) were: pomegranate seed oil
ered to be able to block the UV light, sesame oil is claimed to resist
(Punica granatum) – PSO (65.1% punicic acid, 5% linoleic acid, 4.9%
to 30% of UV rays, whereas coconut, peanut, olive, and cottonseed
oleic acid, 2.5% palmitic acid, 1.9% stearic acid, 0.7% eicosenoic
oils block out about 20% (Korać and Khambholja, 2011). In cosmetic
acid, 0.5% arachidic acid, 0.1% behenic acid, 0.1% lignoceric acid,
advertising, creams based on oils such as pomegranate seed oil,
0.1% linolenic acid), wheat germ oil (Triticum vulgare) – WGO (58%
raspberry seed oil, wheat germ oil and carrot oil are presented to
linoleic acid, 12.2% oleic acid, 8.6% linolenic acid, 1.3% eicosenoic
exhibit SPF values over 20.
acid, 0.6% stearic acid, 0.3% palmitoleic acid, 0.2% euricic acid, 0.2%
The concerns associated with natural antioxidants from oils are
nervonic acid, 0.1% behenic acid, 0.1% arachidic acid, 0.1% margaric
their instability and low bioavailability. This can be improved by
acid, 0.1% myristic acid), blackcurrant seed oil (Ribes nigrum) – BSO
using an encapsulation system that can assure chemical stability
(48% linoleic acid, 27% linolenic acid, 3% stearidonic acid), sesame
and sustained release of the active compounds (Kerdudo et al.,
oil (Sesamum indicum) – SO (44.01% linoleic acid, 39.81% oleic acid,
2014).
8.51% palmitic acid, 5.88% stearic acid, 0.68% arachidic acid, 0.41%
Nanostructured lipid carriers (NLCs), introduced in 1999 as
linolenic acid, 0.17% eicosenoic acid), carrot root extract (Daucus
the second generation of solid lipid nanoparticles (Aditya et al.,
carota sativa) in sunflower oil (Helianthus annuus) – CO (56.5%
2014; Weber et al., 2014), are the most promising encapsulation
linoleic acid, 32.7% oleic acid, 6% palmitic acid, 3.4% stearic acid),
technology developed in the field of nanotechnology with mul-
raspberry seed oil (Rubus idaeus) – RSO (53.1% linoleic acid, 29.1%
tiple applications in the cosmetic and pharmaceutical industries
linolenic acid, 13.1% oleic acid, 2.6% palmitic acid, 0.4% arachidic
(Pardeike et al., 2009; Souto et al., 2004). They are submicron par-
acid, 0.2% palmitoleic acid, 0.2% eicosenoic acid, 0.2% acid behenic,
ticles with a solid matrix composed of a blend of solid lipids and
0.1% margaric, 0.1% lignoceric acid) and rice bran oil (Oryza sativa
oils, which is stabilized by an outer shell of surfactants. These car-
bran) – RBO (44% oleic acid, 32.7% linoleic acid, 16.8% palmitic
rier systems, mostly for lipophilic compounds, have the advantage
acid, 2% stearic acid, 1.1% linolenic acid, 0.8% arachidic acid, 0.6%
of being biodegradable and safe for human use due to their gener-
eicosenoic acid).
ally recognized as safe (GRAS) ingredients (Muller et al., 2000). The
physicochemical properties of NLCs are influenced by a number of
factors, including the type of oil used. 2.2. Preparation of vegetable oil-NLCs
Our continuous research in developing safe and effective pho-
toprotective cosmetics based on natural ingredients (Niculae et al., The NLCs were prepared by modified high pressure homoge-
2013, 2014a,b; Lacatusu et al., 2014; Niculae et al., 2014c), led us nization (HPH) method used in previous studies of the authors
to investigate the potential of several antioxidant vegetable oils (Niculae et al., 2014b; Lacatusu et al., 2014). The aqueous phase
in designing nanostructured lipid carrier formulations, which are consisted in 3% surfactant (wt%) (sodium cholate: Tween 20: polox-
able to provide a broad-spectrum sun protection. In the present amer in a ratio of 49:9:1) and the lipid phase, representing 11%
study, a number of seven vegetable oils were employed to design (wt%), consisted in solid lipids, vegetable oils and UVA filter in a
20 G. Badea et al. / Industrial Crops and Products 67 (2015) 18–24
ratio of 7:3:1. The emulsion was heated under stirring at 85 ◦ C for
15 min, and then homogenized by applying 12,000 rpm for 1 min
(High-shear homogenizer PRO250, Germany), and subjected to six
homogenization cycles at 600 bar (HPH, APV 2000 Lab Homog-
enizer, Germany). To remove the water, the dispersions were
lyophilized using a Christ Alpha 1-2 LD Freeze Dryer (Germany).
2.3. Particle size, polydispersity index and zeta potential
The particle size and polydispesity index (PdI) of vegetable
oil-NLCs were analyzed by dynamic light scattering (DLS) using
Zetasizer Nano ZS (Malvern Instruments Ltd. UK) at a scattering
angle of 90◦ and at a temperature of 25 ◦ C. Samples were prepared
by diluting the NLCs dispersions with ultra-pure water to obtain an
adequate scattering intensity.
Zeta potential (ZP) was measured with the same apparatus by
using electrophoretic light scattering technique.
In order to evaluate the stability of the NLCs over time, they were
stored at room temperature and analyzed after 2 months from the
production time. All measurements were performed in triplicate.
2.4. Entrapment efficiency
The entrapment efficiency (EE%) of DHHB into NLCs was deter-
mined by measuring the amount of free DHHB in the dispersion
medium by UV–Vis spectrometry. The samples were prepared by
uniformly mixing the nanocarriers dispersion with ethanol, fol-
lowed by centrifugation for 30 min at 15,000 rpm. The supernatant
was filtered using a Millipore membrane. The filtrate was col-
lected, diluted with ethanol and measured spectrophotometrically
at  = 354 nm by using UV–Vis-NIR Spectrophotometer type V570
Jasco (Japan). The amount of free DHHB was calculated using the
calibration curve in the concentration range of 0.5 to 7 mg/L DHHB,
with a correlation coefficient of R2 = 0.9996 (n = 6).
The percentage of entrapment efficiency has resulted from the
following equation:
Initial amount of DHHB into NLCs − Amount of Free DHHB
EE% = × 100
Initial amount of DHHB into NLCs
2.5. UV protection properties
The photoprotective properties were evaluated on free veg-
etable oils and on cream formulations based on NLCs. The cream
formulations, which contain 3.5% sunscreen DHHB and 10.5% veg-
etable oil, were developed by incorporating the lyophilized NLCs
into a cream base in a ratio of 1:1.
The UV protection properties of the tested formulations were
assessed from the sun protection factor (SPF) and the UVA pro-
tection factor (UVAPF). The protection factors were determined
in vitro by the TransporeTM test (3M Health Care, U.S.A.), accord-
ing to Diffey–Robson methodology (Diffey and Robson, 1989), by
using the UV–Vis V670 Spectrophotometer Jasco (Japan) equipped
with integrated sphere and adequate soft. Samples were prepared
by evenly spread 2 mg/cm2 of each formulation onto TransporeTM
tape placed onto a quartz plate, and then they were allowed to
dry for 15 min before spectrophotometric measurements. The UV
absorption spectrum of each sample was registered in six different
points, in 5 nm steps from 290 to 400 nm, by using as a reference
support TransporeTM tape without sample placed onto the quartz
plate. The recorded average values were used to calculate the SPF
and the UVAPF, as follows:

400
E B
290
SPF = (2)

400
E B /MPF
290

400
E B
320
UVAPF = (3)

400
E B /MPF
320
where E is the spectral irradiance of terrestrial sun light under
defined conditions; B is the relative erythemal effectiveness; MPF␭
is the monochromatic protection factor at each wavelength incre-
ment.
2.6. Antioxidant activity
The antioxidant activity of the NLCs and of the free oils, which
represents the capacity of the system to scavenge the reactive oxy-
gen species, has been measured in vitro by chemiluminescence
method (Chemiluminometer Turner Design TD 20/20, USA). Lumi-
nol (10−5 M), a light amplifying substance, and H2 O2 (10−5 M) in
Tris-HCl buffer solution (pH 8.6) have been used as a generator
system for free radicals (Lacatusu et al., 2013). The samples were
prepared using lyophilized NLCs, which had a concentration of
7% DHHB and 21% vegetable oils, and ethanol in order to obtain
solutions with a concentration of 107 mg/L DHHB and 320 mg/L
vegetable oils. The percentage of scavenged free radicals was cal-
culated by the following relation:
I0 − Is
%AA = × 100 (4)
I0
where I0 and IS are the chemiluminescence maximum of standard
and of sample at time = 5 s.
(1)
3. Results and discussion
3.1. Physicochemical characterization of lipid nanoparticles
The particle size of NLCs loaded with the UVA filter was assessed
in order to evaluate the potential of the seven vegetable oils in
designing lipid nanostructured formulations as carriers for photo-
protective agents. The lipid matrix of the NLCs was prepared by
using a blend of solid lipids, emulgade and glycerol monostearate,
and one of the seven vegetable oils (NLC 1 ÷ 7) or a mixture of PSO
with one of the other oils in a ratio of 2:1 (NLC 8 ÷ 13). All the
vegetable oils resulted in obtaining appropriate NLCs with a mean
particle size ranging between 108 and 145 nm and with a narrow
size distribution reflected by the small PdI values (0.147 ÷ 0.212)
(Fig. 1). The small size of the developed vegetable oil-nanocarriers
reflects a good compatibility among the solid lipids, the oil and
the incorporated drug. Out of the NLCs prepared with individual
oils, the smallest particle size of 109 nm and with a PdI of 0.16 was
obtained for NLC 2 based on wheat germ oil. In general, according
to Hu et al. (2005), smaller nanoparticles are obtained by using less
viscos oils (Hu et al., 2005). By adding other oils to the pomegranate
seed oil for the development of NLCs, a slow increase in the particle
size was observed for NLC 8 ÷ 13 when compared to NLC 1-PSO.
 G. Badea et al. / Industrial Crops and Products 67 (2015) 18–24 21

Fig. 1. Mean particle size (diameter) and polydispersity index (PdI) of DHHB loaded-NLCs developed based on different vegetable oils and their combinations.

3.2. Physical stability
The physical stability of the nanoparticles was determined by
measuring the zeta potential values, which are shown in Table 1.
The colloidal systems with zeta potential values greater than
±30 mV in absolute value are considered to be physically stable
(Teeranachaideekul et al., 2008). All the vegetable oils and their
combinations have led in obtaining highly stable loaded-NLCs hav-
ing negative zeta potentials values ranging between −34.4 and
−44.3 mV. The negative charge found at the surface of the nanocar-
riers is mainly due to the anionic surfactant sodium cholate. Among
the individual vegetable oils, the best stability was assured by
wheat germ oil –NLC 2 (−44.3 mV). When using pomegranate seed
oil in combination with other oils, a lower stability was obtained
for NLC 9 ÷ 12 and a better stability was obtained for NLC 8 and 13,
by comparing with NLC 1 prepared only with pomegranate seed
oil. Thus, wheat germ oil (NLC 8 with ZP of −38.2 mV) and rice
bran oil (NLC 13 with ZP of −38.1 mV) serve to enhance the electric
charge at the surface of NLC 1 based on pomegranate seed oil (ZP
of −36.7 mV).
3.3. Stability over time of the NLCs
The stability study over time of the NLCs stored at room tem-
perature was performed after 2 months from the production time.
The evaluation of the mean particle size and zeta potential values
is shown in Fig. 1 and in Table 1, respectively. After 2 months, the
particle sizes of NLCs have increased in comparison to their ini-
tial values, but they still possessed the characteristic dimensions
of lipid nanoparticles, by remaining in the submicron range, with
diameters from 131 nm to 190 nm. Also, the NLCs presented a rel-
atively monodispersed population of particles with PdI values less
than 0.25 (data not shown).
With regard to their physical stability, the zeta potential values
are in general decreasing except for the NLCs based on PSO, WGO
and their combination, which are slightly increasing. Overall, after 2
months all the NLCs have remained highly physically stable having
zeta potentials more negative than −35 mV.
3.4. Entrapment efficiency
In order to observe which of the oils or their combinations offer
the best incorporation of the UVA filter and, thus, the best UV block-
ing ability, the entrapment efficiency of DHHB was determined. The
percent of entrapment efficiency of DHHB has varied between 51%
and 70% (Fig. 2). Among individual vegetable oils, the best entrap-
ment was assured by pomegranate seed oil based-NLC 1 (63%). This
value was exceeded by using pomegranate seed oil in combination
with wheat germ oil (EE of 70% for NLC 8) and with rice bran oil (EE
of 67% for NLC 13) in developing the NLCs. According to Muller et al.
(2000), a high drug loading capacity of the lipid matrix of NLCs can
be assured by using lipids with different molecular structures that
could form many imperfections in the lipid lattice of the NLCs. Thus,

Table 1
Zeta potential of DHHB loaded-NLCs developed based on different vegetable oils and
their combinations in the day of preparation (0 month) and after 2 months.

Sample Zeta potential

0 month 2 months

NLC 1-PSO −36.70 ± 1.6 -35.9 ± 2.65

NLC 2-WGO -44.30 ± 1.85 -37.3 ± 2.15
NLC 3-BSO -34.40 ± 1.97 -42.2 ± 1.39
NLC 4-SO -36.10 ± 0.40 -34.2 ± 1.50
NLC 5-CO -37.80 ± 1.17 -41.0 ± 0.40
NLC 6-RSO -40.20 ± 1.31 -43.0 ± 0.55
NLC 7-RBO -38.60 ± 1.07 -44.2 ± 1.57

NLC 8-PSO WGO -38.20 ± 1.56 -37.8 ± 2.46

NLC 9-PSO BSO -36.50 ± 0.51 -44.8 ± 2.76
NLC 10-PSO SO -35.20 ± 1.33 -41.8 ± 1.61
NLC 11-PSO CO -35.80 ± 1.56 -33.7 ± 0.75
NLC 12-PSO RSO -35.70 ± 1.07 -39.4 ± 1.74
Fig. 2. Entrapment efficiency (EE%) of DHHB into NLCs developed based on different
NLC 13-PSO RBO -38.10 ± 0.65 -43.9 ± 0.45
vegetable oils and their combinations.
22 G. Badea et al. / Industrial Crops and Products 67 (2015) 18–24
by using wheat germ oil or rice bran oil together with pomegranate
seed oil leads to the formation of a suitable lipid matrix with more
imperfections in which the drug can be positioned.
3.5. In vitro UV absorptive properties
The efficacy of the photoprotective formulations in prevent-
ing and reducing skin cancer and photoaging depends on their
ability to filter the UVA radiation and mostly UVA short rays
(320–340 nm) (Wang et al., 2010). However, skin also needs to be
protected against UVB radiation, which is responsible for less dam-
age but immediately painful sun burning. Additionally, sunscreen
regulations request that cosmetic sunscreen products should pro-
vide a minimum protection against both UVB and UVA radiation
(Commission Recommendation, 2006). Therefore, besides the syn-
thetic UVA filter encapsulated into NLCs, compounds that absorb in
the UVB range are required. In this sense, the vegetable oils, which
are found in literature to be UVB protective by their antioxidant
activity and, also, by having to some extent UVB filtering properties,
were used to synthetize the NLCs.
In order to evaluate the UV protective performance of the photo-
protective nanostructured formulations developed with different
vegetable oils, their UV protection factors on both UVB and UVA
domain were evaluated by determining the in vitro SPF, which
reflects the protection against UVB and UVA radiation, and the
in vitro UVAPF, which reflects the protection against UVA radiation.
The photoprotective properties of the free vegetable oils were
also evaluated. It was observed from the absorption spectra that all
the seven vegetable oils absorb in the UVB domain (290–320 nm),
but only PSO presents a significant absorbance in UVB (Support-
ing information). Thus, the SPF of PSO was 4.5, while for WGO
and RSO was 1.8, and for BSO, SO, CO and RBO was 1.5. In general,
their UVAPF was 1.1, a little greater value of 1.2 being obtained for
WGO and RSO, which absorbs 26% of the UVA short rays, while PSO
absorbs only 14% of the UVA short rays. Therefore, by using only
vegetable oils without synthetic UV filters in formulating conven-
tional photoprotective products would be ineffective for obtaining
a minimum protection on UVB and mainly on UVA domain.
In the present study, a synthetic UV filter, DHHB, which absorbs
in the UVA domain, was used for the synthesis of photoprotective
nanostructured lipid formulations. The UV protective properties of
the NLCs were evaluated after their incorporation into a base cream
in order to obtain cream formulations with a high content of bioac-
tive vegetable oils (10.5%) and a low concentration of DHHB (3.5%
from the maximum 10% admitted by EU regulations). Having a SPF
of 1, the base cream did not influence the UV protective properties
of the formulations.
The SPFs and UVAPFs of the creams based on NLCs are shown
in Fig. 3. Among the cream formulations based on single vegetable
oil, the best photoprotection was assured by the PSO-NLC 1 based
cream, having a SPF of 4.1 and an UVAPF of 7.8, and it was followed
by raspberry seed oil-NLC 6 based cream (SPF of 3 and UVAPF of
6.8).
Due to the fact that the UV protection performance could be
improved by synergistic interactions that occur between the active
compounds of different oils, new NLCs were developed by using a
mixture of pomegranate seed oil, which had the best photopro-
tective results during the first phase, and one of the other oils.
The results demonstrated that an enhancement of the UV protec-
tive properties, by comparing to the PSO–NLC 1 based cream, were
found for the creams based on NLCs developed with PSO in asso-
ciation with wheat germ oil, raspberry seed oil and rice bran oil.
The cream with PSO WGO–NLC 8 demonstrated the best photo-
protection having a SPF of 5.1 and an UVAPF of 9.5. NLC 8 has a
better absorption in the UVA domain by comparing with the UVA
absorption of its corresponding NLCs prepared with single oil. The
Fig. 3. SPF and UVAPF of DHHB loaded-NLCs developed based on different vegetable
oils and their combinations.
other combinations of PSO with BSO, SO and CO did not lead to an
improved photoprotection for their NLCs based creams.
To sum up, the NLCs prepared with blends of PSO and oils have
shown various UV protective factors, although the oils have similar
UV absorption profiles. This fact could be attributed to the small
differences between their absorption profile in the UVA short rays
and to their different loading capacity of the lipid matrix, which
is affected by the various associations between pomegranate seed
oil and different oils. Thus, the NLC 8-PSO WGO with the best pho-
toprotection contains WGO, which has the best absorption in the
UVA short rays (as it can be seen from the Supporting information)
and, also, it possesses the lipid matrix with the highest entrapment
efficiency of the UVA filter.
3.6. Antioxidant activity
Different parts of plants, and in consequence their oils, contain
antioxidant compounds such as tocopherols and phenolic com-
pounds, which are known to exert strong anti- oxidative activity.
The interest in studying the antioxidant properties of the free veg-
etable oils and of vegetable oil-NLCs is related to their protective
function in human skin against the oxidative stress, which is mostly
produced by UV radiation.
The chemiluminescence method employed in this study has
been widely applied as a sensitive assay for the detection and
measurement of reactive oxygen species involved in the oxidative
processes (Jimenez and Navas, 2002).
The antioxidant activity values of the tested sample are shown
in Fig. 4. The free individual vegetable oils have presented AA val-
ues between 70.8% and 78%. The best antioxidant activity was
obtained for the pomegranate seed oil (78 ± 0.64%) and it was
closely followed by the wheat germ oil and blackcurrant seed oil
(around 76%). Among the free individual oils, the lower antioxidant
activity was obtained for sesame oil (70.8 ± 0.42%). By combining
pomegranate seed oil with other oils, synergistic interactions were
observed for the oils combinations of pomegranate seed oil with
wheat germ oil, blackcurrant seed oil or rice bran oil. Therefore,
enhanced antioxidant activity values were obtained for PSO-BSO
(80.7 ± 0.59%), PSO-WGO (79.4 ± 1.25%) and PSO-RBO (79 ± 2.09%)
when compared to single PSO (78 ± 0.64%) .
The antioxidant activity of the NLCs followed the same profile as
free- vegetable oils, but with improved values ranging between 78%
and 85.7%. The best antioxidant value of 85.7 ± 1.38% was obtained
for NLC 9-PSO BSO. Next was the NLC 8-PSO WGO with an antiox-
idant activity of 84.4 ± 0.05%.
This improvement of the antioxidant activity of NLCs could be
due to the vegetable oils brought to nanometer scale and to the
 G. Badea et al. / Industrial Crops and Products 67 (2015) 18–24
Fig. 4. The antioxidant activity (%) of free vegetable oils and of vegetable oil-NLCs.
synergistic effect between the solid lipids and the oils that form
the lipid matrix of the NLCs.
4. Conclusions
The results presented in this study have shown that vegetable
oils and especially their combinations have a great potential to
be used in the development of antioxidant nanostructured lipid
carriers loaded with UVA filter in order to naturally boost their
photoprotective action. The presence of these vegetable oils has
led to increased photoprotective properties of the NLCs by their
dual action of filtering the UVB radiation and of quenching the
reactive oxygen species generated by the UV rays that were not
absorbed by the UV filters. Among the developed photoprotective
nanostructured formulations, the NLCs based on a combination
of pomegranate seed oil with wheat germ oil has shown the
most effective UV performance, the mixture of these oils hav-
ing significant UVB absorptive properties and a good capacity for
encapsulating the UVA filter. All NLCs, and in particular those
prepared with pomegranate seed oil and its combinations, have
presented a good antioxidant activity that was enhanced when
compared to the unencapsulated vegetables oils, due to the pres-
ence of the oils at nanometer size into NLCs. Furthermore, all
the vegetable oils and their combinations led to the development
of NLCs with appropriate particle size and high physical stabil-
ity. Beside the importance of this research in the development of
nanocosmetics, the results are also useful for conventional cosmet-
ics in order to select appropriate vegetable oils for the development
of photoprotective products with a desirable SPF and improved
safety, by using natural plant sources.
Acknowledgments
The work of Gabriela (Niculae) Badea was supported by Project
SOP HRD–PERFORM/159/1.5/S/138963. This work was also sup-
ported by a grant of the Ministry of National Education (Romania),
CNCS-UEFISCDI, project number PN-II-ID-PCE-2012-4-0111.
Appendix A. Supplementary data
Supplementary data associated with this article can be
found, in the online version, at http://dx.doi.org/10.1016/j.indcrop.
2014.12.049.
23
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