European Journal of Pharmaceutics and Biopharmaceutics 65 (2007) 122125

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In vitro erythemal UV-A protection factors of inorganic

sunscreens distributed in aqueous media using carnauba
waxdecyl oleate nanoparticles
J.R. Villalobos-Hernandez, C.C. Muller-Goymann

Institut fur Pharmazeutische Technologie, Technische Universitat Braunschweig, Braunschweig, Germany

Received 18 May 2006; accepted in revised form 13 July 2006
Available online 26 July 2006

Abstract
This paper describes the in vitro photoprotection in the UV-A range, i.e. 320400 nm obtained by the use of carnauba waxdecyl oleate nanoparticles either as encapsulation systems or as accompanying vehicles for inorganic sunscreens such as barium sulfate, strontium
carbonate and titanium dioxide. Lipid-free inorganic sunscreen nanosuspensions, inorganic sunscreen-free waxoil nanoparticle suspensions and waxoil nanoparticle suspensions containing inorganic sunscreens dispersed either in their oil phase or their aqueous phase
were prepared by high pressure homogenization. The in vitro erythemal UV-A protection factors (EUV-A PFs) of the nanosuspensions
were calculated by means of a sun protection analyzer. EUV-A PFs being no higher than 4 were obtained by the encapsulation of barium
sulfate and strontium carbonate, meanwhile by the distribution of titanium dioxide in presence of waxoil nanoparticles, the EUV-A PFs
varied between 2 and 19. The increase in the EUV-A PFs of the titanium dioxide obtained by the use of waxoil nanoparticles demonstrated a better performance of the sun protection properties of this pigment in the UV-A region.
2006 Elsevier B.V. All rights reserved.
Keywords: Nanoparticles; Nanostructured lipid carriers (NLC); Inorganic sunscreens; In vitro erythemal UVA protection factor

1. Introduction
Modications of solid lipid nanoparticles composed of
binary mixtures of solid lipids and liquid lipids recently
referred to as nanostructured lipid carriers (NLC) constitute suitable vehicles for the dermal delivery of dierent
drugs [1]. These particles constitute not only intrinsic
UV-blockers and scatters in virtue of their crystallinity
and reduced particle size comprising a range between 200
and 400 nm [2], but also carriers capable to integrate
organic and inorganic lters inside of a single structure to
combine or enhance the UV-protection properties of both,
particularly in the case of NLC. Experiments describing
*

Corresponding author. Institut fur Pharmazeutische Technologie,

Technische Universitat Braunschweig, Mendelssohnstr. 1, D-38106,
Braunschweig, Germany. Tel.: +49 531 3915650; fax: +49 531 3918108.
E-mail address: [email protected] (C.C. MullerGoymann).
0939-6411/$ - see front matter 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.ejpb.2006.07.013

this enhancement have been already reported when inorganic sunscreens were enclosed inside or bonded with carnauba waxdecyl oleate nanoparticles [3]. The UV
protection properties of the nanoparticles can be described
in terms of the sun protection factor (SPF) like in the case
of any other sunscreen product. However, the SPF mainly
represents the protection against UV-B [4]. For this reason,
the new developments in sunscreens have to provide a
description of the protection against not only the UV-B
radiation, but also the UV-A one [5,6]. The UV-A region
also called near ultraviolet region is associated with the
wavelengths between 320 and 400 nm. The UV-A radiation
possesses lesser energy than UV-B (290320 nm) or UV-C
radiations (200290 nm); i.e. it is of longer wavelength
and because of this property it penetrates into the skin
and reaches the dermis provoking several damages such
as immediate and delayed tanning reactions, loss of collagen, diminution in the quantity of blood vessels, alteration

J.R. Villalobos-Hernandez, C.C. Muller-Goymann / European Journal of Pharmaceutics and Biopharmaceutics 65 (2007) 122125

of the connective tissue of the dermis and skin photosensitization upon others [79]. To describe the UV-A protection, many in vivo methods have been proposed [5,6].
Nevertheless, in order to avoid expensive and time-consuming in vivo UV-A protection testing, in vitro methods
for UV-A protection assessment based on spectrophotometric analysis have been developed [10,11]. The in vitro
erythemal UV-A protection factor is a parameter derived
from the calculations of the SPF calculations and because
of this reason, it is closely related to the general SPF performance. As described in the literature, the sun protection
factor is a parameter indicating the performance of a sun
protection product or sunscreen formulation [10]. It represents a number derived from the ratio of the time of exposure to a UV spectrum between 290 and 400 nm to produce
erythema in human skin in the presence or absence of a
sunscreen product applied in a specied dose. The in vitro
SPF measured represents an indicator of the UV-A/UVB protective property of a sunscreen product calculated
from the mean monochromatic protection factor, the solar
irradiance and the erythemal constants according to Eq. (1)
,
400
400
X
X
SPF
Ek Bk
Ek Bk =MPFk ;
1
290

290

where Ek is the spectral irradiance of terrestrial sun light

under dened conditions; Bk is the erythemal eectiveness;
and MPFk is the mean monochromatic protection factor.
Ek is dened by the midday midsummer sunlight at
40N with a solar zenith angle of 20. Considering the
UV-A wavelength range, i.e. 320400 nm and using the
terms of Eq. (1), the in vitro erythemal UV-A PF can be
calculated according to Eq. (2).
Erythemal UV-A protection factor
,
400
400
X
X

Ek Bk
Ek Bk =MPFk :
320

320

The aim of this paper was to complete the studies on the

UV protection properties of the nanosuspensions containing carnauba waxdecyl oleate nanoparticles used either
as an encapsulation/bonding media or as accompanying
vehicles for inorganic pigments, i.e. barium sulfate, strontium carbonate and titanium dioxide. The in vitro SPF of
these nanosuspensions have been already reported in a
former paper [3], for this reason, this manuscript was focused only on the description of the in vitro erythemal
UV-A protection factors of those colloidal systems.
2. Materials and methods
2.1. Materials
The inorganic pigments used were aluminastearic acid
surface-treated nanone titanium dioxide having a primary
particle size (PPS) of about 17 nm (Kemira OY, Finland),
nanone barium sulfate (PPS ca. 30 nm) (Solvay GmbH,

123

Germany) and strontium carbonate (PPS ca. 30 nm) (Solvay GmbH, Germany). The lipids applied were carnauba
wax Ph. Eur. and decyl oleate (Caelo GmbH, Germany).
Other excipients used were polysorbate 80 (Atlas Chemie
GmbH, Germany), simethicone antifoamer (SigmaAldrich GmbH, Germany) and methylisothiazolinone
(Brenntag GmbH, Germany). For all preparations bidistilled and ltered water (0.2 lm) was utilized.
2.2. Experimental methods
2.2.1. Manufacture of the nanosuspensions
The chemical compositions of the prepared nanosuspensions as well as the distribution of the pigments in the different phases; i.e., oil phase or aqueous phase, are shown in
Table 1. For the manufacture of the nanosuspensions, a
pre-dispersion of both phases using an Ultraturrax (IKA,
Germany) followed by a high pressure homogenization
performed in a Niro homogenizer (Niro Soavi S.p.A., Italy) was carried out. A detailed description of the preparation of these nanosuspensions can be found in a previous
paper [3]. Four dierent types of nanosuspension were prepared. This includes: nanosuspensions of carnauba wax
decyl oleate matrices (Formulation I); lipid-free inorganic
pigment nanosuspensions (Formulation II); nanosuspensions of carnauba waxdecyl oleate matrices loaded with
inorganic pigments distributed in the lipid phase (Formulation III); nanosuspensions of carnauba waxdecyl oleate
matrices loaded with titanium dioxide distributed in the
aqueous phase (Formulation IV).
2.2.2. In vitro erythemal UV-A protection factor (EUV-A
PF)
The in vitro EUV-A PF was measured by means of a
SPF-290 computer operated analyzer (Optometrics Corp,
UK). One hundred and ten microliters samples were spread
over Transpore tapes (3 M GmbH, Germany) having an
area of 55.5 cm2 with a gloved nger. Radiation attenua-

Table 1
The chemical composition and concentration of the tested formulations
Substances

wt (%)

Formulation
I

II

III

IV

Decyl oleate
Carnauba wax
Pigment*
Tween 80
Simethicone
Methylisothiazoline
Double dist. water

5
5, 10
2, 4, 6
1
0.01
0.0285
q.s. 100

+
+

+
+
+
+



+
+
+
+
+

+
+
+ (LP)
+
+
+
+

+
+
+ (AP)
+
+
+
+

Key: (*), barium sulfate, strontium carbonate or titanium dioxide; (),

absent; (+), present; LP, lipid phase and AP aqueous phase.
Formulation key: I, nanosuspensions of carnauba waxdecyl oleate
matrices; II, lipid-free inorganic pigment nanosuspensions; III, nanosuspensions of carnauba-waxdecyl oleate matrices loaded with pigments in
the lipid phase and IV, nanosuspensions of carnauba-waxdecyl oleate
matrices loaded with titanium dioxide in the aqueous phase.

124

J.R. Villalobos-Hernandez, C.C. Muller-Goymann / European Journal of Pharmaceutics and Biopharmaceutics 65 (2007) 122125

CW:DO (1 :1) =1.6 0; CW:DO (2:1)= 2.55 0.07

Fig. 1. In vitro erythemal UV-A protection factors of the dierent nanosuspensions as function of the pigment and its concentration, the carnauba-wax
decyl oleate ratio and the phase used for its distribution. Key: PC, pigment concentration; CW:DO, carnauba waxdecyl oleate ratio; LP, lipid phase and
AP, aqueous phase.

tion of the applied samples was performed in a wavelength

range of 320400 nm. The radiation was provided by a
compact 125 W xenon arc lamp. Measurements of each
formulation were done in duplicate.
3. Results and discussion
3.1. Erythemal UV-A protection factor (EUV-A PF)
The EUV-A PF is a parameter analog to the SPF and
represents a number derived from the ratio of the time of
exposure to a UV spectrum between 320 and 400 nm to
produce erythema in human skin in the presence or absence
of a sunscreen product. The higher this value, the more
UV-A protection a sunscreen oers. As shown in Fig. 1
(legend), the EUV-A PFs of the pigment-free waxoil
nanoparticles (CW:DO) prepared at ratios 1:1 and 2:1 were
1.6 0.0 and 2.55 0.07, respectively. Hence, from these
results, the EUV-A PFs of the pigment-free waxoil nanoparticles can be labelled as very low. On the other hand, the
use of barium sulfate and strontium carbonate demonstrated to exhibit very low UV-A protection either in encapsulated or in non-encapsulated condition. Although, in these
cases, slight increases probably attributed to higher pigment concentrations combined with dierent waxoil ratios
were observed, EUV-A PFs being no higher than 4 were
detected. Conversely, in the case of titanium dioxide high
EUV-A PFs were seen, particularly after the crystals were
encapsulated with (LP) or dispersed within waxoil nanoparticles (AP). These latter values varied between 2 and
19 although the original protection values of the corresponding lipid-free pigment suspensions were smaller than

three. This indicated that the association with waxoil particles also favoured the UV-A protection properties of the
pigment and thus demonstrated that not only the UV-B
protection character of the titanium dioxide could be
enhanced by the encapsulation process of the pigments as
reported before [3]. Therefore, a sunscreen exhibiting
UV-A and UV-B protection could be referred to as a wide
band sunscreen.
References
[1] R.H. Muller, M. Radtke, S. Wissing, Solid lipid nanoparticles (SLN)
and nanostructured lipid carriers (NLC) in cosmetic and dermatological preparations, Adv. Drug Deliv. Rev. 54 (2002) S131S155.
[2] M. Radkte, Grundlegende Untersuchungen zur Arzneistonkorporation, -Freisetzung und Struktur von SLN und NLC. Ph. D Thesis,
FU Berlin, 2003, Germany.
[3] J.R. Villalobos-Hernandez, C.C. Muller-Goymann, Novel nanoparticulate carrier systems based on carnauba wax an decyl oleate for the
dispersion of inorganic sunscreens in aqueous media, Eur. J. Pharm.
Biopharm. 60 (2005) 113122.
[4] F. Bernerd, C. Vioux, F. Lejuene, D. Asselineau, The sun protection
factor (SPF) innadequately denes broad spectrum photoprotection:
demonstration using skin reconstructed in vitro exposed to UVA,
UVB or UV-solar simulated radiation, Eur. J. Dermatol. 13 (2003)
242249.
[5] D. Moyal, A. Chardon, N. Kollias, Determination of UVA protection factors using the persistent pigment darkening (PPD) as the end
point (part 1) calibration of the method, Photodermatol. Photoimmunol. Photomed. 16 (2000) 245249.
[6] International Sun Protection Factor (SPF) Test Method, COLIPA
Guidelines, Estoril, Portugal, June 2006.
[7] C. Walters, A. Keeney, C.T. Wigal, C.R. Johnston, R.D. Cornaelius,
The spectrophotometric analysis and modeling of sunscreens, J.
Chem. Ed. 74 (1997) 99101.

J.R. Villalobos-Hernandez, C.C. Muller-Goymann / European Journal of Pharmaceutics and Biopharmaceutics 65 (2007) 122125
[8] M.A. Pathak, Photoprotection against harmful eects of solar UVB
and UVA radiation: an update, in: N.J. Lowe, N.A. Shaath, M.A.
Pathak (Eds.), General principles in Sunscreens: Development,
Evaluation and Regulatory Aspects, Marcel Dekker, Inc., New York,
NY, 1997, pp. 59100.
[9] N.J. Lowe, J. Friedlander, Sunscreens: rationale for use to reduce
photodamage and phototoxicity, in: N.J. Lowe, N.A. Shaath, M.A.
Pathak (Eds.), General Principles in Sunscreens: Development,

125

Evaluation and Regulatory Aspects, Marcel Dekker, Inc., New York,

NY, 1997, pp. 3579.
[10] SPF Analyzer Instruction Manual, Optometrics UK Ltd., Leeds, 1997.
[11] N.J. Lowe, Ultraviolet A claims and testing procedures for OTC
sunscreens: A summary and Review, in: N.J. Lowe, N.A. Shaath,
M.A. Pathak (Eds.), General principles in Sunscreens: Development,
Evaluation and Regulatory Aspects, Marcel Dekker, Inc., New York,
NY, 1997, pp. 527558.