Arabian Journal of Chemistry (2017) 10, S1216S1222

King Saud University

Arabian Journal of Chemistry

www.ksu.edu.sa
www.sciencedirect.com

ORIGINAL ARTICLE

Development and stability studies of sunscreen

cream formulations containing three
photo-protective lters
a,*
Slim Smaoui , Hajer Ben Hlima b, Ines Ben Chobba c, Adel Kadri c

a
Department of Life Sciences, Sciences Faculty of Sfax, BP 1171, 3000 Sfax, Tunisia
b
Laboratory of Microorganisms and Biomolecules, Centre of Biotechnology of Sfax, Road of Sidi Mansour Km 6,
P.O. Box 1177, 3018 Sfax, Tunisia
c
Laboratoire de Biotechnologies Vegetales Appliquees a` lAmelioration des Cultures, Faculte des Sciences de Sfax,
University of Sfax, BP 1171, 3000 Sfax, Tunisia

Received 25 December 2012; accepted 27 February 2013

Available online 14 March 2013

KEYWORDS Abstract The present study aimed to formulate and subsequently evaluate sunscreen cream (W/O/
Sunscreen cream; W emulsion) containing three photo-protective lters: benzophenone-3, ethylhexyl methoxycinna-
Photo-protective lters; mate and titanium dioxide at different percentages. Formulations were stored at 8, 25 and 40 C
Organoleptic; for four weeks to investigate their stability. Color, centrifugation, liquefaction, phase separation,
Microbiological stability pH and Sun Protection Factor (SPF) of sunscreen cream formulations were determined. The micro-
biological stability of the creams was also evaluated and the organoleptic quality was carried out for
28 days. Interestingly, the combination of 7% Benzophenone-3, 7% Ethylhexyl methoxycinnamate
and 6% Titanium dioxide preserved physicochemical properties of the product and was efcient
against the development of different spoilage microorganisms as well as aerobic plate counts, Pseu-
domonas aeruginosa, Staphylococcus aureus, and yeast and mold counts. Furthermore, a good sta-
bility was observed for all formulations throughout the experimental period. The newly formulated
sunscreen cream was proved to exhibit a number of promising properties and attributes that might
open new opportunities for the development of more efcient, safe, and cost-effective skin-care, cos-
metic, and pharmaceutical products.
2013 King Saud University. Production and hosting by Elsevier B.V. This is an open access article under
the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

1. Introduction

* Corresponding author. Sunlight is composed of wavelengths ranging from ultraviolet

E-mail address: [email protected] (S. Smaoui). light to visible light. Ultraviolet (UV) is divided into UVA
Peer review under responsibility of King Saud University. (320400 nm), UVB (290320 nm) and UVC (100290 nm)
(Hanson et al., 2006). Exposure to solar radiation has negative
effects on the human skin. Among all, UV is the most harmful
to the skin and causes sunburns and skin cancer after long-
Production and hosting by Elsevier
term exposure(Francis et al., 1998).
http://dx.doi.org/10.1016/j.arabjc.2013.02.020
1878-5352 2013 King Saud University. Production and hosting by Elsevier B.V.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).
Development and stability studies of sunscreen cream formulations containing three photo-protective lters S1217

Organic substances containing chemical groups that can l-

Table 1 Components and concentrations of the formulations.
ter UVA and UVB radiations are used as active ingredients of
sunscreen formulations (Hanson et al., 2006; Nohynek and Ingredients Formulations
Schaefer, 2001; Ibrahim and Brown, 2008). UV absorbers with F1 (%) F2 (%) F3 (%) F4 (%)
an intramolecular hydrogen bridge are widely employed as Benzophenone-3 10 5 5 7
additives against UV radiation (Fluegge et al., 2007; Paterson Ethylhexyl methoxycinnamate 5 10 5 7
et al., 2005). Accordingly, to make skin protection highly effec- Titanium dioxide 5 5 10 6
tive and prevent skin cancer and other types of skin damage, Cetearyl alcohol 3.5 3.5 3.5 3.5
the sunscreens must involve appropriate sun-blocking agents Glycerol monostearate 3.5 3.5 3.5 3.5
and/or preparations that contain combinations of these active Ceteareth-25 1 1 1 1
substances (Hanson et al., 2006; Nohynek and Schaefer, 2001; Isopropyl palmitate 1.5 1.5 1.5 1.5
Palm and ODonoghue, 2007; Pescia et al., 2012). Some of the Caprylic/capric triglyceride 1.5 1.5 1.5 1.5
Polyethylene glycol 1 1 1 1
approved compounds for use in the manufacture formulation
Ethyl paraben 0.5 0.5 0.5 0.5
are benzophenone-3, ethylhexyl methoxycinnamate and tita-
Distillate water 67.5 67.5 67.5 67.5
nium dioxide.
Benzophenone-3 utilizes an excited state intramolecular
proton transfer from a hydroxyl group to dissipate light en- were heated to the same temperature. After that, the rst aque-
ergy. It was supposed to dissipate the absorbed light energy ous phase was added to the oil phase using a mechanical stirrer
in a harmless manner, indeed benzophenone-3 converts the ab- with constant stirring at 2000 rpm for 15 min until the aqueous
sorbed photon energy into heat without chemical damage phase was added completely. Then the (W/O) phase was added
(Schnabel and Kiwi, 1978). Benzophenone-3 has strong to the second phase drop by drop. Finally, for homogeniza-
absorption in 280340 nm (UV-B) range. tion, the speed of the stirrer was decreased to 1000 rpm until
The photochemical behavior of ethylhexyl-p-methoxycin- the emulsion was cooled to room temperature.
namate represents the most widely used sunscreen compound
(Hayden et al., 1998). The photo-induced degradation of eth- 2.2. Physical analysis
ylhexyl-p-methoxycinnamate in emulsion formulations has
been demonstrated by many researchers (De Flandre and The obtained emulsions were submitted to a set of organolep-
Lang, 1988). tic (color, look, feel, thickness) and physical (phase separation
The increasing demand of inorganic UV lters as titanium and creaming) analyses (Akhtar et al., 2011).
dioxide, known to block UVB/UVA sunlight (Serpone et al.,
2007), is related to their low potential for producing irritant 2.3. Stability tests
reactions and to their sunscreen efcacy (Serpone et al., 2007).
The present work was undertaken to investigate the poten-
Stability tests were achieved at different conditions for emul-
tial effects of using three photo-protective chemicals, benzo-
sions to explore the effect of these conditions on the storage
phenone-3, ethylhexyl methoxycinnamate and titanium
of emulsions. These tests were performed on samples kept at
dioxide, with regard to the continuous search for an enhanced
8 C 2 C, 25 C 2 C and 40 C 2 C. Color, phase
formulation of cosmetic and pharmaceutical emulsions and, if
separation and liquefaction of emulsions were observed at var-
any, to submit it to a battery of well-established tests for con-
ious time intervals during 28 days.
sistency and potential industrial application.
Accordingly, the present study was carried out to investi- 2.3.1. Centrifugation tests
gate the physical and microbiological stability of W/O/W
Centrifugal tests were performed for emulsions directly after
emulsions containing three photo-protective lters: benzophe-
preparation. Those tests were repeated after 1 day, 7 days,
none-3, ethylhexyl methoxycinnamate and titanium dioxide at
14 days, 21 days, and 28 days of storage. They were performed
different percentages in order to make a comparative assess-
at 5000 rpm and 25 C for 10 min by placing 10 g of each sam-
ment of these active principles.
ple in centrifugal tubes.

2. Materials and methods 2.3.2. pH determination

The pH value of various emulsions stored at different condi-
2.1. Preparation of emulsions tions was determined using a digital pH Meter. The pH tests
were repeated for multiple emulsions after 1 day, 3 days,
Table 1 shows the components and concentrations of (W/O/ 7 days, 14 days, 21 days, and 28 days of storage.
W) multiple emulsions. The formulations used in this study
are named F1, F2, F3 and F4. Table 2 shows trade name, chem- 2.4. Determination of in vitro SPF of sunscreen cream
ical name and functions of the raw materials which are used in
the formulation preparation. The oil phase consisted of Crod- SPF was determined in the samples in which the previous assay
uret PEG-40, Viatenza Argan PE8, Lipex Shea WL and of organoleptic characteristic evaluation was performed, as
Ethyl Paraben USP24/NF19 heated up to 75 0.5 C. At well on days 1, 2, 5, 8, 12 and 15. To do so, samples were pre-
the same time, two aqueous phases, i.e. Uvinul M40, Uvi- pared according to the method proposed by Dutra et al.
nul MC 80, Micro titanium dioxide JMT-150AO, Lanette (2004). In this manner, 0.5 g of each sample was mixed with
O, Crodamol ICS, Eumulgin B2, Crodamol IPP and Croda- an appropriate amount of distilled water to obtain a nal con-
mol GTCC and distillate water (the second aqueous phase), centration of 0.2 104 g/ml. Briey, samples were dispersed
S1218 S. Smaoui et al.

Table 2 Chemical name, trade name and functions of raw materials used in the preparation of the formulations.
Chemical name Trade name Properties
Benzophenone-3 Uvinul M40 A broad-band UV lter and anti-aging
Ethylhexyl methoxycinnamate Uvinul MC 80 UVB lter
Titanium dioxide Micro titanium dioxide JMT-150AO Negative charging/highly hydrophobic, exhibits water repellency
and surface preparation agent
Cetearyl alcohol Lanette O Viscosity regulation in cosmetic O/W emulsions
Isocetyl stearate Crodamol ICS Emollient with spreading capacity, lubricant and solvent.
Ceteareth-25 Eumulgin B2 Non-ionic emulsier for O/W emulsions.
Isopropyl palmitate Crodamol IPP Non-occlusive eollient and excellent dispersing medium.
Caprylic/capric triglyceride Crodamol GTCC Emollient, lubricant and solvent
Polyethylene glycol Croduret PEG-40 O/W emulsier, eective solubilizer and wetting agent
Argan oil PEG-8 esters Viatenza Argan PE8 Emollient and emulsifying agent
Shea butter oleyl esters Lipex shea WL Oers low viscosity, intermediate spreading and provides
high moisturization
Ethyl p-hydroxybenzoate Ethyl paraben USP24/NF19 Microbiological preservative

in 100 ml of distilled water and were homogenized by ultrason- aeruginosa. Colonies were counted after two days of incuba-
ication for 5 min. The obtained dispersion was ltered with a tion at 25 C (ISO NF- 22717, 2006).
lter paper and the rst 10 ml was rejected. Then 2 ml of l-
tered solution was adjusted to 50 ml using distilled water. 2.5.3. S. aureus
The absorbance of each sample was determined by spectropho- Population of S. aureus was determined by standard plating
tometry in the range of 290320 nm (UVB), with 5 nm inter- methods (ISO NF- 22718, 2008). Colonies of Staphylococcus
vals, using distilled water as blank. A fresh sunscreen sample were selected, gram-stained, and observed for oxidase and cat-
(not submitted to temperature effect) was used as control, in alase reactions to conrm their presence. All microbial counts
order to establish initial SPF. Three replicates of each group were transformed into logarithms of the number of colony-
were performed. The SPF of each sample was determined with forming units (log 10 CFU/g).
the data obtained by spectrophotometric analysis, using the
Mansur equation: 2.5.4. Yeast and mold counts
X
320
The method involved enumeration of colonies on the Sabou-
SPFspectrophotometric CF  EEk  Ik  Absk raud dextrose chloramphenicol agar medium. Enumeration
290
was carried out as a pour plate, surface spread, or membrane
where: CF: correction factor (=10); EE (k): erythemal effect ltration method (ISO NF- 16212, 2008). Microbiological tests
spectrum; I (k): solar intensity spectrum; and Abs (k): absor- were repeated for formulations at 25 C after 0, 7, 14, 21 and
bance of sunscreen product (Mansur et al., 1986). 28 days of preparation.

2.5. Microbiological stability 2.6. Statistical analysis

One gram of emulsion was dispersed in a 4 ml sterile Ringers All measurements were repeated in triplicates and microbial
solution containing 0.25% tween 80. Six dilutions were made counts were transformed into logarithms of the number of
in the same dispersing vehicle, and 0.1 ml was plated out on CFU (log10 CFU/g). Data were subjected to analysis of vari-
the appropriate solid medium using the surface viable method. ance (ANOVA) of the Statistical Analysis System software
Colonies were counted after the incubation and all operations of SAS Institute using the General Linear Models procedure
were carried out in duplicates (ISO NF- 21148, 2000). (SAS, 1990). Differences among the mean values of different
treatments and storage times were achieved by the least signif-
2.5.1. Aerobic plate count icant difference (LSD) test. The signicance was dened at
Aerobic plate counts were determined by inoculating 0.1 ml of P < 0.05 and the differences which are equal to or more than
the homogenate sample onto triplicate sterile plates of pre- the identied LSD values are considered statistically
poured and dried Standard Method Agar. Then, plates were signicant.
incubated for 48 h at 35 C (ISO NF- 21149, 2006). Duplicates
of each dilution (1 ml) of neutralized and non neutralized sam- 3. Results and discussion
ples were pour-plated using Standard Method Agar (Oxoid,
Basingstoke, Hampshire, England) and incubated at 30 C 3.1. Stability of formulated emulsions
for 48 h. Plates containing 25250 colonies were counted and
the average number of CFU/g was calculated. (W/O/W) emulsions are of interest in a number of application
and research areas. In cosmetic research, the emphasis has
2.5.2. P. aeruginosa count been placed on double emulsions as delivery for various activ-
Pseudomonas aeruginosa were enumerated on Pseudomonas ities (Shum et al., 2008).
Agar Base (CM 559, Oxoid) supplemented with fucidin, ceph- In this study, formulations were placed in different storage
aloridine and cetrimide, providing a selective medium for P. conditions (8, 25 and 40 C) for a period of four weeks in
Development and stability studies of sunscreen cream formulations containing three photo-protective lters S1219

Table 3 Physical characteristics of F1 and F2, formulations kept at 8 2 C, 25 2 C and 40 2 C.
Fresh 24 h 3 day 7 day 14 day 21 day 28 day
F1 F2 F1 F2 F1 F2 F1 F2 F1 F2 F1 F2 F1 F2
Liquefaction 8 C              
25 C              
40 C             + +
Color 8 C PY SPW PY SPW PY SPW PY SPW PY SPW PY SPW YW SPW
25 C PY SPW PY SPW PY SPW PY SPW PY SPW PY SPW YW W
40 C PY SPW PY SPW PY SPW PY SPW PY SPW PY W YW W
Phase separation 8 C              
25 C              
40 C           +  + 
Centrifugation 8 C              
25 C              
40 C           +  + 
 = No change; + = Slight change; PY = Pale yellow; SYW = Soft yellowish white; YW = Yellowish white; Y = Yellow; W = White.

Table 4 Physical characteristics of F3 and F4, formulations kept at 8 2 C, 25 2 C and 40 2 C.
Fresh 24 h 3 day 7 day 14 day 21 day 28 day
F3 F4 F3 F4 F3 F4 F3 F4 F3 F4 F3 F4 F3 F4
Liquefaction 8 C              
25 C              
40 C              
Color 8 C W YW W YW W YW W YW W YW W YW W YW
25 C W YW W YW W YW W YW SYW YW SYW YW SYW YW
40 C W YW W YW SYW YW SYW YW SYW YW SYW YW SYW YW
Phase separation 8 C              
25 C              
40 C           +  + 
Centrifugation 8 C              
25 C              
40 C           +  + 
 = No change; + = Slight change; PY = Pale yellow; SYW = Soft yellowish white; YW = Yellowish white; Y = Yellow; W = White.

stability chambers. Color, liquefaction and phase separation begin to contribute to its separation, leading to a decrease in
changes were presented in Tables 3 and 4. viscosity which results in liquefaction increase (Herbert et al.,
1988). As far as the ndings of the present study, no liquefac-
3.1.1. Color tion was observed for the emulsions in any of the storage con-
The ndings revealed that the freshly prepared emulsions were ditions under investigation, i.e., 8, 25 and 40 2 C during the
pale yellow, soft yellowish white, yellowish white and white in 28 days of observation. The absence of liquefaction provided
color for F1, F2, F3 and F4. Little changes in color were ob- strong evidence for the stability of the emulsions under
served for emulsions F1, F2 and F3, as well as the end of storage investigation.
period is characterized by the following colors: soft yellowish
white, white and yellowish white (Tables 3 and 4). For exam- 3.1.3. Phase separation test
ple, for F1, the change in color was observed from the 21st Creaming leads to phase separation and is often attributed to
day. This change was presumably due to the oily phase separa- density differences between the two phases under the inuence
tion promoted at higher temperature. Interestingly, no change of gravity (Derick, 2000). The ndings of this present work re-
in color was observed for F4 at the different storage condi- vealed that all the formulation samples were stable in all stor-
tions: 8, 25 and 40 C 2 C, up to 28 days of observation. age conditions, i.e., 8, 25 and 40 2 C during the 28 days of
the observation period.
3.1.2. Liquefaction
The viscosity of emulsion is often reported to play a vital role 3.1.4. Centrifugation test
in its ow properties (Nasirideen et al., 1998). Starting from No phase separation was observed after centrifugation in any
the emulsion preparation, the temperature and time processes of the samples stored at different conditions up to 21 days. A
S1220 S. Smaoui et al.

weak phase separation was, however, recorded on the 21st day temperature exposure), which was considered to correspond
and up to the 28th day for preparation of F1 and F3 kept at to 100% SPF.
40 C. No other phase separation was observed till the end Generally, SPF values remained stable throughout the
of the experimental period. This was presumably due to the whole period of study. However, when the sunscreen was ex-
proper homogenization speed during emulsion formulation posed to the temperature at 8 2 C, upon 3 days, a slight de-
which might have prevented the breakage of the formulations crease of approximately 5% in SPF was identied (P < 0.05)
during testing (Abdurahman and Rosli, 2006). compared to the initial SPF value. A similar SPF reduction
(6.5%) was perceived in the group of 25 + 2 C, when compar-
3.1.5. pH value determination ing initial SPF with the one measured on day 28 (P < 0.05).
Monitoring the pH value is crucial for determining the emul- Nevertheless, in spite of the statistical signicance of the val-
sions stability. In fact, pH changes indicate the occurrence ues, these determinations do not compromise the general trend
of chemical reactions that can give an idea on the quality of of results, which indicate the maintenance of the SPF.
the nal product. Furthermore, the most important parts of The SPF variation of formulations F1, F2, F3 and F4 at
chemical stability are performances on accelerated testing 8 + 2 C, 25 + 2 C and 40 C + 2 C, upon 28 days of expo-
and kinetics of pH proles (Issa et al., 2000). sition (data not shown), was obtained by comparison with the
The pH of human skin normally ranges from 4.5 to 6.0. fresh sample not subjected to temperature effect, assumed as
Therefore, in order for a formulation to possibly gain admis- 100%. In fact, nal SPF does not display accentuated altera-
sion for industrial application, it should have a pH that is in- tions either when comparing the result of the experimental
cluded into this range (Matousek et al., 2003). Emulsions groups with the initial SPF or when comparing experimental
formulated in this work had a pH value of 6.1, which is close groups themselves. An exception occurs for the maximum
to the neutral pH. Moreover, the pH of the various emulsion average temperature as compared to the initial SPF value, as
samples stored at various storage conditions, i.e. 8, 25 and previously referred, which is signicant (P < 0.05).
40 C, were noted to undergo a continuous decrease up to Although there are many studies concerning the determina-
one month of observation (data not shown). The emulsions tion of SPF in sunscreen of various semisolid dosage forms (lo-
had stable pH values for almost all conditions tested (data tion, milk and cream), most of them do not address the issue of
not shown). In the end of storage, at 40 C, a statistically sig- their behavior when packages are exposed to the effect of high
nicant decrease in the pH of the emulsion was observed. The temperatures. Deccache, describes that a sunscreen in the form
high temperature contributes to the destabilization of the of gel did not exhibit signicant SPF variations during a period
emulsion by hydrolysis, but it did not affect the overall quality of two weeks either at 25 C or at 40 C (Deccache et al., 2010).
of emulsions because the pH values remained around pH 6.0,
which is an acceptable and non skin irritating pH value. 3.3. Microbiological evaluation

3.2. Evaluation of the sun protection factor 3.3.1. Aerobic plate count
The log mean count recorded for the Aerobic plate count of
Since the formulation F4 seemed to have the best properties samples on day 0 was about 2.01 log10 CFU/ g. On day 28
during stability tests, its sun protection factor SPF was calcu- of storage, the log mean count of Aerobic plate count reached
lated in predetermined days by applying Mansur equation 4.33, 4.3, 3.7 and 3.33 for F1, F2, F3 and F4, respectively, which
(Mansur et al., 1986). Fig. 1 represents the variation of SPF did not approximate the maximum limit of 6,9 log10 CFU/g
of the sunscreen emulsion determined upon exposure to differ- for Aerobic plate count recommended by ISO NF- 21149
ent temperatures 8 + 2 C, 25 + 2 C and 40 + 2 C during (2006) in processed cosmetics (Table 5).
the course of study (28 days). An initial SPF determination
was performed in a fresh sample of sunscreen (prior to any 3.3.2. P. aeruginosa and S. aureus counts
The results from the Pseudomonas and S. aureus detection tests
120 were negative, thus conrming that all formulated emulsions
met the conventional standards specied with regard to tness
110 for human consumption (ISO NF- 22717, 2006; ISO NF-
SPF (% of initial value)

22718, 2008). (Table 5).

100
3.3.3. Yeast and mold counts
90 Yeast and molds have been tested in cosmetic products to as-
sess microbiological safety and product quality during process-
80 ing and storage (ISO NF- 16212, 2008). The levels of these
microorganisms were noted to remain under the standard lim-
70 it. In fact, the initial yeast and mold counts recorded for all
treatments were under the detection limit (ISO NF- 16212,
60 2008).
0 3 6 9 12 15 18 21 24 27 30
Time (days)
Moreover, the yeast and mold count values recorded for the
formulated sample F4 were noted to show delayed prolifera-
Figure 1 Sun protection factor (SPF) variation of F4 during tion when compared to F1, F2 and F3 (Table 5).
sunscreen incubation under different conditions: (n) 8 2 C, () In conclusion, the combination of 7% benzophenone-3, 7%
25 2 C; and (m) 40 2 C. ethylhexyl methoxycinnamate and 6% titanium dioxide seems
Development and stability studies of sunscreen cream formulations containing three photo-protective lters S1221

Table 5 Microbial load of aerobic plate count, P. spp, S. aureus and Yeast and molds count of F1, F2, F3 and F4 during 28 days of
storage at 25 2 C.
Days of storage at 25 2 C
0 7 14 21 28
Aerobic plate count
F1 2.0 0.30a 2.43 0.34c 2.85 0.26b 3.19 0.19b 3.43 0.30c
F2 2.02 0.31a 2.15 0.37a 2.36 0.18a 2.98 0.22a 3.03 0.29b
F3 2.04 0.25a 2.28 0.29b 2.88 0.17b 3.15 0.18b 3.40 0.18c
F4 2.01 0.27a 2.11 0.19a 2.34 0.15a 2.96 0.15a 3.33 0.11a
P. spp
F1 <1 <1 <1 <1 <1
F2 <1 <1 <1 <1 <1
F3 <1 <1 <1 <1 <1
F4 <1 <1 <1 <1 <1
S. aureus
F1 <1 <1 <1 <1 <1
F2 <1 <1 <1 <1 <1
F3 <1 <1 <1 <1 <1
F4 <1 <1 <1 <1 <1
Yeast and molds
F1 1.12 0.28a 1.42 0.22a 1.56 0.15a 1.79 0.39a,b 1.8 0.22a
F2 1.14 0.16a 1.55 0.22b 1.78 0.14c 1.88 0.27a 1.98 0.27c
F3 1.11 0.11a 1.4 0.15a 1. 61 0.16a 1.82 0.17a 1.89 0.23b
F4 1.13 0.16a 1.37 0.22a 1.49 0.14b 1.72 0.27b 1.79 0.27a
: Standard deviation of three replicates.
CFU: Colony forming units.
ac
Averages for different microbial analyses with different letters in the same column are different (P < 0.05).

to be very interesting since it preserved physicochemical prop- References

erties of the product and was efcient against the growth of
different spoilage microorganisms. It should be noted that Abdurahman, H.N., Rosli, M.Y., 2006. Stability investigation of
the maximum authorized levels are 7.5% for -Ethylhexyl 4- water-in-crude oil emulsion. J. Appl. Sci. 6, 2895.
methoxycinnamate are 7% for benzophenone-3 according to Akhtar, N., Zaman, S.U., Khan, B.A., Haji, M., Khan, S.,
FDA (FDA, 1999) and 25% for titanium dioxide (Salvador Ahmad, M., Rasool, F., Mahmood, T., Akhtar, R., 2011.
and Chisvert, 2005) in the F4 formulation. Evaluation of various functional skin parameters using a topical
cream of Calendula ofcinalis extract. Afr. J. Pharm. Pharmaco.
5, 199.
4. Conclusion De Flandre, A., Lang, G., 1988. Photostability assessment of sunsc-
reens. Benzylidene camphor and dibenzoylmethane derivatives. Int.
The ndings presented in the current study indicated that sun- J. Cosmet. Sci. 10, 53.
screen cream (W/O/W emulsion) containing three photo-pro- Deccache, D.S., Santos, E.P.D., Cabral, L.M., Rodrigues, C.R., De
tective chemicals: benzophenone-3, ethylhexyl Sousa, V.P., 2010. Development of methodologies for dimethyl-
aminoethanol glycolate assay in association with sunscreens in
methoxycinnamate and titanium dioxide at different percent-
dermocosmetic formulation. Braz. J. Pharm. Sci. 46, 705.
ages yields good physical characteristics and microbiological Derick, R., 2000. Fat crystals and emulsion stability, a review. Food
stability, thus providing a safe and stable emulsion delivery Res. Int. 33, 3.
system. Formulations and subsequent evaluation of the cos- Dutra, E.A., Oliveira, D.A., Kedor-Hackmann, E.R., Santoro,
metic emulsions from the photo-protective lters presented M.I., 2004. Determination of sun protection factor (SPF) of
here, showed no phase separation in emulsions at different sunscreens by ultraviolet spectrophotometry. Braz. J. Pharm.
storage conditions during 28 days except for formulation F1 Sci. 40, 381.
at 40 2 C. Emulsion liquefaction started in the emulsions Fluegge, A.P., Waiblinger, F., Stein, M., Keck, J., Kramer, H.E.A.,
at increased temperatures after the 28th day of storage for for- Fischer, P., Wood, M.G., Debellis, A.D., Ravichandran, R.,
mulations F1 and F2. On centrifugation, the phase separation Leppard, D., 2007. Probing the intramolecular hydrogen bond of
2-(2-Hydroxyphenyl) benzotriazoles in polar environment: a
was noted in both F1 and F2, to start after the 21st day of stor-
photophysical study of UV absorber efciency. J. Phys. Chem. A.
age at 40 C. Furthermore, the multiple emulsions prepared in 111, 9733.
this work had a pH value of 6.5, which is close to the neutral Francis, P.G., Mark, M., Frankash, J., 1998. A review of sunscreen
pH. On the other hand, SPF values of F4, which seemed to be safety and efcacy. Photochem. Photobiol., 68.
the more interesting formulation, remained stable throughout Hanson, K.M., Gratton, E., Bardeen, C.J., 2006. Sunscreen enhance-
the whole period of study. Microbiological assays (Aerobic ment of UV-induced reactive oxygen species in the skin, Free
plate count, P. aeruginosa, S. aureus, and yeast and mold Radic. Biol. Med. 41, 1205.
counts) on elaborated sunscreen cream revealed that the for- Hayden, C.G., Roberts, M.S., Benson, H.A.E., 1998. Sunscreens: are
mulation F4 was stable during storage at 25 2 C. Australian getting the good oil? Aust. NZ J. Med. 28, 639.
S1222 S. Smaoui et al.

Herbert, A.L., Martin, M.R., Gilbert, S.B. (Eds.), 1988. Pharmaceu- Nohynek, G.J., Schaefer, H., 2001. Benet and risk of organic
tical Emulsions Pharmaceutical Dosage Forms: Disperse System, ultraviolet lters. Regul. Toxicol. Pharmacol. 33, 285.
vol. 1. Marcel Dekkar, New York and Basel, pp. 199240, 285288. Palm, M.D., ODonoghue, M.N., 2007. Update on photoprotection.
Ibrahim, S.F., Brown, M.D., 2008. Tanning and cutaneous malig- Dermatol. Ther. 20, 360.
nancy. Dermatol. Surg. 34, 460. Paterson, M.J., Robb, M.A., Blancafort, L., Debellis, A.D., 2005.
ISO NF- 16212, 2008. Cosmetics Microbiology Enumeration of Mechanism of an exceptional class of photostabilizers: A seam of
yeast and mold. conical intersection parallel to Excited State Intramolecular Proton
ISO NF- 21148, 2000. Cosmetics- Microbiology. Instructions for Transfer (ESIPT) in o-Hydroxyphenyl-(1,3,5)-triazine. J. Phys.
microbiological tests. Chem. A. 109, 7527.
ISO NF- 21149, 2006. Cosmetics Microbiology Enumeration and Pescia, A.C., Astol, P., Puglia, C., Bonina, F., Perrotta, R., Herzog,
detection of aerobic mesophilic bacteria. B., Damiani, E., 2012. On the assessment of photostability of
ISO NF- 22717, 2006. Cosmetics- Microbiology. Detection of Pseu- sunscreens exposed to UVA irradiation: from glass plates to pig/
domonas aeruginosa. human skin, which is best? Int. J. Pharm. 427, 217.
ISO NF- 22718, 2008. Cosmetics- Microbiology. Detection of Staph- Salvador, A., Chisvert, A., 2005. Sunscreens analysis. A critical survey
ylococcus aureus. on UV lters determination. Anal. Chim. Acta. 537, 1.
Issa, T.S., Philippe, B., Raymond, H., Michel, H., Jacques, D., 2000. SAS, 1990. SAS/STAT users guide. Statistical Analysis System
Improved kinetic parameter estimation in pH-prole data. Int. J. Institute, Inc, Cary, NC, USA,.
Pharm. 198, 39. Schnabel, W., Kiwi, J., 1978. In: Jellinek, H.H. (Ed.), Aspects of
Mansur, J.S., Breder, M.N.R., Mansur, M.C.A., Azulay, R.D., 1986. Degradation and Stabilization of Polymers. Elsevier Sci. Publ. Co.,
Determinacao do fator de protecao solar por espectrofotometria. Amsterdam.
An. Bras. Dermatol. 61, 121. Serpone, N., Dondi, D., Albini, A., 2007. Inorganic and organic UV
Matousek, J.L., Campbell, K.L., Kakoma, I., Solter, P.F., Schaeffer, lters: their role and efcacy in sunscreens and suncare products.
D.J., 2003. Evaluation of the effect of pH on in vitro growth of Inorg. Chim. Acta. 360, 794.
Malassezia pachydermatis. Can. J. Vet. Res. 67, 5659. Shum, H.C., Lee, D., Yoon, I., Kodger, T., Weitz, D.A., 2008. Double
Nasirideen, S., Kas, H.S., Oner, F., Alpar, R., Hincal, A.A., 1998. emulsion templated monodisperse phospholipid vesicles. Langmuir
Naproxen incorporated lipid emulsion. Formulation and stability 24, 7651.
studies. J. Clin. Pharm. Ther. 23, 57.