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Formulation, characterization and in vitro

evaluation of herbal sunscreen lotion

ARTICLE in ORIENTAL PHARMACY AND EXPERIMENTAL MEDICINE DECEMBER 2012

DOI: 10.1007/s13596-012-0069-z

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Orient Pharm Exp Med (2012) 12:241246
DOI 10.1007/s13596-012-0069-z
RESEARCH ARTICLE
Formulation, characterization and in vitro evaluation
of herbal sunscreen lotion
Arun Rasheed & S. Neelufar Shama &
S. Mohanalakshmi & V. Ravichandran
Received: 7 December 2011 / Accepted: 4 April 2012 / Published online: 11 May 2012
# Institute of Oriental Medicine, Kyung Hee University 2012
Abstract Natural substances extracted from plants have
recently been considered as potential sunscreen resources
owing to high ultraviolet ray absorption and antioxidant
activity. The decrease in the intensity of UV radiation reach-
ing the skin through sunscreens may reduce the risk of sun-
induced skin cancer. The present study attempts to develop
sunscreen lotions, possessing broad spectrum of anti-UV
radiation effectiveness with reduced concentration of chem-
ical UV filters, from the extracts of bioactive products such
as Curcuma longa L. (Zingiberaceae), Aloe vera (Liliaceae)
and Alpinia galanga Willd. The effectiveness of the product
was evaluated using Sun Protection Factor (SPF). Curcumin
was selected as potential bioactive agents due to their phy-
tochemical compositions possessing considerable content of
polyphenolic compounds and Alpinia galanga is used as
key ingredient in various commercial sunscreen lotions as
it has skin protectant action against UV rays and boosts the
activity of conventional sunscreens. The sunscreen lotions
were prepared using three different compositions F1, F2 and
A. Rasheed (*)
Department of Pharmaceutical Chemistry,
Sree Vidyanikethan College of Pharmacy,
A. Rangampet,
Tirupati, Andhra Pradesh, India
e-mail: arunrasheed@rediffmail.com
S. N. Shama : S. Mohanalakshmi
Department of Pharmacognosy,
Sree Vidyanikethan College of Pharmacy,
A. Rangampet,
Tirupati, Andhra Pradesh, India
V. Ravichandran
Faculty of Pharmacy, AIMST University,
Semeling, 08100 Bedong,
Kedah Darul Aman, Malaysia
F3 and evaluated for their stability, safety and SPF. Results
showed that the sunscreen lotions were non-mutagenic, non-
irritant, stable and possess SPF for normal skin. The efficacy
when tested with a standard was observed to be same as that
of a marketed sunscreen with SPF 55 and SPF 20. From the
present study, formulation F2 having curcumin extract was
proved to be stable and effective with high SPF compared to
F1 and F3.
Keywords Skin burn . Bioactive product . Curcuma longa .
Aloe vera . Alpinia galanga . SPF
Introduction
Sunscreens are used to protect the skin from the harmful
effects of the sun, including the appearance of erythema in
the short term and actinic photo-ageing and skin cancers in
the long term. The decrease in the intensity of UV radiation
reaching the skin through sunscreens may reduce the risk of
sun-induced skin cancer. The UV spectrum is divided into
three groups based on wavelength: UVC (100290 nm),
UVB (290320 nm) and UVA (320400 nm). UVA is fur-
ther subdivided into UVA2 (320340 nm) and UVA1 (340
400 nm). Solar UV radiation reaching the Earths surface
approximately consists of 9099 % UVA and 110 % UVB
(COLIPA 2006; Verschooten et al. 2006; Faurschou and
Wulf 2007).
Recently, the development of sunscreens possessing
broad spectrum anti-UV radiation effectiveness with re-
duced concentration of chemical UV filters (Velasco et al.
2008); and bioactive products have been the focus of several
researches due to ecological issues (sustainability), mini-
mum ambient impact and for safe utilization (Banov et al.
2006; Rolim et al. 2006). Various synthetic agents are used
242
as photoprotectives, but their use is limited because of their
potential toxicity in humans and their ability to interfere in
certain selected pathways of multistage process of carcino-
genesis. Phytoconstituents are gaining popularity as ingre-
dients in cosmetic formulations as they can protect the skin
against exogenous and endogenous harmful agents and can
help remedy many skin conditions (Aburjai and Natsheh
2003). Herbal extracts act on these areas and produce heal-
ing, softening, rejuvenating, and sunscreen effects (Deep
and Saraf 2008). Several botanical compounds have been
shown to be antimutagenic, anticarcinogenic and nontoxic
and have the ability to exert striking inhibitory effects on a
plethora of cellular events at various stages of carcinogene-
sis. Few examples include tea polyphenols, curcumin, sily-
marin, garlic compounds, apigenin, resveratrol, ginkgo
biloba, beta-carotenoids, and ascorbic acid (Fguyer et al.
200303). Important categories of beneficial phytoconstitu-
ents include phenolic acids, flavonoids, and high molecular
weight polyphenols (Robbins 2003; King and Young 1999).
Most commonly used herbs in herbal sunscreen lotions
are Aloe vera, basil, green tea, almond, olive, jojoba and
cucumber (Katiyar and Elment 2002; Ashawat et al. 2006;
Ashawat et al. 2005; Shrivastava et al. 2003). Oriented to
the sunscreen development, the use of natural raw materials
that infers UV absorption and skin protection against UVB
and UVA radiation is of great interest, associated with the
benefits of the products and compliance of the consumers
(Tabrizi et al. 2003). Polyphenolic compounds exhibit a
wide number of pharmacological properties such as antial-
lergic, anti-inflammatory, hepatoprotective, vasoactive,
antithrombotic, antioxidant, free radical scavenging, antitu-
mor, antibacterial and antiprotozoa due to their different in
vivo action mechanism (Movileanu et al. 2000; Baby et al.
2006). Due to the structural similarities between polyphe-
nolic compounds such as flavonoids and organic UV filters,
they might exert photoprotection activity in addition to the
antioxidant and absorbance spectrum profiles of these bio-
active compounds (Velasco et al. 2008).
The present study attempts to develop sunscreen lotions,
possessing broad spectrum of anti-UV radiation effective-
ness with reduced concentration of chemical UV filters,
from the extracts of bioactive products such as Curcuma
longa L. (Zingiberaceae), Aloe vera (Liliaceae) and Alpinia
galanga Willd (Sesquiterpenes) at different concentrations.
The effectiveness of the products were evaluated through
various physicochemical properties and biological activities;
the major being Sun Protection Factor (SPF). Curcumin was
selected as potential bioactive agents due to their phyto-
chemical composition possessing considerable content of
polyphenolic compounds and Alpinia galanga is used as
key ingredient in various commercial sunscreen lotions as
having skin protectant action against UV rays and boosts the
activity of conventional sunscreens.
A. Rasheed et al.
Materials and methods
Plant material
The plant materials used in the formulation were collected
from the whole sale supplier of Herbal Crude Drugs, Mumbai,
India. These were authentified by Dr. Madhava Chetty,
Associate Professor, Department of Botany, S.V. University,
Tirupati, India and a specimen was kept in the Herbal
Herbarium, Sree Vidyanikethan College of Pharmacy,
Tirupati, India.
Instruments
Instruments used for analysis were pH meter (Systronic,
India), Brookfield viscometer [DV-I, LV-I spindle,
Brookfield Engineering Laboratories, USA], Muffle furnace
[77 S8HT8,Tempo, India], Micro centrifuge [RM-12CDX,
Remi, India], Deep freezer [RQF 650, Remi, India] and UV
visible spectrophotometer [UV 1700, Shimadzu, Japan].
Animals
Wistar albino rats (150200 g) of five groups, including
control and standard group, each with three animals were
selected. The selected animals were housed in acrylic cages
at standard environmental conditions at 252C, relative
humidity of 4555 %, in a well ventilated room maintained
at 12:12 h light: dark cycle, fed with standard rodent diet
and water ad libitum. All the animals were acclimatized for
a week before experiment. The animal experiments were
carried out according to the guidelines of the Committee for
the Purpose of Control of Experiments on Animals (Reg.
No. 930/a/06/CPCSEA) and approval of the Institutional
Animal Ethics Committee, Sree Vidyanikethan College of
Pharmacy, Tirupati, India was obtained.
Preparation of crude extract
The dried rhizomes of Curcuma longa (Zingiberaceae) and
Alpinia galanga (Zingiberaceae) were finely ground and
separately passed through sieve no. 80. Each powder
(500 g) was macerated for 3 days with 95 % ethanol and
filtered. The filtrates were dried using a vacuum desiccator.
Each extract (45 g) was weighed and dissolved in 150 ml of
ethanol (300 mg/ml) (Fig. 1).
Formulation of herbal sunscreen lotions
Accurate quantities of cetyl alcohol, zinc oxide, stearic acid,
glycerin, and hydroxy propyl methyl cellulose (HPMC)
were weighed (Table 1). Accurate quantity of water was
measured and taken in a 400 ml beaker. 1.0 g of
In vitro evaluation of herbal sunscreen lotion 243

Fig. 1 Absorbance values of

the formulated Sunscreens

triethanolamine was added to water and stirred. The water total of three formulations, F1, F2 and F3 were prepared
solution was heated upto a temperature of 80C to 85C. using various formulas (Table 1).
After the water solution has reached the required tempera-
ture, melted cetyl alcohol, zinc oxide, stearic acid, glycerin, Evaluation of formulated sunscreen lotions
hydroxy propyl methyl cellulose mixture and propyl para-
ben were slowly poured into the water solution a little at a i. Physicochemical studies
time, stirring constantly. Stirring was continued until a
The physicochemical parameters considered for the study
smooth and uniform paste was obtained. The prepared sun-
include color, pH, volatile and nonvolatile content, ash
screen lotion was set aside to cool. Then weighed quantity
value, layer thickness and rheological studies such as vis-
of Aloe gel, ethanolic extract of A.galanga (EEA), ethanolic
cosity (Gaspar and Maia Campos 2003), Spreadability and
extract of C.longa (EEC) and vitamin E (as per Table 1)
stability (Henry 1997; Sagarin 1957). The parameters were
were added and stirred well until all the ingredients mixed
evaluated according to the guidelines of Bureau of Indian
uniformly. Finally rose oil was added as flavoring agent. A
Standard (BIS), World Health Organization [WHO],
European Cosmetic, Toiletry and Perfumery Association
Table 1 Composition of various sunscreen formulations [COLIPA] and Scientific Committee of Cosmetics and
Non-Food Products [SCCNFP]. Viscosity profile of each
S.No Ingredients F 1 (%) F 2 (%) F 3 (%)
formulation was measured using a Brookfield viscometer
1 Aloe gel 5.0 5.0 5.0 at 10 to100 rpm, at a temperature of 25C with 8 ml samples
2 EEA 6.0 and using LV-spindle. Spreadability and layer thickness
3 EEC 6.0 were evaluated as per the procedures of Multimer
4 Olive oil 2.0 2.0 (Multimer 1956). Stability of each formulation was deter-
5 Rose oil 1.0 1.0 mined by centrifugation and freeze thaw method (Butler
6 Rose water 3.0 3.0 2000). The centrifugation was performed at 3500
7 Cetyl alcohol 2.0 2.0 2.0 13500 rpm at intervals of 500 rpm for 10 minutes, and
8 Zinc oxide 12.0 12.0 12.0
further observed for phase separation. In freeze thaw study
9 Stearic acid 4.0 4.0 4.0
all sunscreens were kept alternatively at 20C and 40C,
10 Glycerin 2.0 2.0 2.0
then observe for color change and phase separation. All
evaluations were performed in triplicate.
11 Vitamin E 1.0 1.0 1.0
12 Triethanolamine 1.0 1.0 1.0 ii. Safety evaluation by Mutagenicity assay
13 HPMC 10.0 10.0 10.0
The standard plate incorporation test for Mutagenicity
14 Propyl paraben 0.50 0.50 0.50
was carried out as per Maron and Ames (1983). The strain
15 Distilled water 50.50 50.50 67.50
used for the study was Salmonella typhimurium strain TA
EEA ethanolic extract of A.galanga; EEC ethanolic extract of C.longa; 100 without the S9 mix (Onodera et al. 1998). Positive
HPMC hydroxy propyl methyl cellulose control used for TA 100 was sodium azide (CAS Number:
244 A. Rasheed et al.

Table 2 Physicochemical evaluation parameters

Sunscreen Color pH VM (%) NVM (%) Ash value (g) Spreadability (%) LT (m)

F1 Cream 6.980.03 76.310.6 23.690.6 0.011 980.9 29.212.00

F2 Yellowish orange 7.210.02 80.020.3 19.980.3 0.082 980.7 28.991.55
F3 White 7.760.02 90.070.2 9.930.2 0.051 960.8 32.011.76

All values are represented as Mean SD (n03), p<0.001

VM volatile matter; NVM non-volatile matter; LT layer thickness

26628- 228): 5 g/plate. Sterile distilled water was used as
negative control. Fresh solutions of the reference mutagen
were prepared immediately before the beginning of each
experiment. The samples were dissolved in dimethyl sulf-
oxide (DMSO) and preincubated with the test strain in
phosphate buffer at 37C for 20 minutes. After addition of
the test samples, the plates were incubated at 37C for 48 h.
The assay was performed in triplicate for each sample and
the mutagenic responses of the sunscreen agents were
evaluated.
iii. Irritation test
The skin irritation test was performed on albino rats of
both sexes weighing about 150200 g. The animals were
maintained on standard animal feed and free access to water.
Hair was shaved from the back of rats and an area of 2 cm2
on both sides. One group served as control (5 % SLS in
distilled water), the second group for standard (Marketed
Sunscreen), three more groups served as test (F1, F2 and F3)
(Hiremath et al. 2008). Lotions at 5 ml were applied twice a
day for 3 days and the site was observed for any sensitivity,
edema and erythema. All evaluations were carried out in
triplicate.
iv. Efficacy analysis
Efficacy of herbal sunscreens was determined by In-vitro
method using UV Visible spectrophotometer. 0.10 % solu-
tion (w/v) each of the three formulated sunscreen lotions in
n-propyl alcohol was prepared by dissolving 0.050 g of the
sunscreen lotion in 50.0 ml of n-propyl alcohol. 0.10 %
solution of the two selected commercial sunscreen lotions
(SPF 20 and 55) in n-propyl alcohol was also prepared. The
aliquots of each formulation prepared were scanned between
290 and 320 nm, with 5 nm interval. SPF was calculated by
using the equation derived by Mansaur (Mansaur 1986;
Santo et al., 1999). EE () I () values determined by
Sayre (Sayre et al. 2003; Sayre 1993) was used in below
Equation (1). Each sample observed in triplicate.
X320
SPFspectrophotometric CF290 EE l  I l  Al
where, correction factor, CF010, EE () 0 erythemogenic
effect of radiation of wavelength , I() 0 intensity of solar
light of wavelength , A () 0 spectro photometric absor-
bance values at wavelength .
Statistical treatment
Statistical analysis was carried out by using STATS [70]
software and results were expressed as mean S.D. All
parameters were statistically analyzed at 95 % confidence
level. Statistical result of psychometric evaluation was fur-
ther tested by ANOVA [One way analysis].
Results and discussion
Three formulations F1, F2 and F3 were prepared as per
Table 1. F1 contains EEA, F2 contains EEC and while F3
contains only Aloe gel. The results of physicochemical
properties such as color, pH, volatile and nonvolatile con-
tent, ash value and layer thickness are summarized in
Table 2. pH of formulations range from 6.98 to 7.76 that
complies with that of skin pH. The important chemical

Table 3 Viscosity profile of formulated sunscreens Table 4 Safety and stability evaluation parameters

Sunscreens Viscosity (cps) Sunscreen Stability Erythema

10 rpm 20 rpm 30 rpm 50 rpm 60 rpm 100 rpm Centrifugation 13500 (rpm) Freeze Thaw

F1 181.0 89.2 58.4 28.7 25.4 16.1 F1 unstable stable no irritation

F2 178.0 90.6 64.0 36.0 32.0 18.0 F2 stable stable no irritation
F3 180.5 91.6 60.1 33.8 30.2 19.2 F3 unstable unstable no irritation

CPS Centipoise; RPM rotation per minute RPM rotation per minute
In vitro evaluation of herbal sunscreen lotion 245

Table 5 Absorbance values of

the formulated and marketed Wavelength (nm) F1 F2 F3 Marketed sunscreen Marketed sunscreen
sunscreen (SPF 55) (SPF 20)

290 2.399 2.497 1.986 6.521 2.842

295 2.137 2.229 1.723 6.112 2.574
300 1.704 2.106 1.568 5.873 2.131
305 1.528 1.894 1.393 5.180 1.960
310 1.203 1.502 0.916 4.902 1.380
315 0.881 0.974 0.515 4.767 1.151
320 0.537 0.719 0.486 4.154 0.894

parameters such as non volatile matter (9.93 to 23.69),
volatile matter (76 to 90) and ash value (0.01 to 0.08) were
found to be in controlled range for all formulations, justify-
ing their compatibility with all type of skins. Spreadability
and layer thickness were found to be in the range of 96 to 98
and 28 to 32 respectively confirming good cosmetological
property (Table 2).
The viscosity of the formulated lotions at various speed
(rpm) is given in Table 3. It showed 178 to 181 cp at 10 rpm
indicating good rheology during handling. As the speed of
rotation increases viscosity of the formulations F1, F2 and
F3 decreased, indicating pseudo plastic behavior of the
formulations, a desirable property of sunscreen lotion for
its stability. The rheological properties such as redispersi-
bility and viscosity showed good stability at room temper-
ature. Also, higher viscosity of the lotion reveals more
protection from microbial growth.
The results of stability tests carried out by centrifugation
and freeze thaw method of the formulations are given in
Table 4. For F1 and F3, phase separation at 13500 rpm was
observed indicating the unstability of these formulations at
high stress conditions. During freeze thaw study, water was
separated from F3 formulation revealing that it may not bear
different environmental changes during product transport. The
study showed that F3 requires more attention and protection
from temperature changes and environmental stresses.
The skin irritation test performed in Wistar albino rats
showed no signs of sensitivity, erythema and edema
(Table 4). The mutagenicity studies showed that formula-
tions F1, F2 and F3 are non-mutagenic. So the prepared
formulations were considered to be safe. The psychometric
evaluation suggested that formulation showed good fed
regarding handling, firmness and glowness of skin. The skin
glow effect of formulations F1, F2 and F3 were found to be
long lasting due to removal of dead cells. The formulated
herbal lotion is found to be capable to block the UV radia-
tion and protect the skin from sunburn.
Two marketed formulations having known SPF of 55 and
20 were considered for comparative evaluation with that of
the formulated sunscreen lotions. The absorbance values at
various wavelengths (max) from 290 to 320 nm of both
formulated and marketed sunscreens are given in Table 5.
The SPF values of formulations F1, F2, F3 and the two
marketed sunscreens were calculated and presented in
Table 6. The results showed that F2 have high SPF of
18.2011.6 which may be attributed to the presence of
Curcumin. The formulations F1 and F3 showed medium
SPF (as sunscreens with SPF ranging 1520 are considered
to be medium protection sunscreens) which is sufficient for
protection against sun burn for a period of about 3 h. The
presence of photo shielding flavanoids such as curcumin,
which quench the production of free radicals in the skin,
makes it possible for the formulations to get protection from
UV solar radiation. Determination of SPF of formulated
herbal sunscreens by In-vitro method and comparison with
marketed sunscreens indicates that the method is highly
suitable and reliable.
Conclusion
The study attempted to develop herbal sunscreen lotion
using extracts of Alpinia galanga, Curcuma longa and
Aloe vera and examined their efficacy for preventing sun
burn. The formulations F1, F2 and F3 were prepared by
varying the composition and evaluated for their physico-
chemical properties and SPF. The study showed that formu-
lation F2 having curcumin was found to be more stable with
high SPF value, proving a better sunscreen lotion. As the
formulations with herbal extracts were found to possess SPF
Table 6 SPF of the formulated and marketed sunscreen
S.No. Sunscreens SPF
1 F1 15.0881.2
2 F2 18.2011.6
3 F3 13.0051.0
4 Marketed formulation with SPF 55 53.6852.0
5 Marketed formulation with SPF 20 18.6311.7
All values are represented as Mean SD (n03), p<0.001
246
in the range of 13 to 18, they can be used for normal skin to
prevent sun burns. The sunscreening property of these extracts
may be due to the presence of flavanoids, phenols and terpe-
noids present in herbal drugs Alpinia galanga, Curcuma longa
and Aloe vera. Further the formulations were proved to be
non-mutagenic which differentiates them from the synthetic
sunscreens. It can be concluded that the present research might
hopefully bring advancement in the treatment of sun burns
caused by exposure to UV rays. Further, the study reveals that
UV Spectroscopy is the rapid, acceptable and reproducible
method for the evaluation of efficacy of herbal sunscreens.
The present study can therefore assist the regulatory authori-
ties, scientific organizations and manufacturers in developing
uniform standards for herbal sunscreens.
Acknowledgements The authors express their sincere thanks to
Padmashri Dr. M. MohanBabu, Chairman, Sree Vidyanikethan Educa-
tional Trust, Tirupati, India for providing the necessary facilities to
carry out this work.
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