Determination and comparison of in vitro SPF of topical formulation ...

International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
_________________________________________Research Paper
Determination and comparison of in vitro SPF of topical formulation
containing Lutein ester from Tagetes erecta L. flowers, Moringa oleifera
Lam seed oil and Moringa oleifera Lam seed oil containing Lutein ester
Shantanu Kale*, Megha Gaikwad and Snehal Bhandare
Mahatma Gandhi Vidyamandir’s Pharmacy College, University of Pune, Nashik,
Maharashtra, India.
_________________________________________________________________________________
ABSTRACT
The present study was designed to study the sunscreen activity of herbal formulation containing Lutein ester
extracted from Tagetes erecta L. flowers (Family Asteraceae), Moringa oleifera Lam seed oil (Family-
Moringaceae), Moringa oleifera Lam seed oil containing Lutein ester. The parameters of creams complies with
official acceptance criteria and SPF calculated by COLIPA method. Lutein ester 1.09 with Boots Star Rating 4,
Moringa oleifera Lam seed oil cream is found to be 1.05 with Boots Star Rating 3 and Moringa oleifera Lam
seed oil containing Lutein ester 1.063 with Boots Star Rating 3 and indicating that the cream formulated can be
considered as an efficient validated topical product.
Key Words: Lutein ester, Moringa oleifera Lam, SPF, Moringa oleifera containing Lutein ester, Boot star rating.
INTRODUCTION
Sunscreen is a topical (to apply on the skin)
product that protects the skin against harmful
effects of the sun. Sunscreen has two purposes.
Firstly a good sunscreen protects the skin against
UVA and UVB rays. Secondly, it helps the skin
maintain moisture and its own natural oils which
can be lost through exposure to the sun's radiation 1 .
Sunscreens incorporate a wide variety of chemicals
like Organic Compounds and their derivatives,
Organic Esters, Salt and Inorganic compounds,
(Mineral) which has particular absorbance.
FDA Approved Sunscreen Agents 2 are as follows-
Organic Compounds and their derivatives:
PABA and 4-aminobenzoic acid
derivative (Padimate O),
Phenylbenzimidazole sulfonic acid,
Diethylhexyl butamido triazone etc.
Organic Esters: Cinoxate, Homosalate,
Octyl methoxycinnamate, Octocrylene,
Octyl salicylate etc.
Salt: Salt formed between triethanolamine
and salicylic acid, Trolamine salicylate
In organic compounds (Mineral):
Titanium dioxide, zinc oxide.
________________________________________
*Address for correspondence:
E-mail: shantanu_kale@yahoo.com
In order to get a broad spectrum UV protection,
more than one active sunscreen ingredients are
added in the sunscreens. Inorganic sunscreens are
cosmetically unacceptable because of their opaque
quality and being occlusive. Organic sunscreen
have been increasingly reported for allergic and
contact dermatitis, phototoxic and photo-allergic
reactions, contact urticaria producing skin rash
notable for pale red, raised, itchy bumps and even
solitary cases of severe anaphylactic reactions.
The standardised Pet. Ether 60 0 --80 0 extracts of T.
erecta L. flowers contains Lutein ester. It is
Xanthophyll [3] (carotenoid) containing conjugated
system of alternative double bond with λmax
445nm 3, 4 in Pet. Ether and hence can be good UV
blocking agent. Seeds of Moringa oleifera L.
species (family Moringaceae) yield 30-40% edible
oil (called ben oil containing high concentration of
behenic acid) that can be used medicated
preparations, cosmetics and lubrication 5 . The fatty
acid components of the oil are Palmitic 9.3%,
Stearic 7.4%, Behenic 8.6% and unsaturated oleic
65.7% 6 . Oil have full of radiant boosting fatty acid
& vitamin A & Vitamin C. As Lutein, Lutein ester
are oil soluble , Moringa oil with good lubricating
power and unsaturated compounds can easily
penetrate into skin showing better effect.
Various biologically active botanical agents have
been used in Sun protection. Therefore, we choose
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International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
these Plants. We have already reported data on
“Formulation and in vitro evaluation for sun
protection factor of Lutein ester extracted from
Tagetes erecta Linn. Flowers (Family-Asteraceae)
sunscreen creams” and “Formulation and in- vitro
evaluation of Moringa oleifera Lam Seed oils
sunscreen cream”.
The study was designed with an objective to
determine the in vitro SPF and boot star rating. In
present study we have made an attempt to
formulate the topical cream from Lutein ester M.
oleifera Lam seed oil, M. oleifera Lam seed oil
containing Lutein ester (1%w/w) and evaluation of
the same.
MATERIAL AND METHODS
2.1 Plant Material
Flowers of Tagetes erecta L. (Asteraceae) was
collected from farms of village kasabe- Sukene,
Taluka Niphad, District-Nashik, Maharashtra state,
India. The herbarium was deposited with
(Ref.BSI/WRC/Tech./2011). Voucher no.
SNEBTA4. Plant material of Moringa oleifera
Lam were collected from in-house garden in
Nashik; district Nashik, Maharashtra, India. The
specimens were identified and authenticated by
Botanical Survey of India, Pune, Maharashtra (Ref.
BSI/ WRC/ TECH/ 2011). The herbarium of the
plant was deposited in the BSI against voucher no.
MEGA03.
2.2 Extraction of dried flowers of Tagetes erecta
L. 7 and seeds of Moringa oleifera Lam
Air-dried petals of Tagetes erecta L. and seeds of
Moringa was separately powdered and extracted
with 50 volumes of petroleum ether (60 o -80°C)
using a Soxhlet apparatus. This process of
extraction was repeated for 6hr, the petroleum ether
distilled out by distillation assembly, then
concentrated by hot plate drying and air-drying at
temperature of 40±2 °C.
2.3 Sunscreen cream formulation
Step I: Water phase was prepared by collecting
deionised water (72 %) and then 5 % water was
removed aside from this for final volume makeup.
Water soluble components Disodium EDTA
(0.02%), Sodium Methyl Paraben (0.3%) and
Triethanolamine (0.5%) were dissolved in
deionised water, meanwhile, carbopol (0.5%) was
allowed to swell using an homogenizer and heated
up to 80 0 C.
Step II: Oil phase was prepared by heating Sodium
Propyl Paraben (0.06%), Stearic acid (2%), Cetyl
alcohol (1%), Cetomacrogal-1000 (2%),
Cetostearyl alcohol (5%).Lutein ester, M. oleifera
Lam seed oil, M. oleifera Lam seed oil containing
lutein ester (1%w/w) at 80 O C was incorporated
separately for each cream.
Step III: Oil phase was added in water phase at 80
O C with continuous stirring for 20-25 min and then
it was homogenized till uniform emulsion is
formed. The finished product has dark yellow
colour and semisolid consistency. It was then
poured into the wide mouth container and stored at
temperature not exceeding 37 0 C.
2.4 Determination of physical parameters of
cream
Preparation of herbal cream has always been a
challenging task and the cream is accepted only if it
is tested appropriately for various physical
parameters like ease of spreadability, appearance,
pH, viscosity and pleasant feeling as specified in
Table No.2
Determination of in vitro SPF
This study was performed by Transmittance
measurement of the three creams. The Optometrics
Model SPF-290 Analyzer measures the sun
protection factor of the cream over a wavelength
range from 290nm-400nm. Approximately 110mg
of sample was applied and spread on 56cm 2 area of
Transpore tape to obtain a sample film thickness of
2μl/cm 2 (to get an even film) as suggested in the
operational manual of Optometrics LLC for the
sample application technique. The samples thus
prepared were exposed to Xenon arc lamp for
determining the SPF and Boots Star Rating.
WIN SPF has used the following equation for
calculating SPF value.
Where,
MPFλ = scan MPF value
Eλ = spectral irradiance of terrestrial sunlight under
controlled conditions
Bλ = erythemal effectiveness
RESULT
The topical formulation of Lutein ester extracted
from Tagetes erecta L. flowers, Moringa oleifera
Lam seed oil and Moringa oleifera Lam seed oil
containing Lutein ester were formulated.
Formulated creams were evaluated for all
parameters of cream and in vitro SPF
determination. The results of creams and SPF are
mentioned in Table nos. 2 and 3. Lutein ester with
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International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
Boots Star Rating 4, Moringa oleifera Lam seed oil
cream with Boots Star Rating 3, Moringa oleifera
Lam seed oil containing Lutein ester with Boots
Star Rating 3 indicating that no additive effect on
SPF and Boot star of sunscreen cream was found
due to addition of two active herbal ingredients i.e.
Moringa oleifera Lam seed oil and Lutein ester as
synthetic sunscreen agent shows additive effects in
sunscreen.
DISCUSSION
The Optometrics Model SPF-290 Analyzer is a
computer controlled instrument that is designed to
measure the sun protection factor of sunscreen
preparations. For US-FDA standards the protection
factor is calculated over the wavelength range from
290-400nm. For the calculation of standard
deviation, Diffey's method is used, based on B. L.
Diffey's paper [8] on using Transpore Tape® as the
substrate for SPF measurements.
SPF value for sunscreen above 2 is considered as
having good sunscreen activity. In present study
formulated sunscreen cream was found good
sunscreen activity and hence Lutein ester and
Moringa oleifera L. may be considered as good
candidate for sunscreen or cosmoceutical purposes.
Further these creams can be evaluated for
spreadability, viscosity, microbial testing, water
content, etc.
CONCLUSION
The herbs summarized in above mentioned work
were found to effectively reduce inflammatory UV
radiation through mechanisms related to
conjugation systems. However, an appropriate
delivery system should be developed to impart their
efficacies in addition to the standardization of these
herbs. Furthermore, an optimized and effective
dose should be evaluated prior to the development
of preparations in order to avoid irritation or allergy
in subjects with hypersensitive skin. In addition,
attempt was made by combination treatment of two
active ingredients as it was found to be more
effective than the application of a single product
with regard to synergistic effects on the
pathogenesis of UV radiation but combination of
two herbal sunscreen agents shown no additive
effect.
ACKNOWLEDGEMENTS
We express our sincere gratitude to Mr. Milind
Kataria, Ms. Bharati ,Ms. Priyanka of Rave
Pharmaceuticals, Sinnar, MIDC, Nashik for
guiding us to prepare our formulation, Dr.
Chandrashekhar madam and Ms. Snehal of Kelkar
Education Trust, Mulund (E), Mumbai for their
kind help.
Table 1: Formulation of sunscreen cream
S. No. Ingredients Use Components (%w/w)
1 Cetosteryl alcohol Emulsifier 5
2 Stearic acid Emollient, Coemulsifier 2
3 Cetomacrogal-1000 Emulsifier 2
4 Cetyl alcohol Emollient, Coemulsifier 1
5 Carbopol 940 Gelling agent 0.5
6 Disodium EDTA Chelating Agents 0.02
7 Na Methyl Paraben Preservative 0.3
8 Na Propyl Paraben Preservative 0.06
9 Triethanolamine Surface active agent 0.5
10 Purified Water Vehicle 62
11 Lutein ester Active ingredient 1
12 M. oleifera seed oil Active ingredient 1
13 M. oleifera containing Lutein ester (1%)w/w Active ingredients 1
Table 2: Physical Parameters of 3 sunscreen creams
S. No. Parameters Sunscreen of Lutein ester Sunscreen of M.oleifera
Lam seed oil
Sunscreen of M.oleifera
Lam Seed oil containing
Lutein ester
1. Color Faint orange Faint yellow Dark yellow
2. Odor characteristic Aromatic Aromatic
3. Spreadability Good and uniform Good and uniform Good and uniform
4. pH 6.8 6.7 6.86
5. Viscosity 28000cP 49500cP 37990cP
6. Total microbial count Nil Nil Nil
7. Patch test for irritancy No irritation reaction
persists
No irritation reaction
persists
Table 3: Results of SPF and other Parameters of 3 sunscreen creams
No irritation reaction
persists
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International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
S. No.
Parameter
Sunscreen of
Lutein ester
Sunscreen of M.
oleifera Lam seed oil
Sunscreen of M.
oleifera Lam oil
containing Lutein
ester
1. SPF 1.09 1.05 1.063
2. Standard Deviation 0.02 0.03 0.0231
3. UVA/UVB Ratio 0.843 0.697 0.648
4. Critical Wavelength 389.9 381.3 382.9
5. Boots Star Rating 4 3 3
Table 4: The Boots Guideline for Star Rating
Mean UVA/UVB Ratio Star Rating Category Star Rating Designation
0 to 0.2 - No Claim
0.21 to 0.4 * Minimum
0.41 to 0.6 ** Moderate
0.61 to 0.8 *** Good
0.81 to 0.9 **** Superior
0.91 and above ***** Ultra
Fig. 1: SPF-290 Graph Report of Lutein ester Sunscreen Cream
Fig. 2: SPF-290 Graph Report of Moringa oleifera
Lam seed oil Sunscreen Cream
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International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
Fig. 3: SPF-290 Graph Report of Moringa oleifera Lam seed oil
containing Lutein ester Sunscreen Cream
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