Development of novel herbal cosmetic cream with Curcuma longa extract loaded transfersomes for antiwrinkle effect
ABSTRACT Curcuminoids obtained from Curcuma longa have properties like photoprotection, antiaging, anti-wrinkle, moisturizing, antioxidant, astringent, anti-irritant, antimicrobial and anti-inflammatory activities. Our aim was to develop a stable nano-transfersomes loaded cream which could correct the morphological defects and penetrate deeper to the cellular level of dermis to produce anti-wrinkle effect. Soxhlet extraction of C. longa was carried out with two solvents viz. absolute ethanol and 85% ethanol. The transfersomes were prepared by conventional rotary evaporation technique. Various process variables were studied to optimize the formulation including lecithin: surfactant ratio, surfactants (Tween 20, Tween 80) and different solvent extract (95% ethanol and 85% ethanol). 85% ethanolic extract loaded transfersomes (F4) prepared with 4:1 lecithin: surfactant ratio and Tween 20 as the edge activator was found to have maximum entrapment efficiency of 41±1%. Vesicle size obtained was between 200±2 nm range, zeta potential -30±5 mV and polydispersity index between 0.2 to 0.3. After 12 h study curcumin deposition was found to be 45.9%. Transmission electron microscopy of novel cream revealed the presence of spherical double layered transfersome in intact state thus proving the stability of transfersome in cream. Cutometer studies, showed 30 to 50% improvement in relative and absolute skin parameters. The improvements in overall elasticity, biological elasticity, recovery of deformed skin, firmness and reduction in fatigueless can be correlated with anti-wrinkle properties of cream.
- SourceAvailable from: Mankaran Singh[Show abstract] [Hide abstract]
ABSTRACT: Lipid-based innovations have achieved new heights during the last few years as an essential component of drug development. The current challenge of drug delivery is liberation of drug agents at the right time in a safe and reproducible manner to a specific target site. A number of novel drug delivery systems has emerged encompassing various routes of administration, to achieve controlled and targeted drug delivery. Microparticulate lipoidal vesicular system represents a unique technology platform suitable for the oral and systemic administration of a wide variety of molecules with important therapeutic biological activities, including drugs, genes, and vaccine antigens. The success of liposomes as drug carriers has been reflected in a number of liposome-based formulations, which are commercially available or are currently undergoing clinical trials. Also, novel lipid carrier-mediated vesicular systems are originated. This paper has focused on the lipid-based supramolecular vesicular carriers that are used in various drug delivery and drug targeting systems.ISRN pharmaceutics. 01/2012; 2012:474830.
African Journal of Pharmacy and Pharmacology Vol. 5(8), pp. 1054-1062, August 2011
Available online http://www.academicjournals.org/ajpp
ISSN 1996-0816 ©2011 Academic Journals
Full Length Research Paper
Development of novel herbal cosmetic cream with
Curcuma longa extract loaded transfersomes for
Swarnlata Saraf*, Gunjan Jeswani, Chanchal Deep Kaur and Shailendra Saraf
University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur 492010 (C.G.), India.
Accepted 22 June, 2011
Curcuminoids obtained from Curcuma longa have properties like photoprotection, antiaging, anti-
wrinkle, moisturizing, antioxidant, astringent, anti-irritant, antimicrobial and anti-inflammatory activities.
Our aim was to develop a stable nano-transfersomes loaded cream which could correct the
morphological defects and penetrate deeper to the cellular level of dermis to produce anti-wrinkle
effect. Soxhlet extraction of C. longa was carried out with two solvents viz. absolute ethanol and 85%
ethanol. The transfersomes were prepared by conventional rotary evaporation technique. Various
process variables were studied to optimize the formulation including lecithin: surfactant ratio,
surfactants (Tween 20, Tween 80) and different solvent extract (95% ethanol and 85% ethanol). 85%
ethanolic extract loaded transfersomes (F4) prepared with 4:1 lecithin: surfactant ratio and Tween 20 as
the edge activator was found to have maximum entrapment efficiency of 41±1%. Vesicle size obtained
was between 200±2 nm range, zeta potential -30±5 mV and polydispersity index between 0.2 to 0.3. After
12 h study curcumin deposition was found to be 45.9%. Transmission electron microscopy of novel
cream revealed the presence of spherical double layered transfersome in intact state thus proving the
stability of transfersome in cream. Cutometer studies, showed 30 to 50% improvement in relative and
absolute skin parameters. The improvements in overall elasticity, biological elasticity, recovery of
deformed skin, firmness and reduction in fatigueless can be correlated with anti-wrinkle properties of
Key words: Curcuminoids, cutometer, novel creams, anti wrinkle, transfersomes.
Botanical extracts have always gathered much attention
because of their multi faceted actions (Ahshawat, 2008;
Deep and Saraf, 2008). Properties like photoprotection,
inflammatory activities are all present in a single herb
called Curcuma longa (Bonte et al., 1997; Jayaprakasha
et al., 2006; Deep and Saraf, 2009). To explore the
cosmetic potential utility of C. longa, numerous attempts
have been made including creams, gels, moisturizers etc
(Ahshawat et al., 2008; Nair et al., 2009). To enhance the
effects produced by conventional cosmetic formulations
*Corresponding author. E-mail: firstname.lastname@example.org.
Tel: 0771-2262832. Fax: 0771-2262832.
for anti-wrinkle effect, penetration and deposition at
required depth into the skin have to be considered.
Bangham discovered liposomes in 1963 and since then
vesicular systems have attracted increasing attention
(Bangham, 1963). But recently it has become evident that
classic liposomes are of minor values in terms of
penetration. Confocal microscopic studies have shown
that intact liposomes are not able to penetrate into
granular layer of epidermis but, they rather remain on the
upper layer of stratum corneum. The modification of the
vesicular compositions or surface properties can adjust
the drug release rate and the deposition to the target site.
By using the concept of rational vesicle design,
transfersomes were prepared. As the lipid covers the
curcumin extract it presents the ability to penetrate the
deeper layers of skin (Touitou et al., 2000; Touitou, 1998;
Cevc and Blume, 1992). Transfersomes, due to the
presence of surface active agents which are single chain
surfactants are optimized to overcome the skin transport
barrier spontaneously. Natural transepidermal water
activity gradient enables them to deliver actives to the
deeper epidermal layers through dehydration of the lipid
vesicles within the stratum corneum. Therefore,
transfersome uptake is driven by the hydration gradient
that exists across the epidermis, stratum corneum, and
ambient atmosphere (Cevc and Blume, 1992; Cevc
Recent studies demonstrate that curcumin is excellent
for wrinkles and can curb inflammation and the formation
of free radicals. In particular, curcumin produce anti-
inflammatory and anti- proliferative activity (Saraf and
Kaur, 2010). But pure curcumin was shown to be less
protective than a mixture of curcuminoids, indicating a
possible synergism among curcuminoids (Jayaprakasha
et al., 2006). The polyphenols act as potent antioxidants
and can scavenge reactive oxygen species (ROS), such
as lipid free radicals, superoxide radicals, hydroxyl
radicals, hydrogen peroxide and singlet oxygen (Kaur
and Saraf, 2011).
The reports of Elsayed et al. (2006) seem to indicate
different skin permeation and deposition profiles for
lipophilic and hydrophilic compounds (Elsayed et al.,
2006). The purpose of this study was to investigate the
efficiency of transfersome to deliver the herbal extract to
the levels of epidermis in order to exert its antioxidant
effect. Transfersomes were successfully prepared by thin
film formation method using two different solvent extracts
viz. 95% ethanol and 85% ethanol, having curcumin
concentration of 0.12%.
MATERIALS AND METHODS
Rhizomes of C. longa were procured from a local authentic herbal
distributor of Raipur, Chhattisgarh and were identified (from the
Herbarium catalogue number UIOP/HB/P17) at Department of
Pharmacognosy of Pt. Ravishankar Shukla University, Raipur,
India) and tested phytochemically by spectrophotometric method.
Curcumin was purchased from Himedia Laboratories, Mumbai
(India). Tween 80 and Tween 20 were purchased from Loba Chem
Pvt. Ltd, Mumbai. Soyalecithin and Triton x-100 were purchased
from Himedia Laboratories Pvt. Ltd, Mumbai. All other ingredients
used were of analytical grade. Extracts were chromatographed in
comparison with curcumin external reference standard using Silica
gel G TLC plates (Indian herbal pharmacopoeia, 2002).
Preparation of extract
Extraction of rhizomes of C. longa was done by continuous hot
extraction method (WHO, 2004; Rajpal, 2004). Dried rhizomes of C.
longa were ground to coarse powder separately using a laboratory
mill and packed in the Soxhlet apparatus. Defatting was done with
petroleum ether at 60 to 80°C. Extraction was done with ethyl
alcohol (95%) and then with 85% at temperature of 50 to 60°C until
complete exhaustion of the drug. The extracts were concentrated
and curcumin was determined by UV spectrophotometer
(Shimadzu, Pharmaspec-1700) at maximum wavelength of 425.6
nm (WHO, 2004; Rajpal, 2004; Pothitirat, 2004).
Saraf et al. 1055
Preparation of transfersomes
evaporation technique. The extract, lecithin (PC) and edge activator
(Tween 80 and Tween 20) were dissolved in methanol:chloroform
(1:2) mixture. Organic solvent was removed by evaporation in rotary
vacuum evaporator above lipid transition temperature at 115 rpm. A
thin film was prepared, hydrated and sonicated (Elsayed et al.,
2006). C. longa extract transfersomes were obtained with curcumin
concentration of 0.12% (Martelli, 2004).
Optimization of formulation
The preparation of transfersomes containing C. longa extract
include various process variables such as effect of lecithin:
surfactant ratio (7:1, 6:1 5:1 and 4:1), effect of various surfactants
(Tween 20, Tween 80) and different solvent extract (95% ethanol
and 85% ethanol). Optimization of formulation was done on the
basis of stability at 4±2, 25±2 and 37±2° C. Composition of stable
transfersomes is as shown in Table 1. During the preparation of a
particular system, the other variables were kept constant.
Preparation of novel cream
Cream base was prepared by using a phase inversion technique
(Forster and Tesmann, 1991). Initially, oil and other ingredients
(olive oil, cetyl alcohol, stearic acid, sorbitan monooleate, propylene
glycol and glycerin) were mixed using an overhead stirrer at 200±25
r.p.m. at 65 to75°C on a hot plate (Ahshawat, 2008). After complete
melting and homogenous mixing, temperature was decreased to
30° C and transfersome dispersion (F4) was added at increased
speed (275±25 r.p.m.). For fragrance rose oil was added. Phase
inversion took place and the solution became viscous. Base cream
was prepared similarly instead transfersome dispersion, distilled
water was added as aqueous phase. Contents composition is given
in Table 2.
Characterization of transfersomes
Morphological characterization of optimized vesicle was done using
a digital Labomed microscope (Leica ATC 2000) with camera at 40x
resolution. TEM studies were performed at All Indian Institute of
Medical Sciences, New Delhi, India, (Morgagni 268D Fei Electron
Optics). A drop of the sample was placed on a carbon-coated
copper grid to form a thin film and negatively stained by adding a
drop of 1% w/v phosphotungstic acid. The grid was allowed to air
dry and the samples were viewed and photographed (Kaur et al.,
Entrapment efficiency percentage
Entrapped C. longa extract was estimated by centrifugation
method. Prepared vesicles were placed in centrifugation tube and
centrifuged at 15000 rpm for 30 min. The supernatant (1 ml) was
withdrawn and diluted with phosphate buffer (pH 6.8). The
unentrapped curcumin was determined by UV spectrophotometer
(Shimadzu, Pharmaspec-1700) at 425.6 nm. Samples were diluted
twice before taking absorbance measurement. Free curcumin in the
supernatant gives us the total amount of unentrapped drug.
Encapsulation efficiency is expressed as the percent of drug
trapped. Amount of entrapped curcumin was estimated after fusing
the vesicles with Triton - x 100 by UV spectrophotometer with
were prepared using conventional rotary
1056 Afr. J. Pharm. Pharmacol.
Table 1. Entrapment efficiency and skin deposition data of selected formulations after optimization.
of curcumin (%)
Quantity of skin
For entrapment efficiency all values represent Mean ± SD (n=3), for permeation and deposition data are expressed as % of total curcumin in
formulation after 24 h.
suitable dilution by phosphate buffer (pH 6.8) (Saraf and Kaur,
2010; Kaur et al., 2008).
Vesicle size, size distribution and zeta potential analysis
Vesicle size, size distribution and zeta potential were determined by
Malvern Zetasizer DTS version 5.03 (Malvern, UK). Zeta potential
was analyzed to measure the permeation of transfersome by
studying its colloidal property and stability of the vesicle (Saraf and
Kaur, 2010; Kaur et al., 2008).
Stability of the vesicles was determined by storing the vesicles at
4±0.5° C for six months and then measuring their vesicular size and
In vitro permeation and skin deposition studies
Fresh abdominal skin of goat was collected from slaughter house
and was used after peeling the skin from underlying cartilage (Kaza
and Pitchaimani, 2006; Patel et al., 2009). Skin hairs were shaved
and the preliminary wash of skin was done with normal saline. Skin
was dried between two filter papers and was used directly in study
Experiments were run in modified Franz diffusion cell having
receptor compartment volume of 50 ml. The in vitro study design
used in the present study was similar to that described by Elsayed
et al. (2006). Experiments were performed in two stages. The first
stage was used in determination of the vesicle permeating the skin.
This stage used pH 6.8 phosphate buffers as receptor medium.
Skin membrane was mounted, with stratum corneum side up and
donor compartment dry and open to atmosphere. The skin was
floated on receiver solution for 24 h for equilibrium and pre
hydration. This approach was suggested to maintain transepidermal
hydration gradient which has been proposed to generate the driving
force for skin permeation of transfersomes. Five milliliter of test
formulation was placed on the skin. At appropriate time intervals 1
ml aliquots from the receptor medium was withdrawn and
immediately replaced by an equal volume of fresh pH 6.8
phosphate buffers to maintain sink conditions. The samples were
analyzed by spectrophotometer at 425.6 nm.
At the end of first stage the donor compartment and skin surface
were washed five times with warm receptor medium. The second
stage was employed to determine skin deposition. The receptor
content was completely removed and replaced by 50% (v/v) ethanol
in distilled water and kept for further 12 h followed by spectroscopic
assay. This receiver solution (50% v/v) ethanol in distilled water
was suggested to diffuse through skin, disrupting any vesicle
structure and extracting deposited curcumin from skin, thus giving a
measure of skin deposition (Elsayed et al., 2006).
Characterization of novel cream
Optical microscopy was done with digital Labomed microscope
(Leica ATC 2000) with camera in 40x resolution and transmission
electron microscopy using transmission electron microscope
Morgagni 268D (Fei Electron Optics).
Physicochemical evaluation of cream
(a) Color and odor of the cream noted carefully (Ahshawat et al.,
2008; Saraf et al., 2010).
(b) Net content: Empty container with lid was weighed at the
beginning of analysis. Container was again weighed after adding
cream. Weight of the product was calculated by difference.
(c) pH of cream: 1 g of cream mixed with 9 ml of water and pH of
the resulting mixture taken with a pH meter (Elico LI 610).
(d) Non-volatile matter at 105° C: 1 g of cream was taken in glass
bottle and kept in an oven at 105°C for 2 h.
(e) Ash value: 5 g of cream was weighed in a flat-bottom silica
crucible and heated on a steam bath under a jet of air for 1 h. Then
1 g of ash less cellulose powder was added to it and mixed with a
glass stirring rod. The dish was heated under furnace at 600°C.
(f) Acid value: 5 g of the cream was accurately weighed and
dissolved in 50 ml of a mixture of equal volumes of ethanol (95%)
and ether, previously neutralized with 0.1 M potassium hydroxide
using phenolphthalein solution as indicator. 1 ml of phenolphthalein
solution was then added and titrated with 0.1 M potassium
hydroxide until the solution remains faintly pink after shaking for 30
s. The acid value was calculated from the expression. Acid value =
5.61 n/w, where n = the volume (ml) of 0.1 M potassium hydroxide
required; w = the weight in gram of the substance.
(g) Saponification value: 2 g of cream was accurately weighed and
introduced into a 200 ml borosilicate glass flask fitted with a reflux
condenser. 25 ml of 0.5 M ethanolic potassium hydroxide was
added with a little pumice powder and boiled under reflux on a
water-bath for 30 min. 1 ml of phenolphthalein solution was then
added and titrated immediately with 0.5 M hydrochloric acid (a ml).
The operation was repeated by omitting the substance being
examined (b ml). Saponification value was determined using the
Saponification value =28.05 (b-a)/w, where w is weight of the
substance in gram.
Physical stability of cream
The ability of cream to maintain its consistency was determined by
keeping it at 25±2°C (room temperature, RT) and 4±2° C for 30 days
(Ahshawat et al., 2008).
Saraf et al. 1057
Table 2. Composition of curcumin incorporated transfersome loaded cream.
Transfersome suspension (curcumin content)
Base cream weights (w/w)
Novel cream weights (w/w)
Patch irritation test
0.5 g of cream was applied on the back of volar arm with the help of
surgical gauze of volunteers and the erythemal score was
determined using the scale defined in the Indian Standards.
Average erythemal score was obtained using the expression:
Average erythemal score = Total score of each product/Total
number of volunteers.
The products were compared based on sensory evaluation and
ranking was done as per score obtained according to the hedonic
scale (Horwitz et al., 1999).
Study was done with 6 volunteers (females) having age of 20 to 25
years. Transfersomal cream was applied twice a day (once in
morning and once in evening) at the same time at volar forearm of
the volunteers up to 2 weeks and observations were made by
ranking method. Various questions were asked to volunteers and
according to their answers ranking was done between 0-9 of
hedonic scale, ranking was done as per follows: 8-9 (extremely
liking), 5-7 (medium), 1-3 (dislike), 6 (in between extreme liking and
medium), 4 (in between medium and dislike), verbally for softness,
irritation, stickiness smoothness and after effect of skin. Average
score of each product was determined as follows:
Where E1 = Average score of each product, V1= Total score of
product given by volunteer no.1, V2 = Total score of product given
by volunteer no.2, Vn = Total score of product given by volunteer no.
n, N= No of volunteers.
In vivo studies
Multiple application studies using cutometer was performed during
two weeks on human volunteers after gaining their written informed
consent. Protocol followed was given in Table 3. Transfersomal
cream and base cream were applied non-occlusively to the volar
side of forearm of female volunteers ranging in the age from 20 to
25 years. A dose corresponding to 100 mg was applied to defined
test area. Mechanical properties of epidermis were determined
using non invasive, in vivo suction skin elastic meter equipped with
2 mm measuring probe (Cutometer® MPA 580 Courage and
Khazaka, Köln, Germany). Skin elasticity was measured with the
Cutometer, where the skin was drawn into the aperture of the probe
at a constant negative pressure of 500 mbar. The resulting curve of
each measurement represents the elastic and viscoelastic qualities
of the skin. The first phase (Ue), represents the elastic component
and the second phase (Uv) the plastic component of the skin. The
following parameters (absolute and relative) were analyzed: Ue,
elastic deformation; Uv, visco-elasticity; [R0] or Uf, total
deformation; Ur, retraction; [R2] or Ua/Uf, overall-elasticity of the
skin; [R5] or Ur/Ue, pure elasticity of the skin without viscous
deformation; [R7] or Ur/Uf, biological elasticity, that is, the ratio of
retraction to extension; [R6] or Uv/Ue, the ratio of visco-elasticity to
elastic deformation; and [R8] or (Ua), pliability, that is, ability of the
skin to return into its original state (Kapoor and Saraf, 2009).
Analysis of variance
Statistical analysis of variance for all the data generated was
performed using the SPSS program for Windows. ANOVA test was
applied and p-values less or equal to 0.05 were considered as
RESULTS AND DISCUSSION
In this study, C. longa extract loaded transfersome cream
was evaluated as a carrier for cosmetic application of
natural antioxidant, Curcumin.
transfersomes with various process variables such as
lecithin: surfactant ratio, surfactant, (Tween 80, Tween
20) and different solvent extract (95% ethanol, 85%
ethanol) was prepared. Optimization was done on the
basis of zeta potential, entrapment efficiency, percent
cumulative skin permeation and percent skin deposition
Transmission electron microscopy (TEM) of transfer-
somes confirm three-dimensional structure of vesicles
(Figure 1a and b) in empty transfersomes and extract
loaded transfersomes. Figure 1c confirms the presence
of vesicles in the extract loaded transfersomal cream.
Formulations F1 and F2 prepared with 95% ethanolic
extract produced lower entrapment efficiency, higher
percent cumulative permeated amount of curcumin and
lower skin deposited quantity of curcumin with both the
edge activators. In contrast formulations F3 and F4
For this purpose
1058 Afr. J. Pharm. Pharmacol.
Table 3. Protocol of the in vivo study - multiple applications during 14 days.
Number of human volunteers
Age of volunteers
20-25 years old
Multiple applications, twice a day, on the volar side of the forearm during14 days.
Except on Sundays.
100 mg formulation
Base cream and transfersome cream
After one week and two weeks
Gross elasticity; Net elasticity; Sagginess; Biological elasticity; Pliability; Tiring
effect; Total deformation; Retraction; Elastic Deformation; Viscoelasticity.
(Cutometer MPA 580 Courage and Khazaka, Koln, Germany)
prepared with 85% ethanolic extract produced higher
entrapment efficiency, lower
permeated amount of curcumin and higher skin deposited
quantity of curcumin with both the edge activators. Best
results were with formulation F4. Vesicle size obtained
was in the range 200±2 nm range and zeta potential
between -30±5 mV. There was insignificant difference of
vesicle size and zeta potential by the change of edge
activator or by the change of ratio between soya lecithin
and edge activator (P >0.05). Tween 20 significantly
improved curcumin entrapment into the vesicles while
using 85% ethanol extract. This is in addition to studies
where deoxycholate, Tween 80 and dipotassium
glycyrrhizinate have been used as edge activato (Trotta
et al., 2002). The polydispersity index of all vesicle
formulations was between 0.2 and 0.3, indicating that the
solutions were moderately homogeneous (0.0 is very
homogeneous and 1.0
Entrapment efficiency of transfersome was found to be
higher with 85% ethanol extract (F4) than with 95%
ethanol extract as shown in Table 1.
Results of current study support the existence of the
penetration and deposition enhancing effects of
transfersomes which is contrast to the recent in vivo
electron microscopy study in which intact surfactant
based elastic vesicles partitioned into human stratum
corneum but almost no vesicle could be found beyond
the deepest layer of stratum corneum. The loading
capacity and mean particle size of a colloidal carrier are
important parameters that are able to influence the
percutaneous permeation of an incorporated extract. For
this reason, F4 was selected which was characterized by
highest entrapment efficiency and small vesicle size of
200±2 nm. Transfersomes in size range between 100 to
400 nm can easily penetrate through skin and can form
depot at dermal level as the elastic vesicles are known to
have flexibility and are self adaptable (Benson, 2005;
Cevc et al., 1998).
The percentage of extract permeated through goat
skin over 24 h was higher with 95% ethanolic extract
loaded transfersome than with 85% ethanolic extract
loaded transfersomes due to penetration enhancing
effects of ethanol. Ethanol interacts with the lipid
is very heterogeneous).
molecules in the polar head region, resulting in reduction
in phase transition temperature (Tm) of stratum corneum
lipids increasing their fluidity. The intercalation of ethanol
into polar head group environment can result in an
increase in the membrane permeability (Elsayed et al.,
2007). These findings may explain the result of in vitro
skin deposition studies from different formulas as shown
in Table 1. The percentage of skin deposition from
transfersomal formula with 85% ethanolic extract (F3 and
F4) was higher than percent deposition of transfersomal
formula with 95% ethanolic extract (F1 and F2).
The release of extract in the dermal layers of skin
could be the result of fusion of vesicles with skin lipids
extract release at various points along penetration
pathway. Similarly percent of cumulative permeated
amount of curcumin was lowest for F4 formulation.
Conclusively, formula F4 was considered to the most
suitable formulation to be incorporated into cosmetic for
the delivery of C. longa extract due to its highest
entrapment efficiency, highest percentage deposition to
deep dermal level and lowest skin permeation to deeper
level. The stability of the prepared formulations evaluated
by measuring the zeta potential and the average size
showed no significant changes during the storage time
confirming their stability.
Base cream and extract loaded transfersomal cream
were prepared and evaluated for their stability and
physical characteristics. TEM of transfersomal cream
confirm the presence of vesicles in the cream (Figure 1c).
As shown in Table 4, acid value of transfersomal cream
was greater than of base cream as it contained ethanol
6.4 and 5.6 respectively. Saponification value is an
estimation of free fatty acid esters present in the sample.
It influences the stability, pH and cleansing properties of
formulation Saponification value of the formulation should
be proper, if too high fat may contain too much fatty acid
which are prone to hydrolysis and can cause
rancidification. Saponification value of base cream, was
found to be higher than novel cream; therefore it was
more prone to microbial growth. pH of base cream and
novel cream was 6.3 and 6.2 respectively. Cream
containing 85% ethanolic extract was found to be slightly
acidic. As the pH of skin is between 4.5 to 6.5 pH of
Saraf et al. 1059
Figure 1. Photomicrograph of TEM of (a) Empty transfersomes (F4), (b) Transfersomes (F4) loaded with C.
longa extract, (c) C. longa extract loaded transfersomal cream.
Table 4. Physicochemical evaluation of creams.
Net content (g)
Non-volatile matter (%)
All values represent Mean ± SD (n=3).
No Ash present
No Ash present
Table 5. Percentage efficacy of cream for relative parameter.
Parameter Total deformation (Uf)
Changes in skin mechanical parameters (relative parameters) determined by cutometer after application of formulations expressed as
percent improvement in the values, (n=6).
Elastic deformation Ue
cream should also lie in this range so as to maintain layer
of acidic oils on it, called the acid mantle. Ash test was
conducted to determine inorganic components present in
formulation, that is, borate, chloride, sulphate, which can
cause harm to skin. Patch irritation test for both the
formulations were performed and were found to be non
irritating on skin. Psychometric evaluation of both creams
was done in order to find their acceptance. Novel cream
showed better acceptances in terms of smoothness,
stickiness and shine.
Tables 5, 6, Figures 2 and 3 shows improvement in
relative and absolute parameters after first week itself.
More pronounced improvement was seen in total
deformation (R0), net elasticity (R5) and biological
elasticity (R7) and retraction (Ur). Transfersome cream
significantly improved total resistance to deformation to
27% as compared to base cream, 6% in 2 weeks
duration (P < 0.05). R2 value indicated that overall
elasticity of skin increased 18.37% with novel cream and
only 6% with base cream (P < 0.05), decrease in R2
values with age result due to degradation of elastin fibers
representing inevitable nature of aging. R5 value showed
that pure elasticity increased to 23% with novel cream
and R6, sagginess decreased to 30% with novel cream
(P < 0.05). R7 value, biological elasticity improved to 21%
with novel cream (P< 0.05). R8, pliability that is, ability of
skin to show its original condition after deformation
improved 17% from the original condition (P< 0.05).
Whereas R9, tiring effect improved to 20% (P > 0.05).
Improvement in R8 values indicate that the tendency of
elastin fiber to bind strongly has increased considerably.
This might be due to the curbing of free radicals by
a b c
1060 Afr. J. Pharm. Pharmacol.
Table 6. Percentage efficacy of cream for absolute parameter.
of the skin (R2)
of the skin (R5)
Changes in skin mechanical parameters (absolute parameters) determined by cutometer after application of formulations expressed as percent improvement in the
values, the values are in percentage (n=6).
Figure 2. Percentage efficacy of transfersomes – relative parameters. Changes in skin
mechanical parameters (relative parameters) determined by cutometer after application
of formulations expressed as percent improvement in the values after 1 and 2 weeks
duration, (n=6) [R0] total deformation, [R2] overall-elasticity of the skin, [R5] pure
elasticity of the skin without viscous deformation, [R6] sagginess; [R7] biological
elasticity, [R8] pliability and [R9] tiring effect.
Figure 3. Percentage efficacy of transfersomes – absolute parameters. Changes
in skin mechanical parameters (absolute parameters) determined by cutometer
after application of formulations expressed as percent improvement in the values
after 1 week and 2 week duration , (n=6) Uf = Total deformation, Ur =Retraction, Ue
= Elastic deformation, Uv = Viscoelasticity.
R0 R2R5R6R7 R8R9
1 st Week
2 nd Week
UfUr Ue Uv
1 st Week 2 nd Week
curcumin that also cause wrinkles by activating
metalloproteinases, such as collagenase, that are
responsible for breaking down the skin's connective
tissues (collagen and elastin), thus result in premature
Absolute values, Ur, retraction improves to 9% (P<
0.05) whereas Ue, elastic deformation improves to 28.7%
(P> 0.05). Both Ur and Uv are measure of hydration
effect, thus proving the moisturizing effect of cream due
to the presence of glycerin in formulation. Most prominent
improvement was observed in viscoelasticity, Uv to 55%
(P< 0.05). The results of overall elasticity, biological
elasticity, recovery of deformed skin, firmness and
reduction in fatigueness can be correlated with anti-
wrinkle properties of cream. Therefore, if there is more
improvement in the overall biological elasticity, the anti-
wrinkle property of the product becomes better.
It was possible to prepare stable herbal extract loaded
transfersomal anti- wrinkle
significantly improved curcumin entrapment into the
vesicles and skin deposition with 85% ethanolic extract,
hence Tween 20 could be effectively used as edge
activator. Cutometer results showed improvement in
overall elasticity, biological elasticity, recovery of
deformed skin, firmness and reduction in fatigue which
can be correlated with anti wrinkle properties of cream.
These beneficial effects might have been due to the
synergistic antioxidant, anti-inflammatory, protective
properties of the constituents of extract and hydrant,
moisturizing and lipid components of transfersomes and
cream. Therefore, it may be concluded that proposed
“anti-wrinkle cream” is effective and safe for usage in the
management of facial wrinkles.
Authors are thankful to University Grants Commission
(UGC), (Major Research project, F. No 39-170/2010
(SR)), New Delhi for Instrumental and financial support
for this work and to Director, University Institute of
Pharmacy, Pt. Ravishankar Shukla University, Raipur for
providing laboratory, instrument and all other facilities
required for this work. We are also thankful to All Indian
Institute of Medical Sciences, New Delhi, India, for
providing electron microscope facility and BITS Ranchi
for vesicular size and zeta potential determination.
Ahshawat MS, Saraf S, Saraf
characterization of herbal creams for improvement of skin
viscoelastic properties. Int. J. Cos. Sci., 30: 183-193.
Bangham AD (1963). Physical structure and behaviour of lipids and lipid
creams. Tween 20
S (2008). Preparation and
Saraf et al. 1061
enzymes. Adv. Lipid. Res., 1: 65–104.
Benson HAE (2005). Transdermal Drug Delivery: Penetration
Enhancement Techniques. Current. Drug. Delivery, 2: 23-33.
Bonte F, Noel-Hudson MS, Wepierre J, Meybeck A (1997). Protective
effect of curcuminoids on epidermal skin cells under free oxygen
radical stress. Planta. Medica, 63: 265-66.
Cevc G (1996). Transfersome, liposomes and other lipid suspensions
on the skin: permeation enhancement, vesicle penetration, and
transdermal drug delivery. Crit. Rev. Therap. Drug. Carrier Syst., 13:
Cevc G, Blume G (1992). Lipid vesicles penetrate into intact skin owing
to the transdermal osmotic gradients and hydration force. Biochim
Biophys. Acta, 1104: 226–232.
Cevc G, Gebauer D, Stieber J, Schatzlein A, Blume G (1998).
Ultraflexible vesicles, Transfersomes, have an extremely low pore
penetration resistance and transport therapeutic amounts of insulin
across the intact mammalian skin. Biochim. Biophys. Acta, 1368:
Deep C, Saraf S (2008). Novel approaches in herbal cosmetics. J.
Cosmet. Dermatol., 7: 89-95.
Deep C, Saraf S (2009). Herbal photoprotective formulations and their
evaluation. Open. Nat. Prod. J., 2: 57-62.
Elsayed MA, Abdallah OY, Naggar VF, Khalafallah NM (2007). Lipid
vesicles for skin delivery of drugs: reviewing three decades of
research. Int. J. Pharm., 332: 1Y16
Elsayed MMA, Abdallah OY, Naggar VF, Khalafallah NM (2006).
Deformable liposomes and ethosomes: Mechanism of enhanced skin
delivery. Int. J. Pharm., 322: 60–66.
Forster T, Tesmann H (1991). Phase inversion emulsification. Cosmet.
Toil., 11: 106.
Horwitz E, Pisanty S, Czerninski R, Helser M, Eliav E , Touitou EA
(1999).Clinical evaluation of a novel liposomal carrier for acyclovir in
the topical treatment of recurrent herpes labialis. Oral Surg Oral Med
Oral Pathol Oral Radiol Endod, 87, (6): 700-705.
Indian Herbal Pharmacopoeia (2002). Revised, New edition.
Jayaprakasha GK, Rao LJ, Sakariah KK (2006). Antioxidant activities of
curcumin, demethoxycurcumin and bisdemethoxycurcumin. Food
Chem., 98: 720-724.
Kapoor S, Saraf S (2009). Age dependent studies as various skin
parameters using cutometer. Indian. J. Pharm. Educ. Res., 43, (4):
Kaur CD, Saraf S (2011). Photochemoprotective activity of alcoholic
extract of Camellia sinensis. Int. J. Pharmacol., 7, 3: 400-404.
Kaur CD, Nahar M, Jain NK (2008). Lymphatic targeting of zidovudine
using surface engineered liposomes, J. Drug Targeting., 16: 798-805.
Kaza R, Pitchaimani R (2006). Formulation of transdermal drug delivery
system: matrix type, and selection of polymer- their evaluation. Curr.
Drug. Discov. Technol., 3 (4): 279-285.
Martelli L, Martelli M (2004).
pharmaceutical- dermatological use. US 2009/ 0098226 A1:1-4.
Nair SS, Majeed S, Sankar S, Mathew JM (2009). Formulation of Some
Antioxidant Herbal Creams. Hygeia, 1: 44-45.
Patel R, Singh SK, Singh S, Sheth NR, Gendle R (2009). Development
and characterization of curcumin
transdermal Delivery. J. Pharm. Sci. Res., 1, 4: 71-80.
Pothitirat W, Gritsanapan W (2004). Extraction method for high
curcuminoid content from Curcuma longa. Mahidol University J.
Pharm. Sci., 31: 44-47.
Rajpal V (2004). Standardization of Botanicals. Eastern Publisher, 1,
Saraf S, Kaur CD (2010). Phytoconstituents as photoprotective novel
cosmetic formulations. Pharm. Rev., 4: 1-11.
Saraf S, Sahu S, Kaur CD, Saraf S (2010). Comparative Measurement
of Hydration Effects of Herbal Moisturizers. Pharm. Res., 2: 146-151.
Touitou E (1998). Composition of applying active substance to or
through the skin, US patent, 5: 540-934.
Touitou E, Godin B, Weiss C (2000). Enhanced delivery of drugs into
and across the skin by ethosomal carriers. Drug. Dev. Res., 50: 406–
Trotta M, Peira E, Debernardi F, Gallarate M (2002). Elastic liposome
for skin delivery of dipotassium glycyrrhizinate. Int. J. Pharm., 241:
Compostion for cosmetic or
loaded transfersome for
1062 Afr. J. Pharm. Pharmacol.
WHO (2004). Quality control methods for medicinal plant materials,
World Health Organisation, Geneva, AITBS Publishers and
Distributors, Delhi, p 30.