Formulation and Evaluation of
Antioxidant Face Cream Containing Raspberry Fruit and Grape Seeds Extract
Priya R. Kawarkhe*, Subhash V. Deshmane,
Kailash R. Biyani
Department of Cosmetic technology,
Anuradha College of Pharamcy, Chikhli. Dist-Buldana. 443201. M.S. India
*Corresponding
Author E-mail: kawarkhepriya@gmail.com, subdeshmane@yahoo.co.in,
principal.acpchikhli@gmail.com
ABSTRACT:
Antioxidant neutralizes free radicals,
unstable oxygen molecules that break down skin cells and cause wrinkles, thus
preventing impairment at the cellular level. In the present work, face cream
formulations were prepared using antioxidant from natural source. Raspberry
extract and Grape seed extract was used. Radical scavenging activity of
raspberry, grape seeds was studied with standard ascorbic acid by DPPH method.
Six cream base formulations were prepared and two formulations BF1 and BF4 were
optimized. Six formulations of antioxidant cream were prepared using 2%
extract. The various evaluation test like, pH, viscosity, grittiness,
spreadability, skin irritation and antimicrobial test were carried out. The
stability study over one week was also conducted. Raspberry and grapes seeds
showed very good antioxidant activity with IC50 value 0.033µg/ml and
0.072µg/ml respectively. Face cream formulation shown active zone of inhibition
within range of 9-12 mm, indicating that, all the formulation have good
preservation activity. The study was concluded with that, natural source for
excipient is the more suitable due to easy availability, non-toxicity and
biocompatible properties of excipient. After studying the physical parameters,
stability study evaluation, it is concluded that the product has the best
capacity to reduce free radical and can be used as the provision of a barrier
to protect skin.
KEYWORDS: Antioxidant,
face cream, DPPH, raspberry fruit extract, grape seeds extract.
1.
INTRODUCTION:
Skin facial creams restore a more
youthful complexion, whitening the colour of skin, adds moisture and protective
layer on the skin. The facial creams acts as removal of the flaws from skin,
maintaining the smoothness and moistness of skin.1 The
skin is the largest external organ of body and also known as the cutaneous
membrane that protects against mechanical trauma, UV light and infections.2,3
It protects the underlying structures
from injury and from invasion by microbes. Antioxidant neutralizes free
radicals, unstable oxygen molecules that break down skin cells and cause
wrinkles, thus preventing impairment at the cellular level. They inhibit
inflammation leading to collagen depletion and offer protection against photo
damage and skin cancer.4 It may also reduce the effect of
trace, smoke, and pollution. Some skin expert believes that direct application
of antioxidant cream to the skin might slow down the aging process.5 Now
a day’s antioxidant obtained from natural herbal source is also have wide
applications in preparation of cosmetic preparations because of their easy
availability and non-toxicity.6 Raspberry, perennial
plant species with woody stems in the genus Rubuscrataegifolius of the
rose family.33 The fruit is an aggregate fruit 1 cm diameter, made
up of numerous drupelets. The species grows on forest margins and mountain
slopes, in areas with moist and well-drained soil. Red raspberry contains 31
μg/100 g of folate. Red raspberries have antioxidant effects that play a
minor role in the killing of stomach and colon cancer cells. Grapeseeds
extract, the extract obtained from the seeds of Vitisvinifera,
belonging to the family vitaceae. Many of the other Vitisvinifera (grape)
ingredients are reported to function as skin conditioning agents, and a few are
reported to function as antioxidants.7
2.
EXPERIMENTAL WORK:
Materials: Raspberry
extract and Grape seed extract was procured from Kingvish Kompany, Thane,
India. Other chemicals were used are of analytical grade.
2.1
Evaluation of Antioxidant activity of Raspberry and Grape seed Extract
2,
2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of the ethanol
extract of raspberry and grape seed extracts in alone and combination was
investigated. Stock solution of DPPH (100 mmol/L, 33mg in 1L) was prepared in
methanol and labeled as control. The absorbance of control was measured using
UV-Visible spectrophotometer at 517 nm. 5 ml of control as stock solution was
added to 1ml of test sample dissolved in ethanol was added at different
concentrations (50, 100, 200 μg/mL). Equal
amount of ethanol was added to the control.8 After 30 min.
absorbances of all three test samples were measured at 517 nm
spectophotometrically 9. The scavenging activity was calculated
using the formula:
%
Scavenging activity
=
[(A517control-A517sample)/A517control] x 100.
Control
= stock solution of DPPH, Ascorbic acid was used as a standard.
2.2 Formulation of cream base
Ingredients were used in the preparation
of cream base are mentioned in table 1. The base was prepared by emulsification
method.
2.3 Evaluation of
cream bases
The six formulations of cream base were
subjected to various evaluation tests. The pH of base
was determined by pH meter. The viscosity of cream bases formulation was
determined by using Brookfield viscometer at 100 r/min. Spreadability
of formulations were determined by fabricated apparatus in laboratory and used
for study. The apparatus consist of wooden block, with a fixed glass slide with
one end tied to weight pan rolled on the pulley, which was in horizontal level
with fixed slide. 1.5gm was placed between two glass slide and 100gm weight was
placed on slide for 5 min to compress. The sample to uniform thickness weigh
was added to the pan. The time (seconds) required to separate the two slides
was taken as a measure of spreadability. It was calculated using the formula:
Where,
m = weight tide
to upper slide, l = length moved on the glass slide, and t = time taken (in
sec) to separate the slide completely each other.
Skin irritancy
test was conducted using six human volunteers. An area of 1 cm2
on the dorsal left hand surface was marked. The cream was applied to the
specified area and the time was noted. Irritancy, erythema, edema were checked
for regular intervals up to 24 h and reported. The stability
study of cream base formulations at the room temperature was carried out for
three weeks and all above parameters were checked. The ease of
removal of the cream applied was examined by washing the applied part with tap
water.
2.4 Preparation of antioxidant face
cream
Antioxidant face cream containing
raspberry (RB) and grape seed (GS) extract was prepared with optimized base
cream formulation. The details of components of face cream are mentioned in
table 2. The cream was prepared by emulsification method and water was used as
vehicle up to the volume. The weight of cream was 100 g.
Table 1: Formulation of cream base
|
Formulation
code
|
SA*
(g)
|
AO*
(ml)
|
NaOH
(g)
|
TEA*
(ml)
|
EDTA*
(g)
|
Glycerin
(ml)
|
MP*
(g)
|
Perfume
(ml)
|
|
BF1
|
13
|
1
|
1
|
1
|
0.2
|
6
|
0.1
|
0.5
|
|
BF2
|
14
|
1
|
1
|
1
|
0.2
|
6
|
0.1
|
0.5
|
|
BF3
|
15
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
|
BF4
|
16
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
|
BF5
|
17
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
|
BF6
|
18
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
* SA- Stearic acid, AO- Almond oil, TEA-
Triethanolamine, EDTA-ethylenediamine tetra acetic acid, MP-Methyl paraben
Table 2: Composition of face cream
containing active ingredient
|
Formulation
code
|
Fruit
extract
(2
%)
|
SA*
(g)
|
AO*
(ml)
|
NaOH
(g)
|
TEA*
(ml)
|
EDTA*
(g)
|
Glycerin
(ml)
|
MP*
(g)
|
Perfume
(ml)
|
|
AF1
|
RB
|
14
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
|
AF2
|
GS
|
14
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
|
AF3
|
RB
+ GS
|
14
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
|
AF4
|
RB
|
15
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
|
AF5
|
GS
|
15
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
|
AF6
|
RB
+ GS
|
15
|
0.5
|
1
|
0.5
|
0.2
|
8
|
0.1
|
0.5
|
* SA- Stearic acid, AO- Almond oil, TEA-
Triethanolamine, EDTA-ethylenediamine tetra acetic acid, MP-Methyl paraben
2.5 Evaluation test for antioxidant face
cream
The all formulations of antioxidant
cream were subjected to various primary evaluation tests like pH, viscosity,
spreadability and grittiness, skin irritancy, color and washability. The
anti-microbial assay test for the antioxidant cream formulations was carried
out using disk diffusion method against two human pathogenic bacteria,
including Gram positive (Staphylococcus aureus), Gram-negative bacteria
(E. Coli). The cultures were grown in nutrient broth and incubated at 37
°C for 24 h. A sterile 6mm diameter filter discs (whatman paper no. 42)
impregnated in 6 batches of cream formulation placed on the surface of petri
plates seeded with 0.1 ml of microbial suspension. Soon afterwards the plates
were kept at 10oC for 30 min. After it normalized to room
temperature the plates were incubated at 37 şC for 24 h. After incubation
period the zone of inhibition was measured.13 The
stability study at room temperature was determined for the period of 3 week.
The tests were studied was color, sudden viscosity change and feel properties 10-12.
3. RESULT AND DISCUSSION:
FTIR spectrums of Raspberry, Grape seed
extract and combined Raspberry and Grape seed extract are mentioned in figure
no.1, it was found that the all characteristic functional group were found
between specified band widths and hence identified pure. There is a slight
shifting of peaks, but that is also in specified ranges, hence found both
extract was compatible with each other.
Figure 1: FTIR study of Raspberry
fruit, grape seeds extract and mixture of raspberry and grape seeds
3.1 Antioxidant
activity
The DPPH as a
control was measured with absorbance 0.0775. The highest percent inhibition of
Raspberry fruit extract was found 94.83 % with IC50 value 0.033
µg/ml. Grape seed extract showed 90.96 % inhibition activity with IC50
value 0.072 µg/ml. While, ascorbic acid gives percent inhibition activity was
89.67% with IC50 value 0.150 µg/ml. The combination of Grape seed
and Raspberry extract has 87.09 % inhibition with IC50 value 0.161
µg/ml. The different concentrations showed different percent inhibition which
were mentioned in table no. 3.
Table 3:
Absorbance and % Inhibition activity of raspberry extract.
|
Substance
|
Concentration (μg/ml)
|
Sample absorbance
|
% Inhibition
|
IC50value
(μg/ml)
|
|
Raspberry extract
|
50
|
0.018
|
76.77 %
|
0.148
|
|
100
|
0.014
|
81.93%
|
0.115
|
|
200
|
0.004
|
94.83%
|
0.033
|
|
Grape
seed extract
|
50
|
0.019
|
75.48%
|
0.195
|
|
100
|
0.012
|
84.51%
|
0.123
|
|
200
|
0.007
|
90.96%
|
0.072
|
|
Ascorbic
acid
|
50
|
0.014
|
81.93%
|
0.264
|
|
100
|
0.013
|
83.22%
|
0.245
|
|
200
|
0.008
|
89.67%
|
0.150
|
|
Raspberry
and grape seed extract
|
50
|
0.017
|
78.06%
|
0.274
|
|
100
|
0.016
|
79.35%
|
0.258
|
|
200
|
0.010
|
87.09%
|
0.161
|
Figure 2: Graph
showing antioxidant activity
Table 4: Evaluation of different
parameters of base formulations
|
Formulation
code
|
pH
|
Viscosity
(cps)
|
Spreadability
(cm/sec)
|
Skin
irritation
|
|
BF1
|
5.8
|
32533
|
29±1
|
No
Irritation
|
|
BF2
|
6.2
|
34753
|
27±1
|
No
Irritation
|
|
BF3
|
6.1
|
40200
|
26.33±1.52
|
No
Irritation
|
|
BF4
|
5.9
|
41350
|
25±1
|
No
Irritation
|
|
BF5
|
6.8
|
52400
|
23.66±1.5
|
No
Irritation
|
|
BF6
|
6.9
|
55240
|
22.66±1.52
|
No
Irritation
|
Figure 2,
indicated that single Raspberry fruit extract of 200 µg/ml concentration showed
highest percent free radical scavenging activity (94.83%) as compare to 200
µg/ml ascorbic acid (89.67%). From the observation (fig. 4,5,6 and 7) it was
notice that single Raspberry fruit extract having good antioxidant activity
than Grape seed extract or combined form of Raspberry and Grape seed extract.
Also it was observed that Grape seed extract and combine extract show
antioxidant activity near to ascorbic acid. Hence, both extracts were
considered for formulation of herbal antioxidant face cream.
3.2 Evaluation of base formulations
Six formulation of primary base were
prepared by melting method satisfactorily. All the formulation was subjected to
various evaluation parameters. The results of various tests are mentioned in
table 4 and found satisfactory. All formulations were white in color, feel was
very smooth, viscosity was acceptable and no irritation was cause.
Form the data mentioned in table 4, it
was found that except formulation no. BF5 and BF6 all formulations were shown
desire pH (Desire pH should be 4.5 to 6.2). The viscosity of all formulations
was also passable with good spreadability (22.6-29 cm/s). All formulations were
non-irritant. Remaining parameters were good and/ or desirable, hence at this
stage formulations BF1 to BF4 were found good stable. After 3 week stability
study at room temperature all the cream base formulations were stable upto 1
week. Afterwards slight change in viscosity and feel was found with formulation
BF5 and BF6 respectively. It might be due to increasing concentration of
stearic acid. In 3rd week only formulation BF2 and BF4 was found
stable with white colour, smooth feel and a very specific pH and viscosity
(Table 5). Hence, for the preparation of Antioxidant Face Cream we considered
formulations BF2 and BF4 as optimized batches. This may be due to appropriate
concentration of stearic acid.
Table 5: Stability study of base
formulation
|
Formulation
code
|
Color
|
Sudden
viscosity change
|
Feel
|
pH
|
Viscosity
(cps)
|
|
After
1 Week
|
|
BF1
|
White
|
No
change
|
Smooth
|
5.7
|
32630
|
|
BF2
|
White
|
No
change
|
Smooth
|
6.4
|
34770
|
|
BF3
|
White
|
No
change
|
Smooth
|
6.1
|
40213
|
|
BF4
|
White
|
No
change
|
Smooth
|
6.2
|
41355
|
|
BF5
|
White
|
No
change
|
Smooth
|
6.5
|
52450
|
|
BF6
|
White
|
No
change
|
Smooth
|
6.9
|
55260
|
|
After
2 Weeks
|
|
BF1
|
White
|
No
change
|
Smooth
|
5.7
|
32726
|
|
BF2
|
White
|
No
change
|
Smooth
|
6.1
|
34793
|
|
BF3
|
White
|
No
change
|
Smooth
|
6.0
|
40225
|
|
BF4
|
White
|
No
change
|
Smooth
|
5.8
|
41350
|
|
BF5
|
White
|
No
change
|
Smooth
|
6.6
|
52560
|
|
BF6
|
White
|
Change
|
Tacky
|
6.5
|
55467
|
|
After
3 weeks
|
|
BF1
|
White
|
Change
|
Smooth
|
6.4
|
32742
|
|
BF2
|
White
|
Change
|
Smooth
|
5.8
|
36730
|
|
BF3
|
White
|
No
change
|
Smooth
|
6.1
|
45220
|
|
BF4
|
White
|
No
change
|
Smooth
|
5.9
|
46358
|
|
BF5
|
Yellowish
white
|
Change
|
Tacky
|
6.4
|
53577
|
|
BF6
|
Yellowish
white
|
Change
|
Tacky
|
6.6
|
56489
|
|
|
|
|
|
|
|
Table 6: Evaluation of different
parameters of cream formulations
|
Formulation
code
|
pH
|
Viscosity
(cps)
|
Spreadability
(cm/sec)
|
Skin
irritation
|
Zone
of inhibition
|
|
AF1
|
6.2
|
38590
|
32.6±0.55
|
No
Irritation
|
9
mm
|
|
AF2
|
6.5
|
38166
|
31.21±0.93
|
No
Irritation
|
12
mm
|
|
AF3
|
6.9
|
37459
|
27.96±0.40
|
No
Irritation
|
9
mm
|
|
AF4
|
6.6
|
45470
|
24.3±0.95
|
No
Irritation
|
7
mm
|
|
AF5
|
6.8
|
46598
|
26.3±0.95
|
No
Irritation
|
9
mm
|
|
AF6
|
6.9
|
46577
|
21.42±0.87
|
No
Irritation
|
11
mm
|
3.3 Evaluation of antioxidant cream
formulations
Results shown in table 6, indicated that
all the cream formulations were found within the acceptable limit. The zone of
inhibition was found within range of 9-12 mm, indicating that, all the
formulation have good preservation activity. Stability tests at room
temperature for cream formulation for 3 weeks were conducted satisfactory and
the results were shown in table no.7. Among all the formulas containing active
AF1 showed good consistency and pH around 6 up to 3 week study and it is also
suitable for topical application as the pH of the skin is in between 4.5 to
6.2.
Table 7: Stability study of cream
formulations
|
Formulation
code
|
Color
|
Sudden
viscosity change
|
Feel
|
pH
|
Viscosity
(cps)
|
|
After
1 week
|
|
AF1
|
White
|
No
change
|
Smooth
|
6.2
|
38578
|
|
AF2
|
Faint
brown
|
No
change
|
Smooth
|
5.8
|
38234
|
|
AF3
|
Off
white
|
No
change
|
Smooth
|
6.6
|
37467
|
|
AF4
|
White
|
No
change
|
Smooth
|
6.5
|
45477
|
|
AF5
|
Faint
brown
|
No
Change
|
Smooth
|
6.8
|
45892
|
|
AF6
|
Off
white
|
No
Change
|
Smooth
|
6.9
|
45077
|
|
After
2 weeks
|
|
AF1
|
White
|
No
change
|
Smooth
|
5.9
|
37560
|
|
AF2
|
White
|
Change
|
Tacki
|
6.4
|
38679
|
|
AF3
|
Off
white
|
No
change
|
Smooth
|
6.8
|
38489
|
|
AF4
|
White
|
No
change
|
Smooth
|
6.5
|
45588
|
|
AF5
|
Faint
brown
|
Change
|
Tacki
|
6.7
|
48789
|
|
AF6
|
Off
white
|
No
change
|
Smooth
|
6.5
|
45578
|
|
After
3 weeks
|
|
AF1
|
White
|
No
change
|
Smooth
|
6.1
|
45768
|
|
AF2
|
White
|
Change
|
Tacki
|
6.6
|
38457
|
|
AF3
|
Off
white
|
No
change
|
Smooth
|
6.5
|
39467
|
|
AF4
|
White
|
No
change
|
Smooth
|
6.3
|
47855
|
|
AF5
|
White
|
Change
|
Tacky
|
6.8
|
48765
|
|
AF6
|
Off
white
|
Change
|
Tacky
|
6.6
|
49521
|
4.
CONCLUSION:
Raspberry fruit extract and grape seed
extract showed very good free radical scavenging (antioxidant) activity.
Stearic acid, triethanolamine and other ingredients were used in preparation of
face cream also prove stability of the preparations. Natural source for
excipient is the more suitable due to easy availability, non-toxicity and
biocompatible properties of excipient. After studying the physical parameters,
stability study evaluation, it is concluded that the product has the best
capacity to reduce free radical and can be used as the provision of a barrier
to protect skin.
5.
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