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Assessment of Rose Water and Evaluation of Antioxidant and Anti-inflammatory Properties of a Rose Water Based Cream Formulation

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  • COLORCON LIMITED - ENGLAND
Available online at www.ijpcr.com
International Journal of Pharmaceutical and Clinical Research 2019; 11(1): 43-48
ISSN- 0975 1556
Research Article
*Author for Correspondence: zamahmood@hotmail.com
Assessment of Rose Water and Evaluation of Antioxidant and
Anti-inflammatory Properties of a Rose Water Based Cream
Formulation
Abidi Safia, Zaidi Aamir, Azhar Iqbal, Sultan Rafi, Mahmood Zafar*
Department of Pharmacognosy, Faculty of Pharmacy and Pharmaceutical Sciences, University of Karachi, Karachi
Pakistan
Available Online:25th January, 2019
ABSTRACT
Photo-aging is a universal dilemma that occurs in our population due to constant contact with ultraviolet radiation. The
utilization of antioxidants is a successful approach to avoid symptoms associated to the photo-induced aging of the skin.
In view of this, present study was designed to prepare and evaluate the antioxidant & anti-inflammatory activity of cream
comprising the aqueous petals extract of Rosa damascena for its radical scavenging and protein denaturation activity.
Antioxidant activity assessed using standard ascorbic acid (ferric reducing power assay), and anti-inflammatory activity
assessed using standard diclofenac sodium measuring of the %age inhibition of protein denaturation. The rose water
contains the major phytoconstituents which are polyphenolic compounds flavonoids, tannins, triterpenoids, saponins which
are mainly responsible for the antioxidant and anti-inflammatory properties. Out of three cream formulations (F1, F2, and
F3). F1 cream formulation showed the highest antioxidant (81.55%) and anti-inflammatory activity (80.6%) at 1000µg/ml.
the result noted to be concentration dependent. The IC 50 value of F1 formulation cream was 257.39 while for F2 cream
formulation 374.41. The present results indicate that the Rosa damascena petals extract (Rose water) has a good potential
for cosmetic product development.
Keywords: antioxidant, anti-inflammatory, rose water
INTRODUCTION
Ultraviolet (UV) radiation which directly comes from the
sun divided into three types UVA (320-400nm), UVB
(290-320nm) and UVC (200-290nm)1. Skin is the body
organ that mainly uncovered and bears an extensive
exposure to the ultraviolet radiation. Individual skin
persistently and frequently exposed to potentially unsafe
compounds and rays for the reason that it serves as a
defensive wall between the atmosphere and its in-house
organs, thus making it prone to aging2.Photoaging is a
protracted tenure procedure that fallout as of continual
contact of the skin to solar radiation. Ultraviolet (UV)
radiation has been revealed to cause a variety of lesions in
DNA including pyrimidine dimmers of cyclobutane type
and other photo products of the nucleic acid base. These
photo products are concerned in cellular lethality,
alteration, and further biological effects3. Thickening,
unevenness, coarse wrinkles characterize photoaging,
spotted pigmentation and histological changes together
with injuring to collagen fibers, unnecessary deposition of
abnormal elastic fibers, an increase in
glycosaminoglycans4-6. Ultraviolet radiations aid the aging
procedure and become the reason for the reduction in skin
elasticity. For this cause, the formulation investigations for
the free radical scavenging and anti-inflammatory
activities to know the ability of the formulation to combat
or delay the photoaging process7.
Rosa damascena which furthermore recognized as gul-e-
surkh is one of the mainly immediate sweet-smelling and
therapeutic plants customarily used for various strength
needs. It is an erect shrub up to 2 meters in height.Rose
plant is cultivated all over the world for a reason that of its
attractiveness and scent. It is the most well-known than any
other flower all over the world. It has referred to as a king
of flowers. There are over 200 rose species, and more than
180000 cultivars variety of the plant has been known,
among them the Rosa damascena is one of the most
significant generals of Rosaceae family. Apart from its
uses as a decorative plant in recreational greenhouse and
gardens. They predominantly cultivated for use in
perfume, medicine, and food industry. The plant has
revealed diverse biological and pharmacological actions.It
has used in Unani medicine (Tibb-e-Unani)since ancient
era 8.
Rose plant used for the management of many complains
and described in the ethnobotanical texts, and a range of
uses have been reported such as in aching throat, puffy
tonsillitis, slimming to women and old people, uterine
hemorrhages and urticaria. Locally they are useful to heal
aphthae9. The most beneficial consequence of Rosa
damascena in ancient medicine are together with the
healing of abdominal chest pain, strengthening the heart,
cure of menstrual bleeding and digestive disorders and
decrease inflammation particularly of the neck. This plant
Abidi et al. / Assessment of Rose…
IJPCR, Volume 11, Issue 1: January-February, 2019 Page 44
Table 1: Phytochemical Analysis of Rose Petal Extract
(Rose Water).
Phytoconstituents
Presence
Tannins
+
Triterpenoids
++
Saponins
++
Fixed oil
+
Flavanoids
++
(+) indicates the presence of components in a low
amount
(++) indicates the presence of components in a moderate
amount
Table 2: Percentage of Ferric Reducing Power Capacity
of Cream Formulation F1, F2, and F3.
Concentratio
n µg/ml
% of reduction power capacity
Equivalent to Ascorbic Acid
F1
formulatio
n
F2
formulatio
n
F3
formulatio
n
25
11.41
8.05
7.38
100
41.30
28.69
14.56
500
74.36
65.38
10.89
1000
81.55
72.82
9.12
also used as a mild laxative10. Flowers of the plant are
good
for eyes. Relieve a pain, toothache, stomatitis,
reimbursement the lungs, kidney and liver. It is also used
in reheat of the body, chronic fever, inflammation and
intestinal affection and to decrease too much
perspiration11. Rose water forms a satisfying vehicle for
the grounding of lotions and collyrium12.
MATERIAL AND METHODS
Chemicals
All chemicals and solvents used in the study were of
analytical grade and they were purchased locally
Collection and Identification
10 kilogram of Rosa damascena flowers purchased from
the local market care was taken to select healthy flowers
and identified by the expert, and its herbarium No.
deposited in the Department of Pharmacognosy.The petals
were separated from other parts of the plant and washed
carefully with water to remove dust and foreign material
and dried under shade.
Extraction
Extraction of rose water was carried out by the method
reported by Verma et al .,201113. 60 g of dried rose petals
were hydrodistilled with 1.5 liters of distilled water for 4
hours and obtained 800 ml of rose water.The collected
extract was stored in airtight container in the refrigerator at
4 oC for further study.
Qualitative Phytochemical Analysis
The extract subjected to preliminary phytochemical
screening of saponin, tannins, triterpenoids, flavonoids and
fixed oils.
Formulation of Rosewater cream
Rosa damascena petals extract (rose water),50 g and 30 g
were used for the preparation of cream formulation F1 and
F2 respectively; the F3 formulation was of placebo cream.
The procedure for the rose water cream formulation was as
follow. The oil part of the cream formulation that is
cetostearyl alcohol, cetomacrogol-100, lanolin, stearic
acid, glycerin was collectively uniform at 75 OC ± 2 with
continuous stirring using the hot plate. While for the
grounding of aqueous part purify water was heated
separately in 2000 ml volume beaker at 80 OC ± 2. Upon
constant mixing put in methylparaben, propylparaben and
borax and the heat brought to 75 OC ± 2. The two phases
mixed with continuous stirring for about 1-2 minutes
finally rose water was added with constant stirring till
cream produced. The temperature was additionally
reduced to 45 OC using cold water bathtub.The cream
stored in an airtight amber colored bottle at room
temperature for further studies14.
In Vitro Antioxidant Activity of Cream Formulation
In vitro antioxidant activity of cream formulations F1, F2,
and F3 performed by the method described as ferric
reducing power assay used by Henneberger et al. (2006)15.
Preparation of Extract Solutions
Accurately weighed 200 mg of extract and dissolved in 50
ml ethanol to obtain solutions of 2000 µg/ml
concentration. This solution was serially diluted separately
to achieve the lower concentrations25 .100, 500, 1000
µg/ml. The same procedure is adopted for the preparation
of standard ascorbic acid dilutions 25,100, 500, 1000µg/ml
in distilled water.
S.no
Ingredient
Used as
Amount ( gram)
01
Cetostearyl alcohol
Emulsifying agent
12.5
02
Cetomacrogol-100
Emulsifying agent
2.5
03
Lanolin
Emollient
20
04
Stearic acid
Emollient
10
05
Glycerin
Humectants
50
06
Liquid paraffin
Barrier
15
07
Methylparaben
Preservative
2
08
Propylparaben
Preservative
0.2
09
Borax
Emulsifier / Preservative
Buffering agent
1
10
Active (rose water)
Sun protecting agent
30 g and 50 g
11
Distill water
Vehicle
q.s to make 500 g
12
Total weight
500 gram
Abidi et al. / Assessment of Rose…
IJPCR, Volume 11, Issue 1: January-February, 2019 Page 45
Table 3: Anti-Inflammatory Activity of Cream Formulations F1, F2, F3 and Standard Drug Diclofenac Sodium.
Concentration
µg/ml
% Inhibition of Protein Denaturation
F1 Formulation
F2 Formulation
F3 Formulation
Standard Drug
%
inhibition
Viscosit
y
(cps)
%
inhibition
Viscosity
(cps)
%
inhibition
Viscosity
(cps)
%
inhibition
Viscosit
y
(cps)
50
52.2
0.69
41.4
0.63
-0.375
0.23
35.33
0.98
150
63.9
0.73
42
0.69
17.14
0.09
38.53
1.03
300
65.3
0.82
53.01
0.70
32.41
0.34
64.29
1.04
750
70.3
0.91
57.7
0.78
14.51
0.30
78.49
1.08
1000
80.6
0.95
65.2
0.80
43.67
0.42
86.32
1.10
Mean
±S.D
66.46
±10.31
0.82
±0.11
51.86
±10.24
0.72
± 0.06
21.47
±17.00
0.27
±0.12
60.59
±23.02
1.04
±0.04
Figure-1: Ferric Reducing Power of F1, F2, F3 Cream Formulation Equivalent to Ascorbic Acid.
Figure-2: percentage inhibition of diclofenac sodium and formulated creams against denaturation of the protein.
0
10
20
30
40
50
60
70
80
90
25 100 500 1000
% of reduction power
Concentration µg/ml
ANTIOXIDANT ACTIVITY
F1
F2
F3
-20
0
20
40
60
80
100
50 150 300 750 1000
% inhibition of protein denaturation
Concentration µg/ml
IN-VITRO ANTI-INFLAMMATORY ACTIVITY
F1
F2
F3
F4
Abidi et al. / Assessment of Rose…
IJPCR, Volume 11, Issue 1: January-February, 2019 Page 46
Ferric reducing power assay
2.5 ml of the F1 cream formulation (ranges 25- 1000
µg/ml) were individually assorted using 2.5 ml of 0.2 M
phosphate buffer pH 6.6 and 2.5 ml of (1%,w/v) potassium
ferricyanide. The combination incubated at 50 OC for 20
minutes.At the ending of incubation, 2.5 ml of (10 %,
W/V) trichloroacetic acid (TCA) added to the mixture then
centrifuged at 3000 rpm for 10 minutes. Afterward, 2.5 ml
of supernatant mixed with 2.5 ml of distilled water and 0.5
ml of FeCl3 (0.1 %, W/V), the absorbance was recorded at
700 nm using UV-VIS spectrophotometer. Ascorbic acid
used as a reference standard. The percentage reduction
calculated by using the following formula16.
Percentage of reduction power (%) =1- [1- As/Ac] x 100
Where: Ac absorbance of the standard at the different
concentration tested
As: absorbance of the sample
The similar procedure was adopted to calculate the
reduction power of F2 and F3 formulations.
In vitro anti-inflammatory activity of cream formulations
Inhibition of albumin denaturation method as described by
Gautam et al., 201317. Moreover, previously reported by
Mizushima and Kobayashi 196818. was adopted to confirm
the anti-inflammatory activity of cream formulations
Preparation of stock solution
250 mg of extract was weighed and transferred into 100 ml
volumetric flask to make 2500µg/ml stock solution further
dilution (50,150,300,750, 1000 µg/ml) prepared from that
solution.
Antiinflammatory activity method
2ml of egg albumin from the fresh hen egg, 28ml
phosphate buffer pH 6.4 and 20ml
(50ug/ml,150ug/ml,300ug/ml,750ug/ml and 1000ug/ml)
dilutions of F1 ,F2 ,F3 and Standard) .Incubated the
Figure-3: Relationship between Ferric Reducing Power Antioxidant Activity and Percentage Inhibition of Protein
Denaturation of Cream Formulation F1.
Figure 4: Relationship between Ferric Reducing Power Antioxidant Activity and Percentage Inhibition of Protein
Denaturation of Cream Formulation F2.
y = 0.023x + 55.92
R² = 0.856
IC50=257.39
0
10
20
30
40
50
60
70
80
90
0 200 400 600 800 1000 1200
Percentage
Concentration µg/ml
% age of
reduction power
% Inhibition of
protein
denatured
y = 0.024x + 40.99
R² = 0.923
IC50=375.41
0
10
20
30
40
50
60
70
80
0 200 400 600 800 1000 1200
Percentage
Concentration µg/ml
% age of
reduction
power
% inhibition
of protein
denaturation
Abidi et al. / Assessment of Rose…
IJPCR, Volume 11, Issue 1: January-February, 2019 Page 47
mixture at 37oc for about 30min and then heated at 70oC
for about 15 min, cooled the mixture and observed their
absorbance at 517, 660 and 700 nm using the vehicle as
blank. The percentage inhibition of protein denaturation
was calculated from the control using the following
formula 19.
% inhibition =Abs control- Abs test x 100
Abs control
Whereas, Abs: absorbance
RESULTS AND DISCUSSION
Qualitative Phytochemical Studies
The qualitative phytochemical analysis of rose petals
extract (rose water) showed the presence of flavonoids,
tannins, saponin, fixed oil, and triterpenoids.Represented
in Table-1
Ferric Reducing Power Ability (FRPA)
Antioxidant activity of cream formulation F1, F2, and
F3(placebo) calculated by using ferric reducing power
assay, and the result shown in Table-2.The highest
reducing power ability seen at 1000µg/ml F1 (81.55%)
and F2(72.82%) formulations.It is seen that by increasing
the amount of rose water and concentration of the solution
reducing the power of the cream also increases as shown
in Figure-1
% inhibition of protein denaturation
In-vitro anti-inflammatory activity of cream formulations.
F1, F2, and F3 calculated by using in-vitro protein
denaturation method.The result is shown in Table-3.It
noticed that anti-inflammatory activity was concentration
dependent as we increase the concentration the inhibitory
effect also increased the highest value recorded at
1000µg/ml F1 (80.6%), F2 (65.2%) and F3
(43.67%).Moreover, when compared with standard drug
diclofenac sodium, diclofenac sodium had higher anti-
inflammatory activity as compared to F1, F2, and F3 with
(86.32%) at 1000µg/ml.The extracts were found to be less
active compared to the standard diclofenac sodium, as
shown in Figure-2. Thus the result can conclude that the
present study proposed that the rose water cream
formulation F1 and F2 possess the high potential of
antioxidant and anti-inflammatory properties. The
observed antioxidant and anti-inflammatory effects can be
attributed majorly to the presence of polyphenolic
compounds in the rose water. A comparison graph of % of
reducing power and % inhibition of protein denaturation is
presented in the Figure-3 for F1 and Figure-4 for F2. The
50% inhibitory concentration (IC50) of cream formulation
F1 was 257.39 and for F2 375.41.
CONCLUSION
The Rosa damascena petal extract commonly known as
rose water widely used for its medicinal value in the
traditional system of medicine. The rose water contained
four major polyphenolic compounds which are flavonoids,
tannins, saponin and triterpenoids which are responsible
for the antioxidant and anti-inflammatory properties4. The
antioxidant creams are extensively used nowadays as they
appear to be an attractive way to defend the skin in
opposition to oxidative stress caused by a variety of
extrinsic sources. A numeral of therapeutic plants used in
emergent countries for the management of some disease
condition together with pain and inflammatory condition.
The validation of folkloric claims of these medicinal plants
will present a scientific foundation for the conservation of
tropical medicinal resources. The use of rose water as an
anti-inflammatory agent also reported (Maleev et
al.,1972). As rose water contains a polyphenolic
compound which is helpful in reducing inflammation. To
sustain the efficiency of antioxidant and anti-inflammatory
activity of cream formulation against free radicals and
inflammation, it is vital to stabilizing the final formulation
on its properties. As a part of synergistic effects, the
current practice moves towards in the formulation of
different combinations of active ingredient instead of
single products. The present study revealed that by
increasing the concentration of rose water its antioxidant
and anti-inflammatory activities were also increased.
Moreover, our study presented that formulation of F1
(50g) is more efficient as compared to F2 (30g). The
research work suggests that to ensure the quality and purity
of the cream, it must have the consistency and uniformity
in the ingredients of the herbal antioxidant cream. The
trend of using herbal skin cream is becoming in demand
since it is proven that topical application of anti-oxidant
cream will be effective against UV radiation and protect
the skin from the major consequence of UV damage. In
conclusion, the topical application of the formulated cream
rose water will help in reducing oxidative damage and give
the antioxidant effect to our skin due to its high antioxidant
values.
ACKNOWLEDGEMENT
The authors are thankful to the entire staff of the
Department of Pharmacognosy, Faculty of Pharmacy and
Pharmaceutical Sciences, University of Karachi for the
technical assistance.
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Gul-e-Surkh (rose) is the flower of the plant Rosa damascena Mill that is one of the most important aromatic and medicinal plants traditionally used for various health needs. It is an erect shrub, up to 2 m in height. This plant is cultivated throughout the world because of its beauty and fragrance. It is the most famous than any other flower throughout the world. It has been referred to as the king of flowers. At present time, over 200 rose species and more than 18000 cultivars form of the plant has been identified, among them Rosa damascena is one of the most important species of Rosaceae family. Apart from its use as ornamental plants in parks, gardens, and houses, they are principally cultivated for use in perfume, medicine, and food industry. Rose oil or otto or attar of roses is freely used as perfumes by rich classes. Otto is seldom used medicinally except for perfuming emollients and medicinal soap. The plant has shown diverse biological and pharmacological activities. It has been used in Unani Medicine (Tibb-e-Unani) since ancient era. Keeping in view the high medicinal importance of the plant in Unani Medicine, this review provides available information on its therapeutic uses, phytochemistry and pharmacological properties.
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