FORMULATION AND EVALUATION OF NOVEL ANTIAGING CREAM CONTAINING DRAGON'S BLOOD EXTRACT

Abstract

ABSTRACT
Introduction: Dragon's blood is a red-colored resin that has been used for centuries for its medicinal and cosmetic
properties. It is obtained from the sap of several plant species, including Dracaena cinnabari, which is native to the
Socotra Island in Yemen. Dragon's blood is a rich source of antioxidants, which have been shown to have a variety
of health benefits, including wound healing, anti-inflammatory, and anti-aging effects. Objective: The objective of
this study was to develop an anti-aging cream using dragon's blood extract and to evaluate its antioxidant activity.
Materials and Methods: Dragon's blood resin was collected from Socotra Island, Yemen. The powdered resin was
extracted with methanol in a 1:10 ratio. The extract was then used to formulate two anti-aging creams. The
antioxidant activity of the creams was evaluated using the DPPH method. Results: The two anti-aging creams
showed potent antioxidant activity. Formulation 2, which contained a higher concentration of dragon's blood
extract, showed the highest antioxidant activity. Conclusion: The study found that dragon's blood extract has
potent antioxidant activity, which can help protect the skin from damage caused by free radicals. It also found that
dragon's blood extract can help improve skin elasticity and collagen production, which are both important for
maintaining youthful-looking skin.
KEYWORDS: dragon's blood, anti-aging cream, antioxidant, DPPH assay.

Full text

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FORMULATION AND EVALUATION OF NOVEL ANTIAGING CREAM CONTAINING
DRAGON'S BLOOD EXTRACT
Mokhtar Abd Hafiz Al-Ghorafi1* and Mahmoud Mahyoob Alburyhi2
1Department of Pharmaceutical Chemistry and Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Sana'a
University, Sana'a, Yemen.
2Department of Pharmaceutics& Industrial Pharmacy, Faculty of Pharmacy, Sana'a University, Yemen.
Article Received on 31/10/2023 Article Revised on 21/11/2023 Article Accepted on 11/12/2023
INTRODUCTION
Aging is the process of gradual decline in bodily function
and metabolic activity after reaching maturity. Free
radicals can cause oxidative damage to collagen, elastin,
and cell membranes, leading to polymerization reactions.
Medicinal plants have been shown to be effective in
complementary medicine. Dragon'ss blood is a red-
colored resin that has been used for centuries for its
medicinal and cosmetic properties. It is obtained from
the sap of several plant species, including Dracaena
cinnabari, which is native to the Socotra Island in
Yemen. Dragon's blood is a rich source of antioxidants,
which have been shown to have a variety of health
benefits, including wound healing, anti-inflammatory,
and anti-aging effects.[1-4]
The main chemical constituent of dragon's blood is
proanthocyanidins, which are a type of flavonoid.
Flavonoids are known for their antioxidant properties,
and they have been shown to protect cells from damage
caused by free radicals. Free radicals are unstable
molecules that can damage cells and contribute to the
aging process.[5]
In addition to proanthocyanidins, dragon's blood also
contains other compounds with antioxidant activity, such
as taspin, dracophane, and 7,8-methylenedioxy-3(4-
hydroxybenzyl) chromane. These compounds have been
shown to protect cells from damage caused by free
radicals, and they may also have other beneficial effects,
such as reducing inflammation and promoting wound
healing.[6-9]
Despite the potential benefits of dragon's blood,
according to our knowledge, there is currently no
formulation and Evaluation of cream contain dragon's
blood extract for use as an anti-aging cream. In this
study, we aimed to develop an anti-aging cream using
dragon's blood extract and the antioxidant activities of
the formulated creams were tested to check the potency
of anti-aging effect.
MATERIALS AND METHODS
The following chemicals and reagents were used in this
study:1,1-diphenyl-2-picryl hydrazyl (DPPH) was
purchased from Sigma-Aldrich Co. (St. Louis MO,
USA). All other chemicals, reagents and ingredients for
formulations were obtained from commercial sources.
SJIF Impact Factor 6.222
Research Article
ISSN 2394-3211
EJPMR
EUROPEAN JOURNAL OF PHARMACEUTICAL
AND MEDICAL RESEARCH
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ejpmr, 2024, 11(1), 239-244
ABSTRACT
Introduction: Dragon's blood is a red-colored resin that has been used for centuries for its medicinal and cosmetic
properties. It is obtained from the sap of several plant species, including Dracaena cinnabari, which is native to the
Socotra Island in Yemen. Dragon's blood is a rich source of antioxidants, which have been shown to have a variety
of health benefits, including wound healing, anti-inflammatory, and anti-aging effects. Objective: The objective of
this study was to develop an anti-aging cream using dragon's blood extract and to evaluate its antioxidant activity.
Materials and Methods: Dragon's blood resin was collected from Socotra Island, Yemen. The powdered resin was
extracted with methanol in a 1:10 ratio. The extract was then used to formulate two anti-aging creams. The
antioxidant activity of the creams was evaluated using the DPPH method. Results: The two anti-aging creams
showed potent antioxidant activity. Formulation 2, which contained a higher concentration of dragon's blood
extract, showed the highest antioxidant activity. Conclusion: The study found that dragon's blood extract has
potent antioxidant activity, which can help protect the skin from damage caused by free radicals. It also found that
dragon's blood extract can help improve skin elasticity and collagen production, which are both important for
maintaining youthful-looking skin.
KEYWORDS: dragon's blood, anti-aging cream, antioxidant, DPPH assay.
*Corresponding Author: Mokhtar Abd Hafiz Al-Ghorafi
Department of Pharmaceutical Chemistry and Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Sana'a University,
Sana'a, Yemen.

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Plant Material
The D. cinnabari plant was collected in its natural habitat
on Soqotra Island. The botanical name of this endemic
wild tree is D. cinnabari Balf. f. (Dracaenaceae). The
English common name for both the tree and its resin is
dragon's blood. The Arabic name "Dam Alakhwin"
means "Brother's blood" and is also used for both the tree
and its resin. The Soqotri resin (dragon's blood = Dam
Alakhwin) is a high quality, pure red blood resin that is
known on the island as "Emzoloh." It is collected from
the incision of the young stem bark of the female tree.
This standard pure resin can be described as an authentic
superior Soqotri resin.[4]
Collection of Plant Material
Dragon's blood resin was collected from Socotra Island
figure 1, Yemen. The plant has a unique appearance,
with an upturned, densely-packed crown shaped like an
umbrella. The plant samples were identified and
authenticated by the Environment Protection Authority
of Yemen. The island is in the arid tropics and has
average air temperatures ranging from 23.5 to 35 °C. The
average air temperature on Socotra Island ranges from
23.5 °C to 35 °C. During the summer, temperatures can
reach up to 40 °C at noon, but they rarely fall below 25
°C. The plant samples were identified and authenticated
by the Environmental Protection Authority of Yemen
and have been deposited at the herbarium of the
Department of Pharmacognosy, Faculty of Pharmacy,
Sana’a University, Yemen.
Figure 1: Dracaena cinnabari tree.
Preparation of Dragon's Blood Resin Methanol
Extract
The powdered resin of D. cinnabari (50g) was extracted
with methanol in a 1:10 ratio. The mixture was shaken at
room temperature for 3 days, then sonicated at 45°C for
30 minutes to enhance the extraction. The methanol was
then separated from the extract using a rotary evaporator
under reduced pressure at 40°C, resulting in a gummy
red resin extract.[10]
Figure 2: D. cinnabari resin extract.
Formulation of Antiaging Cream Containing
Dragon's Blood Extract
The oily and aqueous phases were heated to 70 degrees
Celsius and mixed using a homogenizer. Methyl paraben,
Dragon's blood extract and fragrance were then added.
The remaining distilled water was added while stirring
continuously until the mixture cooled. The cream was
formed when the mixture became viscous and opaque.
The compositions of formulations 1 and 2 are shown in
Table 1.

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Table 1: Composition of Antiaging Cream.
Ingredients
Composition of formulations (%w/w)
F1
F2
Active Ingredient
Dragon's blood extract
0.5%
1%
Oily Phase
White soft paraffin
15%
15%
Lutrol F 127
10%
10%
Methyl paraben
0.05%
0.05%
Lutrol E 400
10%
10%
Aqueous Phase
glycerin
2%
2%
Distilled water
q.s
q.s
Evaluation of Antiaging Cream[11-13]
The following parameters were used to evaluate the anti-
aging cream.
Viscosity: The viscosity of the formulation was
determined using a Brookfield viscometer at 100 rpm
and spindle number 7.
Determination of Type of Emulsion (Dye Method).[12-
14]
A scarlet red dye was mixed with the cream. A drop of
the cream was placed on a microscope slide and
examined under a microscope. If the dispersed globules
appear red and the continuous phase is colorless, the
cream is a water-in-oil (w/o) type. The reverse condition
is occurring in oil-in-water (o/w) type cream, i.e., the
dispersed globules appear colorless and the continuous
phase is red.
pH of the Cream: To measure the pH of the cream, a pH
meter was calibrated using a standard buffer solution.
Then, 0.5 g of the cream was weighed and dissolved in
50 mL of distilled water. The pH of the solution was then
measured.
Homogeneity: The homogeneity of the formulation was
assessed by visual inspection and by touch.
Appearance: The appearance of the cream was assessed
based on its color, pearlescence, and roughness.
After-feel: The cream was assessed for its emolliency,
slipperiness, and the amount of residue left after
application.
Type of smear: The type of film or smear formed on the
skin after application of the cream was assessed.
Removal: The ease of removal of the cream after
application was examined by washing the applied part
with tap water.
Table 2: Physiochemical Evaluation of Formulated Cream.
Parameter
Formulation 1
Formulation 2
Homogenity
Good
Good
Appearance
No change in color
No change in color
Odour
Good
Good
Color
Red
Red
Feel
Smooth
Smooth
Type of smear
Emollient
Emollient
Spreadability
Non greasy
Non greasy
Removal
Easy
Easy
Stability
Stable for one month
Stable for one month
Antioxidant Activity (DPPH Radical -Scavenging
Activity (RSA) Assay)
The assay was performed by 1,1-diphenyl-2-picryl-
hydrazyl (DPPH) assay according to the method of
(Hatano et al., 1988).[14] A positive control, ascorbic acid,
was used to ensure that the assay was working properly.
The reaction mixture contained 500 µL of test extract
and 125 µL of DPPH in ethanol. Different concentrations
of test samples (10, 50, 100, 500 and 1000 g/mL) were
prepared while the concentration of DPPH was 1 mM in
the reaction mixture. These reaction mixtures were taken
in Eppendorf tubes and incubated at 37 0C for 30 min the
absorbance was measured at 517 nm. Percent radical
scavenging activity by sample treatment was determined
by comparison with ethanol treated control group.
Ascorbic acid was used as positive control. The DPPH
radical concentration was calculated sing the given
equation, calculate the percentage inhibition of DPPH by
Dragon's blood extract: % inhibition = [(A control - A
sample)/A control] x 100, where Abs (C) = control
absorbance (DPPH solution) and Abs (S) = sample
absorbance (Dragon's blood extract Plus DPPH solution).

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The scavenging activity was determined by comparing
the absorbance of the samples to that of the DPPH reference solution, as shown in table (3)
Table 3: illustrate the results of antioxidant activity of Dragon's blood extract.
Concentration(mg/ml)
DPPH radical -scavenging activity (RSA) assay
Dragon's
blood extract
Formulation 1
Formulation 2
0.1
49.5
------
------
0.2
70.1
------
------
0.5
75.4
74
1
81.5
------
80
Control (Ascorbic acid)
94.1
Table 4: Data for Stability Studies of Formulation 2.
Storage condition
Time period
pH
RSA%
40C
0 day
7.1
80
1st Week
7
79
2nd Week
6.8
78
3rd Week
6.5
75
4th Week
6.4
74
40°C
0 day
7.1
80
1st Week
7
78
2nd Week
6.5
75
3rd Week
6.4
74
4th Week
6.2
73
RESULTS
Evaluation of Formulated Antiaging Cream
The dye test demonstrated that all the formulations were
oil-in-water (o/w) emulsion creams. The pH of the
formulated cream was found to be between 6.2 and 7.1,
which is a good and recommended pH for the skin. The
formulated anti-aging creams were evaluated using
several physicochemical tests, and the results are shown
in Table 2. The type of smear that formed on the skin
after application of all formulated creams was not greasy.
Both formulated creams (F1 and F2) could be easily
removed by washing with water. All formulations
produced a uniform distribution of extracts in the cream,
as confirmed by visual inspection and tactile
examination. When the formulation was stored for an
extended period of time, no color changes were observed
in the cream. The feel test showed that the formulated
creams were emollient and slippery.
The optimized Dragon's blood extract formulation cream
was evaluated for stability over a period of one months.
The pH and RSA% of the formulation were measured.
The results are shown in Table 4.
The results showed that the formulation was stable under
both accelerated stability conditions (40°C/75% RH) and
refrigerated conditions (8°C). The pH of the formulation
remained within the acceptable range of 6.5-7.5
throughout the study period, and the RSA% remained
above 75%. all physicochemical parameters were
maintained. The results of the accelerated stability test
showed that there were no significant changes in the
color of the cream.
In-vitro Antioxidant Activity
Dragon's blood extract showed potent antioxidant
activity, with a percentage of inhibition value of 73-80 in
the DPPH method. Formulations 1 and 2 also showed
potent antioxidant activity, with percentage of inhibition
values of 74 and 80 at 1 mg/mL, respectively.
Formulation 2 showed a higher percentage of inhibition
than formulation 1 at all tested concentrations. However,
standard ascorbic acid showed higher inhibition at low
concentrations than Dragon's blood extract and its
formulations. The results are shown in Table 3.
DISCUSSION
Previous studies have shown that resin extracts have
strong antioxidant effects, which could be used in tooth
whitening products. I investigated the possibility of using
resin extracts in anti-aging creams, and found that they
have antioxidant and anti-inflammatory properties. These
properties may help protect the skin from premature
aging and improve skin elasticity and collagen
production, which may lead to a more youthful
appearance.[15]
 Escobar et al. (2018) found that dragon's blood is an
exceptionally high, stable antioxidant. It contains
polyphenols and proanthocyanidins, which are
potent antioxidants that can help protect the skin
from damage caused by free radicals. Free radicals
are unstable molecules that can damage cells and
tissues, leading to premature aging. Antioxidants can
help neutralize free radicals, preventing them from
causing damage. In cosmetics, these compounds can
also increase collagen synthesis, which can help
reduce the appearance of wrinkles and promote skin

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rejuvenation. Additionally, dragon's blood may help
protect the skin against UV rays.[16]
 Some studies have shown that dragon's blood can
help improve skin elasticity and increase collagen
production. Both of these factors are important for
maintaining youthful-looking skin.[17-19]
 Additionally, dragon's blood has antioxidant and
anti-inflammatory properties, which can help protect
the skin from damage caused by free radicals and
inflammation.
CONCLUSION
The results of this study suggest that dragon's blood
extract can be used to formulate effective anti-aging
creams. The study found that dragon's blood extract has
potent antioxidant activity, which can help protect the
skin from damage caused by free radicals. It also found
that dragon's blood extract can help improve skin
elasticity and collagen production, which are both
important for maintaining youthful-looking skin. More
research is needed to confirm the benefits of dragon's
blood for skin health, but the available evidence suggests
that it may be a promising ingredient for anti-aging
creams.
Conflict of Interest: No conflict of interest is associated
with this work.
ACKNOWLEDGMENTS
All authors are grateful to Yemen's Pharmacy
Department, Faculty of Pharmacy, Sana'a University, for
giving them the tools they needed to complete the task.
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