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Hair tonic formulation of clove leaves (Syzygium aromaticum) ethanol extract and the effectiveness on rabbit hair growth


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Objective: The purpose of this study was to determine the physical stability and effectiveness of hair growth in hair tonic preparations containing ethanol extract of clove leaves (Syzygium aromaticum) with a concentration variation of 5, 10, and 15%. Methods: Formulations were evaluated, including organoleptic observation, pH checking, and preparation stability tests using the Climatic Chamber. The effectiveness of clove leaf extract as a hair tonic was tested by looking at hair growth in test animals. Results: The results showed that the preparation was stable at high temperatures (40±2 °C) where the odor, color, clarity, and pH remained constant. Effectiveness test on rabbit hair growth showed significantly different results after measuring hair growth on the 21st day, where the results were 11.83+0.06 mm for positive control, 1.71+0.06 mm as blank, 1.69+0.06 mm for the group without treatment, and successively the results of the group given clove leaf extract 5, 10, and 15% were 4.00+0.06; 4.80+0.06; and 5.46+0.08 mm. Conclusion: Hair tonics containing clove ethanol extract can increase rabbit hair growth.
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1Faculty of Pharmacy, Institut Kesehatan Helvetia, Medan 20124 Indonesia, 2Study Program of Diploma III of Pharmacy, Sekolah Tinggi
Ilmu Kesehatan Indah, Medan 20226 Indonesia
Received: 13 Jul 2020, Revised and Accepted: 11 Sep 2020
Objective: The purpose of this study was to determine the physical stability and effectiveness of hair growth in hair tonic preparations containing
ethanol extract of clove leaves (Syzygium aromaticum) with a concentration variation of 5, 10, and 15%.
Methods: Formulations were evaluated, including organoleptic observation, pH checking, and preparation stability tests using the Climatic
Chamber. The effectiveness of clove leaf extract as a hair tonic was tested by looking at hair growth in test animals.
Results: The results showed that the preparation was stable at high temperatures (40±2 °C) where the odor, color, clarity, and pH remained
constant. Effectiveness test on rabbit hair growth showed significantly different results after measuring hair growth on the 21st day, where the
results were 11.83+0.06 mm for positive control, 1.71+0.06 mm as blank, 1.69+0.06 mm for the group without treatment, and successively the
results of the group given clove leaf extract 5, 10, and 15% were 4.00+0.06; 4.80+0.06; and 5.46+0.08 mm.
Conclusion: Hair tonics containing clove ethanol extract can increase rabbit hair growth.
Keywords: Ethanol extract, Hair tonic, Syzygium aromaticum
© 2020 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license ( y/4.0/)
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Hair loss is a condition where the hair is detached from the surface
of the skin, such as the head and body. Head loss can occur
physiologically or pathologically, which is influenced by external and
internal factors in the body. In general, hair loss can be caused by
heredity and the influence of hormones. It also can be caused by a
lack of nutrient intake into the hair, free radicals, side effects of
drugs, stress, and an unhealthy diet [13].
One way to prevent hair loss is by doing hair care. In treating hair
needs a variety of cosmetics, ranging from hair cleaning cosmetics,
hair conditioner, cream bath, and hair tonic. An easy way to treat
hair loss is to do hair care using hair tonic as an ingredient to
nourish hair [47].
Hair growth stimulants (hair tonic) are preparations that contain
ingredients needed by hair, hair roots and scalp [1, 2]. Currently,
hair tonic preparations are widely available in the market both from
chemicals and herbal ingredients [8, 9]. The use of chemicals in
cosmetics products is considered less safe because it can cause side
effects on long-term use. One of the synthetic chemicals commonly
used in hair tonic as a nutritious agent is Minoxidil which has
allergic skin side effects, headaches, vertigo, weakness and edema
The clove (Syzygium aromaticum) belongs to the Myrtaceae family and
is one of the native Indonesian spice plants [13]. Clove is one type of
spice plant that has high antioxidant activity due to the high content of
eugenol. Clove is a spice plant that has long been used in the clove
cigarette industry, food, beverages and medicines. Parts of plants that
can be utilized are flowers, flower stalks and clove leaves [1416].
The antioxidant activity of Syzygium aromaticum leaf extract is quite
good. The highest total antioxidant activity and reducing power were
obtained from methanol extract and both were closely related to the
total phenol content of clove leaves [17, 18]. Based on the results of
TLC analysis it is estimated that the compounds contained in clove
leaves are alkaloids, phenolics, flavonoids, triterpenoids, terpenoids
and saponins. Saponin, phenolic and flavanoid compounds in each
plant have a role to stimulate hair growth [19-21].
Empirically the people of Nias Island Indonesia, use clove leaves to
trigger hair growth by using clove leaf oil which is applied to the
scalp regularly morning and evening. In this article we will discuss
the activity of clove leaf extract in triggering hair growth or as a hair
The tools used in this study include rabbit maintenance equipment
(rabbit cages, feed containers, and drinking containers), shoves to
measure the length of rabbit hair, analytical balance, scissors and
razor blades, pH meters, drop pipettes, spatulas, tools glassware,
and extraction equipment (blenders, filter paper, funnels, rotary
The clove leaf (Syzygium aromaticum) was obtained from the island
of Nias, Indonesia. The solvent used was 96% ethanol, and the other
chemicals were sodium metabisulfite, methyl paraben, propyl
paraben, menthol, tween 80, propylene glycol and distilled water.
The clove leaves were washed with running water. Clove leaves are
dried in a drying cabinet to form a simplicia. Simplisia clove leaves
are mashed using a blender then extracted using maceration method
using 96% ethanol solvent. Maceration is done for 5 d. Then maserat
is filtered using filter paper to produce filtrate and residue. The
resulting residue was remaserated for 2 d while stirring
occasionally. Then the filtrate from the results of the first maceration
and remaseration was collected and concentrated with a rotary
evaporator until a thick extract was obtained [22, 23].
The formula used is a standard gel formula that is modified with the
addition of the extract as the active substance. The formula used in
making hair tonic preparations (24) can be seen in table 1.
Dissolve tween 80 in a portion of distilled water (solution 1), then
dissolve the extract in solution no. 1 (solution 2). Dissolve sodium
metabisulfite in distilled water (saolution 3). Dissolve each
ingredient of nipagin, nipasol and menthol in ethanol then mix with
solution No. 3 (solution 4). Add propylene glycol into solution 4
while stirring homogeneously (solution 5). Mix solutions 2 and 5,
add distilled water to the volume limit.
- 0975-7058 Vol 12, Issue 6, 2020
Abadi et al.
Int J App Pharm, Vol 12, Issue 6, 2020, 245-248
Table 1: Hair tonic formula
Clove Leaf Extract (%)
Ethanol 96% (ml)
Sodium Metabisulfite (g)
Methylparaben (g)
Propylparaben (g)
Menthol (g)
Tween 80 (ml)
Propylene glycol (ml)
Distilled Water up to (ml)
Evaluation of the formula includes organoleptic observation, pH
checking, and preparation stability test using the Climatic Chamber. Hair
tonic is observed organoleptic changes, including: consistency, odor and
color [25, 26]. The pH was measured by means of a pH meter. The
instrument is first calibrated using a neutral pH standard buffer solution
(pH 7.01) and an acidic pH buffer solution (pH 4.01) until the instrument
shows the pH value, the electrodes are washed with distilled water, then
dried with tissue paper. Then the electrode is dipped in a hair tonic
preparation solution, until the instrument shows a constant pH value.
The number shown by the pH meter is the pH value of the formula [25,
26]. Physical stability test was carried out by storing hair tonic at 40±2 °C
using Climatic Chamber. Organoleptic observations and pH
measurements were carried out once a week for 1 mo [27].
Animals that are used as research objects are rabbits (Oryctolagus
cuniculus). Before the research began, rabbits were acclimated for 2
w so that the rabbits could adapt to their new environment. During
the acclimation process, rabbits are given standard feed. Before
testing all the rabbits to be used shaved hair on the back first. Then a
depilatory cream is applied so that the hair roots are thoroughly
clean. After that, 6 treatment boxes were made with each box area of
4 cm2 (2 cm x 2 cm) and each box was spaced 1 cm apart. The
treatment box is demarcated using permanent markers to
differentiate between the locations of the treatments [28, 29]. The
illustration can be seen in fig. 1.
Hair tonic is applied to each treatment box on the back of a rabbit as
much as 1 ml twice a day for 3 w. Determination of the length of hair
that grows on the back of the rabbit after being shaved is done on
the 7th, 14th and 21st days. Pull hair up to the root of the hair using
tweezers. A total of 10 longest rabbit hair lengths were measured
using calipers. Data on average hair length obtained were processed
statistically to see whether there were significant differences
between the test areas and positive control [30].
Fig. 1: Illustration of the location of tonic formula hair tonic on rabbit, Explanation-M: Positive Control, X: Without Treatment, F0:
Negative Control (Blank), F1: Clove Leaf Extract 5%, F2: Clove Leaf Extract 10%, F3: Clove Leaf Extract 15%
Fig. 2: Clove leaves extract hair tonic
Abadi et al.
Int J App Pharm, Vol 12, Issue 6, 2020, 245-248
Determination of hair weights is done to determine the effectiveness
of hair tonic preparations against hair dense. Hair weight
measurement is done after 21 d by shaving the hair that grows in the
test area then weighed. The results obtained were calculated
statistically to see whether there were significant differences
between the test areas and positive controls [24, 30].
Hair tonic with the addition of clove leaf extract, each 5% light
brown, 10% dark brown, and 15% blackish brown. Clove hair tonic
preparations of clove leaves have a distinctive clove odor. Hair tonic
F0 (blank) is transparent (clear) and has a distinctive smell of
menthol. The formulated hair tonic can be seen in fig. 2.
The results of checking the pH value on hair tonic are still in the
range of pH values that do not irritate the skin (pH balance). The pH
value obtained is 4.5-6.0 where the desired pH range is 4.5-6.5. So it
can be concluded that each formula has the characteristics of a good
pH value. The pH value of hair tonic can be seen in table 2.
Table 2: pH value of hair tonics
Average of pH
The Data given in mean+SD, number of each formula (F0-F3) was 3 times of test.
Hair tonic preparations stored in a Climatic chamber with a high
temperature of 400C±20C for 2 w did not show any changes in
shape, color, or aroma. This shows that the preparation is stable. The
pH check is one of the testing parameters to determine whether the
preparation is included in the skin's pH range or not, which is 4.5-
6.5. Where the pH of the preparations used can affect absorption of
the skin. If the pH of the preparation is too acidic, it can cause skin
irritation. And if the pH of the preparation is too alkaline it can cause
scaly skin [31, 32].
Based on the results of measurements of rabbit hair length for 21
d, each concentration of the formula has effectiveness on the
growth of rabbit hair. The results of hair growth in rabbits can be
seen in table 3.
Table 3: The results of hair growth
Hair growth value
Positive Control
Formula 1 (5%)
Formula 2 (10%)
Formula 3 (15%)
Formula 0 (Blank)
Without Treatment (X)
The Data given in mean+SD with number of each formula (F0-F3) was 3 times of test.
Based on the fig. of the statistical test results, each concentration of
the clove leaf formula showed results that were further lower than
the positive control, the formula containing Minoxidil 2%. This is
influenced by the mechanism of action of minoxidil in stimulating
hair growth by extending the anagen phase and increasing the size
of hair follicles. Histology shows that minoxidil therapy can increase
the proportion of hair follicles in the anagen phase and decrease hair
follicles in the telogen phase [33].
Hair growth is influenced by compounds contained in clove leaves,
including phenolic, flavonoids, and saponins. Saponin, phenolic and
flavonoid compounds in each plant have a role to stimulate hair
growth. Flavonoid compounds are found in many plant tissues
which act as antioxidants. Phenolic compounds in leaves are also
responsible for antioxidant activity. Saponin compounds are
secondary metabolites produced by mainly dicotyledonous plants.
Saponins in the human body function to increase blood flow to the
hair follicles, if blood flow to the hair follicles decreases it will affect
the hair follicles and cause hair loss [29, 34-36].
The results of this study concluded that hair tonic clove leaf ethanol
extract showed relatively good physical stability, where the high
temperature stability test of odor, color, clarity, and pH still showed
constant results so that the preparation was safe for use. Hair tonic
extract of clove leaf ethanol can increase rabbit hair growth.
All the authors have contributed equally.
No conflict of interest associated with this work. All authors
guarantee that the contents of this article are known and approved
by all parties
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... The measurement results showed that the skin evenness of all groups of volunteers before using sheet mask was in normal to rough category (32-51) and (52-100) respectively. After using sheet mask for 4 weeks, the evenness of the volunteers increased from rough to normal (32-51) and smooth skin (0-31) 16,17 . Formula F4 is more effective in smoothing skin compared to formulas F0, F1, F2 and F3 with a significant difference (p<0.05). ...
... Organoleptic Results pH of the preparation was measured before and during cycling test (accelerated stability) with 3 repetitions in each cycle16 . Result can be seen in table 5. ...
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Lady finger (Abelmoschus esculentus (L.) Moench) pod contain polyphenolic compounds, carotene, folic acid, thiamine, riboflavin, niacin, vitamin C, oxalic acid, and amino acids. Lady finger seed contain polyphenolic compounds, mainly oligomeric catechins and flavonol derivatives, protein (i.e., high lysine levels) and oil fraction (in particular, its derived oil is rich in palmitic, oleic, and linoleic acids). This study aimed to formulate a sheet mask of Lady Finger extract, and examining its moisturizing effect on volunteers' skin. This was an experimental study, which conducted by maceration of Lady Finger simplicia, then formulated the essence into sheet mask sheet preparation. Lady Finger extract was varied in 3%, 5% and 7% concentration. Evaluation of preparation include of the quality of the preparation (organoleptic, homogeneity, pH value, and stability). Sheet mask was then examined for its irritation test and effectiveness as moisturizer on volunteers skin. Moisturizing effect was seen each week for 4 weeks treatment by using it twice a week. The results showed that all sheet masks were homogeneous, had pH of 5.6-6.1, stable for 12 days of storage by using cycling test method, and did not irritate the skin. Moisturizing effect of Lady Finger extract sheet mask at 7% concentration was more effective than other concentrations in terms of increasing the water content by 37.36% and smoothness by 40.02%. It can be concluded that Lady Finger (Abelmoschus esculentus (L.) Moench) extract can be formulated as an moisturizing sheet mask and 7% concentration of Lady Finger extract sheet mask preparation showed the best moisturizing activity. Keywords: Moisturizing, Sheet Mask, Lady Finger, Extract
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Objective: Previous research has proven that the water fraction of the Angiopteris evecta root has hair growth activities. The objective of this research was to determine the formula of gel and hair tonic preparations of pakis munding (Angiopteris evecta L.) which met the requirements as a pharmaceutical preparation, good and effective in stimulating hair growth. Methods: Formulation of hair tonic and gel preparations were made using 7.5, 10.0, and 12.5 % of A. Evecta root water extracts as active compounds. The formulation of the two preparations was based on the standard formulation method. The evaluation and formulation test were carried out by organoleptic examination and homogeneity, pH, viscosity, scattering power test, stability test, and safety test based on the standard preparations including Indonesian pharmacopeia. Statistical tests were carried out on both formulas against rabbits based on the modification of the Tanaka method. Comparison of statistics on both formulas was also carried out. Results: It was found both formulas fulfill standard requirements as pharmaceutical preparations. Statistically, the best activity in hair tonic preparations was at a concentration of 12.5% and for gels at a concentration of 10.0% ethanol extract seen from measurements of hair length. Statistically, using the Independent T-test to find out the significant differences in the average hair length on hair tonic and gel, it was found there was no significant difference between the two formulas. It was found that the best formula for hair tonic and gel was in the addition of 10.0% and 12.5% extracts, respectively. Conclusion: The results obtained in this research work clearly indicated that both hair tonic and gel formulas of A. evecta root water extract may be used as stimulating hair growth.
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Medicinal plants are generating an ever-increasing amount of interest due to the effectiveness, low cost and minimal side-effects associated with drugs derived from them. Clove (Syzygium aromaticum (L.) (Family Myrtaceae) is one of the most important herbs in traditional medicine, having a wide spectrum of biological activity. Phytoconstituents of clove comprise of various classes and groups of chemical compounds such as monoterpenes, sesquiterpenes, phenolics and hydrocarbon compounds. The major phytochemicals found in clove oil is mainly eugenol (70-85%) followed by eugenyl acetate (15%) and β-caryophyllene (512%). Their derivatives result in biological benefits such as antibacterial, antifungal, insecticidal, antioxidant, anticarcinogenic capacities. In addition to clove oil’s worldwide use as a food flavoring agent, it has also been employed for centuries as a topical analgesic in dentistry. This review presents an overview and details of the phytochemical and pharmacological investigations on the S. aromaticum.
Objectives: This work was aimed to formulate, characterize and determine hair growth promoting activity of the Carthamus tinctorius (safflower) florets extract-loaded nano structured lipid carriers (NLC) in C57BL/6 mice. Methods: Safflower florets were extracted by maceration with ethanol, and then incorporated into NLC formula. Safflower extract-loaded NLCs were assessed for their physical properties, stabilities and hair growth promoting activity in C57BL/6 mice. Results: Safflower-loaded NLCs had particle size around 100 nm, zeta potential in the range of -40 to -49 mV. The data from DSC and XRD suggested that this NLC occurred as an amorphous type NLC. Safflower extract-loaded NLC promoted hair growth in the mice better than minoxidil. Safflower yellow, the principle phytochemical in safflower extract, along with synergistic activity between other phytochemicals may account for hair growth promoting activity observed in mice. The further investigations in human volunteer should be conducted for the confirmation. Conclusion: Safflower-loaded NLC that provided good physical properties and stabilities, exerted hair growth promoting activity in C57BL/6 mice. © 2015, International Journal of Pharmacy and Pharmaceutical Sciences. All rights reserved.
clove (Syzygium aromaticum) is a rich source of bioactive compounds. The goal of this study was to test different extracts of clove in the term of their content of phenolics, their antioxidant potential and their antibacterial action against pathogenic bacteria.
The volatile compounds originating from fresh clove ( Syzygium aromaticum ) buds cultivated in a medicinal plant garden located in southern Tokyo were investigated. The volatile compounds originating from fresh clove buds were adsorbed onto polydimethylsiloxane‐coated devices and then analysed using gas chromatography‐mass spectrometry. Sampling was performed at three different development stages of clove buds, i.e., phase I (green), phase II (pink), and phase III (red). In thermal desorption‐gas chromatography‐mass spectrometry procedure volatile compounds: eugenol, β‐caryophyllene, α‐humulene, α‐farnesene, and eugenol acetate were the predominant constituents in most cases. Methyl salicylate, α‐copaene, β‐ocimene, 2‐nonanone, ethyl hexanoate, and acetophenone were additionally found as the buds ripened. Volatile compounds obtained from clove leaves were analysed using the same method, and β‐caryophyllene, eugenol, α‐humulene, 3‐hexen‐1‐ol, 3‐hexen‐1‐yl acetate, and hexyl acetate were identified. The antioxidant activity of both water and methanol extracts of clove buds and leaves collected from the medicinal plant garden was confirmed using electron spin resonance spectrometry. Scavenging activities against superoxide anion radical and hydroxyl radical were evaluated. Copyright © 2015 John Wiley & Sons, Ltd.