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Short Communication
HAIR TONIC FORMULATION OF CLOVE LEAVES (SYZYGIUM AROMATICUM) ETHANOL
EXTRACT AND THE EFFECTIVENESS ON RABBIT HAIR GROWTH
HAFIZHATUL ABADI1*, HANAFIS SASTRA WINATA1, PARHAN1, VIVI EULIS DIANA1, ADEK CHAN1, RENY
HARYANI2
1Faculty of Pharmacy, Institut Kesehatan Helvetia, Medan 20124 Indonesia, 2Study Program of Diploma III of Pharmacy, Sekolah Tinggi
Ilmu Kesehatan Indah, Medan 20226 Indonesia
Email: abadihafizhatul@gmail.com
Received: 13 Jul 2020, Revised and Accepted: 11 Sep 2020
ABSTRACT
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 (http://creativecommons.org/licenses/b y/4.0/)
DOI: http://dx.doi.org/10.22159/ijap.2020v12i6.39027. Journal homepage: https://innovareacademics.in/journals/index.php/ijap
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 [1–3].
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 [4–7].
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
[10–12].
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 [14–16].
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
tonic.
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
evaporators).
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.
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ISSN
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Abadi et al.
Int J App Pharm, Vol 12, Issue 6, 2020, 245-248
246
Table 1: Hair tonic formula
No.
Chemicals
F0
F1
F2
F3
1.
Clove Leaf Extract (%)
-
5
10%
15%
2.
Ethanol 96% (ml)
30.00
30.00
30.00
30.00
3.
Sodium Metabisulfite (g)
0.01
0.01
0.01
0.01
4.
Methylparaben (g)
0.25
0.25
0.25
0.25
5.
Propylparaben (g)
0.025
0.025
0.025
0.025
6.
Menthol (g)
0.10
0.10
0.10
0.10
7.
Tween 80 (ml)
2.0
4.0
6.0
8.0
8.
Propylene glycol (ml)
15.00
15.00
15.00
15.00
9.
Distilled Water up to (ml)
100
100
100
100
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
247
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
Hair tonic
Average of pH
F0
6.00±0.057
F1
4.70±0.057
F2
4.60±0.057
F3
4.60±0.057
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
Groups
Hair growth value
Day-7
Day-14
Day-21
Positive Control
0
6.35±0.042
11.83±0.06
Formula 1 (5%)
0
2.27±0.056
4.00±0.06
Formula 2 (10%)
0
2.86±0.048
4.80±0.06
Formula 3 (15%)
0
3.16±0.042
5.46±0.08
Formula 0 (Blank)
0
0±0
1.71±0.06
Without Treatment (X)
0
0±0
1.69±0.06
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.
FUNDING
Nil
AUTHORS CONTRIBUTIONS
All the authors have contributed equally.
CONFLICT OF INTERESTS
No conflict of interest associated with this work. All authors
guarantee that the contents of this article are known and approved
by all parties
REFERENCES
1. Randall VA. Androgens and hair growth. Dermatol Ther
2008;21:314–28.
2. Powell BC, Rogers, GE. Cyclic hair loss and regrowth in
transgenic mice overexpressing an intermediate filament gene.
EMBO J 1990;9:1485–93.
3. Phillips III JH, Smith SL, Storer JS. Hair loss: common
congenital and acquired causes. Postgrad Med
1986;79:207–15.
4. Kapoor VP. Herbal cosmetics for skin and hair care. Indian J Nat
Prod Resour 2005;4:306–14.
5. T Chiu CH, Huang SH, D Wang HM. A review: hair health,
concerns of shampoo ingredients and scalp nourishing
treatments. Curr Pharm Biotechnol 2015;16:1045–52.
6. Goluch Koniuszy ZS. Nutrition of women with hair loss problem
during the period of menopause. Prz Menopauzalny
2016;15:56–61.
7. Dureja H, Khaushik D, Gupta M, Kumar V, Lather V.
Cosmeceuticals: an emerging concept. Indian J Pharmacol
2005;37:155–9.
8. Baumann LS. Less known botanical cosmeceuticals. Dermatol
Ther 2007;20:330–42.
9. Joshi LS, Pawar HA. Herbal cosmetics and cosmeceuticals: an
overview. Nat Prod Chem Res 2015;3:1–8.
Abadi et al.
Int J App Pharm, Vol 12, Issue 6, 2020, 245-248
248
10. Shorter K, Farjo NP, Picksley SM, Randall VA. Human hair
follicles contain two forms of ATP-sensitive potassium
channels, only one of which is sensitive to minoxidil. FASEB J
2008;22:1725–36.
11. Sawaya ME, Shapiro J. Androgenetic alopecia: new approved
and unapproved treatment. Dermatol Clin 2000;18:47–61.
12. Savin RC, Atton AV. Minoxidil: update on its clinical role.
Dermatol Clin 1993;11:55–64.
13. Mittal M, Gupta N, Parashar P, Mehra V, Khatri M.
Phytochemical evaluation and pharmacological activity of
Syzygium aromaticum: a comprehensive review. Int J Pharm
Pharm Sci 2014;6:67-72.
14. Chomchalow N. Spice production in asia-an overview. AU J
Thechnol 2001;5:27–8.
15. Prajneshu M, Gupta VK. Patenting on clove. J Intellect Prop
Rights 2002;7:24–32.
16. Kaur J, Kaur S, Mahajan A. Herbal medicines: possible risks and
benefits. Am J Phytomed Clin Ther 2013;1:226–39.
17. Gulcin I, Elmastas M, Aboul Enein HY. Antioxidant activity of
clove oil–a powerful antioxidant source. Arab J Chem
2012;5:489–99.
18. Abo El-Maati M, Mahgoub SA, Labib SM, Al-Gaby AMA,
Ramadan MF. Phenolic extracts of clove (Syzygium
aromaticum) with novel antioxidant and antibacterial activities.
Eur J Integr Med 2016;8:494–504.
19. Fitrilia T, Bintang M, Safithri M. Phytochemical screening and
antioxidant activity of clove mistletoe leaf extracts
(Dendrophthoe pentandra (L.) Miq). IOSR J Pharm 2015;5:13–8.
20. Nassar MI, Gaara AH, El-Ghorab AH, Farrag A-RH, Shen H, Huq
E, et al. Chemical constituents of clove (Syzygium aromaticum,
fam. myrtaceae) and their antioxidant activity. Rev Latinoamer
Quin 2007;35:47–57.
21. Kasai H, Shirao M, Ikegami Kawai M. Analysis of volatile
compounds of clove (Syzygium aromaticum) buds as influenced
by growth phase and investigation of antioxidant activity of
clove extracts. Flavour Fragr J 2015;31:178–84.
22. Azwanida NN. A review on the extraction methods use in
medicinal plants, principle, strength and limitation. Med
Aromat Plants 2015;4:1-6.
23. Lemaire C, Plenevaux A, Aerts J, Del Fiore G, Brihaye C, Le Bars
D, et al. Solid phase extraction-an alternative to the use of
rotary evaporators for solvent removal in the rapid
formulation of PET radiopharmaceuticals. J Label Compd
Radiopharm J Int Isot Soc 1999;42:63–75.
24. Nanda B, Lina S, Shivam L. Formulation and evaluation of
herbal hair tonic. Res J Top Cosmet Sci 2015;6:86–90.
25. Patil A, Bhide S, Bookwala M, Soneta B, Shankar V, Almotairy A,
et al. Stability of organoleptic agents in pharmaceuticals and
cosmetics. AAPS PharmSciTech 2018;19:36–47.
26. Kusuma SAF, Abdassah M, Valas BE. Formulation and
evaluation of anti acne gel containing Citrus Auratifolia fruit
juice using carbopol as gelling agent. Int J Appl Pharm
2018;10:147-52.
27. Chen XQ, Antman MD, Gesenberg C, Gudmundsson OS.
Discovery pharmaceutics-challenges and opportunities. AAPS J
2006;8:E402–8.
28. Tanaka S, Saiyo M, Tabata M. Bioassay of crude drugs for hair
growth promoting activity in mice by a new simple method.
Planta Med 1980;40:84–90.
29. Roy RK, Thakur M, Dixit VK. Development and evaluation of
polyherbal formulation for hair growth–promoting activity. J
Cosmet Dermatol 2007;6:108–12.
30. Hwang SL, Kim JC. In vivo hair growth promotion effects of
cosmetic preparations containing hinokitiol-loaded poly(ε-
caprolacton) nanocapsules. J Microencapsul 2008;25:351–6.
31. Wendtner S, Korting HC. The pH of the skin surface and its
impact on the barrier function. Ski Pharmacol Appl Ski Physiol
2006;19:296–302.
32. Soppimath KS, Kulkarni AR, Aminabhavi TM. Chemically modified
polyacrylamide-g-guar gum-based crosslinked anionic microgels as
pH-sensitive drug delivery systems: preparation and
characterization. J Controlled Release 2001;75:331–45.
33. Messenger AG, Rundegren J. Minoxidil: mechanisms of action
on hair growth. Br J Dermatol 2004;150:186–94.
34. Patel S, Sharma V, Chauhan NS, Thakur M, Dixit VK. Hair
growth: focus on herbal therapeutic agent. Curr Drug Discovery
Technol 2015;12:21–42.
35. Mustarichia R, Gozali D, Wicaksono IA, Nafis FDR. A
comparison study of hair tonic and gel formulation or
Angiopteris evecta as a hair growth stimulant. Int J Appl
Pharm 2019;11:199-205.
36. Kumar N, Chaiyasut C. Hair growth promoting activity of
Carthamus tinctorius florets extract-loaded nanostructured
lipid carriers. Int J Pharm Pharm Sci 2015;7:252-7.
