Formulation and Evaluation of Hair Shampoo Containing Tea Tree ( Melaleuca alternifolia ) Oil and Virgin Coconut ( Cocos nucifera ) Oil

Abstract

Tea Tree oil (TTO) contains beneficial properties such as antibacterial, antimicrobial, antiviral and anti-fungal. Whereas, the medium chain fatty acids in Virgin Coconut oil (VCO) able to protect hair follicles from heat, restoring hair’s moisture and other damage. This paper describes the physical properties of seven hair shampoo formulations containing differing amount of TTO and VCO. The essential oils (TTO) applied in these formulations were extracted from fresh tea trees using steam distillation method and the VCO was produced from fermentation of fresh mature kernel coconut. Gas Chromatography-Mass Spectrometry (GC-MS) analysis was conducted to determine the essential oil components of TTO and fatty acid composition of VCO. The shampoo formulations were subjected to evaluation of several parameters namely organoleptic, pH, viscosity, total solid content, foam stability, and dirt dispersion. The results show that the TTO was composed of terpene hydrocarbons with terpinene-4-ol as the major component; meanwhile lauric acid is major component of VCO. All the shampoo formulations were acid-balanced with pH range between 6.23 – 6.43; total solid contents were between 29.92 – 35.61%; stable foaming with the same foam volume for 4 minutes and no dirt was observed. Rheological test showed formulation with 6% TTO (0% VCO) has pseudo-plastic behavior and relatively lower total solid content which are desirable attributes in hair shampoo. Overall, TTO- and VCO-containing shampoo formulations showed ideal physicochemical properties for hair cleansing and treatments.

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Formulation and Evaluation of Hair Shampoo Containing Tea Tree
(Melaleuca alternifolia) Oil and Virgin Coconut (Cocos nucifera) Oil
To cite this article: Henzilenah Kinjuit and Noumie Surugau 2019 J. Phys.: Conf. Ser. 1358 012022
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12th Seminar on Science and Technology
Journal of Physics: Conference Series 1358 (2019) 012022
IOP Publishing
doi:10.1088/1742-6596/1358/1/012022
1
Formulation and Evaluation of Hair Shampoo Containing Tea
Tree (Melaleuca alternifolia) Oil and Virgin Coconut (Cocos
nucifera) Oil
Henzilenah Kinjuit1,2 and Noumie Surugau1*
1 Faculty of Science and Natural Resources, Universiti Malaysia Sabah, Jalan UMS,
88400 Kota Kinabalu, Sabah, Malaysia.
2 Knowledge and Technology Management Division, Sabah Economic Development &
Investment Authority (SEDIA), 88873, Kota Kinabalu, Sabah, Malaysia.
*Corresponding author: e-mail: lnoumie@ums.edu.my
Abstract. Tea Tree oil (TTO) contains beneficial properties such as antibacterial, antimicrobial,
antiviral and anti-fungal. Whereas, the medium chain fatty acids in Virgin Coconut oil (VCO)
able to protect hair follicles from heat, restoring hair’s moisture and other damage. This paper
describes the physical properties of seven hair shampoo formulations containing differing
amount of TTO and VCO. The essential oils (TTO) applied in these formulations were extracted
from fresh tea trees using steam distillation method and the VCO was produced from
fermentation of fresh mature kernel coconut. Gas Chromatography-Mass Spectrometry (GC-
MS) analysis was conducted to determine the essential oil components of TTO and fatty acid
composition of VCO. The shampoo formulations were subjected to evaluation of several
parameters namely organoleptic, pH, viscosity, total solid content, foam stability, and dirt
dispersion. The results show that the TTO was composed of terpene hydrocarbons with
terpinene-4-ol as the major component; meanwhile lauric acid is major component of VCO.
All the shampoo formulations were acid-balanced with pH range between 6.23 – 6.43; total
solid contents were between 29.92 – 35.61%; stable foaming with the same foam volume for 4
minutes and no dirt was observed. Rheological test showed formulation with 6% TTO (0%
VCO) has pseudo-plastic behavior and relatively lower total solid content which are desirable
attributes in hair shampoo. Overall, TTO- and VCO-containing shampoo formulations showed
ideal physicochemical properties for hair cleansing and treatments.
1. Introduction
The concept of incorporating natural-based ingredients in cosmetic industries is widely grown in the
market due to the major concern and consciousness on producing safe and environmentally friendly
products [1]. Active ingredients from natural sources (plants, animals, microorganisms) have been
broadly used for centuries as main substances in cosmetics preparations [2]. Natural sources may contain
compound such as polyphenols, fatty acids, terpenes and vitamin which promote several bioactivities
and can be incorporated in cosmetic preparation [3]. In this paper, TTO and VCO were incorporated in
haircare preparations due to variety of benefits offered from both oils. TTO has been used largely in
various industries for its beneficial properties [4], whereas virgin coconut oil has been used since ancient
time as multipurpose oils and nowadays it’s become a trend in cosmetic application [5].
Tea tree oil has been used for many years as a component in medicinal products especially in
treating cutaneous infections due to its antimicrobial and anti-inflammatory properties [6]. The essential
oil of tea tree is obtained by using steam distillation process, originated from Australia and also known
as Melaleuca alternifolia [7]. The chemical composition contains approximately 100 components,
which are mostly consists of monoterpenes, sesquiterpenes and related alcohols. Major compound of

12th Seminar on Science and Technology
Journal of Physics: Conference Series 1358 (2019) 012022
IOP Publishing
doi:10.1088/1742-6596/1358/1/012022
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tea tree oil is Terpinen-4-ol, which has long been considered as the main anti-microbial properties to
TTO [8].
Virgin coconut oil was extracted from fresh and mature kernel without the use of heat and also
without undergoing chemical refining [9]. It was previously demonstrated by protein loss and water
retention measurements, that coconut oil can prevent cuticular damage which usually occur during
combing process [10]. The hydrophobicity properties of coconut oil, reduces the water penetration into
the fiber which give beneficial effect by providing a lubricating film on the hair [11]. Ruetsch et al. [12]
have shown that coconut oil penetrates into the hair cortex and reduces the swelling of the hair
fiber, a factor that can cause hair damaged. VCO also acts by filling the gap between the cuticle and
prevent the penetration of aggressive substances such as surfactants into the hair follicle [13].
As TTO and VCO were proven to have beneficial properties, incorporating these two oils in a
formulation of shampoo will benefit in overcoming hair and scalp problem. Therefore, this research was
designed to formulate and to study the characteristics by evaluating the physicochemical properties of
shampoo containing TTO and VCO.
2. Materials and Methods
2.1 Preparation of Raw Material
Tea tree leaves were collected from tea tree field at the Demonstration Plot located at Mile 30 Kimanis,
Papar, Sabah, Malaysia. Approximately 1.0 kg of tea tree leaves were subjected to steam distillation
system to obtain the essential oil [8]. VCO was produced using fermentation method [14]. Sodium
lauryl ether sulfate (SLES), natrium chloride, ethylenediaminetetraacetic acid (EDTA),
cocamidopropyl betaine, polysorbate-20, DMDM hydantoin, citric acid, potassium hydroxide, sulfuric
acid, methanol and hexane were United States Pharmacopeia (USP) grade which purchased from
Sigma Aldrich (USA).
2.2 GC-MS analysis of TTO and VCO
The resulted TTO was subjected to methanol dilution of 1:10 for analysis. GC-MS analysis were carried
out on Agilent 5997A. Helium was used as a carrier gas and the column used was an HP-5MS UI (5%
diphenyl 95% dimethylpolysiloxane), 30m × 0.25mm × 0.25 um. Temperature program was
from 50°C to 280°C at 3°C/min. The temperatures of the transfer line and the injector were 280oC and
250oC, respectively, with MS parameters were as follows: ion source filament voltage 70eV and the
mass range was 40–400 m/z [15].
Similarly, the obtained TTO was subjected to two-step methylation process according to the
method described by Wang et al. [16]. Exactly 40 μL of VCO was placed into 10 mL centrifuge tubes
to which 0.7 mL of 10 M potassium hydroxide solution and 5.3 mL of methanol were added. The
reaction was performed at 55°C for 1.5 h with mixing for 5 s every 20 min. After cooling to room
temperature, 0.58 mL of sulfuric acid (10 M) solution was added and the reaction was continued at
55°C for 1.5 h with mixing for 5 s every 20 min. After cooling to room temperature, 3 mL of n-hexane
was added and mixed for 5 min. Finally, the tubes were centrifuged for 5 min and subjected to GC-MS
analysis. Helium was used as a carrier gas and the column used was an HP-5MS UI (5% diphenyl 95%
dimethylpolysiloxane), 30m × 0.25mm × 0.25 um. The temperature program was held at 180oC for
40mins. The temperatures of the transfer line and the injector were 230oC and 200oC, respectively.
MS parameters were operated with the electron impact mode at 70eV and in the scan range 35 – 600
m/z [17]. The identification is based upon the mass spectra matching with libraries (NIST and
WILEY) and mass spectra from literatures.
2.3 Formulation of Hair Shampoo
Seven shampoo formulation were prepared (w/v) and tested in this study. As can be seen in Table 1,
formulations F1 to F7 are containing increasing amount of TTO (0 – 6 %, w/v) and vice versa for the
amount of VCO. The rest of the ingredients are the same for all the seven formulations

12th Seminar on Science and Technology
Journal of Physics: Conference Series 1358 (2019) 012022
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doi:10.1088/1742-6596/1358/1/012022
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Table 1: The composition of ingredients used for the formulation (%w/w)
Ingredient
F1
F2
F3
F4
F5
F6
F7
Distilled water
55.5
55.5
55.5
55.5
55.5
55.5
55.5
SLES
20
20
20
20
20
20
20
Natrium Chloride
1.8
1.8
1.8
1.8
1.8
1.8
1.8
EDTA
0.2
0.2
0.2
0.2
0.2
0.2
0.2
Cocamidopropyl Betaine
10
10
10
10
10
10
10
Polysorbate-20
6
6
6
6
6
6
6
Virgin coconut oil
6
5
4
3
2
1
0
Tea tree oil
0
1
2
3
4
5
6
DMDM Hydantoin
0.5
0.5
0.5
0.5
0.5
0.5
0.5
Citric Acid
0.1
0.1
0.1
0.1
0.1
0.1
0.1
2.4 Evaluation of Shampoo Formulation
The prepared shampoo formulations were evaluated for its organoleptic characteristic, pH, foaming
ability, total solid contents, viscosity and dirt dispersion.
2.4.1. Organoleptic characteristic
The formulations prepared were evaluated for the organoleptic characteristics, such as color, smell and
texture as described by Krunali et al. [18].
2.4.2. pH measurement.
Each formulation was diluted with distilled water to 10% (v/v) shampoo solution [19]. Readings of pH
were taken using Mettler Toledo FE20-ATC Kit FiveEasy™ Benchtop pH Meter.
2.4.3. Foaming ability.
Foaming ability was evaluated by using cylinder shake method as reported by Badi & Khan [20]. At
room temperature, 1% of 50 ml shampoo solution was put into a 250 ml graduated cylinder, which was
then covered by hand and shaken 10 times. The total volume of the content after 1 and 4 minutes of
shaking were recorded.
2.4.4. Total solid contents.
Percentage of solid content was carried out based on SM 2540 C standard [21]. The samples were placed
into an oven at 105oC for 16 hours or until a constant oven dry weight was achieved and then was placed
into desiccator to cool. The weight of samples were recorded for total solid calculation.
2.4.5. Viscosity measurement
Rheology of product determine the requisite stability of formulation in terms of consistency and flow
characteristics of each formulation. The viscosity measurement was performed at room temperature
using Brookfield DV3T Rheometer by varying the rotational speeds (100 rpm, 150rpm and 200rpm)
[22].
2.4.6. Dirt dispersion
For this test, 10 ml of distilled water was placed into a large test tube. Two drops of the shampoo were
added into the test tube followed by one drop of Indian ink. Test tube was shaken for ten times. The
amount of ink stained in the foam was recorded as none, light, moderate or heavy [19].

12th Seminar on Science and Technology
Journal of Physics: Conference Series 1358 (2019) 012022
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doi:10.1088/1742-6596/1358/1/012022
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2.5 Statistical analysis
Data were analyzed using SPSS general linear model and one way ANOVA with p values <0.05
were considered as significant.
3. RESULTS AND DISCUSSION
3.1 GC-MS Analysis on TTO and VCO
Figures 1 and 2 show the GC-MS chromatograms of TTO and VCO, respectively. Meanwhile, Tables 2
and 3 show the components of TTO and VCO, respectively, identified by GC-MS analysis. The major
component of TTO is terpinene-4-ol (42.54%); an organic compound possesses strong antimicrobial and
anti-inflammatory properties [22]. From previous study, it is suggested that TTO treatment can treat
fungal infections of the skin and mucous membranes as well as in the treatment of dandruff [23].
Figure 1: GC-MS chromatogram for chemical composition of TTO
Figure 2: GC-MS chromatogram of VCO

12th Seminar on Science and Technology
Journal of Physics: Conference Series 1358 (2019) 012022
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doi:10.1088/1742-6596/1358/1/012022
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Table 2: Chemical composition for TTO
Table 3: Fatty Acid Composition of VCO from GC-MS Analysis
The composition of fatty acid showed that the coconut oil is rich in saturated fatty acids, with
high proportion of lauric acid (43.83%). Lauric acid is the triglyceride component in coconut oil with
short chain fatty acid that has a high affinity for hair proteins and its straight linear chain along with low
molecular weight makes them easily to penetrate and absorb deeper into the hair shaft [24]. When VCO
penetrates the hair it reduces the amount of water absorbs in the hair and leading to lowering of swelling
propensity of the cuticle, which limits the upward curving of the surface cuticle [13]. This reduces the
chipping away of the cuticle cells, which leads to protein loss prevention [25].
3.2 Evaluation of Shampoo Formulation
From the parameters evaluated (Table 4), the pH level for all formulations were acid-balanced which
range between 6.23 to 6.43 and it was observed there was a significance differences among the
formulations (p<0.05). The higher pH level of shampoo the harsher the shampoo on human’s hair. As
previous study [26] shown that lower pH of shampoos may cause less frizzing for generating less
negative static electricity on the fiber surface.
Components identified
Molecular
weight (g/mol)
Retention
time (min)
Peak area
(%)
MS
Matching
quality (%)
Alpha-phellandrene
136.238
6.70
1.17
91
Alpha-pinene
136.23
6.89
2.83
95
Alpha terpinene
136.238
9.46
10.89
96
p-cymene
134.21
9.70
2.49
95
Sabinene
136.23
9.84
2.19
87
Eucalyptol
154.249
9.91
3.07
96
Gamma-terpinene
136.238
10.91
19.60
95
Terpinolene
136.23
11.87
4.24
97
Terpinene-4-ol
154.253
15.09
42.54
95
Alpha-terpineol
154.253
15.44
4.19
86
Valencene
204.357
23.61
1.65
90
Bicylogermacrene
204.357
25.38
2.78
98
Delta-cadinene
204.357
26.18
2.36
99
Components
identified
Molecular
weight (g/mol)
Retention
time (min)
Peak area
(%)
MS
Matching
quality (%)
Methyl caprylate
158.241
13.11
2.54
95
Methyl caprinate
186.295
19.92
2.39
96
Capric acid
172.268
21.55
1.88
97
Methyl laurate
214.349
26.19
10.56
97
Lauric acid
200.322
27.98
43.83
98
Methyl myristate
242.403
31.84
4.26
98
Myristic acid
228.376
33.09
14.07
99
Methyl isopalmitate
298.511
36.98
5.32
98
Methyl oleate
296.495
41.07
4.40
99
Methyl stearate
298.511
41.67
3.54
98

12th Seminar on Science and Technology
Journal of Physics: Conference Series 1358 (2019) 012022
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doi:10.1088/1742-6596/1358/1/012022
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Table 4: Physical properties of the hair shampoo formulations
From the results, all formulations have good foaming ability and there was no significance
difference observed among the different formulations. Height of foam produced only shows slight
changes for 4 minutes observation which showed the stability of foaming for the shampoos. Total solid
content in shampoo indicates their capability in cleaning action. Study by Badi & Khan [20], revealed
the best range are between 20 – 30% as it is easy to apply and rinse out from hair. Statistically, there
were significance difference between all formulations (p<0.05). The results showed F7 (6 % of TTO)
has the capability to be washed away on human hair more easily than the other formulations.
Table 5 shows rheological evaluations for the shampoo formulations, with only F7 (6% TTO 0%
VCO) showed pseudo plastic behavior. Pseudo-plastic behavior is a desirable attribute for rheological
properties in shampoos. Pseudoplastic behavior is when the rheological properties will show high
viscosity at low rotational speed and decrease in viscosity at high rotational speed. Low viscosities at
high shear rates (e.g. spreading the product on the hair) make the product easier to apply. High viscosities
at low shear rates may also help suspend materials in the product.
Table 5: Rheological evaluations by measuring viscosity at different rotational speed
Formulation
Organoleptic
characteristics
pH
Foam Height (ml)
Total Solid
content
(%)
Dirt
Dispersion
1min
2mins
3mins
4mins
F1
Clear light-yellow
liquid, VCO odour
6.43±0.02
90
90
90
88
34.87±0.39
None
F2
Clear light-yellow
liquid, VCO and
TTO odour
6.26±0.01
102
102
102
102
35.61±0.43
None
F3
Clear light-yellow
liquid, VCO and
TTO odour
6.33±0.03
100
100
100
98
35.49±0.25
None
F4
Clear light-yellow
liquid, strong TTO
odour
6.39±0.05
94
94
94
93
32.29±0.34
None
F5
Clear light-yellow
liquid, strong TTO
odour
6.23±0.02
110
110
108
108
33.42±0.58
None
F6
Clear light-yellow
liquid, strong TTO
odour
6.28±0.03
96
96
96
95
31.59±0.35
None
F7
Cloudy white
liquid, strong TTO
odour
6.36±0.01
102
102
102
102
29.92±0.22
None
Formulation
Viscosity (cP)
100rpm
150rpm
200rpm
F1
55.60±0.80
63.73±0.27
70.27±0.31
F2
58.37±1.10
69.23±0.59
76.60±0.20
F3
86.40±1.20
89.45±0.39
92.73±0.50
F4
58.83±0.85
70.76±0.15
80.47±0.12
F5
57.20±1.04
70.53±0.23
81.67±0.42
F6
90.33±0.70
111.23±0.25
130.87±0.12
F7
815.33±3.06
796.00±4.16
774.67±2.00

12th Seminar on Science and Technology
Journal of Physics: Conference Series 1358 (2019) 012022
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doi:10.1088/1742-6596/1358/1/012022
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It is considered as poor quality shampoo if the ink were concentrated in the foam, as the dirt
should stay in water. As reported by Saad et al. [19], dirt that remains in the foam will be difficult to
rinse away and will be redeposited on the hair. From the observations of dirt dispersion test (Figure 3),
the ink was not dispersed into the foam for all the formulations, therefore prepared formulations are
satisfactory. This might be due to same amount of surfactants which acts as a cleaning agents in all of
the seven formulations.
Figure 3: Dirt dispersion test on shampoo formulation
From the physical properties (pH, foaming ability and dirt dispersion), all seven formulations show
results which indicate good quality shampoos. The results show high solid content and non-
pseudoplastic behavior on formulations containing 6 %VCO which are not desirable quality for
shampoo. The combination between surfactants and VCO might not suitable in this formulation and
affecting the stability of the emulsion. Yani et al. [20] suggested that to improve the stability of VCO
emulsions in cleansing formulations, more suitable emulsifier and additives, better operations and
conditions are required.
4. Conclusion
This research aimed to prepare and evaluate shampoo formulation containing TTO and VCO. These oils
were analyzed for its major component and the results obtained were in accordance with other previous
study. Terpinene-4-ol from TTO benefits against bacteria which can cause scalp infection and fatty acids
of VCO helps in reducing swelling and protein loss from hair fiber. TTO and VCO plays an important
role and benefits in the treatment of human hair and scalp due to its own important characteristics and
beneficial properties. However, further research and development are still required to improve and
enhance the physical properties of the shampoo in order to produce more quality products.
5. Acknowledgements
The authors would like to thank the management of Sabah Economic Development and Investment
Authority (SEDIA) for their continuous support on this research.
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