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Formulation and In Vitro Characterization of Tea Tree Oil Anti-Dandruff Shampoo


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Background Dandruff is a common scalp condition affecting half of the population of the world. Objective The current study aimed at developing anti-dandruff shampoos containing tea tree oil, which is believed to be effective against Malassezia furfur, a fungus involved in dandruff production. Methodology Various shampoos containing tea tree oil in 0.5 to 3% concentration were prepared after careful selection of various shampoo ingredients. The formulated shampoos were subjected to various quality tests such as pH, viscosity, foam production, dirt dispersion, wetting time, surface tension, solid contents, and antimicrobial activity against a model fungal strain, namely Candida albicans. The formulated shampoos were also compared with the marketed shampoos for quality attributes. Results The results revealed that tea tree oil shampoos had pH values in the range of 5 – 6, which is close to the slightly acidic skin’s pH and considered as good for hair. All other quality attributes were comparable to the marketed products. The marketed shampoos had superior antifungal activity due to the presence of zinc pyrithione or a higher concentration of salicylic acid or selenium sulfide. Notwithstanding, the tea tree oil shampoos demonstrated an appreciable antifungal activity due to synergistic effects of tea tree oil, sodium lauryl sulphate, and salicylic acid. Furthermore, the tea tree oil shampoos were stable during two months-long stability testing. Conclusion Thus, tea tree oil anti-dandruff shampoos have the potential to address the dandruff problem.
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Formulation and In Vitro Characterization of Tea Tree Oil Anti-Dandruff
Humra Umar1, Tariq Mahmood1, Talib Hussain2,3, Rabia Aslam1, Yasser Shahzad2 and Abid
Mehmood Yousaf2,*
1Faculty of Pharmacy, University of Central Punjab, Lahore, Pakistan; 2Department of Pharmacy, COMSATS Univer-
sity Islamabad, Lahore Campus, Lahore, Pakistan; 3Provincial Pharmacovigilance Centre, Provincial Drug Control
Unit, Lahore, Pakistan
Abstract: Background: Dandruff is a common scalp condition affecting half of the population of the
Objective: The current study aimed at developing anti-dandruff shampoos containing tea tree oil, which
is believed to be effective against Malassezia furfur, a fungus involved in dandruff production.
Methodology: Various shampoos containing tea tree oil in 0.5 to 3% concentration were prepared after
careful selection of various shampoo ingredients. The formulated shampoos were subjected to various
quality tests such as pH, viscosity, foam production, dirt dispersion, wetting time, surface tension, solid
contents, and antimicrobial activity against a model fungal strain, namely Candida albicans. The formu-
lated shampoos were also compared with the marketed shampoos for quality attributes.
Results: The results revealed that tea tree oil shampoos had pH values in the range of 5 6, which is
close to the slightly acidic skin’s pH and considered as good for hair. All other quality attributes were
comparable to the marketed products. The marketed shampoos had superior antifungal activity due to the
presence of zinc pyrithione or a higher concentration of salicylic acid or selenium sulfide. Notwithstand-
ing, the tea tree oil shampoos demonstrated an appreciable antifungal activity due to synergistic effects
of tea tree oil, sodium lauryl sulphate, and salicylic acid. Furthermore, the tea tree oil shampoos were
stable during two months-long stability testing.
Conclusion: Thus, tea tree oil anti-dandruff shampoos have the potential to address the dandruff problem.
Received: November 09, 2020
Revised: January 04, 2021
Accepted: January 10, 2021
Keywords: Anti-dandruff, antifungal, formulation, shampoo, tea tree oil, marketed shampoos.
Dandruff is a non-inflammatory chronic dermal disorder
that affects almost half of the world’s population [1]. Dan-
druff is characterized as an unpleasant post-puberty problem
of hair and scalp, especially in males, which is often associ-
ated with itching due to flaky hair and scalp [2, 3]. A lipo-
philic yeast, namely Malassezia furfur, is associated with
dandruff production in sebum-rich areas such as the scalp
[4]. Dandruff is generally treated with shampoos containing
synthetic or natural ingredients. Synthetic ingredients such
as selenium sulfide or antifungals containing azole ring are
often used in chemical-based shampoos; however complete
eradication of dandruff is difficult with this regime [5].
Shampoos containing major components of synthetic origin
are also responsible for adverse reactions such as skin
*Address correspondence to this author at the Department of Pharmacy,
COMSATS University Islamabad, Lahore Campus, Lahore, Pakistan;
irritation, allergy, hair breakage, skin and hair discoloration
[6]. On the other hand, herbal shampoos have shown prom-
ising results in addressing the issue of dandruff. Herbal
shampoos containing natural plant extracts are usually com-
patible with the skin and may not elicit side effects that are
often associated with synthetic ingredients [7].
Many natural extracts from plants have beneficial ef-
fects on skin and hair due to the presence of vitamins,
amino acids, sugars, glycosides, phytohormones, bioflavo-
noids, fruit acids, and essential oils and are commonly
used in shampoos [8]. Various plants such as Azadirachta
indica (Neem), Ocimum sanctum (Tulsi), Aloe vera (aloe),
Terminalia chebula (harda, haritaki), Terminalia bellirica
(bahera), Sapindus mukorossi (Reetha), Phyllanthus embli-
ca (Amla), and Acacia concinna (Sheekakai) have been
traditionally used for hair cleansing, conditioning, and
removing dandruff in sub-continent including Pakistan and
India [9]. Although these plants have been used for centu-
ries for hair care, yet commercial herbal shampoos mainly
2 Current Cosmetic Science, XXXX, Vol. X, No. X Umar et al.
consist of synthetic ingredients boosted with natural prod-
ucts [10].
Essential oil from the plant Melaleuca alternifolia, (tea
tree oil) has recently shown promising results against skin
infections because of its strong antibacterial, anti-fungal,
and anti-viral activities[11]. Tea tree oil contains a number
of hydrocarbons and terpenes; however, terpene–4–ol is the
major component.Terpene–4–ol is believed to have anti-
microbial activity and could possibly be effective against
Malassezia furfur[12]. Satchell and co-workers have
demonstrated that 5% tea tree oil shampoo was effective
against dandruff without any skin irritation [13]. However,
they did not disclose the complete composition of the sham-
poo they used in their study.
Here, we have described a new shampoo formulation
containing tea tree oil as a dandruff remedy. The prepared
shampoos were extensively characterized for various quality
attributes. More importantly, the prepared shampoos were
compared with five commercially available shampoos in
Pakistan, namely Vatika, Clear for Men, Head & Shoulder,
Selsun blue, and Garnier. The anti-dandruff activity of tea
tree oil shampoo and marketed shampoos was determined
by anti-microbial activity against Candida albicans as a
model fungal strain [14].
2.1. Materials
Tea tree oil was procured from Xpel marketing Ltd
(England). Sodium lauryl sulphate, salicylic acid, sodium
EDTA, guar gum, Tween 80, sodium hydroxide, urea, and
xanthan gum were purchased from Sigma-Aldrich (Germa-
ny). Lanolin was sourced from Suru Chemicals & Pharma-
ceuticals Private Ltd. (India). Triethanolamine was procured
from Merk (Germany). The purified water that was prepared
at an in-house facility was used throughout the studies.
2.2. Formulation of Anti-dandruff Shampoo
Initially, a total of eight formulation trials were conduct-
ed in order to find an optimal shampoo formulation. In all
eight trials, we fixed the quantities of SLS, lanolin, sodium
EDTA, Tween 80, urea, and salicylic acid, whilst we sys-
tematically studied the effect of xanthan gum, guar gum,
and gelatin, as given in Table 1. The shampoo formulations
were prepared by mixing the ingredients in distilled water
using a magnetic stirrer operating at 500 RPM. The final pH
of the shampoo was adjusted between 5 6 either by 0.4N
NaOH solution or 2% triethanolamine solution. The pre-
pared shampoo trials were evaluated for physical appear-
ance, and the most stable formulation was selected for mak-
ing the anti-dandruff shampoo by adding tea tree oil at vari-
ous concentrations, namely 0.5%, 1%, 1.5%, 2%, 2.5%, and
3% designated by S1, S2, S3, S4, S5, and S6, respectively.
2.3. Characterization of Shampoos
2.3.1. Organoleptic Properties
The prepared anti-dandruff shampoos were compared
with the marketed anti-dandruff shampoos based on sensory
evaluation. The shampoos were inspected for color, clarity,
odor, andtexture [15].
2.3.2. Determination of pH
The pH values of prepared shampoos and marketed se-
lected shampoos were determined using a digital pH meter
at ambient temperature 25 ± 0.2°C.
Table 1. Various trials of shampoo formulations.
Ingredients (Grams)
Sodium EDTA
Xanthan gum
Guar gum
Tween 80
Sodium hydroxide
Salicylic acid
Distilled water
q.s. to make 100 mL
Formulation and In Vitro Characterization of Tea Tree Oil Current Cosmetic Science, XXXX, Vol. X, No. X 3
2.3.3. Viscosity Measurements
The viscosity of shampoos was determined using a
Brookfield viscometer (BDV-8S) equipped with an L3 spin-
dle, and the spindle rotation speed was set at 1.5, 3, 6, 12,
30, and 60 RPM. Viscosity measurements were performed
in triplicate at ambient temperature (25 ± 0.2°C).
2.3.4. Solid Contents Determination
The percentage of solid contents was measured by the
loss-on-drying method. Briefly, 5 g of each shampoo formu-
lation was poured in clean, dry, and pre-weighed Petri dish-
es, and the final weight was recorded. This was followed by
placing Petri dishes in a convection oven set at 50°C for 1 h
or until shampoos were completely dried. The driedPetri
dishes were weighed again, and the solid content after dry-
ing was estimated using the following formula [9]:
Where,Wo is the initial weight of the sample while W1 is the
weight of solid contents.
2.3.5. Surface Tension Determination
Surface tension was measured using a stalagmometer by
making a 10% dilution of each shampoo in distilled water at
ambient temperature (25 ± 0.2°C). The surface tension of
shampoos was determined in triplicate using the formula
given below:
Where,γ1 is the surface tension of shampoo, n1is the number
of drops of shampoo, n2 is the number of drops of water, ρ1
is the density of shampoo, ρ2 is the density of water, and γ2 is
the surface tension of water.
2.3.6. Foam Volume
The foam volume of the prepared shampoos and the
marketed shampoos was determined by the shake cylinder
method, as described previously in a study[9]. Briefly, 50
mL of 10% shampoo solution was added to a 250mL gradu-
ated cylinder and shook 10 times. The total volume of foam
after 1, 2, 3, and 4 min of shaking was recorded immediate-
ly using the graduates of the cylinder. The process was re-
peated three times.
2.3.7. Dirt Dispersion
Two drops of shampoo were added to 10 mL of distilled
water in a test tube, followed by the addition of one drop of
Indian ink. The test tube was stoppered and gently shaken10
times. The amount of Indian ink was visually estimated in
the foam as none, light, moderate and heavy. Shampoos that
caused the color to stay in the foam were considered low
2.3.8. Wetting Time
The wetting time of shampoos was determined by drop-
ping 50 mL of 1% aqueous shampoo solution on 1g of wool
yarn in a 100 mL beaker [17]. The time when wool yarn
started to float at the surface of the shampoo solution and
when it started to sink was recorded carefully by a stop-
watch. The mean values and standard error of at least three
replicates were reported.
2.3.9. Detergency Power
Detergency power (DP) of each shampoo was estimated
by soaking 5 grams of wool yarn in grease, weighing it, and
then placing it in 100 mL of 1% aqueous solution of sham-
poo in a 250 mL capacity flask. The flask was then fixed in
a shaking water bath that was set at 35°C and was shaken
for 4 min at the rate of 50 agitations per min. Afterward, the
solution was removed from the flask, and the wool yarn was
allowed to dry. After drying, the wool yarn was weighed,
and the detergency power was calculated by using the fol-
lowing equation [18]:
Where, C is the initial weight of grease and T is the weight
of grease after removing it from the shampoo solution.
2.3.10. Antimicrobial Activity of Shampoos
Anti-microbial activity of formulated shampoos and the
marketed shampoos was determined using the agar well
diffusion method against Candida albicans culture[19, 20].
Briefly, Sabouraud Dextrose Agar (SDA) media was pre-
pared and poured into sterilized Petri dishes. After solidifi-
cation of media in the Petri dishes, 4 mm diameter hole was
made using a sterilized cork borer. This was followed by
adding 3 mL of Candida albicans culture and was spread
using a sterilized glass spreader. The holes were filled with
150 µL of shampoo, and Petri dishes were covered immedi-
ately with a lid and sealed with parafilm. The sealed Petri
dishes were placed in an incubator operating at 37°C for 24
h. After 24 h, zones of inhibition were measured and report-
ed as the mean and standard deviation of at least three repli-
cates. Tea tree oil solutions of respective concentrations (as
used in formulated shampoos) were used to compare zones
of inhibition of each formulated tea tree oil shampoo. This
was further compared with the antifungal activity of the
marketed shampoos.
2.3.11. Stability Studies
Stability studies on shampoos were conducted by plac-
ing them in an incubator at 45°C and 75% relative humidity
for 2 months and were regularly evaluated for a physical
appearance at 1, 4, 7, 15, 30, and 60 days.
2.3.12. Statistical Analysis
All the experiments were conducted in triplicate, and
their mean ± standard deviation was reported. One-way
ANOVA was applied to check the significance of the differ-
ence, and the p values<0.05 were considered significant.
The data was statistically analyzed using Origin software
(version 8.5, OriginLab, USA).
Here we have described a new anti-dandruff shampoo
formulation containing tea tree oil at various concentrations.
Initially, we have performed various trials to find a suitable
shampoo base to incorporate tea tree oil. Trial shampoos
4 Current Cosmetic Science, XXXX, Vol. X, No. X Umar et al.
were formulated using different natural polymers such as
xanthan gum, guar gum, and gelatin and were evaluated for
their physical appearance, pH, and day-long stability.
Among eight trial formulations, F2 appeared to be more
stable than any other formulation after keeping it at 45°C for
24 h, and it had a pH value of 5.8, which is very close to the
skin pH. Additionally, the physical appearance of shampoo
with a formulation containing guar gum (F2) was more ap-
pealing in terms of texture and clarity. Furthermore, sham-
poo formulation with triethanolamine as pH adjuster was
found to have a pungent smell as compared to the formula-
tion containing sodium hydroxide as a pH adjuster; thus
sodium hydroxide was deemed suitable. Finally, based on
initial observations, F2 formulation was chosen for incorpo-
ration of tea tree oil at 6 concentrations, namely 0.5%,1%,
1.5%, 2%, 2.5%, and 3%. The prepared tea tree oil sham-
poos were also compared with marketed shampoos, namely
Vatika, Clear for Men, Head & Shoulder, Selsun blue, and
Garnier Fructis.
3.1. Organoleptic Properties
All the formulated shampoos and marketed shampoos
were evaluated for color, clarity, odor, and texture. The re-
sults of organoleptic evaluations are summarized in Table 2.
In the formulated shampoos, we did not add any color or
fragrance; however, the presence of lanolin resulted in pale
yellow colored shampoo formulations. On the other hand,
all the marketed shampoos were perfumed and colored for
their aesthetic appeal. The tea tree oil shampoos appeared
cloudy with gel-type texture whilst the marketed shampoos
appeared shiny with gel-type texture. While the formulated
shampoos were acceptable, the marketed shampoos had a
clear edge over the tea tree oil shampoos in terms of appear-
ance and odor.
3.2. pH and Viscosity Measurements
Our prepared shampoos had pH in the range of 5.4 5.7,
which is close to the pH of the skin, as given in Table 2.
However, the marketed shampoos showed a markedly vari-
able pH value from 3.9 to 7.4. Hair consists of tiny scales
known as cuticles, which are sensitive to pH, especially al-
kaline pH, which opens up these scales, ultimately leading
to hair damage. Thus a slightly acidic pH is favourable [21].
Furthermore, the slightly acidic pH of shampoos increases
the hair’s quality, decreases eye irritation, and maintains the
ecological balance of the scalp [22].
The viscosity of tea tree oil shampoos and marketed
shampoos was determined at various spindle speeds. As can
be seen from Fig. (1A), the viscosity of formulated sham-
poos decreased from 4170.5cP to 1114.8cP with increasing
tea tree oil concentrations from 0.5% to 3%at a spindle
speed of 1.5 rpm. It was not clear why the viscosity of
shampoos decreased with increasing tea tree oil concentra-
tion; one possibility could be the modification in the gelling
ability of natural hydrophilic polymer by adding the essen-
tial oil. This would, in turn, had reduced the overall viscosi-
ty of the shampoo formulation. Previous research has re-
vealed an alteration in pH by increasing the tea tree oil con-
tents, thereby reducing the viscosity [23], however, this was
not true in our study, and the pH values remained fairly sta-
ble with increasing tea tree oil contents (Table 2). Neverthe-
less, the important factor is pseudo-plastic behaviour which
is a desirable attribute in shampoo formulations [24]. Re-
sults revealed that the viscosity of each of the tea tree oil
shampoo was decreased with increasing spindle speed from
Table 2. Organoleptic properties and pH of anti-dandruff shampoos.
Tea Tree Oil Anti-Dandruff Shampoos
5.7 ± 0.0
5.6 ± 0.0
5.7 ± 0.1
5.6 ± 0.1
5.4 ± 0.0
5.5 ± 0.1
Marketed Shampoos
Head & Shoulder
7.4 ± 0.2
Selsun Blue
3.9 ± 0.0
Garnier Fructis fortifying shampoo
4.9 ± 0.6
5.4 ± 0.1
6.3 ± 0.1
Abbreviations: PY=Pale Yellow, C=Cloudy, GT=Gel Type, W=White, BG=Blue-Green, SB=Sky Blue, PB=Pale Blue, Y=Yellow, S=Shiny, PL=Pleasant.
Formulation and In Vitro Characterization of Tea Tree Oil Current Cosmetic Science, XXXX, Vol. X, No. X 5
1.5 rpm to 60 rpm, which confirmed the pseudo-plastic be-
haviour of prepared shampoos [25]. On the other hand, the
marketed shampoos had significantly higher(p<0.05) viscos-
ity as compared with that of the formulated tea tree oil
shampoos (Fig. 1B), possibly because of compositional dif-
ference in formulated and marketed shampoos. Neverthe-
less, a similar trend of decreasing viscosity with increasing
spindle speed was observed for the marketed shampoos.
Generally, a decrease in viscosity with increasing shear rate
is a favorable activity, which helps in the easy spreading of
shampoo on hair [26].
Fig. (1). Viscosities of (A) Tea Tree Anti-Dandruff shampoos and
(B) marketed shampoos. (A higher resolution / colour version of
this figure is available in the electronic copy of the article).
3.3. Percentage Solid Contents
For good quality anti-dandruff shampoos, solid content
should ideally be between 20 30%. This allows for easy
rinsing and removal from the hair [7]. Fig. (2A & B) depicts
solid contents in the prepared tea tree oil anti-dandruff
shampoos and the marketed shampoos. The solid contents in
formulated shampoos ranged from 20.03 22.25%, whilst
the solid contents in marketed shampoos ranged from 21.96
32.20%. All the tea tree oil shampoos had a lower range of
solid contents, thus deemed excellent. On the other hand,
marketed shampoos, namely Garnier Fructis and Clear had
slightly higher solid content, whilst Head & Shoulder, Sel-
sun Blue, and Vatika shampoos had solid content within an
ideal range.
Fig. (2). Percentage of solid contents in (A) tea tree anti-dandruff
shampoos and (B) marketed shampoos. (A higher resolution / col-
our version of this figure is available in the electronic copy of the
3.4. Measurement of Surface Tension
Surface tension is linked with the cleaning ability of
shampoos. Surfactants have the ability to reduce the surface
tension and the lesser the surface tension, the higher the
cleaning ability. Good shampoos generally reduce the sur-
face tension of water, which is approximately 72.28
dyn/cm[25]. The measured surface tensions of tea tree oil
and marketed shampoos are listed in Table 3. As can be
seen from Table 3, all the shampoos had surface tension in
the range of 24.02 36.27 dyn/cm, which is sufficiently
below the surface tension of pure water. Thus, all shampoos
were able to significantly reduce the surface tension of wa-
ter and were deemed good for their cleaning ability. How-
ever, the surface tension of marketed shampoos was signifi-
cantly less (p<0.05) than that of the tea tree oil shampoos.
Furthermore, all tea tree oil shampoos reduced the surface
tension significantly, yet no correlation existed between tea
tree oil concentration and reduction in surface tension.
3.5. Foam Volume
From the customer perspective, foaming has paramount
importance in the quality of shampoos, however, there is no
correlation existed between foaming and cleaning [25]. Fig.
6 Current Cosmetic Science, XXXX, Vol. X, No. X Umar et al.
(3A & B) depicts the volume of foam produced by tea tree
oil shampoos and marketed shampoos in distilled water at
different time points. All the shampoos (formulated and
marketed) resulted in foam in higher than 100 mL volume,
and the foam was stable throughout the testing time. The
foam volume produced by tea tree oil shampoos and mar-
keted shampoos was comparable and no significant difference
existed. The foaming ability of shampoos is correlated with
the surface tension reduction by the surfactants. Without the
presence of surfactants, there will be enough surface tension
that could pull the bubbles closed, resulting in limited or no
foam at all. However, the presence of surfactants at the
air/water interface reduces the surface tension to a point
where the pull caused by the tension becomes negligible and
air bubbles stabilize for a longer duration of time[27]. Since
our formulated shampoos and marketed shampoos had surfac-
tants, a reduced surface tension resulted in bubble formation
as revealed by the foaming of shampoos.
3.6. Dirt Dispersion
Dirt dispersion is an important quality attribute of sham-
poos and it is linked with the cleansing action of shampoos.
Generally, shampoos that cause the ink to stay in the foam
instead of the liquid portion are considered poor quality prod-
ucts because of difficulty in washing away the ink, conse-
quently, the dirt would be difficult to remove from the hair
[16]. Among the prepared tea tree oil shampoos, S2, S3, S5,
and S6 had light ink in their foam section while no ink was
detected in the S1 and S4 shampoo formulation (Table 3). On
the other hand, all marketed shampoos except Selsun Blue
shampoo showed light ink distribution in the foam. Thus, tea
tree oil shampoos were similar to marketed shampoos, while
S1 and S4 shampoo formulations were found to be superior in
terms of cleansing action on the basis of dirt dispersion.
3.7. Wetting Time
The wetting ability of shampoos describes their efficacy
as cleansing products and it depends on the surfactant used.
The marketed shampoos had wetting time in the range of
4.24 6.03s (Table 3). In comparison, the formulated tea
tree oil shampoos showed wetting time in the range of 4.54
7.55s (Table 3). The results revealed that there was no
significant difference (p>0.05) in the wetting time of formu-
lated tea tree oil shampoos and the marketed shampoos.
Fig. (3). Foam Volume of (A) tea tree oil anti-dandruff shampoos
and (B) marketed shampoos. (A higher resolution / colour version
of this figure is available in the electronic copy of the article).
Table 3. A comparison of surface tension, dirt dispersion, and wetting time of tea tree oil and marketed shampoos.
Surface Tension (dyn/cm)
Dirt Dispersion
Wetting Time (Seconds)
35.61 ± 1.12
5.46± 0.09
36.27 ± 0.98
4.65± 0.11
34.73 ± 0.66
4.54± 0.32
36.09 ± 0.32
7.30± 0.15
35.15 ± 0.11
7.55± 0.12
34.54 ± 1.09
6.98± 0.50
Head & Shoulder
27.24 ± 0.87
4.40± 0.08
Selsun Blue
32.96 ± 0.54
4.24± 0.57
Garnier Fructis fortifying shampoo
24.02 ± 0.21
5.29± 0.03
27.03 ± 0.33
6.03± 0.12
29.28 ± 1.99
5.04± 0.85
Formulation and In Vitro Characterization of Tea Tree Oil Current Cosmetic Science, XXXX, Vol. X, No. X 7
3.8. Cleaning Action
The prepared tea tree oil shampoos and the marketed
shampoos were subjected to evaluation of detergency power
or cleaning action. Although experimental assessment of
detergency power is difficult to standardize, yet it is a gen-
eral agreement that good quality shampoos should effective-
ly remove the grease or oil from the hair [18]. Our results
showed that the cleaning action of tea tree oil shampoos was
in the range of 23.0 30.7% as compared to 28.6 30.5%
for the marketed shampoos, as depicted in Fig. (4A & B).
With little or no difference in cleaning action, we can say
that our formulated shampoos were as good as the marketed
Fig. (4). Cleaning action of (A) tea tree oil anti-dandruff shampoos
and (B) marketed shampoos. (A higher resolution / colour version
of this figure is available in the electronic copy of the article).
3.9. Antimicrobial Activity of Tea Tree Oil Anti-
Dandruff Shampoos
Antifungal activity of tea tree oil shampoos and the mar-
keted shampoos was studied against a model fungal strain,
namelyCandida albicans. Moreover, pure tea tree oil solu-
tions were also studied to compare the shampoo formula-
tion. In this case, pure tea tree oil solutions were considered
as a control to compare the effect of other ingredients of
formulations for their antifungal activity, and it was found
that the solutions had the same concentration as used in the
shampoo formulations. The antifungal activity in terms of
zones of inhibitions of pure tea tree oil, tea tree oil sham-
poos, and the marketed shampoos is listed in Table 4.
As can be seen from Table 4, pure tea tree oil solutions
had antifungal activity against Candida albicans, as demon-
strated by zones of inhibition. The inhibition of fungal
growth was concentration-dependent for pure tea tree oil
solutions. The fungal growth inhibition was more pro-
nounced in the formulated tea tree oil shampoos as demon-
strated by the values. Although no concentration-dependent
inhibition was observed, yet the inhibition zones were larger
than those produced with pure tea tree oil solution. The en-
hanced antifungal activity of tea tree oil shampoos was at-
tributed to the presence of SLS and salicylic acid, which
have some antifungal activity and contributed synergistical-
ly against the fungal strain [28, 29]. On the other hand, the
marketed shampoos had significantly higher (p<0.05) zones
of inhibition, thus had the higher antifungal activity against
Candida albicans. However, Selsun Blue had the lowest
antifungal activity as compared to other marketed shampoos
and its antifungal activity was insignificantly different
(p>0.05) from our formulated shampoos (Table 4). Overall,
the higher antifungal activity of the marketed shampoos was
attributed to a combined effect of zinc pyrithione, SLS, and
salicylic acid. Conclusively, the tea tree oil shampoos had
demonstrated an appreciable antifungal activity and could
serve as an excellent alternative to the marketed shampoos.
3.10. Stability Studies
Finally, the tea tree oil shampoos and the marketed
shampoos were compared in terms of their stability at vari-
ous temperatures, such as 4°C, 25°C, and 40°C (75% RH).
The shampoos were stored at each of the temperatures for 2
Table 4. Zone of inhibition of tea tree oil shampoos and their Standards.
Standard (Tea Tree
Oil Solution)
Zone of Inhibition
Tea Tree Oil
Zone of Inhibition
Zone of Inhibition
14.2± 0.4
23.8± 1.1
Head & Shoulder
33.35 ± 2.1
16.2± 0.6
22.3± 0.8
Selsun Blue
20.45 ± 0.9
17.4± 0.2
24.8± 0.3
Garnier Fructis
41.85 ± 2.8
18.6± 0.8
26.9± 1.4
35.5 ± 1.9
22.9± 0.9
20.1± 0.7
35.8 ± 2.6
21.7± 0.4
25.7± 1.7
8 Current Cosmetic Science, XXXX, Vol. X, No. X Umar et al.
months, and the physical appearance and pH values were
determined at various time points, as shown in Table 5-7.
As can be seen from the stability data at various conditions,
no change in physical appearance was observed, however,
an insignificant change (p>0.05) in pH values was observed
at all storage conditions. Since the pH change was small, we
can say that all the formulations were sufficiently stable
over the 2 months stability testing period.
Table 5. Stability studies on tea tree oil shampoos and the marketed shampoos at 4°C.
Stability Studies at 4°C
Shampoo Formulations
At 1st Day
At 4th Day
At 7th Day
At 15th Day
At 1 Month
At 2 Months
5.70± 0.01
5.73± 0.07
5.75± 0.21
5.78± 0.42
5.81± 0.00
5.88± 0.13
5.60± 0.18
5.61± 0.04
5.64± 0.08
5.66± 0.00
5.70± 0.02
5.76± 0.09
5.70± 0.14
5.70± 0.21
5.73± 0.07
5.75± 0.03
5.82± 0.08
5.88± 0.00
5.60± 0.00
5.63± 0.04
5.65± 0.03
5.69± 0.07
5.73± 0.00
5.76± 0.00
5.40± 0.04
5.42± 0.18
5.48± 0.22
5.52± 0.11
5.59± 0.10
5.50± 0.03
5.50± 0.00
5.53± 0.17
5.56± 0.2
5.60± 0.15
5.67± 0.06
Marketed Shampoos
Head & Shoulder
7.41± 0.01
7.43± 0.01
7.46± 0.01
7.49± 0.01
7.54± 0.01
7.59± 0.01
Selsun Blue
3.90± 0.00
3.91± 0.11
3.95± 0.16
4.01± 0.03
4.08± 0.08
4.12± 0.10
Garnier fructis
4.92± 0.00
4.95± 0.02
4.97± 0.10
5.02± 0.20
5.09± 0.19
5.14± 0.12
5.43± 0.17
5.47± 0.05
5.49± 0.00
5.54± 0.02
5.60± 0.01
5.64± 0.09
6.30± 0.04
6.33± 0.11
6.36± 0.03
6.41± 0.01
6.46± 0.08
6.52± 0.00
Table 6. Stability studies on tea tree oil shampoos and the marketed shampoos at 25°C.
Stability Studies at 25°C
Shampoo Formulations
At 1st Day
At 4th Day
At 7th Day
At 15th Day
At 1 Month
At 2 Months
5.70± 0.05
5.75± 0.00
5.77± 0.13
5.81± 0.05
5.84± 0.03
5.91± 0.09
5.60± 0.00
5.63± 0.07
5.67± 0.01
5.68± 0.21
5.73± 0.11
5.79± 0.06
5.70± 0.02
5.73± 0.09
5.76± 0.10
5.77± 0.11
5.85± 0.16
5.92± 0.05
5.60± 0.06
5.65± 0.03
5.68± 0.14
5.72± 0.12
5.76± 0.04
5.79± 0.07
5.40± 0.02
5.44± 0.07
5.49± 0.03
5.51± 0.05
5.54± 0.11
5.62± 0.00
5.50± 0.09
5.52± 0.14
5.55± 0.11
5.59± 0.12
5.63± 0.07
5.69± 0.03
Marketed shampoos
Head & Shoulder
7.41± 0.00
7.45± 0.04
7.48± 0.07
7.53± 0.03
7.57± 0.00
7.63± 0.15
Selsun Blue
3.90± 0.19
3.93± 0.11
3.97± 0.08
4.04± 0.05
4.11± 0.07
4.16± 0.10
Garnier fructis
4.92± 0.03
4.96± 0.00
4.99± 0.05
5.05± 0.02
5.09± 0.03
5.18± 0.08
5.43± 0.01
5.49± 0.05
5.52± 0.21
5.57± 0.11
5.63± 0.13
5.67± 0.09
6.30± 0.22
6.36± 0.11
6.38± 0.06
6.43± 0.02
6.49± 0.03
6.56± 0.04
Formulation and In Vitro Characterization of Tea Tree Oil Current Cosmetic Science, XXXX, Vol. X, No. X 9
Table 7. Stability studies on tea tree oil shampoos and the marketed shampoos at 40°C.
Stability Studies at 40°C and 75% RH
Shampoo Formulations
At 1st Day
At 4th Day
At 7th Day
At 15th Day
After 1 Month
After 2 Months
5.70± 0.07
5.78± 0.02
5.80± 0.11
5.85± 0.21
5.86± 0.06
5.95± 0.00
5.60± 0.02
5.66± 0.08
5.71± 0.04
5.73± 0.11
5.76± 0.04
5.84± 0.05
5.70± 0.11
5.76± 0.05
5.79± 0.02
5.79± 0.08
5.88± 0.03
5.96± 0.13
5.60± 0.06
5.68± 0.03
5.72± 0.11
5.75± 0.09
5.79± 0.00
5.83± 0.04
5.40± 0.01
5.47± 0.04
5.52± 0.13
5.54± 0.07
5.57± 0.11
5.65± 0.05
5.50± 0.01
5.55± 0.00
5.57± 0.01
5.63± 0.07
5.67± 0.02
5.73± 0.11
Marketed shampoos
Head & Shoulder
7.41± 0.14
7.47± 0.21
7.51± 0.08
7.56± 0.19
7.62± 0.07
7.66± 0.02
Selsun Blue
3.90± 0.06
3.95± 0.11
4.02± 0.09
4.09± 0.04
4.18± 0.01
4.23± 0.03
Garnier fructis
4.92± 0.03
4.98± 0.05
5.04± 0.18
5.09± 0.10
5.15± 0.02
5.26± 0.01
5.43± 0.08
5.52± 0.03
5.56± 0.00
5.62± 0.09
5.67± 0.11
5.71± 0.06
6.30± 0.07
6.39± 0.11
6.42± 0.15
6.48± 0.09
6.54± 0.03
6.62± 0.00
The current study demonstrated successful preparation
of tea tree oil anti-dandruff shampoos at various concentra-
tions of tea tree oil. The formulated shampoos had excellent
properties in terms of pH, viscosity, wetting time, detergen-
cy and cleaning action, dirt dispersion, and antifungal activi-
ty. Furthermore, the formulated shampoos had comparable
properties with the marketed shampoos. In conclusion, tea
tree oil can be used to formulate anti-dandruff shampoos at
various concentrations, and different concentrations may be
advantageous to various skin and hair types. However, fur-
ther studies are warranted to establish the efficacy of formu-
lated shampoos in human volunteers with different skin and
hair types.
Not applicable.
Not applicable
Not applicable.
Not applicable.
The authors have no conflicts of interest to declare.
The authors would like to thank Professor Muhammad
Jamshid, Dean of Faculty of Pharmacy, the University of
Central Punjab, for providing laboratory facilities to conduct
this research.
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Full-text available
Most of the traditional strategies used for facing the management of insect pest and diseases have started to fail due to different toxicological issues such as the resistance of target organism and the impact on environment and human health. This has made mandatory to seek new effective strategies, which minimize the risks and hazards without compromising the effectiveness of the products. The use of essential oils, their components and semiochemicals (pheromones and allelochemicals) has become a promising safe and eco-sustainable alternative for controlling insect pest and pathogens. However, the practical applications of this type of molecules remain rather limited because their high volatility, poor solubility in water and low chemical stability. Therefore, it is required to design strategies enabling their use without any alteration of their biological and chemical properties. Oil-in-water nano/microemulsions are currently considered as promising tools for taking advantage of the bioactivity of essential oils and their components against insects and other pathogens. Furthermore, these colloidal systems also allows the encapsulation and controlled release of semiochemicals, which enables their use in traps for monitoring, trapping or mating disruption of insects, and in push-pull strategies for their behavioral manipulation. This has been possible because the use of nano/microemulsions allows combining the protection provided by the hydrophobic environment created within the droplets with the enhanced dispersion of the molecules in an aqueous environment, which favors the handling of the bioactive molecules, and limits their degradation, without any detrimental effect over their biological activity. This review analyzes some of the most recent advances on the use of emulsion-like dispersions as a tool for controlling insect pest and pathogens. It is worth noting that even though the current physico-chemical knowledge about these systems is relatively poor, a deeper study of the physico-chemical aspects of nanoemulsions/microemulsions containing essential oils, their components or semiochemicals, may help for developing most effective formulations, enabling the generalization of their use.
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Background Dandruff is a major problem of hair, which cannot be fully cured with the aid of chemicals. An attempt has been made to prepare a poly-herbal anti-dandruff hair care formulation and it is standardized to ensure its stability and quality aspects. Objective The present work is oriented at the synthesis of herbal anti-dandruff hair formulation and the estimation of its various aspects for its valuability. Materials and Methods The herbal preparation was prepared in the laboratory, on the basis of decided proportion, with the help of all the herbal contents. The pack was gauged for its various parameters. Results The results were concluded to be sufficiently ample for the assessment of herbal formulation. The findings of different aspects vindicated the worth of the formulation. Conclusion Herbal hair formulation has been formulated and assessed using the various aspects. It serves as a good alternate, devoid of any side effects. The results can be unified while evolving the pharmacopoeial standards.
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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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Hair is an important part of the body appeal and its look is a health indicator. Accordingly, recent advances in hair science and hair care technologies have been reported in literature claiming innovations and strategies for hair treatments and cosmetic products. The treatment of hair and scalp, primarily, involved the use of shampoo for an effective, but gentle cleansing; however, for years, the shampoo is considered not only as a cosmetic product having the purifying purpose, but it is also responsible for maintaining the health and the beauty of hair, imparting gloss and improving manageability. For meeting the needs of a multitasking formulation, following also the recent marketing-trend addressed to the "natural world", new challenges for cosmetic technology are aimed towards the research of natural ingredients, as well as new techniques for shampoo formulation. Regarding the recent development of solid shampoos, little information is available about their use, formulation and advantages. This review is largely focused on the description of solid shampoos, mainly based on the use of clays, herbs or flours as washing bases alternative to the traditional ones, consisting of a combination of synthetic surfactants, together with other usual ingredients expected in a shampoo formulation.
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This study aimed to investigate Malassezia furfur inhibitory activity of the fermented product from Ocimum sanctum and develop an antidandruff shampoo. The fermented product was obtained by the fermentation process of the aerial part of O. sanctum. Total soluble protein was detected in the fermented product with the amount of 65.32 ± 0.14 mg/100 mL, whereas there was no organic acid. The inhibitory activity against four strains of M. furfur (No. 133, 656, 6000, and 7966) of the fermented product and shampoos containing the fermented product were investigated by broth dilution and agar diffusion method, respectively. The fermented product possessed high antifungal activity with the minimum inhibitory concentrations for 50% (MIC50) of M. furfur 133, 656, 6000, and 7966 of 0.125, 0.25, 0.125, and 0.125 mg/mL, respectively. Interestingly, the antifungal activity against M. furfur 656 was comparable to that of ketoconazole. Shampoo formulation C, which was the best formulation in terms of characteristics and stability, obtained a high level of satisfaction scores in terms of hair smoothness, hair shine, ease in combing, frizz reduction, and triboelectric reduction while brushing. Additionally, the shampoo containing 2% (w/w) of the fermented product of O. sanctum also possessed inhibitory activity against M. furfur 133, 656, 6000, and 7966 with inhibition zones of 13.2 ± 1.6, 12.8 ± 1.1, 18.7 ± 0.3, and 17.0 ± 1.1 mm respectively. Therefore, this shampoo was suggested for use as an antidandruff shampoo.
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Objective: This study aimed to formulate tea tree oil into a nanoemulsion gel dosage form and evaluate its physical stability and antibacterial activity.Methods: Nanoemulsion gels were formulated with various concentrations of tea tree oil, namely, 5%, 7%, and 9%, using Tween-80 as a surfactantand propylene glycol as a cosurfactant. The tea tree oil nanoemulsion gels showed a stable physical appearance over 8 weeks of storage at lowtemperature (4±2°C) and room temperature (25±2°C), cycling test, and centrifugation test.Results: The best formula was nanoemulsion gel formulation 1 (F1), which contained 5% tea tree oil, due to its good stability, smaller globule size,and greater viscosity. The results for antibacterial activity, determined by in vitro study, showed that the tea tree oil nanoemulsion gels exhibitedantibacterial activity against Propionibacterium acnes through the formation of an inhibition zone.Conclusion: Higher concentrations of tea tree oil in nanoemulsion gels (5%, 7%, and 9%) showed greater mean inhibition zones (28.33±0.88 mm,30.33±0.33 mm, and 31.67±0.33 mm, respectively).
Darunaka is a disease concerned to hair root which can be correlate with Dandruff, which is the most common cause for hair loss. It is more of a social problem and affects aesthetic value as a person with Dandruff is quite likely to feel down on the social ring. Acharya Sushruta has mentioned Darunaka under Kshudraroga’s with symptoms like rukshata, kandu, twak sphutana etc. Dandruff can occur due to air pollution, water pollution, changed life style, irregular daily regimen, poor hygiene and immune system, sweating, mental stress etc which can lead to several bacterial and fungal infections. It is a common scalp disorder affecting almost half of the population at the pre-pubertal age and of any gender and ethnicity. No population in any geographical region would have passed through freely without being affected by dandruff at some stage in their life. Shiroabhyanga, Nasya, Shirodhara , Shirobasti,Shirolepa and Siravyadha is the line of treatment for Darunaka in the classics. In traditional medicine paste of leaves of Elaeocarpus serratus and Psidium guajava are using for thousand years as a home remedy for dandruff in Sri Lanka .In spite of the present medical solutions Dandruff relapses quite often. A permanent and holistic solution is the need of the hour. Hence, this work is intended to study the efficacy of Elaeocarpus serratus and Psidium guajava leaves paste for dandruff. Thirty patients were selected; advised to apply the paste covering whole scalp area for one hour and rinse it, for a period of one month every alternative day. Signs and symptoms were assessed by using grading system and observed the significant reduction of symptoms. Considering the total treatment assessment more than 40% patients gain complete remission and 30% got marked improvement. No adverse reaction reported in this study period. It is concluded that Elaeocarpus serratus and Psidium guajava leaves paste is very beneficial to manage Dandruff.
Aim A recent approach in shampoo research has been to find a natural alternative for synthetic detergents that have deteriorative effects on hair follicles. This study aimed at the formulation of a completely herbal shampoo containing a natural foaming agent, in addition to having conditioning, antioxidant, and antimicrobial effects. Material and Methods The leaves of Salix babylonica L., Ziziphus spina-christi L. (Willd), and Glycyrrhiza glabra rhizomes were extracted with 70% methanol then quantified for their phenolic and flavonoid contents using colorimetric assays that were qualitatively identified by Ultra-performance liquid chromatography (UPLC) with tandem mass spectrometry (MS-MS). The mineral content was also estimated. The radical scavenging activity was estimated using 1,1-diphenyl-2-Picrylhydrazyl (DPPH) and the half maximal inhibitory concentration (IC50) was determined. Additionally, the antimicrobial activity was tested using agar diffusion assay and compared to vancomycin and ketoconazole. Four formulations, consisting of the different plant extracts and a combination of the extracts, were prepared and evaluated for several physicochemical properties. The best formula was evaluated for its conditioning effects using scanning electron microscope and blind touch tests by asking volunteers for grading the formulations. Results and Discussion UPLC-MS-MS analysis of S. babylonica and Z. spina-christi allowed tentative identification of 12 phytoconstituents in each. Z. spina-christi showed the highest phenolic content and a high copper, zinc, and manganese content beside the best antioxidant activity, whereas G. glabra had a high potency against Bacillus cereus and Candida albicans. The polyherbal shampoo formulation (F4) was selected as an optimized formulation because of a high foam stability after 4 min, low wetting time (2 s), surface tension reduction, and comparable results for percent solid content. F4 showed good conditioning effect and consumer contentment. Conclusion The formulated polyherbal shampoo is chemical free, extra-nourishing shampoo with excellent conditioning, cleansing, and antimicrobial effects.
Objective Dandruff is a complex skin condition characterized by unpleasant itching and flaking of the scalp. It is primarily attributed to the over colonization of Malassezia yeasts such as M. globosa and M. restricta. Some studies also suggest the involvement of Staphylococci bacteria in dandruff disease pathogenesis. We aimed to access the effectiveness of anti‐dandruff treatments by determining the efficacy of the active antifungal agents alone or in commercial shampoo formulations against Malassezia and Staphylococcus. Methods The minimum inhibitory concentrations of three anti‐dandruff shampoo antifungals (zinc pyrithione, ketoconazole and ciclopirox) and the witch hazel extract, hamamelitannin were tested against commensal Malassezia and Staphylococcus species using broth microdilution methods. In experiments simulating shampoo exposure and washing conditions on the scalp, we also tested the ability of the above agents in shampoo formulation (Head and Shoulders®, Ketomed®, Sebiprox®, Erol Healthcare Hair Shampoo®, respectively) along with a generic over‐the‐shelf shampoo to inhibit microbial growth. Results Ketomed® and Head and Shoulders® shampoo were the most effective treatments against Malassezia in in vitro assays and washing simulation. experiments. Erol Healthcare Hair Shampoo® was less effective against Malassezia as it required a longer contact time to achieve growth inhibition for some species. Sebiprox® showed variable efficacy in washing and contact time experiments while the generic over‐the‐shelf shampoo was the least effective in inhibiting Malassezia and Staphylococcus growth. Conclusion From these findings, it is reasonable that patients with dandruff may benefit from applying specific antifungal shampoo although results may vary with microbial species, time of contact and shampoo formulation components. This article is protected by copyright. All rights reserved.
Surfactants form the core of all shampoo formulations, and contribute to a wide range of different benefits including cleansing, foaming, rheology control, skin mildness and the deposition of benefit agents to the hair and scalp. The purpose of this review is to assist the design of effective, modern, shampoo surfactant technologies. The mechanisms through which surfactants help deliver their effects are presented, along with the appraisal techniques through which surfactant options can be tested and screened for product development. The steps that should be taken to select the most appropriate blend of surfactants are described, and useful information on the most widely used surfactants is provided. The review concludes with an examination of recent developments in ‘greener’ surfactants, ‘sulphate-free’ technologies and structured liquid phases for novel sensory properties and for suspending benefit agents.