Content uploaded by Nattaya Lourith
Author content
All content in this area was uploaded by Nattaya Lourith on Feb 17, 2017
Content may be subject to copyright.
Fax +49 761 4 52 07 14
Information@Karger.com
www.karger.com
Accessible online at:
www.karger.com/fok
Original Article · Originalarbeit
Forsch Komplementmed 2015;22:226–229
DOI: 10.1159/000432407
Anti-dandruff Hair Tonic Containing Lemongrass
(Cymbopogon flexuosus) Oil
Wannee Chaisripipat Nattaya Lourith Mayuree Kanlayavattanakul
School of Cosmetic Science, Mae Fah Luang University, Chiang Rai, Thailand
Schlüsselwörter
Schuppen · Klinische Studie · Kosmetische Wirksamkeit ·
Haarerkrankungen · Pflanzenkosmetik ·
Traditionelle Medizin
Zusammenfassung
Hintergrund:
Zur Behandlung von Schuppen erfreuen
sich natürliche Heilmittel immer größerer Beliebtheit.
Material und Methoden:
Es wurde eine randomisierte,
doppelblinde, Placebo-kontrollierte Studie an 30 Teilneh-
mern (20–60 Jahre) aus Thailand mit Schuppen-Level 3
auf der D-Squame
®
-Skala durchgeführt. Sie wurden mit
einem ein einfach anwendbaren Haartonikum mit dem
ätherischen Öl Zitronengras (Cymbopogon flexuosus)
behandelt, das insbesondere gegen lipophile Hefepilze
wirksam ist. Die in einem Prätest am häufigsten bevor-
zugte Variante des Basistonikums (p < 0,05) war mit
einem Ätherischölanteil von 5, 10 bzw. 15% angerei-
chert. Angewendet wurde das Präparat zweimal täglich,
wobei jeder Teilnehmer auf der einen Kopfhälfte das
Basistonikum (Placebo) und auf der anderen Kopfhälfte
das verumhaltige Tonikum anwendete. An Tag 7 und 14
nach Anwendungsbeginn wurde die Wirksamkeit mittels
der D-Squame
®
-Skala bewertet.
Ergebnisse:
Die Reduk-
tion der Schuppen mit den 5, 10 bzw. 15%igen Zitronen-
grasöl-Tonika war nach 7 Tagen signifikant (p < 0,005;
33, 75 bzw. 51%). Am Tag 14 stieg der Effekt weiter an
(p < 0,005; 52, 81 bzw. 74%).
Schlussfolgerung:
Das Prä-
parat mit dem 10%igen Ätherischölanteil war am wirk-
samsten.
Keywords
Dandruff · Clinical trial · Cosmetic efficacy ·
Hair disorder ·
Herbal cosmetics · Traditional medicine
Summary
Background: Natural remedies for treating dandruff are
becoming popular. Materials and Methods: A rand-
omized, double-blind, placebo-controlled, split-head ef-
ficacy evaluation was conducted 30 Thai volunteers aged
20–60 years experiencing dandruff measured at level 3
on D-Squame
®
scale. An easy to use hair tonic contain-
ing essential oil of lemongrass (Cymbopogon flexuosus)
active against lipophilic yeasts was developed and then
evaluated for efficacy and preference. The base formula-
tion with the significantly highest preference (p < 0.05)
was stowed with the oil at 5, 10 or 15%. Subjects applied
the formulation twice a day, and an efficacy assessment
with D-Squame
®
scale was conducted on days 7 and 14
of application. Results: The application of lemongrass oil
hair tonics with 5, 10, or 15% reduced dandruff signifi-
cant (p < 0.005) at day 7 (33, 75, and 51%) and increased
the effect even more (p < 0.005) at day 14 (52, 81, and
74%). Conclusion: The hair tonic formulation with 10% of
lemongrass oil seems to be the most effective prepara-
tion.
© 2015 S. Karger GmbH, Freiburg
Published online: July 14, 2015
Nattaya Lourith
School of Cosmetic Science
Mae Fah Luang University
Chiang Rai 57100, Thailand
nattayal@mfu.ac.th
© 2015 S. Karger GmbH, Freiburg
1661–4119/15/0224–0226$39.50/0
Downloaded by:
Siriraj Medical Library, Mahidol University
198.143.39.97 - 9/21/2015 5:10:32 AM
Anti-dandruff Hair Tonic Containing Lemongrass
(Cymbopogon flexuosus) Oil
Forsch Komplementmed 2015;22:226–229
227
Introduction
Dandruff is a cluster of corneocytes detached from the stratum
corneum of the scalp accumulating with parakeratotic cells. This
shedding of cells is influenced by seborrhea in addition to microor-
ganisms on the scalp. Normally, pathogen density on the healthy
scalp reaches 10
3–
10
5
organisms per mm
2
. Malassezia spp. is the
primary flora [1] found on the scalp; in dandruff scalp, this yeast
might multiply to 1.5
–
2 folds greater amount.
Therefore, many cosmetic-management products, particularly
shampoos, contain antifungal agents [2]. Scalp flake production
and parakeratosis decrease when treated, however Malassezia spp.
increases to the initial level within a couple of weeks after stopping
the treatment. In addition, apart from the inflammatory reaction
caused by the yeasts, it exacerbates the severity of dandruff that an-
tifungal treatments may not be able to minimize. Furthermore, the
contact time of anti-dandruff shampoo with the scalp is short;
rinse-off shampoos therefore are insufficient to treat the lipophilic
yeast Malassezia spp. In addition, sebum reduction would limit
itching and facilitate dandruff treatment. But even with a low
sebum excretion, the secreted compounds alter the inflammatory
response and worsen the dandruff [3].
Therefore, alternative treatments, particularly natural remedies,
are becoming a popular choice among consumers [4]; specifically,
these treatments are intended to be used in such a way that contact
time is prolonged, thus enhancing treatment efficacy [3].
The essential oil of West or East Indian lemongrass Cymbopo-
gon citratus or C. flexuosus is widely used in pharmaceutical prod-
ucts on the basis of its therapeutic and psychological effects [5]. C.
flexuosus not only functions as an anti-bacterial herb against sev-
eral human pathogens, but also exhibits antioxidant activity [6],
confirming its frequent use as a common traditional medicine
and an important culinary herb in Thailand and other Southeast
Asian countries. When used as a scalp treatment, C. flexuosus oil
inhibits Malassezia furfur, the major yeast associated with dan-
druff, at the minimum inhibitory concentration (MIC) of 6.70
ppm. Therefore, in the present study a hair tonic containing lem-
ongrass oil is evaluated for efficacy in dandruff-affected
volunteers.
Materials and Methods
Formulations; Stability and Preference Tests
A base hair tonic containing cremaphor RH-40, menthol, salicylic acid,
ethyl alcohol, propylene glycol, glycerin (Niamsiang, Thailand) and water was
formulated. We developed this hair tonic to have a less greasy formulation than
standard hair tonics used to treat dandruff. Physicochemical characteristics of
the formulated products were determined by pH-meter (B200, QiS, Oosterhout,
The Netherlands). We obtained 3 base formulas that were stable following ac-
celerated stability test by means of centrifugation assays (3,000 rpm for 15 min)
and 6 cycles of acceleration tests (heating and cooling at 4°C and 45°C for 48
h, each cycle); these formulations were included in our user preference test.
We recruited 10 healthy adult volunteers (5 males and 5 females) with dan-
druff to test these 3 formulations. Preference tests were carried out by all volun-
teers and were scored using a 5-point Likert scale (1 = greatly dislike; 5 = great
preference) using interview questionnaires developed by the research team. The
most preferred base formulation was selected for further development [8].
Commercial lemongrass oil (HH-CP lemongrass 100% of C. flexuosus) pur-
chased from Charabot (Grasse Cedex, France; Lot number: 0009003281) was
incorporated at 5, 10, or 15% into the most preferred stable basefor further
clinical evaluation.
Clinical Evaluation
Inclusion Criteria
A total of 30 healthy Thai volunteers between 20 and 60 years of age experi-
encing dandruff measured at level 3 on the D-Squame
®
(fig.1) were included in
the efficacy study described below. All recruited subjects were informed about
the study both in writing and verbally, and they signed a written consent form
that was approved by the ethical committee of the Mae Fah Luang University
prior to enrollment. All aspects of this study involving human volunteers were
in accordance with the most recent Declaration of Helsinki.
Irritation Test
A closed patch test using the base hair tonic (0.02 ml, 2% in alcohol) was
comparatively tested against the product containing lemongrass oil at different
concentrations for 2 h of contact time. Water was used as a negative control,
whereas 0.25% sodium lauryl sulfate (Namsiang) was the positive control. Ob-
servation was undertaken immediately and at 24, 48 and 72 h following Finn
chamber
®
(8 mm, SmartPractice, Phoenix, AZ, USA) removal. Mean Irritation
Index (MII) was calculated [9].
Efficacy Evaluation
All 30 subjects participating in the efficacy evaluation were asymptomatic
for 1 week, and had not used steroids or an anti-dandruff product for 4 weeks
prior to study enrollment. Subjects who were pregnant, lactating, or dieting
were excluded from the study. Any skin treatments on the scalp were not al-
lowed, as were smoking and consumption of alcohol. In addition, any partici-
pants who used the assigned product less than 90% of the required time were
not included in the final study analysis.
Fig. 1.
Dandruff scaling using D-squame® a), representative reduction of
dandruff from the group treated with lemongrass hair tonics, b) D0, c) D7, and
d) D14.
1 2 3 4 5
a)
5% 10% 15%
b)
5% 10% 15%
c)
15%
5% 10%
d)
Downloaded by:
Siriraj Medical Library, Mahidol University
198.143.39.97 - 9/21/2015 5:10:32 AM
Chaisripipat/Lourith/KanlayavattanakulForsch Komplementmed 2015;22:226–229
228
The subjects were asked not to apply any products to the scalp the night be-
fore starting the randomized, double-blind, placebo-controlled study. The vol-
unteers were asked to apply the test products twice daily (morning and evening)
by a split-head procedure for intra-individual comparative trials [8] in 3 groups.
Each group of the volunteers was therefore assigned 2 bottles of the test prod-
ucts; one was the base without lemongrass oil and one was lemongrass hair
tonic at different concentrations. The volunteers were randomly directed to use
5 drops (0.13 g) of the lemongrass hair tonic on one side of the head and the
base formulation on the other side at the same quantity. The same site of the
palm used for rubbing the product into the scalp and the same site of the head
were fixed throughout the study period. The control cleansing shampoo (No
More Tears, Johnson & Johnson, Thailand) was provided and used for hair
wash (10 g/washing) for a 2-day interval. The anti-dandruff efficacy evaluation
was conducted on day 7 (D7) and day 14 (D14) by means of an adhesive disc.
Participants’ hair was spread out as much as possible prior to use of the D-
Squame® disc, and the examined score (fig 1 A) of each side of the head was
recorded in each volunteer. The reduction (%) of dandruff was calculated by R
= (Dt-D0)/D0 ×100 (R = reduction efficacy; Dt = dandruff scale at D7 or D14;
D0 = dandruff scale at D0).
Statistical Analysis
The parameters were compared and analyzed using the Wilcoxon signed test
for stability evaluation, ANOVA for preference, and an independent paired t-
test for efficacy evaluations among the groups with the best and the worst results
at a reliability of 95%, and were expressed as mean ± standard deviation (SD).
Results
The 3 hair tonic base formulations developed for this study
were all stable following accelerated stability test under heating-
cooling for 6 cycles. Using these formulations, the preference test
was further evaluated in the initial group of 10 Thai male and fe-
male volunteers (5 each). Formulation B achieved the highest satis-
faction in odor and greasiness of the tonic, although adsorption
was not differed. Overall preference of formulation B was further
tested via questionnaire, and was found significantly (p = 0.000)
superior compared to the others (table1). The formula with the
highest preference was further incorporated with lemongrass oil at
5, 10, or 15% (formulation B1, B2, and B3).
A single closed patch test in 10 of the healthy volunteers was per-
formed as an irritation test. All formulations were found safe (MII =
0). We then used clinical evaluations to examine these 10 volunteers.
Dandruff scaling via D-Squame
®
with different severity ranging
from <10, 10–30, 31–50, 51–70, and >70% that was also interpreted
using a scale of 1–5 [4], as shown in fig.1, was used to assign the
volunteers. Of the initial group of volunteers, only those with 31–
50% or a D-Squame
®
scale 3 level were included. They were ran-
domly and double-blind assigned to apply base and lemongrass
Table 1.
Stability and preference of hair tonic formula
Formula
Physical property Preference
pH color odor adsorption greasiness overall preference
A
initial 4.65 ± 0.05 clear colorless 27.00 ± 1.06 38.00 ± 0.79 34.00 ± 0.97 34.00 ± 1.26
heating-cooling 4.36 ± 0.05 clear colorless
Binitial
heating-cooling
4.53 ± 0.08
4.26 ± 0.06
clear colorless
clear colorless
29.00 ± 0.32*37.00 ± 0.82 39.00 ± 0.72*43.00 ± 0.82*
B1
initial
heating-cooling
4.03 ± 0.12
3.66 ± 0.05
clear yellow
clear yellow
not available
B2
initial
heating-cooling
3.99 ± 0.05
3.67 ± 0.03
clear yellow
clear yellow
B3
initial
heating-cooling
3.85 ± 0.07
3.64 ± 0.13
clear yellow
clear yellow
C
initial
heating-cooling
4.38 ± 0.06
4.14 ± 0.07
clear colorless
clear colorless
27.00 ± 0.82 35.00 ± 1.08 35.00 ± 0.85 38.00 ± 0.79
* p < 0.05
Group Treatment
Reduction scale Reduction, % p value
D7-D0 D14-D0 D7-D0 D14-D0 D7-D0 D14-D0
1B
B1
0.22 ± 0.44
1.60 ± 0.60
0.89 ± 0.60
2.20 ± 0.97
8.33 ± 0.40
33.33 ± 0.60
38.10 ± 0.60
52.38 ± 0.97
0.084
0.000
0.025
0.002
2B
B2
0.44 ± 0.73
2.67 ± 1.00
2.00 ± 0.87
2.89 ± 0.78
12.90 ± 0.73
75.00 ± 1.00*
58.06 ± 1.00
81.25 ± 1.00*
0.052
0.000
0.000
0.000
3B
B3
0.22 ± 0.44
1.78 ± 1.20
1.33 ± 0.71
2.56 ± 0.88
7.41 ± 0.40
51.61 ± 1.20
44.44 ± 0.71
74.19 ± 0.80
0.084
0.001
0.000
0.000
*p = 0.000 of B1-B3 following 1 and 2 weeks treatment, respectively.
Table 2.
Efficacy of
lemongrass oil hair
tonic against dandruff
exhibited in D-squame®
scale and percentage of
reduction
Downloaded by:
Siriraj Medical Library, Mahidol University
198.143.39.97 - 9/21/2015 5:10:32 AM
Anti-dandruff Hair Tonic Containing Lemongrass
(Cymbopogon flexuosus) Oil
Forsch Komplementmed 2015;22:226–229
229
tonic on different sides of the head. Group 1 used the tonic base
and 5% lemongrass tonic, group 2 the base with 10% lemongrass
oil tonic, and group 3 the base with 15% lemongrass oil tonic. Fol-
lowing the first week of treatment, the tonics significantly (p <
0.05) exhibited anti-dandruff efficacy further increasing by the end
of the study (table2). In particular, the 10% lemongrass oil prepa-
ration significantly (p = 0.000) reduced dandruff, superior to the 5
and 15% formulas. The hair tonic base itself also showed some
anti-dandruff activity due to the presence of salicylic acid (0.2%)
and alcohol (70%) (table2).
No volunteer reported any scalp irritation followings 2 weeks of
application
Discussion
Lemongrass oil with antifungal activities, particularly dandruff-
associated pathogens [5–7, 10], was selected as active ingredient of
a hair tonic, with prolonged contact time on the sculp [11]. The
hair tonics passing centrifugation assay, particularly those with the
lowest amount of oil, were formulated prior to comparative exami-
nation of their stability and preference in volunteers.
The addition of lemongrass oil decreased pH of the formulation
due to the acidic nature of the oil constituents, particularly those of
antifungal oxygenated volatile compounds [5, 6]. Although the
hair tonics became acidified following an accelerated stability test,
pH was not significantly shifted (p > 0.05).
Dandruff scoring was monitored on the basis of squamometry, an
easy to apply and reproducibl non-invasive method using an adhe-
sive disc [3, 13, 14]. The disc was implied in this clinical intra-indi-
vidual comparative trail by means of a split-head application. The
therapeutic effect of lemongrass oil against dandruff was smaller than
that of tea tree oil at the same concentration, as 5% tea tree oil sham-
poo insignificantly reduced 78.57% of dandruff following 2 weeks of
application [4]. However, the lemongrass oil hair tonic, even at the
lowest concentration, significantly suppressed scalp flakes. This is the
primary benefit of a dosage format with prolonged scalp contact. In
addition, the efficacy was increased with higher lemongrass oil con-
tent. Yet, the activity of hair tonic containing 10% of lemon grass oil
was higher as compared to the 15% formula, which might be related
to the higher dandruff level at baseline (fig.1).
Considering the efficacy and essential oil content, hair tonic for-
mulation with 10% of lemongrass oil seems to be more effective in
dandruff treatment and easier to manufacture on a large scale than
the other formulations. This study demonstrated the clinical effi-
cacy of dandruff treatment using lemongrass oil hair tonic for the
first time. Further studies of the effectiveness of lemongrass oil hair
tonic should include research in the cosmetic effects associated with
combing and hair quality/appearance. In addition, scalp hydration
should be examined in accordance with the reduction of desquama-
tion of scalp stratum corneum, as this also helps to relieve itching
and to reduce scalp greasiness. The present study has some limita-
tions: . Although we intended to limit the interference factor of this
trial by means of a split-head application, the volunteers might have
been able to detect differences in the test tonic odor.
The hair formulation containing cremaphor and alcohol is easy
to spread on the scalp and accelerates evaporation of the oil, leav-
ing a bio-active film on the scalp. Therefore, greasiness did not cur-
tail the preference of the volunteers who were not familiar with the
oily hair care product. The anti-dandruff lemongrass oil hair tonic
was assessed positively, meets the consumers’ needs for natural
products, and its daily use at least for 2 weeks seems to be safe. .
Acknowledgments
The authors acknowledge Mae Fah Luang University for providing facility
during the manuscript preparation.
Disclosure Statement
The authors have nothing to disclose.
References
1 Hay RJ: Malassezia, dandruff and seborrhoeic dermati-
tis: an overview. Br J Dermatol 2011; 165: 2–8.
2 Schwart JR, Shah R, Krigbuam H, Sacha J, Vogt A,
Blume-Peytavi U: New insights on dandruff/sebor-
rhoeic dermatitis: the role of the scalp follicular infun-
dibulum in effect treatment strategies. Br J Dermatol
2011; 165(suppl 2):18–23.
3 Piérard-Franchimont C, Xhauflaire-Uhoda E, Piérard
GE: Revisiting dandruff. Int J Cosmet Sci 2006; 28:
311–318.
4 Satchell AC, Saurajen A, Bell C, Barnestson R: Treat-
ment of dandruff with 5% tea tree oil shampoo. J Am
Acad Dermatol 2002; 47: 852–855.
5 Attokaran M: Lemongrass; in Attokaran M (ed): Natu-
ral Food Flavours and Colorants. Ames, Wiley-Black-
well, 2011, pp 267.
6 Khunkitti W: In vitro antimicrobial and antioxidant
activities of some Cymbopogon species; in Akhila A
(ed): Essential Oil-bearing Grasses: The Genus Cym-
bopogon. New York, CRC Press, 2010, pp 167–184.
7 Galuppo R, Aureli S, Bonoli C, Ostanello F, Gubellini
E, Tampieri MP: Effectiveness of essential oils against
Malassezia spp.: comparison of two in vitro tests. Mik-
ologia Lekarska 2010; 17: 79–84.
8 Chuarienthong P, Lourith N, Leelapornpisid P: Clini-
cal efficacy comparison of anti-wrinkle cosmetics con-
taining herbal flavonoids. Int J Cosmet Sci 2010; 32:
99–106.
9 Schnuch A, Aberer W, Agathos M, Becker D, Brasch J,
Elsner P, Frosch PJ, Fuchs T, Geier J, Hillen U, Löffler
H, Mahler V, Richter G, Szliska C: Performing patch
testing with contact allergens. J Dtsch Dermatol Ges
2008; 6: 770–775.
10 Chao SC, Young DG: Screening for inhibitory activity
of essential oils on selected bacteria, fungi and viruses.
Journal of Essential Oil Research 2000; 12: 639–649.
11 Saint LD: Dandruff (pityriasis capitis simplex): of yeast
and men; in Bouillon D, Wilkinson J (eds): The Sci-
ence of Hair Care, ed 2. Boca Raton, Taylor and Fran-
cis, 2005, pp 609–631.
12 Robinson MK, Mills KJ: Noninvasive clinical aessment
of skin inflammation; in Barel AO, Paye M, Maibach
HI (eds): Handbook of Cosmetic Science and Technol-
ogy, ed 4. Boca Raton, CRC Press, 2014, pp 375–383.
13 Schatz H, Altmeyer PJ, Kligman AM: Dry skin and
scaling evaluated by D-squames and image analysis; in
Serup J, Jemec GBE, Grove GL (eds): Handbook of
Non-invasive Methods and the Skin, ed 2. Boca Raton,
CRC Press, 2006, pp 375–379.
14 Sparavigna A, Setaro M, Caserini M, Bulgheroni A: As-
sessment of the antidandruff activity of a new sham-
poo: a randomized, double-blind, controlled study by
clinical and instrumental evaluations. Skinmed 2013;
11: 85–91.
Downloaded by:
Siriraj Medical Library, Mahidol University
198.143.39.97 - 9/21/2015 5:10:32 AM
