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The antioxidant activity of Clitoria ternatea flower petal extracts and eye gel

Article  in  Phytotherapy Research · November 2009

DOI: 10.1002/ptr.2832 · Source: PubMed

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 PHYTOTHERAPY RESEARCH
Phytother. Res. 23: 1624–1625 (2009)
1624 Published online 15 April 2009 in WileyN.InterScience
KAMKAEN AND J. M. WILKINSON
(www.interscience.wiley.com) DOI: 10.1002/ptr.2832

SHORT COMMUNICATION
The Antioxidant Activity of Clitoria ternatea
Flower Petal Extracts and Eye Gel

N. Kamkaen1 and J. M. Wilkinson2*

1
Faculty of Pharmacy, Srinakharinwirot University, Ongkharak District, Nakhon-Nayok Province 26120, Thailand
2
School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW 2678, Australia

Extracts of Clitoria ternatea (butterfly pea) flowers are used in Thailand as a component of cosmetics and the
chemical composition of the flowers suggest that they may have antioxidant activity. In this study the potential
antioxidant activity of C. ternatea extracts and an extract containing eye gel formulation was investigated.
Aqueous extracts were shown to have stronger antioxidant activity (as measured by DPPH scavenging
activity) than ethanol extracts (IC50 values were 1 mg/mL and 4 mg/mL, respectively). Aqueous extracts incor-
porated in to an eye gel formulation were also shown to retain this activity, however, it was significantly less
than a commercial antiwrinkle cream included for comparison. The total phenolic content was 1.9 mg/g extract
as gallic acid equivalents. The data from this study support the use of C. ternatea extracts as anti-
oxidant inclusions in cosmetic products. Copyright © 2009 John Wiley & Sons, Ltd.

Keywords: antioxidant; Clitoria ternatea; flower; butterfly pea.

by their inclusion in food to prevent deterioration or

INTRODUCTION
Clitoria ternatea Linn (Family Fabaceae) is a highly
nutritious legume used a livestock forage plant in many
countries (Gomez and Kalamani, 2003; Hall, 1992). The
plant is also known as a medicinal plant in many parts
of the world (International Legume Database & Infor-
mation Service, 2005) where it is known by a number
of common names including butterfly pea. All parts of the
plants (roots, seeds and leaves) are used medicinally
and are purported to have a range of actions including
effects to improve cognitive function and reduce demen-
tia, treatment of respiratory conditions such as asthma
and bronchitis, treatment of inflammation and laxative
and diuretic actions (Devi et al., 2003; Jain et al., 2003).
The medicinal properties of this plant have been the
subject of several studies with much of this research
being focused towards the investigation of the neuro-
logical action of C. ternatea. For example, Jain et al.
(2003) found that extracts of the aerial parts of the plant
had a number of actions on the central nervous system
of mice including antidepressant and anxiolytic effects.
Other studies on this plant have investigated the anti-
microbial and insecticidal potential of extracts of the
seed (Kelemu et al., 2004). Little is known, however,
about other activities of these extracts, or the activity
of the flower. The flowers are a source of anthocyanins
and other flavonoids (Kazuma et al., 2003; Terahara
et al., 1996) which may have medicinal value particu-
larly as antioxidants.
Many herbs and medicinal plants have been shown
to have antioxidant activity and this has been exploited
* Correspondence to: Dr J. M. Wilkinson, School of Biomedical Sciences,
Locked Bag 588, Charles Sturt University, Wagga Wagga, NSW 2678,
Australia.
E-mail: jwilkinson@csu.edu.au
Copyright © 2009 John Wiley & Sons, Ltd.
Copyright © 2009 John Wiley & Sons, Ltd.
as nutraceuticals, as medical products and as ingredi-
ents in cosmetics (Aburjai and Natsheh, 2003; Frankel,
1999; Rotblatt and Ziment, 2002). In this study the anti-
oxidant activity of aqueous and ethanol extracts of
Clitoria ternatea flowers was determined. As the flower
extracts are also used cosmetically in Thailand the study
also assayed the antioxidant activity of aqueous extract
when incorporated into a gel base for use as an eye gel.
MATERIALS AND METHODS
Preparation of flower extract and eye gel. C. ternatea
was collected from the Prachinburi Province, Thailand
in October 2006. Taxonomic identification was performed
by Dr Supaporn Pitiporn, Chaopraya Abhaibhubejhr
Hospital, Prachinburi and a voucher specimen (CT)
deposited in the Herbarium of the Department of Phar-
macognosy, Faculty of Pharmacy, Srinakharinwirot Uni-
versity, Nakhon-nayok province, Thailand.
Fresh petals from the flowers were air-dried in shade
at room temperature (37 °C). Following drying, ethanol
and aqueous extracts were prepared by macerating
200 mg of dry material with either 100 mL of distilled
water or 95% ethanol in an Erlenmeyer flask. The mixture
was agitated using an automatic shaker (250 rpm) at
room temperature (37 °C) for 15 min and the crude
preparations were then filtered using cotton wool and
filter paper and concentrated under reduced pressure
using a rotary vacuum evaporator (Buchi, Germany)
to provide a crude extract. The final weight of the crude
extract was 50 mg. Extracts were stored at -20 °C until
use.
To prepare the eye gel, a gel base was prepared by
adding 0.8 g Carbopol 940 (Noveon, USA) to 40 mL
distilled water. The gel base was allowed to stand for
Phytother.Received 3 December(2009)
Res. 23: 1624–1625 2007
Revised 14DOI:
December 2007
10.1002/ptr
Accepted 17 December 2007
 ANTIOXIDANT ACTIVITY OF CLITORIA TERNATEA EXTRACTS
1 h after which 2 mL triethanolamine and a butterfly
pea flower extract mix comprising 30 mL of the water
extract (equivalent to 2 mg/mL). 0.9 g citric acid, 0.01 g
disodium EDTA, 0.2 g concentrated paraben and 15 g
propylene glycol was added. The final product was
adjusted to 100 g by the addition of purified water.
Antioxidant assay. The antioxidant activity of the
butterfly pea extracts and the eye gel were evaluated
spectrophotometrically using the method reported
by Bors et al. (1992). Briefly, 100 mL of a methanol
solution containing the test compound in varying con-
centrations from 0.25 to 5 mg/mL was added to 100 mL
of a 100 mM methanol solution of 2,2-diphenyl-1-
picrylhydrazyl (DPPH) (Sigma) and the absorbance at
517 nm was determined after 30 min of incubation at
25 °C, protected from light. Ethanol alone was used as
a negative control while the potent antioxidant Trolox®
(6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic
acid) (Sigma) was used as a positive control. A com-
mercial antiwrinkle, firming eye cream was also included
in the assay.
The absorbance (A) of the control and samples was
measured, and the percentage DPPH scavenging acti-
vity was determined as:
DPPH° scavenging activity (%) =
[(Acontrol - Asample)/Acontrol] ¥ 100
Each assay was performed in triplicate and the concen-
tration required for a 50% reduction (IC50) in DPPH°
radical activity was determined graphically.
The total phenolic content was determined accord-
ing to the Folin-Ciocalteu method (Chang et al., 2001).
The concentrations of total phenols in extracts were
measured by a UV-Vis spectrophotometer (Shimadzu
UV-1601, Japan), according to a colorimetric oxida-
tion/reduction reaction. Briefly, 0.5 mL of extract
solution was mixed with 0.5 mL of 1 N Folin-Ciocalteu
reagent (Carlo Erba). After 2–5 min, 1.0 mL of 20%
Na2CO3 was added and the mixture was incubated for
10 min at ambient temperature (25 °C). The mixture
was centrifuged for 8 min and the absorbance of the
supernatant measured at 730 nm. The results were
REFERENCES
Aburjai T, Natsheh FM. 2003. Plants used as cosmetics. Phytother
Res 17: 987–1000.
Bors W, Saran M, Elstner EF. 1992. Screening for plant anti-
oxidants. In Modern Methods of Plant Analysis. Plant Toxin
Analysis, Linskens MF, Jackson JF (eds). Springer: Berlin,
277–295.
Chang ST, Wu JH, Wang SY, Kang PL, Yang NS, Shyur LF.
2001. Antioxidant activity of extracts from Acacia confusa
bark and heartwood. J Agric Food Chem 49: 3420–3424.
Devi DP, Boominathan R, Mandal SC. 2003. Anti-inflammatory,
analgesic and antipyretic properties of Clitoria ternatea root.
Fitoterapia 74: 345–349.
Frankel EN. 1999. Food antioxidants and phytochemicals: present
and future perspectives. Fett/Lipid 101: 450–455.
Gomez SM, Kalamani A. 2003. Butterfly pea (Clitoria ternata): A
nutritive multipurpose forage legume for the tropics – an
overview. Pak J Nutr 2: 374–379.
Copyright © 2009 John Wiley & Sons, Ltd.
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1625
expressed as gallic acid equivalents (GAE) in milligrams
per gram of extracts. Each test was repeated three times,
and the results were averaged.
RESULTS AND DISCUSSION
It was demonstrated that a water extract of C. ternatea
has stronger antioxidant activity, as assayed by the re-
duction of DPPH, than that of an ethanol extract. The
IC50 values were 1 mg/mL and 4 mg/mL, respectively.
As antioxidant activity is often attributed to phenolic
compounds within plants the total phenolic content
within the aqueous extract and gel were measured. The
total phenolic content, expressed as GAE, was 1.9 mg/
g for the aqueous extract and 2.1 mg/g for the extract +
gel. As the water extract had the stronger antioxidant
activity it was used in the formulation of an eye gel and
it was found that the inclusion of the extract into the
gel base did not significantly reduce the antioxidant
activity of the extract. At 0.25 mg/mL the percentage
inhibition of DPPH reduction was 32% for the extract
and 28% for the gel + extract; at 0.5 mg/mL the per-
centage inhibition for both the extract and for the gel +
extract was 34%. However, the inhibition caused by
the gel containing extract was significantly less than
that of a commercial eye antiwrinkle firming cream. A
0.1 mg/mL sample of commercial eye cream produced
a 93% inhibition of DPPH reduction compared with
only 9% inhibition for 0.1 mg/mL of the gel + extract.
This suggests that while the extract maintains its anti-
oxidant activity within the gel base, the concentration
required to do so is higher than that of a commercial
product.
This study supports the traditional Thai use of C.
ternatea as an ingredient in cosmetic products as an
antioxidant. Activity was best when extracted into
water mirroring traditional extraction practices and the
data for total phenolics suggest that the flowers of C.
ternatea may be a good source of natural antioxidants
which may be incorporated into a range of cosmetic
and other products.
Hall TJ. 1992. Clitoria ternata L. (butterfly pea) cv. Milgarra.
Aust J Exp Agric 32: 547–548.
International Legume Database & Information Service. 2005.
Clitoria ternata. http://www.ildis.org/LegumeWeb/.
Jain NN, Ohal CC, Shroff SK et al. 2003. Clitoria ternatea and
the CNS. Pharmacol Biochem Behav 75: 529–536.
Kazuma K, Noda N, Suzuki M. 2003. Flavonoid composition
related to petal color in different lines of Clitoria ternatea.
Phytochemistry 64: 1133–1139.
Kelemu S, Cerdona C, Segura G. 2004. Antimicrobial and insec-
ticidal protein isolated from seeds of Clitoria ternatea, a
tropical forage legume. Plant Physiol Biochem 42: 867–873.
Rotblatt M, Ziment I. 2002. Evidence-based Herbal Medicine.
Hanley & Belfus: Philadelphia.
Terahara N, Oda M, Matsui T et al. 1996. Five new anthocyanins,
ternatins A3, B4, B3, B2, and D2, from Clitoria ternatea
flowers. J Nat Prod 59: 139–144.
Phytother. Res. 23: 1624–1625 (2009)
DOI: 10.1002/ptr