Napagoda et al.

BMC Complementary and Alternative Medicine (2016) 16:479

DOI 10.1186/s12906-016-1455-8

RESEARCH ARTICLE Open Access

Photoprotective potential in some

medicinal plants used to treat skin
diseases in Sri Lanka
Mayuri Tharanga Napagoda1*, Benthota Malavi Arachchige Shamila Malkanthi1,
Subasinghe Appuhamillage Kaumudi Abayawardana1, Mohomed Mallique Qader2 and Lalith Jayasinghe2

Abstract
Background: The constant exposure to solar ultraviolet radiation (UV) has a variety of harmful effects on human
health. Although synthetic sunscreen products have been introduced as a preventive/therapeutic strategy, with the
realization of their adverse side effects, the recent trend is to search for human friendly alternative formulations
especially of plant origin. Therefore, the present study focuses on evaluation of photoprotective activity of aqueous
extracts (1 mg/ml) of eleven medicinal plants in Sri Lanka that have been widely employed in traditional medicine
as treatment options for various skin diseases and to improve the complexion.
Methods: For the determination of UV filtering potential of the extracts, UV absorption was measured and the sun
protection factor (SPF) was calculated according the Mansur equation. The antioxidant activity was evaluated by
DPPH and ABTS assays.
Results: Among the extracts, Atalantia ceylanica, Hibiscus furcatus, Leucas zeylanica, Mollugo cerviana, Olax zeylanica
and Ophiorrhiza mungos have displayed SPF value ≥ 25, which are even higher than two commercial photoprotective
creams used as reference compounds. L. zeylanica and O. mungos have displayed a high UV absorbance in
260–350 nm range indicating their potential of being broad spectrum sunscreens. In addition, the extract of
O. mungos was found to be photostable, without any significant reduction in the SPF after exposure to direct
solar radiation for 21 days. DPPH assay and the ABTS assay revealed that the extracts possess high antioxidant activity.
Conclusion: The results of the present study suggest that the presence of secondary metabolites with antioxidant
property could be responsible for the high UV absorbance. Our findings would offer an exciting avenue for further
research towards the development of herbal cosmetics.
Keywords: Photoprotective, Sunscreen, Antioxidant, Medicinal plants

Background depending on the wavelength; UV-A (320–400 nm)

During the past few decades, the anthropogenic activ- UV-B (290–320 nm) and UV-C (200–290 nm). In-
ities had led to a substantial damage to the protective tense or overexposure to UV-A and UV-B leads to
ozone layer and resulted in a significant increase of the sun burn, erythema, inflammation, hyperpigmenta-
solar radiation reaching the earth. As a consequence, the tion, wrinkling, hyperplasia, local immunosuppres-
incidence of various diseases and disorders related with sion, photoaging and photocarcinogenesis. Although
the excessive exposure to solar ultraviolet (UV) radiation UV-C is most biologically damaging, it gets effect-
has alarmingly increased over the recent years [1]. Solar ively filtered by the ozone layer [2–4].
UV radiation is classified into three major categories The UV exposure to the skin leads to the generation
of free radicals/reactive oxygen species which exert dele-
* Correspondence: [email protected]
terious effects by oxidizing biologically essential mole-
1
Department of Biochemistry, Faculty of Medicine, University of Ruhuna, cules and induced oxidative damage. The excess of free
Galle 80000, Sri Lanka radicals results in a cascade of events for example,
Full list of author information is available at the end of the article

© The Author(s). 2016 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Napagoda et al. BMC Complementary and Alternative Medicine (2016) 16:479 Page 2 of 6

mitogen-activated protein kinase cascade, induction of example, the oral administration or topical application of
heme oxygenase-1 and matrix metalloproteinase in plant extracts such as green and black tea, coffee, Aloe
the skin and thereby mediating progressive deterior- vera, cucumber is speculated to be protective against UV-
ation of cellular structure and function as well as induced erythema, early aging and irradiation-induced
modification of DNA and abnormal expression of cel- cancer [9].
lular genes. Although the skin possesses an elaborate Plants and their products have been systematically
antioxidant system to deal with the UV induced oxi- used in Sri Lanka for treating illnesses for over thousand
dative stress, the extensive and chronic exposure to years. Among the native flora of Sri Lanka more than
UV could exceed the cutaneous antioxidant capacity, 1400 plants are used in indigenous medicine [10] and
hence leads to oxidative damage causing variety of the literature reveals that large number of plants are ex-
harmful effects in the skin [5]. tensively used to treat various dermatological diseases as
As it would be impractical to reduce the excessive ex- well as to improve complexion [11]. These alternative
posure to solar radiation, a novel approach called “photo- medications seem promising, although their true effects
chemoprevention”/“photoprotection” has been introduced are not scientifically proven, thus further investigations
to overcome the detrimental effects caused by UV ra- should be performed to assess the clinical benefits. How-
diation. It involves the use of various photochemopre- ever, only a handful of scientific evidences are available
ventive/photoprotective agents which functions via on bioactivity studies of medicinal plants in Sri Lanka
prevention of the damage caused by UV radiation that could lead towards the development of herbal cos-
and/or modulation of different cellular responses to metics. Apart from the study on photoprotective proper-
UV radiation to prevent, stop or correct tumour promo- ties of Sri Lankan black tea [12], there has been hardly
tion and progression. The most popular strategy in the any report on photoprotective potential of Sri Lankan
present day practice to reduce the amount of UV radiation plants. In order to fulfill this knowledge gap, the present
penetrating the skin is the topical application of sunscreen study has focused on evaluation of sunscreening and
products that contain UV absorbing, reflecting or scat- antioxidant activity of eleven medicinal plants that have
tering active molecules. Sunscreens with a sun protect- been extensively utilized in Sri Lanka for improving
ive factor (SPF) value of 15 or greater are highly complexion and as dermatological therapeutics. Thereby
recommended and these compounds are incorporated the ethnopharmacological usage of these plant species
in several cosmetic products such as creams, gels, oils and could also be rationalized.
lotions [6]. Octylmethoxycinnamate, benzophenone-3,
mexenone, provatene, avobenzone are a few examples of Methods
synthetic sunscreen agents, however their usage is limited Plant material
owing to the adverse side effects such as development of Leaves of Aporosa lindleyana (Euphorbiaceae) Atalantia
irritant dermatitis, hypersensitivity, allergies and even ceylanica (Rutaceae), Hibiscus furcatus (Malvaceae), Olax
melanoma [7]. Therefore the usage of natural/herbal zeylanica (Olacaceae), Ophiorrhiza mungos (Rubiaceae)
sunscreens has gained considerable attention over the re- and whole plants of Argyreia populifolia (Convolvulaceae),
cent years and several natural compounds with UV ab- Ipomoea mauritiana (Convolvulaceae), Lasia spinosa
sorption property have been used to substitute for or to (Araceae), Leucas zeylanica (Lamiaceae) and Plectranthus
reduce the quantity of synthetic sunscreen agents [8]. zeylanicus (Lamiaceae) were collected in Gampaha Dis-
However it was revealed that the protection is not fully trict - Western Province of Sri Lanka in 2013, while seeds
achieved even if the sunscreen is effective in blocking of Mollugo cerviana (Aizoaceae) was purchased from
harmful UV rays, thus the high SPF value alone is not Ayurvedic retail outlet at the Market Place, Nittambuwa,
adequate for an effective photoprotection. Over the last Sri Lanka. Based on the application in folklore medicine,
few years, a significant number of evidence has emerged, the above plant parts were specifically selected for the
indicating that chemically diverse classes of naturally oc- study. The plants were identified by the author (MTN), a
curring substances are potent in the treatment of several botanist, and confirmed based on the books “A Revised
dermatological conditions caused by the chronic expos- Handbook to the Flora of Ceylon: volume – 1-XIII, M.D.
ure to the UV radiation. Polyphenols, flavonoids, ter- Dassanayake & F.R. Fosberg” and “Medicinal plants
penes, catechins and alkaloids are compound classes (indigenous and exotic) used in Ceylon: Volume 1–5
that have been targeted for photochemopreventive activ- by D.M.A. Jayaweera” and authenticated by compari-
ity and curcumine, resveratrol, caffeic acid and quercetin son with the herbarium specimens at the National
are a few examples of such compounds. It has been sug- herbarium, Royal Botanical Garden, Peradeniya, Sri
gested that the beneficial effects of these antioxidants Lanka. A voucher specimen of each plant is deposited
might be a successful strategy for diminishing UV- at the Department of Biochemistry, Faculty of Medicine,
radiation mediated oxidative damage of the skin. For University of Ruhuna, Sri Lanka.
Napagoda et al. BMC Complementary and Alternative Medicine (2016) 16:479 Page 3 of 6

Preparation of crude extracts ABTS assay

The plant materials were thoroughly washed and dried ABTS (2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic
in shade (30 °C) for six days. Dried plants were pow- acid) assay was employed as described by Re et al. 1999
dered using a domestic grinder (Singer model KA- [15]. Briefly, ABTS.+ solution (2.5 mM) and potassium
MIXEE). The powdered materials (10–15 g) were ex- persulfate (2 mM) solution in PBS buffer were prepared,
tracted in 300 ml of 70% methanol–water by heating for mixed in 1:1 ratio and the mixture was allowed to react
2 h at 60 °C. The extracts were evaporated into dryness for 12 h at room temperature in the dark. The absorb-
with the use of rotary evaporator (BÜCHI, R-114, ance of the ABTS radical solution was adjusted at 0.8 at
Germany) or a vacuum centrifuge (Thermo, Germany). 734 nm. The ABTS radical solution (150 μl) was mixed
with plant sample (50 μl) and the absorbance was mea-
Evaluation of UV filtering potential sured at 734 nm. All the measurements were carried out
For the determination of UV filtering potential the in triplicate. Trolox was used as the antioxidant standard
UV absorption of each extract (1 mg/ml) was mea- and the antioxidant potential was expressed as milimolar
sured between 260 and 400 nm using UV-visible of Trolox equivalents per milligram.
spectrophotometer (Shimadzu, UV_1800) and the sun
protection factor (SPF) was calculated according the Statistical analysis
Mansur equation [13]. All the above experments were perormed in triplicate
and the values were given as mean ± S.D.
X
320
SPF spectrophootometric ¼ CF  ΕΕ ðλÞ  Ι ðλÞ  AbsðλÞ Results
290
UV filtering potential
Among the eleven extracts Atalantia ceylanica, Hibiscus
Where: EE(λ) – erythemal effect spectrum; I(λ) – solar
furcatus, Leucas zeylanica, Mollugo cerviana, Olax zey-
intensity spectrum; Abs(λ) – absorbance of sunscreen
lanica and Ophiorrhiza mungos have displayed SPF ≥ 25,
product; CF – correction factor (=10)
which were even higher than two commercial photopro-
The plant extracts were exposed to direct sunlight for
tective creams used as the standard reference substances
21 days and the UV absorbance was measured at 7th 14th
(Table 1). SPF values of L. zeylanica, O. mungos, M.
and 21st day to determine the photostability of the extracts.
cerviana and H. furcatus were found to be even higher
Two commercially available sunscreen products SC1
than the SPF of well-known medicinal plant, Aloe vera.
(benzophenone-4 and TiO2 as active ingredients) and
Interestingly, the real SPF value of the two commer-
SC2 (Aloe, Sandlewood, Ficus as active ingredients)
cial photoprotective creams were found to be much
were used as reference substances. In addition, an ex-
lower than their labelled SPF value. The most potent
tract prepared with Aloe vera (leaves) following the
six extracts were further subjected to detailed
same procedure was used as a control to compare
with the UV filtering potential of the selected plant
extracts. Table 1 Measured SPF values of the plant extracts (listed in
descending order) and commercial sunscreens
Radical scavenging capability Extract Measured SPF Value
DPPH assay Leucas zeylanica 39.8 ± 0.35
The radical scavenging capability of the extracts was Ophiorrhiza mungos 39.2 ± 0.92
assessed by measuring the reduction of the stable free Mollugo cerviana 29.5 ± 0.30
radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) as de-
Hibiscus furcatus 29.4 ± 0.40
scribed by the method of Blois 1958 [14]. The ab-
Atalantia ceylanica 26.8 ± 0.16
sorbance was recorded at 517 nm after 30 min
incubation of test extracts and DPPH solution under Olax zeylanica 24.5 ± 0.47
gentle shaking in the dark. The percentage antioxi- Aporosa lindleyana 21.4 ± 0.90
dant activity (AA) was calculated using the following Argyreia populifolia 12.5 ± 0.61
formula and the EC50 was determined using Graph- Plectranthus zeylanicus 11.5 ± 0.96
Pad Prism version 6.01.
Ipomoea mauritiana 11.3 ± 0.70

ðAbsorbance of the control−Absorbance of the sampleÞ Lasia spinosa 8.9 ± 0.57

AA ¼  100
Absorbance of the control SC1 (Standard reference) 10.7 ± 0.07 (labeled SPF : 30+)
SC2 ((Standard reference) 18.6 ± 0.01 (labeled SPF : 45)
BHA was used as the positive control and all the mea-
Aloe vera (control) 28.86 ± 0.11
surements were carried out in triplicate.
Napagoda et al. BMC Complementary and Alternative Medicine (2016) 16:479 Page 4 of 6

Fig. 1 Variation of SPF overtime

investigations to determine their suitability as com- Discussion

ponents of herbal sunscreens. Sri Lanka lies within the equatorial belt, a region which
Furthermore O. mungos did not display a reduction in receive ample amount of solar radiation throughout the
the SPF after exposition to direct solar radiation for year. The strength of sunburn-producing UV radiation
21 days. Similarly, Aloe vera extract also displayed photo- at a particular place and a time is explained in terms of
stability with SPF values of 24.61 ± 0.82, 23.65 ± 0.08 and the UV Index that is defined as the amount of skin dam-
22.84 ± 0.16 on 7th, 14th and 21st day respectively. aging UV radiation expected to reach the earth’s surface
Moreover L. zeylanica and O. mungos have exhibited at the time when the sun is highest in the sky. In Sri
high UV absorbance throughout the range of 260– Lanka, the UV index varies from 6 to 12 throughout the
350 nm (Fig. 1 and Fig. 2) - UV absorbance of the ex- year [16], that falls within the “high”, “very high” and
tracts). In addition, the maximum absorbance of H. fur- “extreme” risk categories for which the usage of sun-
catus lies within the UV- B range while the other screens with SPF of 30+ are highly recommended.
extracts have displayed the maximum absorption in the SPF value is an indicator that is mentioned in sun-
UV-C range. screens which indicates that how much photoprotection
provides against UV radiation by sunscreen when it is
Radical scavenging capability applied thickness of 2 mg/cm [3] skin [17]. Sunscreen
The radical scavenging potential was extremely high in products can be categorized according to their SPF
H. furcatus L. zeylanica, M. cerviana, O. zeylanica and values as minimal (SPF < 12), moderate (SPF 12–30) and
O. Mungos extracts according to the DPPH assay, while high sun protection products (SPF ≥ 30) [18]. According
all the extracts have displayed strong antioxidant activ- to the SPF value of the sunscreen product, the protec-
ities proving their capacity to scavenge the ABTS radical tion percentage from UV radiation is different. When
cation (Table 2). SPF is 15, it provides > 93% protection against UV- B

Fig. 2 UV absorbance of the extracts

Napagoda et al. BMC Complementary and Alternative Medicine (2016) 16:479 Page 5 of 6

Table 2 Antioxidant activity of the most potent plant extracts defence system against the formation of UV induced free
Extract EC50 for DPPH Antioxidant activity as expressed radicals, thus resulting in superior photoprotection. Fur-
assay as mM trolox eq/mg thermore, these observations suggest that the extracts
Atalantia ceylanica 527 ± 0.83 108 ± 0.003 might contain secondary metabolites such as polyphe-
Hibiscus furcatus 32.32 ± 0.79 110 ± 0.01 nols and flavonoids that are capable of absorbing UV ra-
Leucas zeylanica 30.54 ± 0.93 116 ± 0.01 diation as well as quenching the reactive oxygen species
generated in the body. Therefore, the secondary metabo-
Mollugo cerviana 98.72 ± 0.80 100 ± 0.005
lites in the bioactive extracts will be identified by liquid
Olax zeylanica 28.05 ± 0.94 86 ± 0.01
chromatography coupled mass spectrometric (LC-MS)
Ophiorrhiza mungos 32.49 ± 0.94 109 ± 0.007 analysis while formulation of a topical sunscreen by in-
BHA (positive control) 23.12 ± 0.96 – corporation of the most active extracts is currently in
progress. Thereafter, more detailed cell based assays are
and SPF +30 provides 97% protection from UV-B. Most planned for the future to evaluate their suitability for the
of the commercially available sunscreen products are development of herbal cosmetics and we hope that these
highly effective against UV-B, but not against UV-A [18, findings may further support the ethnopharmacological
19]. significance of these plant species as well.
The present investigation reveals that Sri Lankan plant
species have a high sunscreening potential that falls Conclusions
within “moderate” and “high” sun protective categories. The preliminary findings of this study reveals that Sri
Thus a sunscreen product developed by these plant ex- Lankan medicinal plant preparations have a high poten-
tracts would be of high importance specially for the tial to be used as natural skin care agents due to the
people living in tropical countries. Furthermore, the high UV absorption properties and the strong antioxi-
study reveals that real SPF value was found to be much dant activities. This would offer an exciting avenue for
lower than the labelled value in two commercial sun- further research towards the development of herbal
screen products and these observations agreed with the cosmetics.
previous literature data where the labelled SPF did not
correspond to the actual SPF value [20]. Since most che- Abbreviations
ABTS: 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid; DPPH: 2,2-
micals capable of blocking only a narrow region of the diphenyl-1-picrylhydrazyl; LC-MS: Liquid chromatography coupled mass
UV light, incorporation of several chemicals with each spectrometry; SPF: Sun protective factor; UV: Ultraviolet
one blocking a different region of UV light would be ne-
Acknowledgments
cessary to offer a broad spectrum UV protection. There-
“Faculty of Medicine- Research Grant 2015” from the University of
fore the characteristic absorption band exhibited by the Ruhuna, Sri Lanka is gratefully acknowledged.
extracts of L. zeylanica and O. mungos in the UV-B and
UV-A suggest a possible broad spectrum sunscreening Funding
“Faculty of Medicine- Research Grant 2015” from the University of
potential, a desirable character of an ideal sunscreen. Ruhuna, Sri Lanka.
Usually a sunscreen UV filters may degraded or
destroyed over the time or when get exposed to sunlight, Availability of data and materials
thus the development of a sunscreen with photostability The datasets generated and analysed during the current study are not
publicly available as the authors intend to develop a commercial product,
is one of the dilemma in the cosmetic industry. In this but are available from the corresponding author on reasonable request.
respect, the photostability displayed by the extract of O.
mungos indicates that the plant is having a great promise Authors’ contributions
MTN designed the experiments. BMASM, SAKA and MMQ carried out the
to be developed as a potent topical sunscreen. experiments under the supervision of MTN and LJ. MTN, MMQ and LJ wrote
Usually the herbal cosmetics contain one or more ac- the manuscript. All authors read and approved the final manuscript.
tive sunscreening agents with antioxidant properties in
order to achieve a good photoprotective effect. As UV Competing interests
The authors declare that they have no competing interest.
rays absorbed by the skin leads to the production of free
radicals such as O−2 , H2O2, OH., ROO. etc. and deacti- Consent for publication
vate the antioxidant enzymes, thus the incorporation of This information is not relevant.
antioxidants is now widely recommended in sunscreens.
Ethics approval and consent to participate
Therefore the evaluation of most potent extracts for the This information is not relevant.
antioxidant activity would be important for the develop-
ment of more effective and efficient sunscreens. As the Author details
1
Department of Biochemistry, Faculty of Medicine, University of Ruhuna,
selected extracts have displayed antioxidant activities, a Galle 80000, Sri Lanka. 2National Institute of Fundamental Studies, Hantana
sunscreen developed with those could boost the body’s Rd, Kandy 20000, Sri Lanka.
Napagoda et al. BMC Complementary and Alternative Medicine (2016) 16:479 Page 6 of 6

Received: 14 March 2016 Accepted: 31 October 2016

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