The Sunscreen activities of ethanol, ethyl acetate, n-hexane, and water
fractions from papaya (Carica papaya l.) leaf extract
Sri Wardatun*, Siti Mahyuni, Putra Setiawan
.
Department of Pharmacy, Faculty of Mathematics and Natural Sciences Universitas Pakuan
Jl. Pakuan PO Box 452 Bogor 16143 Indonesia
Submitted: 26/04/2023, Revised: 19/07/2024, Accepted: 20/12/2024, Published: 24/01/2025
ABSTRACT
The application of sunscreen products containing natural compounds is one of
alternative popular ways to protect the skin from the harmful effects of sun
exposure. The aim of this study was to evaluate the sunscreen activity of papaya
leaf extract. The ethanol extract of papaya leaf was fractionated to obtain ethanol,
n-hexane, ethyl acetate, and water extracts. The value of SPF 15, percent
transmission of erythema (1% Te ), and the percent transmission of pigmentation
40 (% Tp) of each extract were calculated in vitro by using spectrophotometric
method. among the extract examined, the ethyl acetate extract was found to be the
most effective to reach Sun Protection Factor (SPF) 15 (ultra-protection category)
and 1% Te (sunblock category) at a minimum concentration of 90.08 ppm and
63.49 ppm. Meanwhile n-hexane extract was the most effective to reach 40% tp
(sunblock category) at a minimum concentration of 30.68 ppm. The minimum
concentration of papaya leaf extracts required to reach SPF 15, 1% Te and 40% Tp
were far below the concentration of sunscreen compounds allowed in commercial
cosmetic products. It can be concluded that papaya leaf extract have significant
sunscreen property for use as cosmetic ingredient.
Keywords: Carica papaya, SPF, Transmission of erythema, Transmission of
pigmentation
Vol. 5, Issue 2, 2023 (422-431)
http://journal.unpad.ac.id/IdJP
*Corresponding author,
e-mail: sri.wardatun@unpak.ac.id (S. Wardatun)
https://doi.org/10.24198/idjp.v5i2.46557
© 2023 S. Wardatun et al
423
1. Introduction
Excessive exposure to sunlight is
considered to be the main cause of many
types of skin cancer including melanoma,
the most violent form of skin cancer (1).
The prolonged exposure to direct sunlight
also triggers the skin aging and other skin
damage such as dry skin, wrinkles, over
pigmentation, actinic keratosis marked
by scaly and rough skin texture, freckles
and abnormal skin discoloration (2).
Despite the negative effects, exposure to
sunlight cannot be eliminated. Lack of
sunlight led to a deficiency of serotonin
and vitamin D in the body and increase
the risk for depression, osteoporosis, heart
disease, cancers, infectious diseases,
fatigue, and tiredness (3).
The application of sunscreen
products was used to overcome these
harmful effects of sunlight
exposure. Ingredients in sunscreen
products protect skin from UV light in
two different ways. Inorganic compounds
like titanium dioxide and zinc oxide, act
as a physical barrier that reflects UV light
whereas organic compounds provide
chemical structures that absorb UV
radiation (4). The sunscreen material
derived from natural sources such as plant
phenolic, flavonoids, and carotenoids
gain its popularity as a natural sunscreen
related to their capacity to absorb UV ray
and minimizes the negative effect of
sunlight on the skin (5). The sunscreen
capacity of plant compounds is obtained
from the structure of the chromophore
that absorbs sunlight and transform it into
thermal or fluorescent/phosphorescent
energy. The protection activity of
secondary plant metabolites against
sunlight spans a wide spectrum range
from UV irradiation to visible and
infrared light (6).
Papaya plant including its leaf,
seeds, ripe and unripe fruits, and juice are
commonly used as a traditional medicine.
The leaves of papaya are knowns to
contain many active compounds such as
papain, chymopapain, cystatin,
tocopherol, ascorbic acid, flavonoids,
cyanogenic glucosides and glycosylates.
These compounds proved to have
immunomodulatory and antioxidant
effects, reduce lipid peroxidation, exhibit
anti-tumor activity and prevent
inappropriate proteolysis. The papain
enzyme from papaya leaves is used
topically to remove dead cells from skin’s
surface to help the skin rejuvenate. (7, 8).
Many reports of C. papaya leaf
application to improve skin health have
been published but more specific studies
to find out about the capacity of papaya
leaves to protect skin from sunlight
exposure are still lacking. Thus the aim of
this study was to evaluate the potential
role of papaya leaf extract as a sunscreen
agent.
2. Method
Plant material: papaya leaf var.
calina was collected from Indonesian
Research Institute for Medicinal Spices
(BALITRO) and authenticated at the
Indonesian Institute of Sciences (LIPI),
Botanical Gardens, Bogor. The papaya
leaf was washed and dried in electric
oven at a temperature of 500-600C and
then grinded to produce fine powder
material. The dried papaya leave powder
stored in a vacuum flask at room
temperature for further usage.
2.1. Reagents and instruments
Reagents and instruments used in
this study were of analytical grade i.e.
ethanol, methanol, n-hexane and ethyl
acetate (Merck); rotary evaporator
(IKA®) and UV-Visible
spectrophotometer (Jasco V-730®).
S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
424
2.2.2 Extraction and fractionation
The sample was macerated using
96% ethanol (1: 10 w/w) for 24 hours,
filtered and dried in rotary evaporator to
obtain condensed crude extract. The
condensed crude extract was diluted with
hot double distilled water (20:100 w/v)
prior to fractionation. The liquid-liquid
fractionation method based on the
polarity of the solvent was applied with
hexane, ethyl acetate and ethanol
successively to fractionate the papaya leaf
extract. Furthermore, the sunscreen
activity of each fraction was determined
using UV-VIS spectrophotometer.
2.3. Determination of sunscreen activity.
A series of concentrations for each
extract were prepared in….(please
mentioned the solvent applied) prior to
the determination of sunscreen activity
using a spectrophotometer as listed in
table 1.
S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
Table 1 Concentration of Papaya Leaf Extract Fractions
Sampel Concentration (ppm)
Ethanol extract 25; 50; 75; 100
n-heksan extract 10; 25; 50; 75
ethyl acetate extract 25; 30; 35; 40
Water extract 25; 50; 75; 100
2.4. Determination of SPF 15, % TE
AND % TP
Sunscreen activity of papaya leaf
extracts were determined in vitro using
the spectrophotometry method. The
absorbance data to calculate SPF value
was read at wavelength 290-320 nm
every 5 nm interval meanwhile the
percent transmission of erythema (% Te )
and the percent transmission of
pigmentation (% Tp) was read at
wavelength 292,5 -372,5 every 5 nm
interval. The value of SPF was calculated
according to Mansur equation (9) as
follows:
Where:
CF = correction factor (10),
EE (λ) = erythemal effect of radiation at wavelength λ,
Abs (I) = spectrophotometric absorbance of sample at wavelength λ.
SPF = CF EE󰇛󰇜 󰇛󰇜  󰇛󰇜
310
290 
(1)
The SPF value obtained was extrapolated
to set up the linear equation and the linear
equation will be used to calculate the
value of SPF 15.
Value of % Te and % Tp was calculated
using the following equations:
425
S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
3. Result and Discussion
The sunscreen activities of
papaya leaf extracts were displayed in
Figure 1. The effectiveness of papaya
leaves extracts as sunscreen was
evaluated from the concentration of
extract required to reach SPF 15, 1% Te,
and 40% Tp. The chart in Fig. 1
indicates a negative correlation between
the concentration of extracts and its
sunscreen capacity. The lower the
concentration required to reach SPF 15,
1% Te and 40% Tp, the higher sunscreen
capacity of papaya leaf extracts.
A = -log T (Equation 1) (2)
Where:
A = Absorbance of sample
T = Transmition value
The T value obtained from equation 1 put into Balsam equation (10) to obtain (%) Te
and (%) Tp.
(% ) Te= 
 󰇛󰇜
 (2) (3)
(%) Tp = 
 󰇛󰇜
 (3) (4)
Where:
T = Transmission value
Fe = erythema flux at wavelength λ
Ee = The erythema flux passed by sunscreen
Fp = Pigmentation flux at wavelength λ
Ep = Pigmentation flux passed by sunscreen
The % Te and Tp value obtained from each concentration was extrapolated to set up the
linear equation to calculate the 1% Te and 40% Tp.
426
S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
SPF is a measurement that shows
how much the sunscreen will protect the
skin from the burning radiation of UV
rays. The SPF value according to
European Commission (EC)
Recommendation (11) categorized as
follows: SPF 6-10 (low protection), SPF
15-25 (medium protection), SPF 30-50
(high protection), SPF 50+ value (very
high protection). The SPF 15 evaluated
in this study fall into medium protection
category which block about 93% of UV
B rays. The concentration of papaya leaf
extracts required to reach SPF 15 are
shown in table 2.
Figure 1: Sunscreen activities from different fractions of Carica papaya leaves extract
0
50
100
150
200
250
300
350
400
450
Water
Fraction Ethyl Acetate
Fraction n-Hexane
Fraction Papaya Leaves
Ethanol
Extract
Quercetin
Concentration (ppm)
SPF 15 1% Erythema Transmission 40 % Pigmentation Transmission
Table 1. Concentration of papaya leaf extracts at SPF 15
Sample Concentration (ppm)
Water extract
278.29
Ethyl acetate extract
90.08
n-hexane extract 424.83
Ethanol extract
305.03
Quercetin (control)
46.33
Table 1. shows that the ethyl
acetate extract has the most effective
sunscreen activity with a minimum
concentration to reach the SPF 15 occur
at concentration of 90.08 ppm. This
concentration is quite low compared to
the compounds allowed in cosmetics
products such as glyceryl PABA (3%),
escalol 75A (5%), 2ethylhexyl salicylate
(5%), sunarome WMO (5%), digalloyl
trioleate (3%), homomenthyl salicylate
(8%), propylene glycol salicylate 4%
and other natural compounds (12, 13).
427
S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
The sunscreen capacity of papaya
leaf ethyl acetate extract was supported
by study of Marliani et al., (14) and
Sachin et al. (15). Previous data reveal it
was required 50 μg / mL to 7 0 μg / mL
papaya peel extract to reach SPF 2
(minimal protection category), and
14.2% papaya fruit extract to reach SPF
16.
The sunscreen activity of ethyl
acetate extract could be associated with
the present of the phenolic compounds
(65.12 mg GAE/g) in its dry leave
powder (16). Phenolic well known as
main a compound that are responsible for
the antioxidant activity as demonstrated
by various natural ingredients (17, 18).
Many researches also confirm the strong
correlation between antioxidant activity
and the sunscreen activity of phenol
compounds (19, 20). For example, some
Brazilian medicinal plants which contain
phenolic compounds up to 3.77 to
57.14 mg GAE/g showing sunscreen
activity ranging from SPF = 20.12
to SPF = 26.82 (21). Among the various
phenolic compounds, the flavonoids are
perhaps the most important group. The
sunscreen activity of flavonoids against
UV radiation were provided by the
presence of double bonds structure and
hydroxyl groups which act as strong UV
absorbing agent (22). Flavonoids are
suspected as a responsible compound for
sunscreen activity also due to property of
ethyl acetate as one of the best
flavonoids’ solvent (23). Other
compounds possible for sunscreen
activity in the ethyl acetate extract come
from natural benzophenones group which
is soluble in ethyl acetate (24).
Benzophenones are a naturally occurring
compounds in some flowering plants that
block ultraviolet (UV) rays and widely
used as sunscreen ingredients in cosmetic
products.
Percent transmission of
erythema (% Te) and transmission of
pigmentation (% Tp) are the amount of
sunlight transmitted to the skin surface
after passing through the sunscreen
which causes erythema and
pigmentation. Erythema was a
hypersensitivity reaction of the skin to
UV radiation characterized by redness
and inflammation symptoms, and the
pigmentation was a darkening
(tanning) of certain skin areas because of
the formation of new melanin in the skin
surface. The value of (%) Te and (%) Tp
were used to determine the sunscreen
category of a material i.e. % Te <1 and %
Tp 3-40 is categorized as sunblock, Te%
1-6 and% Tp 42-86 is categorized as
extra protection, % Te 6 -12 and % Tp
45-86 is categorized as suntan, and Te%
10-18 and% Tp 45-86 and is categorized
as tanning. Table 2 and Table 3 show the
concentrations of papaya leaf extracts
required to reach 1 % Te and 40% Tp at
the extra protection category.
Tabel 2. The concentration of papaya leave extracts at 1% Te
Sample Concentration (ppm)
Water extract 161.91
Ethyl acetate extract 63.49
N-hexane extract 188.55
Ethanol extract 152.87
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S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
Table 2 show that ethyl acetate extract
was the most effective to prevent the
incidence erythema less than one % at a
concentration of 63,49 ppm. Similar to
the evaluation of SPF 15, the compounds
responsible for blocking sunlight and
reducing the incidence of erythema in
ethyl acetate extract are suspected to
come from the phenolic group. Apart
from phenolic, alkaloids are the possible
compounds that act as a sunscreen
because the alkaloids dissolve in ethyl
acetate. Several studies have confirmed
the positive correlation between plant
alkaloids and sunscreen activity.
Tabel 3. Concentration of papaya leave extracts at 40% Tp
Quercetin (control) 20.88
Sample Concentration (ppm)
Water extract 72.57
Ethyl acetate extract 45.21
N-hexane extract 30.68
Ethanol extract 84.96
Quercetin (Control)
5.48
Different from the determination
of SPF 15 and 1 % Te, table 3 shows
that n-hexane extract was the most
effective to reach 40% Tp. N-hexane
extract reduce 40% pigmentation at the
concentration of 30,68 ppm. The
difference may occurs due to the
different cause of erythema and
pigmentation. Erythema was induced by
exposure of skin to UV-B rays while
pigmentation was induced by exposure to
UV-A rays, therefore the determination
of %Te was applied to show protection
against UV-B rays and determination of
% Tp was applied to show protection
against UV-A rays.
The sunscreen compounds which
inhibit transmission of pigmentation n-
hexane extracts probably comes from
terpenoids group especially carotenoids.
The presence of carotenoids in n-hexane
extract is associated with n-hexane
property as a non-polar solvent suitable
for extracting the terpenoid/carotenoid
compounds from fruit and plant. Study
of Ugo et al. (25) revealed the presence
of beta carotene in papaya leaves extract
up to 303.5 mg/100g. The carotenoids in
plants act to protect the photosynthetic
elements from damage by absorbing the
excess of light energy. The application
these compounds in skin are expected to
possess similar effect to protect the skin
from negative effects of excessive
sunlight (26, 27). The sunscreen property
of carotenoids was confirmed in a recent
study of Darvin et al. (28). The study
revealed that application beta carotene
topically effectively neutralized free
radicals produced on the skin surface
after IR irradiation. Other study also
confirmed the positive effects of
combined oral/topical antioxidant
treatment with lutein and zeaxanthin on
human skin (29).
429
S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
4. Conclusion
The results of this study
confirmed that papaya leaf extracts have
a significant sunscreen property. The
ethyl acetate and n-hexane extracts
proved to reduce the occurrence of
erythema and pigmentation at low
concentrations of 63.49 ppm and 30.68
ppm respectively. A combination of two
extracts has the potential for use as an
active ingredient in sunscreen cosmetic.
Further study should be conduct to
elucidate the compounds responsible for
sunscreen activity and how to utilize as a
safe cosmetic ingredient.
5. Acknowledgements
The authors thank to Faculty of
Sciences and Mathematic and Natural
Sciences, Universitas Pakuan, Indonesia
for funding this study.
5. References
[1] Rhee H, Vries E, Coomans C, Velde
P, Coebergh J. Sunlight: For Better
or For Worse? A review of positive
and negative effects of sun exposure.
Cancer Research Frontiers. 2016;
2(2): 156-183, 10.17980/2016.156.
[2] Taylor CR, Sober AJ. Sun exposure
and skin disease, Annu Rev Med.
1996; 47:181-191.
doi:10.1146/annurev.med.47.1.181.
[3] Mead MN. Benefits of sunlight: a
bright spot for human
health. Environmental Health
Perspectives 2008; 116(4): A160–
A167.
https://doi.org/10.1289/ehp.116-a160
[4] Serpone N, Dondi D, Albini A.
Inorganic and organic UV filters:
their role and efficacy in sunscreens
and suncare product. Inorganica
Chimica Acta 2017; 360(3): 794-
802. 10.1016/j.ica.2005.12.057.
[5] Pandey KB, Rizvi SI. Plant
polyphenols as dietary antioxidants
in human health and
disease. Oxidative Medicine and
Cellular Longevity 2009; 2(5): 270–
278.
https://doi.org/10.4161/oxim.2.5.949
8.
[6] Korkina L, Vladimir K, Potapovich
A, Mayer W, Talib N, De Luca C.
Secondary Plant Metabolites For Sun
Protective Cosmetics: From Pre-
Selection to Product Formulation.
Cosmetics 2018; 5(32): pp. 1-20.
doi:10.3390/cosmetics5020032
[7] Otsuki N, Dang NH, Kumagai E,
Kondo A, Iwata S, Morimoto C.
Aqueous extract of Carica papaya
leaves exhibits anti-tumor activity
and iimmunomodulatory effects. J
Ethnopharmacol. 2010; 127(3): 760-
767. doi:
10.1016/j.jphotobiol.2018.10.013.
[8] Seigler DS, Pauli GF, Nahrstedt A,
Leen R. Cyanogenic allosides and
glucosides from Passiflora edulis and
Carica papaya. Phytochemistry 2002
60(8): 873-882. doi: 10.1016/s0031-
9422(02)00170-x.
[9] Mansur JS, Breder MNR, Mansur
MCA, Azulay RD. Determination
of sun protection factor by
spectrophotometry. An. Bras.
Dermatol. 1986; 61:121-124.
[10] Balsam MS, Sagarin E. Cosmetics :
Science and Technology 2nd Ed.,
Vol. 1-3. Interscience Publishers,
Inc: New York, 1972.
430
S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
[10] Balsam MS, Sagarin E. Cosmetics :
Science and Technology 2nd Ed.,
Vol. 1-3. Interscience Publishers,
Inc: New York, 1972.
[11] Official Journal of the European
Union. COMMISSION
RECOMMENDATION of 22
September 2006 on the efficacy of
sunscreen products and the claims
made relating thereto. Notified under
document number C (2006) 4089.
[12] Lowe NJ, Shaath NA, Pathak MA.
Sunscreens: Development:
Evaluation, and Regulatory
Aspects, 2nd ed., pp. 744, Marcel
Dekker, New York, 1997.
[13] Cefali LC, Ataide JA, Foglio P,
Mazzola G. Plantbased active
photoprotectants for
sunscreens.International Journal of
Cosmetic Science 2016; 38(4): 346-
353.
doi: 10.1111/ics.12316.
[14] Marliani L, Velayanti R, Roni A.
Antioxidant and sunscreen activities
on Carica papaya L. fruit peel
extract, Prosiding SNaPP Universitas
Islam Bandung, 2015; 1(1): 319-324.
eISSN 2477-2356.
[15] Sachin GL, Salunkhe SD, Shenekar
PN, Sutar ST, Ukirade PS, et al. In
vitro evaluation of SPF,
photostability and chemical stability
of Carica papaya L. fruit extract gel.
Pelagia Research Library Der
Pharmacia Sinica 2013; 4(6): 43-47.
[16] Asghar N, Naqvi SA, Hussain Z,
Rasool N, Khan ZA, Shahzad SA,
Sherazi TA, Janjua MR, Nagra SA,
Zia-Ul-Haq M, Jaafar HZ.
Compositional difference in
antioxidant and antibacterial activity
of all Parts of Carica papaya using
different solvents. Chemistry Central
Journal 2016: 10: 5,
https://doi.org/10.1186/s13065-016-
0149-0
[17] Fidrianny I, Paramitha KA,
Kusmardiyani S. Antioxidant
activities from various leaves
extracts of three cultivars Papaya
from West Java-Indonesia. Asian J
Pharm Clin Res. 2016; 9(4): 299-
303.
[18] Saric, S. & Sivamanni, R.K.,
Polyphenol and Sunburn, Intl J Mol
Sci 17(9), pp. 1521, 2016.
https://doi.org/10.3390/ijms1709152
1
[19] de Alencar Filho JM, Sampaio PA,
Pereira EC, de Oliveira Júnior RG,
et al. Flavonoid as a photoprotective
agents: a systematic review. Journal
of Medicinal Plant Research 2016;
10(47): 848-864,
DOI: 10.5897/JMPR2016.6273
[20] Ebrahimzadeh MA, Reza E, Khalili
M, Ghaffarloo M, Saeedi M, Charati
JY. Correlation between sun
protection factor and antioxidant
activity, phenol and flavonoid
contents of some medicinal plants.
Iran J Pharm Res. 2014; 13(3):
1041–1047.
[21] Nunes AR, Moreira AL,
RodriguesALM, Queiróz D, Vieira
IGP, Neto JFC, et. al.
Photoprotective potential of
medicinal plants from Cerrado
biome (Brazil) in relation to phenolic
content and antioxidant activity.
Journal of Photochemistry and
Photobiology 2018; 189:119-123.
doi: 10.1016
431
S. Wardatun et al / Indo J Pharm 5 (2023) 422-431
[22] Mambro VMD, Fonseca MJV.
Assays of physical stability and
antioxidant activity of a topical
formulation added with different
pant extracts. J Pharm Biomed Anal.
2005; 37(2): 287-295.
[23] Thavamoney N, Sivanadian L, Tee
LH, Khoo HE, Prasad KN, Kong
KW. Extraction and recovery of
phytochemical components and
antioxidative properties in fruit parts
of Dacryodes rostrata influenced by
different solvents. Journal of Food
Science and Technology
2018; 55(7); 2523–2532.
https://doi.org/10.1007/s13197-018-
3170-
[24] Winarno H, & Katrin
WE.Benzophenone glucoside
isolated from the ethyl acetate
extract of the Bark of mahkota dewa
(Phaleria macrocarpa (scheff.) boerl)
and its inhibitory activity on
leukemia l1210 Cell line. Indo J
Chem. 2009; 9(1):142-145. [
[25] Ugo NJ, Adesanmi R, Ade, AR,
Tochi JA, Nutrient composition of
Carica papaya leaves extracts. J
Food Sci Nutr Res. 2019; 2(3): 274-
282.
[26] Tendulkar PA, Surve MA, Deodhar
MA. Sun protective formulation
from carotenoids extracted from
thermotolerant genera Synechocystis
pevalekii. Int J Pharm Sci Res. 2018:
9(8); 3223-3234. doi:
10.13040/IJPSR.0975-
8232.9(8).3223-34.
[27] Martinez AJ, Brahm P, Gonzalez A,
Stinco CA. Comprehensive review
on colorless carotenoids phytoene
and phytofluene. Archives of
Biochemistry and Biophysics 2015
572: 188-200.
[28] Darvin ME, Fluhr JW, Meinke MC,
Zastrow L, Sterry W, Lademann J.
Topical beta-carotene protects
against infra-red-light-induced free
radicals. Exp Dermatol. 2011; 20(2):
125-129.
[29] Palombo P, Fabrizi G, Ruocco V,
Ruocco E, Fluhr J, Roberts R, et al.
Beneficial long-term effects of
combined oral/topical antioxidant
treatment with the carotenoids lutein
and zeaxanthin on human skin: a
double-blind, placebocontrolled
study. Skin Pharmacol Physiol.
2007;20(4):199-210.

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