Industrial Crops & Products 130 (2019) 208–215

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Industrial Crops & Products

journal homepage: www.elsevier.com/locate/indcrop

In vitro antioxidant and photoprotective activity of five native Brazilian T

bamboo species
Katarzyna Barbara Wróblewskaa, André Rolim Babyb, Maria Tereza Grombone Guaratinic,
Paulo Roberto Hrihorowitsch Morenoa,
⁎

a
Institute of Chemistry, University of São Paulo, Av. Prof. Lineu Prestes 748, 05508-000, São Paulo, SP, Brazil
b
Faculty of Pharmacy, University of São Paulo, São Paulo, Brazil
c
Institute of Botany, São Paulo, SP, Brazil

ARTICLE INFO ABSTRACT

Keywords: Nature provides an abundant source of antioxidant substances as well as potential compounds for sunscreen
Aulonemia products. Asian bamboos, well known and widely cultivated, are already used in cosmetic and pharmaceutical
Chusquea industries thanks to their rich phenolic contents which is related to their antioxidant activity. This study aimed
Merostachys to evaluate the photoprotective and antioxidant activities of five native Brazilian bamboo species. Firstly, hy-
Flavonoids
droalcoholic extracts from culms and leaves of Aulonemia aristulata (Döll) McClure., Chusquea bambusoides Rupr.
Sun protection factor
ex Döll, Chusquea capituliflora Trin. var. pubescens McClure, Chusquea meyeriana Rupr., Merostachys pluriflora
Photostability
Munro ex E.G. Camus were evaluated for their DPPH scavenging activity and total contents of flavonoids and
phenolic compounds. Secondly, cosmetic formulations containing the extracts in a combination with three
different synthetic ultraviolet (UV) filters (avobenzone, octyldimethyl PABA, and octyl methoxycinnamate) were
developed to evaluate their Sun Protection Factor (SPF), critical wavelength (cλ) and photostability by diffuse
transmittance analysis to determine the interactions involving the extracts and the anti-UV active ingredients.
The bamboos’ antioxidant potential, expressed in IC50, varied between 137.55 and 260 μg/mL. Phenolic contents
ranged from 43.64 to 87.81 mg of gallic acid equivalents (GAE) per g of plant material. The extract richest in
flavonoids was that from C. bambusoides leaves with 6.44 mg of equivalents of quercetin (EQ) per g of dried
leaves. The SPF of the formulations with bamboo extracts varied between 34 and 86 before the irradiation, and
the obtained UV absorption profile allowed to classify them as broad spectrum. After irradiation, the SPF values
diminished to 14–44, whereas, the area of the absorbed wavelengths remained equal. This study showed that the
addition of bamboo extracts to commercial UV filters increased significantly their SPF and photostability.

1. Introduction Although the skin possesses an elaborate defense system to deal

Solar radiation was one of the crucial factors for the existence of life on
Earth. However, besides its beneficial effects, it also causes deleterious
impact on humans, mostly due the exposure to ultraviolet radiation (UV).
The shorter UV wavelengths (290–320 nm) are related to the vitamin D
and melanin production, as a sunscreen function. The longer wavelengths
(320–400 nm) can penetrate to deeper skin layers, generating reactive
oxygen and nitrogen species which moderate cell metabolism and func-
tions, and induce genetic changes (Juzeniene and Moan, 2012). In hu-
mans, intense overexposure to UVA and UVB radiation provokes erythema
(or sunburn), an inflammatory reaction of the skin to UV radiation
(Hönigsmann, 2002), oxidative stress, photoaging, immunosuppression
and photocarcinogenesis (Polefka et al., 2011).
with the UV-induced oxidative stress, humans still need to use addi-
tional photoprotective measures to avoid the harmful effects of UV ir-
radiation, such as diminishing sun exposure, wearing protective
clothing and using topical sunscreens (Balogh et al., 2011). An optimal
sunscreen must be multifunctional being able to reflect, absorb and
scatter UV radiation, giving protection throughout the whole UV range
and, preferentially, exerting antioxidant activity (Morabito et al.,
2011). As the perfect sunscreen substance has not yet been discovered,
the need for further research remains the same nowadays.
Most sunscreen formulations are composed of several chemicals
with each one absorbing at different regions of the UV radiation.
However, several of these ingredient mixtures are not photostable, and
their unknown photoproducts may be toxic for human skin cells (Butt

⁎
Corresponding author.
E-mail address: prmoreno@iq.usp.br (P.R.H. Moreno).

https://doi.org/10.1016/j.indcrop.2018.12.081
Received 4 September 2018; Received in revised form 14 November 2018; Accepted 26 December 2018
Available online 29 December 2018
0926-6690/ © 2019 Elsevier B.V. All rights reserved.
K.B. Wróblewska et al.
and Christensen, 2000). For these reasons, the use of natural products
has gained interest over the last years, mainly the polyphenols due to
their broad UV absorption spectrum, including the UVB and UVA re-
gions. In addition, these compounds can decrease oxidative stress and
anti-inflammatory processes, and they can also stimulate DNA-repair
and prevent skin damage (Radice et al., 2016).
Bamboo is a group of genera of evergreen plants belonging to
Poaceae, or grass family. These species are recognized for their multiple
biological effects, such as antioxidant, anti-free radical, anti-aging, and
antibacterial effects, as well as prevention of cardiovascular diseases
(Hu et al., 2000; Kweon et al., 2001; Fujimura et al., 2005; Wang et al.,
2012). The biological effects were related to the presence of different
phenolic compounds, like flavonoids, cinnamates (Hu et al., 2000),
lignophenol derivatives (Akao et al., 2004), anthocyanins, poly-
saccharides, catechins (Jin et al., 2006) and various volatile compounds
(Fu et al., 2002; Takahashi et al., 2010).
Although Eastern Asian bamboo species have already been well
studied, little is known about the biological activities and chemical
composition of the native Brazilian ones. Bamboo species share some
common characteristics in phenolic types, accumulating several gly-
cosylated flavones whose aglycones are represented by orientin,
homoorientin, vitexin, isovitexin, and tricin (Zhang et al., 2005, Jiao
et al. 2007). As natural phenols have been shown to perform topical
photoprotective action (Saija et al., 2000; Martorana et al., 2013), the
purpose of our study was to verify the potential use of Brazilian bamboo
extracts as natural ingredients in sunscreen formulations. Therefore, we
evaluated the in vitro antioxidant and photoprotective potential of
ethanol extracts from culms and leaves of five native Brazilian bamboo
species: Aulonemia aristulata (Döll) McClure., Chusquea bambusoides
Rupr. ex Döll, Chusquea capituliflora Trin. var. pubescens McClure,
Chusquea meyeriana Rupr., Merostachys pluriflora Munro ex E.G. Camus.
2. Material and methods
2.1. Chemicals
Ethanol and methanol provided by Synth® was used for the extrac-
tion process and chemical analyses. Standards - quercetin, gallic acid,
and the reagents: 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and
Folin-Ciocalteu used for the assays were obtained from Sigma Aldrich®.
Aluminum chloride used for flavonoids quantification was provided by
Merck®. Ingredients of photoprotective formulations contained:
Aristoflex® AVC and Phenonip® by PharmaSpecial®, butylated hydro-
xytoluene (BHT) by Vital®, Avobenzone and Propyleneglycol by All
Chemistry®, Octyldimethyl PABA by Deg®, Octylmethoxycinnamate and
Parrafinum liquidum by Volp®.
2.2. Plant material
Culms and leaves of Chusquea bambusoides Rupr. ex Döll and
Chusquea capituliflora Trin. var. pubescens McClure were collected at
Municipal Natural Springs Park of Paranapiacaba, Santo André
(23°46′41″S and 46°18′16″W), whereas those from Aulonemia aristulata
(Döll) McClure Chusquea meyeriana Rupr. and Merostachys pluriflora
Munro ex E.G. Camus at the Fontes do Ipiranga State Park in São Paulo-
SP, Brazil (23°38′08″-23°40′18″S and 46°36′48″-46°38′00″W). All spe-
cimens were identified by Dr. Tarcísio Filgueiras from Botany Institute
of São Paulo and deposited at the herbarium in the same institution.
Voucher species numbers are: Shirasuna et al. 672 (SP), Shirasuna, RT
& Suzuki, R. 1580 (SP), Vinha, D. s/n. 398161 (SP), Grombone, MT s/n
412118 (SP) and Shirasuna, RT 2619 (SP), respectively.
2.3. Crude extract preparation
Culms and leaves were separated, dried at room temperature, pro-
tected from light and grounded in a knives mill (Tecnal TE-580, Brasil)
209
Industrial Crops & Products 130 (2019) 208–215
with a sieve of 200 μm. Extracts were prepared in a Soxhlet apparatus
with 60% ethanol-water solution, in the proportion of 1.5 L solvent for
each 100 g of plant, until exhaustion (until no residue was detected in
the extractor solvent). The extracts were concentrated using rotary
evaporator under vacuum at 50 °C, dried being lyophilized for 72 h and
stored in a freezer at −20 °C. The yield was calculated based on the dry
weight basis and expressed as percentage (%).
2.4. Scavenger radical activity
Potential antioxidant activity was evaluated using a colorimetric
method based on the scavenging of the DPPH radical described by
Brand-Williams et al. (1995), adapted for 96-well plates. Bamboo ex-
tract samples were dissolved in methanol to obtain final concentrations
in microplates: 500, 250, 200, 100, 50 e 25 μg/mL. Quercetin was used
as a standard in five different concentrations between 2.5 and 0.25 μg/
mL. To the microplates, were added subsequently 50 μL of the sample/
standard and 150 μL of 200 μmol/L methanol solution of DPPH. Me-
thanol together with DPPH was utilized as a negative control. Sample
blanks were prepared by diluting the extracts solutions with methanol,
without the reagent. After 30 min of reaction protected from light, the
absorbances were measured at λ = 515 nm in a microplate reader (LGC
Biotecnologia, LM-LGC). Values of antioxidant activity were expressed
by the half maximum inhibitory concentration (IC50, mean ± standard
deviation), concentration needed to quench 50% of the DPPH radical,
using the linear regression curves of the samples’ radical inhibition.
2.5. Total phenolic contents
Total amount of phenols in bamboo extracts was determined by a
spectrophotometric method developed by Ainsworth and Gillespie,
(2007) using Folin-Ciocalteu reagent. Bamboo extracts were dissolved
in methanol to obtain a solution of 1 mg/mL, whereas the final con-
centration in the plates was 100 μg/mL. Gallic acid in different con-
centrations (10 – 2.5 μg/mL) was used as reference compound for
quantification. The assay was performed by adding to the microplates
20 μL of the sample, 30 μL of distilled water, 50 μL of Folin-Ciocalteu
reagent and 100 μL of solution of sodium carbonate (700 mmol/L).
Sample blanks were prepared to discount the absorbance of the ex-
tracts’ solutions using methanol in place of the reagent. After 2 h of
reaction in dark, the absorbance of the samples was measured in
spectrophotometer (SpectraMax M4), at λ = 760 nm. The results were
expressed in milligrams of gallic acid equivalents per g of plant material
(mg GAE/g, mean ± standard deviation).
2.6. Total flavonoid contents
Another spectrophotometric method (Woisky and Salatino, 1998)
was used to estimate total flavonoid contents. Test samples in con-
centration of 5.0 mg/mL (400 μL/mL in the well) were prepared by
dissolving dry bamboo extracts in 80% solution of methanol. Quercetin,
in five different concentrations between 15 and 2.5 μg/mL in the same
solvent solution, was used as quantification reference. For the assay,
20 μL of sample, 210 μL of 80% methanol and 20 μL of 2% aluminum
chloride solution were added to the microplates. As blank, some mi-
croplate wells received only the sample and 80% methanol. After
30 min in darkness, absorbances were measured at λ = 425 nm in
spectrophotometer (SpectraMax M4). The results were expressed in
milligrams of quercetin equivalents per g of plant material (mg QE/m,
mean ± standard deviation).
2.7. Preparation of photoprotective formulations
Sunscreen formulations (20 g) were prepared using m/m propor-
tions. The bamboo extracts were added in concentration of 10%.
Aristoflex® AVC (ammonium acryloyldimethyltaurate/VP copolymer), a
K.B. Wróblewska et al. Industrial Crops & Products 130 (2019) 208–215

synthetic polymer, which was an emulsifier and gelling agent was c

A ( ) d = 0.9
400 nm
A ( )d
added in 3%. All formulations contained chemical UVA and UVB filters 290 nm 290 nm (2)
in the following concentrations: 3.0% avobenzone; 8,0% octyldimethyl The results were presented as mean ± standard deviation.
PABA, and 7.5% octylmethoxycinnamate. Additionally, BHT was used
as antioxidant (0.1%), Paraffinum liquidum (mineral oil) as emollient
2.9. Photostability assay
and solvent of the oil phase of the formulation (5%). Propylene glycol
was added as humectant (5%) and mixture of 2-phenoxyethanol, me-
Formulations had their functional photostability evaluated using the
thyl 4-hydroxybenzoate, ethyl 4-hydroxybenzoate, propyl 4-hydro-
same procedure as described above with an additional step of irra-
xybenzoate and butyl 4-hydroxybenzoate called Phenonip® as pre-
diating the samples with a fixed dose of artificial solar light (580.08 W/
servative (0.75%). Distilled water (q.s.) was a vehicle of the aqueous
m2 - 2088 J/m2) in a photostable chamber for one hour at 35 °C. The
(gel) phase.
sample and control absorbances were measured using the same diffuse
In the first step, anti-UV agents were dissolved together by mixing
reflectance spectrophotometer with the identical reading program. The
with BHT and mineral oil at 70 °C in a beaker. Dry bamboo extracts
results were also converted to SPF and cλ values and presented as
were weighed and pulverized in a mortar. One half of total distilled
mean ± standard deviation. Nine readings were recorded for each
water volume was used to dissolve bamboo extracts in a mortar with
plate.
propylene glycol and then, the solution obtained was transferred to a
beaker with Aristoflex® AVC. The remaining quantity of water was
added, and gel was obtained by mixing the water solution with a glass 2.10. Statistical tests
rod. Finally, the UV filter solution was incorporated by parts to the
formed gel. The formulation was then homogenized with the pre- To compare the differences among all the samples in the measure-
servative in mechanical stirrer (IKA RW 20) at 1000 rpm for 5 min. ments one-way ANOVA and Tukey test were used. Changes in SPF and
In total, 11 sunscreen formulations were prepared, 10 of them were cλ after irradiation in photoprotective and photostability assessments
test samples prepared with leaf or culm bamboo extracts plus the UV were detected by t-Student test. All the statistical analyses we per-
filters, numbered as formulations F1 to F10, and one control formula- formed in Minitab® 17.
tion (F11), containing only the filters, as presented in Table 1.
3. Results and discussion

2.8. In vitro SPF evaluation 3.1. Crude extract analyses

In vitro photoprotective efficacy of bamboo extracts was estimated 3.1.1. Extraction yield
by diffuse reflectance spectrophotometer with integrating sphere Since the bamboos are known to possess glycosylated flavonoids as
(Labsphere®UV2000S). For the assay, formulations containing the one of the main compounds, 60% ethanol was chosen as a solvent for
bamboo extracts and the control were weighed and uniformly spread as the extraction process because of its higher polarity. The crude extracts
a thin film (1.3 mg/cm2) on the surface of a polymethyl-methacrylate yield varied between 7.6 and 22.2% (m/m), with the leaves affording
plate (PMMA, HD Helioplate® 6 HelioScreen) according to the higher values for all species, as it is usual in plant materials (Table 2). C.
Cosmetics Europe guidelines (2011), previously known as COLIPA (The capituliflora leaves presented almost the double of extractable material
European Cosmetic and Perfumery Association). After 40 min of drying, (22.2%) than those from the other species, where the yields ranged
sample absorbances were measured between 290 and 400 nm, with from 10 to 14%. In general, culm extracts yields were alike to those
progression rate of 1.0 nm. Nine readings were recorded for each plate. obtained for leaves, with the lowest results for M. pluriflora culms.
Plates without the film were used as blanks, and two of them were used Comparing these values with Asian bamboo species, the native Brazi-
for the control formulation (F11). Absorbance values were converted to lian bamboos presented similar yields (Wang et al., 2012; Macwan
in vitro sun protection factor (SPF) and critical wavelength (cλ) using et al., 2010).
the Eqs. 1 and 2 (Cosmetics Europe, 2011; Diffey, 2002; Springsteen
et al., 1999), where: Eλ is spectral irradiance effectiveness accordingly
3.1.2. Antiradical activity
to CIE (Commission Internationale de l'Eclairage); Sλ is solar spectral
Due to their antioxidant effects (Hu et al., 2000), associated with the
irradiance; Tλ is spectral transmittance of the sample; dλ is range of
high polyphenol contents, bamboos are currently proposed as pro-
wavelengths and A (λ) is spectral absorbance of the sample.
mising anti-UV agents, once there is a correlation between reactive
400 nm
E S d oxygen species (ROS) and UVB exposure (Dinkova-Kostova, 2008).
290 nm
SPF = 400 nm These factors lead to an initial screening for antioxidant properties,
290 nm
E S T d (1) expressed as radical scavenger potential, and the evaluation of the total
phenol and total flavonoid contents in the Brazilian bamboo extracts.
Table 1 Although, DPPH method does not directly reflect the biological anti-
List of sunscreen formulations developed for the photoprotective assay sorted oxidant activity, it is commonly used, especially for screening purposes,
by bamboo species and part of the plant used. because it is quick, reliable and reproducible.
The bamboo extracts presented DPPH scavenging activity (IC50)
Formulation Bamboo species Part of the plant used
ranging from 137 to 260 μg/mL, where the most potent extract was C.
F1 A. aristulata Culm capituliflora leaves (Table 2). Despite the wide activity range, no sig-
F2 Leaves nificant difference was observed among the culm and leaf samples. In
F3 C. bambusoides Culm
general, all the native Brazilian bamboos presented an antioxidant
F4 Leaves
F5 C. capituliflora Culm potential approximately 10 times higher than those observed for the
F6 Leaves Asian bamboos Phyllostachys pubescens and P. nigra (Park and Jhon,
F7 C. meyeriana Culm 2010). The antioxidant potential observed for the native Brazilian
F8 Leaves bamboos were alike to reported for turmeric roots with
F9 M. pluriflora Culm
IC50 = 104.91 μg/mL in the DPPH assay, which are well known for
F10 Leaves
F11 Control NA their antioxidant properties (Sukandar et al., 2015). These results in-
dicate a promising antioxidant potential for the bamboo extracts,

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Table 2
Yield (%), antioxidant activity assay (IC50), total phenolic compounds (GAE) and total flavonoid content (QE) tests obtained for native Brazilian bamboo species.
Bamboo species Yield DPPH IC50 GAE QE
[%] [μg/mL] [mg/g] [mg/g]

Culm Extracts
A. aristulata 9.9 205.68 ± 31.22b,c 55.02 ± 0.47c,d,e 3.16 ± 0.38f,g,h,i
C. bambusoides 12.2 236.59 ± 42.22c 43.64 ± 2.24e 2.22 ± 0.25i
C. capituliflora 8.6 244.36 ± 50.30c 46.31 ± 0,95d,e 4.83 ± 0.25b,c,d
C. meyeriana 7.8 168.18 ± 21.79a,b 87.81 ± 2.14a 4.60 ± 0.16c,d,e
M. pluriflora 7.6 174.17 ± 12.88a,b 54.40 ± 2.30c,d,e 4.01 ± 0.33d,e,f,g
Leaf Extracts
A. aristulata 10.1 260.17 ± 42.14c 49.54 ± 0.74d, e
3.65 ± 0.40d, e, f, g, h

C. bambusoides 14.2 218.16 ± 34.56b,c 59.53 ± 4.83b, c, d

6.44 ± 0.25a
C. capituliflora 22.2 137.55 ± 15.62a 57.00 ± 4.49b, c, d, e
3.87 ± 0.31d, e, f, g, h

C. meyeriana 14.2 205.48 ± 38.14b,c 71.03 ± 3.61b 4.08 ± 0.04d, e, f

M. pluriflora 13.9 222.52 ± 20.05b,c 47.34 ± 0.62d, e

5.93 ± 0.27a, b

IC50 – Inhibition Concentration – concentration needed to inhibit 50% of the radical formation; GAE – Gallic Acid Equivalents; QE – Quercetin Equivalents. Statistic
tests were performed separately for each assay. Small letters represent results of one-way ANOVA test with Tukey comparison. Values in the same column that do not
share the same letter are statistically different.

although the IC50 values were much higher than that from pure quer-
cetin (1.76 μg/mL), the positive control.
3.1.3. Quantification of phenolic compounds and flavonoids
As the DPPH scavenging activity might be related with the presence
of polyphenols (Dudonné et al., 2009), the contents of total phenolic
compounds and flavonoids were measured in the extracts. The total
phenolic compounds varied from ˜44 to 88 mg GAE/g of extract as per
Table 2. The highest amounts of phenolics were found in the C.
meyeriana culm and leaf extract (87.81 and 71.03 mg GAE/g, respec-
tively). All five Brazilian bamboo species showed higher amounts of
phenolic than those reported for the Asian species. B. arundinacea
containing only 14.6 mg GAE/g (Macwan et al., 2010). Similarly, total
phenolic content of Schizostachyum lumampao (Blanco) Merr were 76.7
and 13.5 mg of GAE per 100 g of air-dried sample for the ethanolic and
aqueous extracts, respectively (Tongco et al., 2014).
Flavonoids are a special group of phenolic compounds which are
also involved in the antioxidant properties of plant extracts (Molay
et al., 2010). The total amount of flavonoids found in native Brazilian
bamboo species, expressed in milligrams equivalents of quercetin (QE),
ranged from 2.22 to 6.44 QE per g of extract (Table 2). Leaf extracts
were richer in flavonoids than those from culms. The results obtained
for other bamboo species varied, for example, in Dendrocalamopsis
oldhami (Munro) Keng f. total flavonoid content was higher – 74 mg
QE/g of bamboo leaf extract (Lv et al., 2012). On the other hand,
aqueous and methanolic extracts from Sasa senanensis (Franch. & Sav.)
Rehder gave only 1.18 and 1.85 mg Q/100 g of leaf powder (Khatun
et al., 2013), whereas S. lumampao had 70.2 and 17.86 mg QE per
100 g air-dried leaves for the ethanolic and aqueous extracts, respec-
tively (Tongco et al., 2014) which is much less than the Brazilian
bamboos. Nevertheless, seasonal variations can be responsible for the
differences observed in the chemical composition of the bamboos (Ni
et al., 2012).
3.2. Photoprotection and photostability of bamboo formulations
3.2.1. Formulations
All the formulations containing bamboo extracts were emulsified
systems. Their individual characteristics are listed in Table 3. The
emulsions started to separate small amounts of the oil phase up to 2–3 h
after the preparation. Formulations F2, F4, F6 and F8 were defined as
more homogeneous due to a uniform aspect. The color of formulations
varied from yellowish, light to dark green or brown with a specific
herbal odor. The control formulation (F11) was homogenous and with
creamy color and a characteristic odor of the gelling base. Difficulties
during gel formation can be caused by diverse chemical compounds
found in bamboo extracts that may interact differently with other oil
phase ingredients, such as synthetic UV filters. Also, the variations
between each formulation properties resulted from distinctive features
of each extract.
3.2.2. In vitro photoprotection and photostability assays
SPF is the worldwide parameter used to evaluate the efficacy of
sunscreens and its general definition is the ratio of the energy of UV
radiation required to produce erythema in the protected skin (with
sunscreen) to that required to produce erythema in the unprotected skin
(without sunscreen), obtained through in vivo assays (Ebrahimzadeh
et al., 2014). However, recently some methods using diffuse reflectance
spectroscopy (DRS), a non-invasive assay to determine the UV protec-
tion, have demonstrated good correlation with in vivo methodologies
(Junior et al., 2014).
In our study, the photoprotection of sunscreen formulations con-
taining bamboo extracts and well-known UV filters (Avobenzone,
Octyldimethyl PABA and Octylmethoxycinnamate) was evaluated by
measuring the SPF using DRS before sunlight irradiation and compared
with a control containing only the solar agents. All formulations with
the bamboo extracts presented SPF values between 34.52 (F3) and
86.15 (F1), showing higher values than the control (F11), SPF 13.00, as
can be seen in Table 4. Although the standard deviations for the sam-
ples with the extracts were considered high, in some cases more than
50%, statistical analysis showed a significant difference with the con-
trol. According to Tukey test, only the C. bambusoides formulations (F3
and F4) were not different from the control. The high standard devia-
tions observed for the formulations containing bamboo extracts might
be related to lack of homogeneity of the samples, that might affect the
equipment reflectance optic readings. The experiment was performed 3
times, since application of the formulations over the PMMA plates is
done manually, and the film characteristics might have been different
among the exposed area of analysis, influencing the result variations. In
addition, formulations containing only bamboo extracts did not present
a considerable SPF (data not shown), but when combined with syn-
thetic filters they were able to increase measured photoprotection.
As the SPF alone is an indicator only for a protection against er-
ythemally effective solar UV, largely, confined to the UVB
(290–320 nm) and partially short-wavelength UVA (320–340 nm) ra-
diation, it does not provide information regarding protection against
long-wavelength UVA1 (340–400 nm) (Hojerová et al., 2011), which
can lead to several cumulative damages, such as immunosuppression,
skin aging, and various other photodisorders (Velasco et al., 2008). For
this reason, critical wavelength (cλ) is another parameter that describes
if the photoprotective product achieved broad spectrum of action. It is
defined as the wavelength at which the area under the curve gives 90%

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Table 3
Formulations received from bamboo extracts and synthetic solar filters.
Formulation number Aspect Organoleptic properties Formulation Aspect Organoleptic properties
number

F1 Partially homogeneous emulsion, brown F6 Homogeneous emulsion, green color and herbal
color and herbal odor odor

F2 Homogeneous emulsion, brown color and F7 Partially homogeneous emulsion, yellowish color
herbal odor and herbal odor

F3 Partially homogeneous emulsion, brown- F8 Homogeneous emulsion, green-brown color and

yellowish color and herbal odor herbal odor

F4 Homogeneous emulsion, dark green color and F9 Partially homogeneous emulsion, yellowish color
herbal odor and herbal odor

F5 Partially homogeneous emulsion, yellowish F10 Partially homogeneous emulsion, dark orange-
color and herbal odor brown color and herbal odor

F11 Homogeneous emulsion, clear yellowish color and

(control) specific odor of filters/cream base

Extract type used in formulations: F1 – A. aristulata culms; F2 – A. aristulata leaves; F3 – C. bambusoides culms; F4 –C. bambusoides leaves; F5 – C. capituliflora culms;
F6 – C. capituliflora leaves; F7 – C. meyeriana culms; F8 – C. meyeriana leaves; F9 – M. pluriflora culms; F10 – M. pluriflora leaves; F11: water (control).

of the total absorbance of the sample between 290–400 nm. The Food
and Drug Administration (FDA) considers 370 nm as the minimal cλ for
broad spectrum sunscreens (Polonini et al., 2014). Additionally, an
ideal sunscreen must have a minimal SPF value of 15, in accordance
with the FDA (Food and Drug Administration, 2007). As can be seen in
Table 4, all bamboo formulations can be defined as good efficacy
samples (cλ higher than 370 nm and SPF more than 15), whereas the
control sample presented broad spectrum of action (cλ higher than
370 nm) but low photoprotective effect (SPF less than 15).
Photostability is another essential property describing the effec-
tiveness and safety of sunscreen products, since UV radiation can lead
to reduction of the photoprotective capacity of UV filters and can also
generate free radicals (Gonzalez et al., 2007). Avobenzone is a widely
used UVA filter that is known to be a rather unstable substance, loosing
much of its protective capability after irradiation through tautomer-
ization, fragmentation and photoproduct formation. The photolysis
mechanism involves singlet oxygen formation that may induce irre-
versible modifications in skin proteins, leading to tissue photo-
degradation and other harmful effects to the skin cells (Abid et al.,
2017; Paris et al., 2009). Additionally, it is known that octylmethox-
ycinnamate in the presence of avobenzone undergoes a photolysis
process, rather than the expected E/Z photoisomerization, producing

Table 4
Sun Protection Factor (SPF) and critical wavelength (cλ) of sunscreen formulations containing native Brazilian bamboo extracts, before and after sunlight irradiation.
Formulation SPF (mean ± standard deviation) cλ [nm]

a b
Before irradiation After irradiation Before irradiationx After irradiationx

F1 86.15 ± 40.81A,B 32.30 ± 15.55E,F,G 380X 378X

F2 61.41 ± 37.36A,B,C 22.04 ± 12.16G,H,I 381X 379X
F3 34.52 ± 10.39C,D 14.33 ± 4.17H,I 380X 378X
F4 36.61 ± 10.68C,D 35.22 ± 13.48E,F 381X 380X
F5 78.22 ± 53.15A,B 44.44 ± 37.31E,F 381X 379X
F6 61.59 ± 35.99A,B,C 25.93 ± 11.86F,G,H 383X 381X
F7 84.37 ± 40.98A,B 28.78 ± 20.12E,F,G,H 381X 379X
F8 51.41 ± 25.84B,C 20.41 ± 13.53G,H,I 381X 379X
F9 83.19 ± 41.11A,B 41.33 ± 24.91E,F 380 X 379X
F10 61.48 ± 32.26A,B,C 41.00 ± 27.33E,F 380X 380X
F11 (control) 13.00 ± 2.50D 6.00 ± 1.6I 381X 379X

SPF - Sun Protection Factor; cλ – critical wavelength. Small letters show the results of t-Student test; differences between the values obtained before and after
irradiation. Capital letters show the differences between all the samples (one-way ANOVA + Tukey comparison test). Values not sharing the same letter are sta-
tistically different. Extract type used in formulations: F1 – A. aristulata culms; F2 – A. aristulata leaves; F3 – C. bambusoides culms; F4 –C. bambusoides leaves; F5 – C.
capituliflora culms; F6 – C. capituliflora leaves; F7 – C. meyeriana culms; F8 – C. meyeriana leaves; F9 – M. pluriflora culms; F10 – M. pluriflora leaves; F11: water
(control).

212
K.B. Wróblewska et al.
Fig. 1. Average UV absorbance profiles for formulations F1-F11 before (A) and after (B) irradiation with artificial sunlight for 1 h. Extract type used in formulations:
F1 – A. aristulata culms; F2 – A. aristulata leaves; F3 – C. bambusoides culms; F4 –C. bambusoides leaves; F5 – C. capituliflora culms; F6 – C. capituliflora leaves; F7 – C.
meyeriana culms; F8 – C. meyeriana leaves; F9 – M. pluriflora culms; F10 – M. pluriflora leaves; F11: water (control was recorded twice, as F11_1 and F11_2).
long lived radicals in the sunscreen film (Sayre et al., 2005). For these
reasons, these filters were used in the formulations to test the photo-
stabilization capacity of the Bamboo extracts.
The results obtained after irradiation with the artificial sunlight
(Table 4) showed that the formulations continued to be broad spectrum
sunscreens as the cλ did not change. However, SPF decreased sig-
nificantly for all the samples, except of F4 (C. bambusoides leaf). The
photoprotection efficacy was still good after irradiation (FPS > 15) for
most formulations, with exception of F2, F3 and F8 that were not sig-
nificantly different from the control. The differences observed in the
absorbance profiles after irradiation can be seen in Fig. 1.
Sunscreens could have antioxidants added in their composition to
complement UV filter photoprotection by reducing the damage from
the free radicals generated after solar exposure and to stabilize che-
mical UV filters (Lim et al., 2017; Badea et al., 2015). Natural extracts
have been one of the strategies used to broaden the photoprotection
spectrum and increase the photostability of sunscreens, mainly those
containing polyphenolic compounds due to their ability to absorb the
solar spectrum and react with free radicals (Cefali et al., 2016; Korać
and Khambholja, 2011). A positive correlation has been found for the
phenolic contents, like flavonoids and hydroxycinnamic acids, in an
extract and the increase in SPF (Ebrahimzadeh et al., 2014). Flavonoids,
besides their UV absorption, may contribute to stabilize the UV filters
by multiple mechanisms including oxygen radical scavenging, inhibi-
tion of lipid peroxidation and metal ion chelation. It has been demon-
strated that rutin stabilization effect on two UV filters cannot be at-
tributed only to its antioxidant activity (Velasco et al., 2008).
Additional mechanisms contribute to quercetin effectiveness such as
antiradical property, chelation of metal ions acting as catalyst in reac-
tions leading to free radical formation and quenching reactive oxygen
species, like the excited triplet states from the organic UV filters butyl
methoxydibenzoylmethane and octyl methoxycinnamate (Scalia and
Mezzena, 2010). In this sense, the photoprotection and photostability
conferred to the formulation by the native Brazilian bamboo species
might be related to their phenolic and/or flavonoid contents, but
213
Industrial Crops & Products 130 (2019) 208–215
mechanisms involved and the compounds responsible for these inter-
actions still need to be elucidated.
Several plant extracts have already demonstrated a photoprotective
activity by increasing the formulations SPF or other mechanisms.
However, acceptable SPF values are only achieved with high amounts
of the plant extract, like with the oil fraction of spent coffee grounds
and green coffee beans that needed 35% of the oils to give SPF of 51.9
and 82.3 (Marto et al., 2016), or in the presence of inorganic filters, as
was observed for formulations containing 1.68% (w/w) Passiflora in-
carnata L. dry extract with 7.0% (w/w) ethylhexyl methoxycinnamate,
2.0% (w/w) benzophenone-3 and 2.0% (w/w) TiO2 generated an SPF of
20 (Velasco et al., 2008). Comparing with our results, extracts from
native Bamboo species at 10% (w/w) concentration could improve the
photoprotective activity from formulations using only organic filters,
reaching SPF values up to 86, and they also promoted the filters sta-
bility as SPF’s remained 2.4 to 7.4 times higher than the control after
irradiation.
4. Conclusions
In this study, screening of crude extracts from Brazilian bamboo
species for potential antioxidant activity, phenolic and flavonoid con-
tents indicated that the phenolic contents are higher than in Asian
species and a good radical scavenging activity. These results led to the
preparation of sunscreen formulations, with the same plant extracts, in
a combination with commercial UV filters. The results showed that the
addition of the biological samples increased the SPF and photostability
of the synthetic UV filters. According to our observations, due to their
rich in phenolic substances composition, the Brazilian bamboo species
can provide a promising source of biologically active agents increasing
the efficacy of synthetic solar filters. The next step of this research will
be to isolate and identify the substances responsible for these properties
and their mechanism of action for the most active species.
K.B. Wróblewska et al.
Acknowledgements
This work was realized thanks to the financial support of Brazilian
National Council for Scientific and Technological Development (CNPq,
Conselho Nacional de Pesquisas) and São Paulo Research Foundation
(FAPESP / process 2016/24360-4). We would like to acknowledge the
provision of plant material and great technical support of the personnel
from Botany Institute of São Paulo for as well as extremely friendly
cooperation of the staff of Cosmetology Department of Pharmacy at
University of São Paulo.
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