Jiratchayamaethasakul et al.

Fisheries and Aquatic Sciences (2020) 23:0

https://doi.org/10.1186/s41240-020-00149-8

RESEARCH ARTICLE Open Access

In vitro screening of elastase, collagenase,

hyaluronidase, and tyrosinase inhibitory
and antioxidant activities of 22 halophyte
plant extracts for novel cosmeceuticals
Chanipa Jiratchayamaethasakul1, Yuling Ding1, Ouibo Hwang1, Seung-Tae Im1, Yebin Jang1, Seung-Won Myung1,
Jeong Min Lee2, Hyun-Soo Kim2, Seok-Chun Ko3 and Seung-Hong Lee1*

Abstract
Background: Halophyte plant (HPs), a salt-resistant flora, has been reported to provide several health benefits, but
the knowledge of its cosmeceutical potential is still ambiguous. Here, 70% ethanol extracts of 22 HPs collected from
along the coast of South Korea were investigated for their potentials of antioxidant, anti-aging, and whitening
properties for use as materials in novel cosmeceuticals.
Methods: Antioxidant activities were determined by DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical and
hydrogen peroxide scavenging assays, and skin aging-related enzyme activities (anti-elastase, anti-collagenase, anti-
hyaluronidase, and anti-tyrosinase) were evaluated by using the spectrophotometric method.
Results: Among the 22 HPs, we found that Ischaemum antephoroides f. coreana and Atriplex gmelinii extracts
presented the strongest scavenging effects against DPPH free radical and hydrogen peroxide, respectively. Our
finding additionally suggested that Salicornia europaea extract might provide a major source of anti-elastase and
anti-hyaluronidase; meanwhile, Rosa rugosa extract showed the highest anti-collagenase effect. Furthermore, the
highest tyrosinase inhibitory activity was possessed by Spartina anglica extract.
Conclusion: These findings may suggest that halophyte plants showing biological activities may be potent
inhibitors of tyrosinase, elastase, collagenase, and hyaluronidase and could be useful for application in
cosmeceuticals.
Keywords: Halophyte plants, Antioxidant, Skin aging-related enzyme activities, Cosmeceuticals

Introduction undergone by the passage of time (Thring et al. 2009),

Skin is the most visible part in the human body which whereas extrinsic aging is an event resulting from the
plays an essential role as a barrier protecting an internal exposure to external factors, predominantly by ultravio-
organ against physical, chemical, and biological detrac- let (UV) radiation known as photoaging (Yang et al.
tors (Kendall and Nicolaou 2013). In recent decades, an 2016). Solar radiation, or UV radiation, is the major
awareness of the aging skin has become one of the most stimulator that accelerates the overproduction of react-
highlighting issues for scientists, and the number of skin ive oxygen species (ROS) which leads the endogenous
aging studies are continually increasing. Skin aging can oxidative stress in the epidermis (Kim et al. 2016). More-
be identified into intrinsic and extrinsic aging processes; over, the excess oxidative stress is so harmful that it in-
intrinsic aging is an inevitable process or natural aging duces unhealthy and aging skin contributing as wrinkle,
roughness, dryness, elasticity loss, and uneven pigmenta-
* Correspondence: seunghong0815@gmail.com tion due to the degradation of extracellular matrix
1
Department of Pharmaceutical Engineering, Soonchunhyang University, (ECM) (Horng et al. 2017). In addition, the exalted levels
Asan 31538, Republic of Korea of ROS can cause not only the senescence and damage
Full list of author information is available at the end of the article

© The Author(s). 2020 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.
Jiratchayamaethasakul et al. Fisheries and Aquatic Sciences (2020) 23:0 Page 2 of 9

of biological functions such as lipid membrane peroxida- the UV radiation and hormonal factors, like the cytokines
tion, DNA deterioration, cell death but also human’s dis- (Asanka et al. 2018; Chatatikun et al. 2019; Tu and Tawata
ease, namely, cancer, stroke, Parkinson’s disease, heart 2015). However, uncontrolled and overproduction of mel-
disease, arteriosclerosis, infection, and autoimmune dis- anin may cause skin disorders, including freckles, mel-
ease (Popoola et al. 2015). It has been carried out that asma, age spots, senile lentigines, and post-inflammatory
an antioxidant provided a great potential to defend ROS hyperpigmentation leading to flaw and premature aging
and deplete the oxidative stress, thus, a compound with appearance. Tyrosinase is a melanogenic enzyme that
the strong antioxidant activity additionally facilitates the plays a crucial role as a rate-limiting step during melanin
skin protecting against the oxidative damages along with pigmentation (Chatatikun et al. 2019). Therefore, the
delaying the skin aging process (Chanda et al. 2015; downregulation or inhibition of tyrosinase activity is a
Palmer and Kitchin 2010). common approach that is recommended to deal with dis-
ECM, the outermost part of the skin, consists of fibro- order pigmentation and used as a whitening agent in aes-
blasts and proteins including collagen and elastin (Fulop thetic purpose (Kang et al. 2018).
et al. 2012; Ndlovu et al. 2013). Collagen is the most abun- Since the lesser adverse reactions were mentioned by a
dant protein structure in the human dermis layer provid- natural product, the use of natural cosmetic ingredient in-
ing the tensile strength of the skin; meanwhile, elastin, a cluding botanic plants has received attention and become
fiber network located in the connective tissue, is respon- target for investigation (Liyanaarachchi et al. 2018). Halo-
sible for the elastic recoil property. Indeed, collagen and phyte is a saline-tolerant plant that can grow under the ex-
elastin are necessary to the skin which play major role for treme environment; the recent data mentioned that it
the plumpness, flexibility, integrity, and elasticity keeping might provide a good biological potential for human health
skin youthful and healthy (Horng et al. 2017; Siedle et al. due to its great resistance surviving in stressful conditions
2002; Thring et al. 2009; Varani et al. 1998). However, the (oxidative stress, UV radiation, salinity, and extreme
accumulated ROS after skin exposure to photoaging temperature; Jdey et al. 2017). Moreover, several investiga-
stressors can indirectly activate dermal enzymes such as tions have been carried out, its health benefits used as folk
collagenase and elastase which basically break down and drug such as antioxidant, anti-inflammatory, antinocicep-
degrade collagen as well as elastin, respectively (Chatati- tive, anti-cancer, and antimicrobial properties (Dudonné
kun and Chiabchalard 2017; Popoola et al. 2015; Sahasra- et al. 2011; Küpeli et al. 2006; Meot-Duros et al. 2008).
budhe and Deodhar 2010). Thereby, the synthesis of However, there have rarely been many previous reports of
elastase and collagenase promotes premature skin aging HP application in terms of cosmetic material. Here, bio-
as evidenced by signs such as wrinkles, freckles, sallow- logical activities on skin health of 22 halophytic plants lo-
ness, deep furrows or severe atrophy, laxity, and leathery cated from South Korea were challenged by screening their
appearance (Ding et al. 2018; Peres et al. 2011). antioxidant activity and anti-elastase, anti-collagenase, anti-
Hyaluronic acid or hyaluronan (HA), a glucose-based hyaluronidase, and anti-tyrosinase effects as candidates for
polymer, can be commonly found in tissues and fluids of cosmeceutical applications.
the body, but it is most bountiful in the dermal compart-
ment of skin and the epidermal layer. HA mainly pro- Materials and methods
motes skin rejuvenation, contains moisture, increases Chemicals and reagents
viscosity, and reduces the permeability of extracellular 1,1-Diphenyl-2-pricrylhydrazyl (DPPH), peroxidase, 2,2-
fluid (Leach et al. 2003). Owing to the excellence of water- azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS),
holding capability, the HA-rich area contributes emolli- peroxidase from horseradish, tyrosinase from mushroom, L-
ence, smoothness, and youngness together with wrinkle tyrosine, collagenase from Clostridium histolyticum, Azo
diminution of the skin (Jegasothy et al. 2014). Unfortu- dye-impregnated collagen, N-succinyl-Ala-Ala-Ala-p-nitroa-
nately, HA is naturally decreased during the aging process, nilide (AAAPN), elastase from porcine pancreas, 4-
whereas hyaluronidase is synthesized. Hyaluronidase is an (dimethylamino)benzaldehyde (DMAB), potassium tetrabo-
HA-destructive enzyme which leads to loss of strength, rate tetra-hydrate (K2B4O7·4H2O), phosphate buffered saline
flexibility and moisture, and subsequently, skin aging (PBS) were purchased from Sigma-Aldrich Chemical Co.
(Ndlovu et al. 2013). Accordingly, one of the anti-wrinkle (St. Louis, MO, USA). All chemicals and reagents used in
approaches is to prolong skin moisture by preserving HA this experiment were analytical grade.
contents underneath the skin.
Melanin, the black or brown pigment, is a major com- Sample preparation
ponent of the skin, hair, and eye color which is synthe- Seventy percent ethanol extract of 22 HPs used in the
sized by melanogenesis, a mechanism of the melanocyte. present study were kindly provided by the National Mar-
Melanin pigmentation mainly enhances the skin’s protect- ine Biodiversity Institute of Korea (MABIK) as shown in
ive barrier against various environmental factors especially Table 1.
Jiratchayamaethasakul et al. Fisheries and Aquatic Sciences (2020) 23:0 Page 3 of 9

DPPH free radical scavenging activity Collagenase inhibitory assay

The DPPH scavenging activity was performed as de- Collagenase inhibitory activity was evaluated based on
scribed by Nanjo et al. (1996). Briefly, DPPH reagent the method described by Wang et al. (2018) with some
was dissolved in methanol for a solution concentration modifications. A fixed weight of 1 mg of Azo dye-
of 1.5 × 10-4 M. One hundred microliters of DPPH re- impregnated collagen was measured in the test tubes
agent was mixed with 100 μl sample in 96-well plates. and then the homogenization was proceeded after the
After incubation at room temperature for 30 min, the addition of an 800 μl of 0.1 M Tris-HCl (pH 7) and a
absorbance was measured 517 nm using a microplate 100 μl of sample into each of test tubes. A 100 μl colla-
reader (Sunrise TW, Tecan Trading AG, Männedorf, genase (200 units/ml) was immediately mixed into the
Switzerland). mixture and incubated at 43 °C for 1 h. Afterward, the
test tubes were centrifuged at 3000 rpm for 10 min. The
supernatant section of each test tube was transferred
Hydrogen peroxide scavenging activity into 96-well plates and the absorbance of each super-
Hydrogen peroxide scavenging activity was examined ac- natant was measured at 550 nm (Sunrise TW, Tecan
cording to Müller (1985) with slight modification. A Trading AG, Männedorf, Switzerland).
100 μl of 0.1 M PBS buffer (pH 5) was added into a 96-
well plate. Each 20 μl of sample and 20 mM hydrogen Elastase inhibitory effect assay
peroxide (H2O2) were added to mix with the buffer, and Elastase inhibition was determined by measuring the in-
then incubate 37 °C for 5 min. After the incubation, a tensity of the solution color assayed using the method of
30 μl of 1.25 mM ABTS and peroxidase (1 unit/ml) were Tu and Tawata (2015) with slight modifications. The mix-
added to the mixture and then incubated at 37 °C for 10 ture of AAAPVN elastase substrate in 0.1232 M Tris-HCl
min. The absorbance was read with a microplate reader buffer solution (pH 8) was prepared to obtain a concentra-
at 405 nm (Sunrise TW, Tecan Trading AG, Männedorf, tion of 1.015 mM. The elastase substrate was mixed with
Switzerland). the 10 μl of sample in the 96-well plates, and preincubated
at 25 °C for 10 min. After the preincubation, the reaction
was initiated by adding 10 μl of elastase from porcine pan-
Table 1 Scientific name of HPs used in this study creas (7.5 units/ml) in Tris solution buffer to the preincu-
Halophyte plants Scientific name (Korean name) bated mixtures. Finally, the absorbance was measured at
HP. 1 Salsola komarovii (수송나물) 410 nm using a microplate reader (Sunrise TW, Tecan
HP. 2 Salicornia europaea (퉁퉁마디) Trading AG, Männedorf, Switzerland).
HP. 3 Sonchus brachyotus (사데풀)
Hyaluronidase inhibitory assay
HP. 4 Triglochin maritimum (지채)
Hyaluronidase inhibitory effect was evaluated as the
HP. 5 Suaeda japonica (칠면초) method described by Sahasrabudhe and Deodhar (2010)
HP. 6 Argusia sibirica (모래지치) with few modifications. A 0.1 M of acetate buffer at pH 3.6
HP. 7 Artemisia scoparia (비쑥) was prepared and used as the buffer solution in this assay.
HP. 8 Vitex rotundifolia (순비기나무) Firstly, each 10 μl of 8 mg/ml hyaluronidase in buffer solu-
HP. 9 Spartina anglica (갯끈풀)
tion and sample were mixed in the test tube, and incubate
at 37 °C for 20 min. A 20 μl of 12.5 mM calcium chloride
HP. 10 Artemisia princeps (쑥)
was, then, treated to the mixture, and incubated again at
HP. 11 Elymus mollis (갯그령) 37 °C for 20 min. After the incubation, the activated Ca2+
HP. 12 Calystegia soldanella (갯메꽃) mixture was treated with a 50 μl of 2.4 μg/ml hyaluronic
HP. 13 Rosa rugosa (해당화) acid in buffer and incubated at 37 °C for 40 min. Next, the
HP. 14 Asparagus oligoclonos (방울비짜루) mixture reaction developed the color by adding 2 μl of 0.4
HP. 15 Glehnia littoralis (갯방풍)
N sodium hydroxide (NaOH) and 20 μl of 0.4 N potassium
tetraborate tetra-hydrate and then incubated in the water
HP. 16 Suaeda maritima (해홍나물)
bath at 100 °C for 3 min. The DMAB solution containing
HP. 17 Suaeda glauca (나문재) 0.4 g of DMAB dissolved in 35 ml of 100% acetic acid and
HP. 18 Spergularia marina (갯개미자리) 5 ml of 10 N hydrochloric acid (HCl) were prepared.
HP. 19 Chenopodium glaucum (취명아주) Lastly, 600 μl of DMAB was added into the mixture solu-
HP. 20 Atriplex gmelinii (가는갯능쟁이) tion after cooling to room temperature and incubated at
HP. 21 Peucedanum japonicum (갯기름나무)
37 °C for 20 min. The absorbance was measured at a
wavelength of 585 nm (Sunrise TW, Tecan Trading AG,
HP. 22 Ischaemum antephoroides f. coreana (갯쇠보리)
Männedorf, Switzerland).
Jiratchayamaethasakul et al. Fisheries and Aquatic Sciences (2020) 23:0 Page 4 of 9

Tyrosinase inhibitory assay with 62.24 ± 0.55% and 74.47 ± 0.18% of A. sibirica
Tyrosinase inhibitory effect of mushroom tyrosinase extract together with 52.86 ± 0.50% and 52.86 ± 0.50%
assay was determined as the previous reported by No of C. glaucum extract, respectively. Among 22 HPs, S.
et al. (1999). In brief, the mixture solution contained europaea extract performed the highest elastase
the following reagents: 110 μl of 0.1 M sodium phos- inhibitory effect with 74.88 ± 4.84% whereas anti-
phate buffer (pH 6.8), 10 μl of sample, 10 μl of tyro- collagenase activity could not be found in the concen-
sinase in phosphate buffer (1500 units/ml), and 20 μl tration at 1 mg/ml. Notably, a good inhibition of
of 1.5 mM L-tyrosinase were added into the 96-well elastase activity was illustrated by S. anglica extract
plates. The reaction mixtures were incubated at 37 °C (71.63 ± 2.2%) with the minor anti-collagenase effect
for 12 min and the reaction was stopped afterward by observation (34.99 ± 0.55%).
incubating on ice for 1 min. The microplate reader at
490 nm was used to measure the absorbance of the
mixture. Anti-hyaluronidase activity of HP extracts
The inhibitory effects of 22 HPs on hyaluronidase were
Results evaluated as illustrated in Table 3. Among examined
Antioxidant capacities of HPs extracts HPs, three of 22 extracts (V. rotundifolia, A. oligoclonos,
In this study, there are two antioxidant methods to assess and S. glauca) did not show any inhibition at 1 mg/ml.
antioxidant activities which are DPPH free radical and In contrast, S. europaea extract possessed the highest hyal-
hydrogen peroxide scavenging assays. As depicted in uronidase inhibition up to 72.70 ± 1.24% and the significant
Table 2, the antioxidant activities were performed and di- inhibitions were demonstrated by A. princeps, S. marina,
luted to achieve the final concentration at 0.1 mg/ml for
both assays. All 22 HP extracts were possessed antioxidant Table 2 DPPH free radical and hydrogen peroxide scavenging
activities to a varying degree ranging from 11.39 ± 5.99% activities of 22 HP extracts
to 93.32 ± 0.46% and 3.95 ± 2.65% to 79.28 ± 1.40% of Sample DPPH radical Hydrogen peroxide
DPPH free radical and hydrogen peroxide assays, respect- scavenging scavenging
ively. Among 22 HPs extracts, I. coreana extract exhibited activity (%) activity (%)
the highest DPPH free radical scavenging effect (93.32 ± Salsola komarovii 82.90 ± 0.23 33.46 ± 0.32
0.46%) whereas of which 51.14 ± 3.86% hydrogen peroxide Salicornia europaea 60.02 ± 5.39 50.56 ± 2.71
scavenging activity was observed. Furthermore, the next Sonchus brachyotus 76.30 ± 2.09 34.62 ± 2.80
strongest antioxidant capacities (> 80%) expressed by
Triglochin maritimum 24.58 ± 6.64 32.47 ± 1.04
DPPH free radical assay were 82.22 ± 0.23% of S. komaro-
Suaeda japonica 20.97 ± 0.34 9.24 ± 3.15
vii extract, 82.90 ± 0.23% of C. soldanella extract, 84.52 ±
1.39% of R. rugosa extract, and 84.84 ± 1.05% of A. prin- Argusia sibirica 32.92 ± 0.76 6.98 ± 3.53
ceps extract. Meanwhile, the highest hydrogen peroxide Artemisia scoparia 39.16 ± 0.79 22.76 ± 2.83
scavenging effect was found by A. gmelinii extract at Vitex rotundifolia 51.64 ± 0.68 23.04 ± 4.71
79.28 ± 1.40% and the comparative scavenging effect (> Spartina anglica 21.60 ± 0.79 21.17 ± 4.20
60%) were found by S. marina and C. glaucum extracts
Artemisia princeps 84.84 ± 1.05 30.56 ± 4.76
showing 65.91 ± 3.91% and 64.32 ± 8.30%, respectively.
Elymus mollis 28.65 ± 0.46 15.43 ± 1.33
The percentages against hydrogen peroxide of all HP ex-
tracts showed the lower scavenging effects when com- Calystegia soldanella 82.22 ± 0.23 29.78 ± 3.97
pared with that of DPPH free radical except S. marina, C. Rosa rugosa 84.52 ± 1.39 37.38 ± 2.48
glaucum, and A. gmelinii (Table 2). Asparagus oligoclonos 33.20 ± 0.77 11.97 ± 3.88
Glehnia littoralis 56.94 ± 1.88 9.18 ± 2.03
Anti-collagenase and anti-elastase activities of HP extracts
Suaeda maritima 11.39 ± 5.99 3.95 ± 2.65
The collagenase and elastase inhibition effects of all
Suaeda glauca 24.62 ± 0.97 21.90 ± 7.05
HP extracts at the final concentration of 1 mg/ml
were determined and elucidated as shown in Table 3. Spergularia marina 34.28 ± 1.05 65.91 ± 3.91
Firstly, it was notable that the highest collagenase in- Chenopodium glaucum 44.30 ± 1.52 64.32 ± 8.30
hibitory effect was possessed by R. rugosa extract Atriplex gmelinii 30.41 ± 2.50 79.28 ± 1.40
(90.31 ± 0.05%) and its potential also employed good Peucedanum japonicum 38.68 ± 1.39 19.76 ± 0.99
effect in anti-elastase activity (60.76 ± 3.58%). Apart
Ischaemum antephoroides 93.32 ± 0.46 51.14 ± 3.86
from R. rugosa extract, there were other two of all f. coreana
examined HP extracts that exhibited fairly high activ- The values are expressed as the mean ± SD in triplicate experiments. The final
ities in both collagenase and elastase inhibitory effects concentration of tested samples was 0.1 mg/ml.
Jiratchayamaethasakul et al. Fisheries and Aquatic Sciences (2020) 23:0 Page 5 of 9

and C. glaucum extracts represented at 51.71 ± 0.33%, which is deleterious to the DNA, followed by cell damage
57.73 ± 4.09%, and 57.73±2.36%, respectively. and cell death. Therefore, the depletion of ROS generations
defended by the antioxidant activity may integrate the post-
Anti-tyrosinase activity of HP extracts ponement of skin aging problems (Thring et al. 2009; Wit-
The summarized inhibitory potential of HPs against tenauer et al. 2015).
mushroom tyrosinase at the final concentration of 1 mg/ Halophyte is a plant with unique adaptive mechanisms
ml was demonstrated in Table 4. S. anglica extract pre- that naturally survives in saline environments such as
sented the highest tyrosinase inhibitory as 58.62 ± 6.08% tidal flats, sand dunes, and coasts (Lee et al. 2018a,
in the comparison with the other HP extracts at 1 mg/ 2018b). Regarding the successful tolerance in hostile
ml. Moreover, there were 5 HPs that exhibited good in- conditions, for example, salinity, UV radiation, and ex-
hibitory effects (> 50%) noticed by A. sibirica (58.16 ± treme temperature, HP is believed to allow valuable sec-
7.24%), followed by P. japonicum (54.74 ± 0.54%), S. ondary metabolites including polyphenol, carotenoid,
glauca (52.85 ± 0.00%), S. maritima (52.03 ± 3.45%), and and vitamins to cope with those stress-overwhelmed en-
A. gmelinii extract (50.77 ± 0.00%). vironments (Jdey et al. 2017). It is also suggested that
HP may offer a great unexplored source of bioactivity
Discussion for the medicine or cosmetic development with eco-
Skin aging is one of the most concerned issues to all of us nomic benefit (Kim et al. 2016). In our present study, 22
especially facial skin due to its unavoidable process. In daily HPs harvested along the coast of South Korea were
life, the skin exposure to sunlight is considered to be the screened and evaluated whether to provide antioxidant,
most significant factor that accelerates premature skin anti-aging, as well as whitening potentials for cosme-
aging appearance called photoaging. Photoaging can trigger ceutical applications.
the extrinsic mechanism of ROS synthesis in the cells, and First of all, antioxidant capabilities of HPs were chal-
the overproduction of ROS may cause lipid peroxidation, lenged, performed by DPPH free radical and hydrogen

Table 3 Skin aging-related enzyme inhibitory effects of 22 HP extracts

Sample Collagenase inhibition (%) Elastase inhibition (%) Hyaluronidase inhibition (%)
Salsola komarovii 29.69 ± 0.17 17.68 ± 2.10 45.68 ± 5.85
Salicornia europaea - 74.88 ± 4.84 72.70 ± 1.24
Sonchus brachyotus - 32.65 ± 11.63 41.62 ± 1.62
Triglochin maritimum 11.94 ± 1.84 6.00 ± 2.62 39.74 ± 1.24
Suaeda japonica 42.61 ± 1.22 - 12.52 ± 5.00
Argusia sibirica 62.24 ± 0.55 74.47 ± 0.18 11.13 ± 2.60
Artemisia scoparia 15.21 ± 0.55 10.38 ± 7.87 11.74 ± 2.44
Vitex rotundifolia 31.28 ± 1.79 3.96 ± 10.38 -
Spartina anglica 34.99 ± 0.55 71.63 ± 2.21 18.00 ± 2.90
Artemisia princeps 23.45 ± 0.05 5.41 ± 2.15 51.71 ± 0.33
Elymus mollis 25.05 ± 3.34 - 16.12 ± 1.66
Calystegia soldanella 19.60 ± 3.08 - 48.01 ± 1.66
Rosa rugosa 90.31 ± 0.05 60.76 ± 3.58 48.67 ± 1.51
Asparagus oligoclonos 24.89 ± 0.05 0.09 ± 7.45 -
Glehnia littoralis 13.06 ± 0.05 - 1.70 ± 1.99
Suaeda maritima 36.53 ± 0.05 6.19 ± 1.95 9.43 ± 1.50
Suaeda glauca 42.90 ± 0.05 15.49 ± 3.44 -
Spergularia marina 31.60 ± 2.81 31.60 ± 2.81 57.73 ± 4.09
Chenopodium glaucum 52.86 ± 0.50 52.86 ± 0.50 57.73 ± 2.36
Atriplex gmelinii 7.73 ± 0.67 7.73 ± 0.67 40.91 ± 3.36
Peucedanum japonicum 36.89 ± 1.86 36.89 ± 1.86 30.45 ± 0.68
Ischaemum antephoroides f. coreana 24.87 ± 1.10 24.87 ± 1.10 36.14 ± 3.15
The values are expressed as the mean ± SD in triplicate experiments. The final concentration of tested samples was 1 mg/ml.
Jiratchayamaethasakul et al. Fisheries and Aquatic Sciences (2020) 23:0 Page 6 of 9

Table 4 Tyrosinase inhibitory effect of 22 HP extracts found that the ethanol extract of Artemisia princeps
Sample Tyrosinase inhibition (%) (HP. 10) demonstrated the effective scavenging activities
Salsola komarovii 16.70 ± 1.84 by DPPH and ABTS assays along with a good anti-
Salicornia europaea 21.04 ± 2.04
obesity effect on 3T3-L1 preadipocyte cells. Notably, in
the investigation of Lee et al., R. rugosa (HP. 13) was
Sonchus brachyotus 14.31 ± 4.83
found to provide a strong DPPH scavenging activity, and
Triglochin maritimum 20.02 ± 4.68 another study by Zheng et al. also indicated that R. ru-
Suaeda japonica 21.03 ± 13.17 gosa possessed the most powerful antioxidant potential
Argusia sibirica 58.16 ± 7.24 evaluated by DPPH and ABTS assays among 65 edible
Artemisia scoparia 39.85 ± 2.65 flowers (Kim et al. 2016; Zheng et al. 2018). Moreover,
Vitex rotundifolia 49.81 ± 4.65
there have been few investigations referring to the anti-
oxidant activities of S. komarovii, A. princeps, and R. ru-
Spartina anglica 58.62 ± 6.08
gosa (Carvalho et al. 2011; Kim et al. 2018; Lee et al.
Artemisia princeps 43.64 ± 2.77 2012; Youwei and Yonghong 2007). On the other hand,
Elymus mollis 31.18 ± 2.94 A. gmelinii extract would be recommended as hydrogen
Calystegia soldanella 12.16 ± 7.99 peroxide free radical defense. However, there has been
Rosa rugosa 38.07 ± 5.53 no previous study related to the hydrogen peroxide scav-
Asparagus oligoclonos 19.37 ± 4.98
enging effect of A. gmelinii.
ECM is the infrastructure foundation of skin which
Glehnia littoralis 46.75 ± 3.45
consists of various components such as microfibril, pro-
Suaeda maritima 52.03 ± 3.45 teoglycan, collagen, elastin fiber, as well as HA. During
Suaeda glauca 52.85 ± 0.00 the maturation process, the transformations of those
Spergularia marina 18.08 ± 13.60 ECM structural components (the exiguous distribution
Chenopodium glaucum 20.90 ± 2.72 and decrease of collagen, the shortening and absence of
Atriplex gmelinii 50.77 ± 0.00
elastin including loss of HA content) are represented by
atrophied skin, dryness, wrinkling, and sagging skin ap-
Peucedanum japonicum 54.74 ± 0.54
pearance (Bauman 2004; Paliwal et al. 2014). Tyrosinase,
Ischaemum antephoroides f. coreana 21.54 ± 9.79 an enzyme in melanogenesis, plays a key role in melanin
The values are expressed as the mean ± SD in triplicate experiments. The final production. The overproduction of melanin is respon-
concentration of tested samples was 1 mg/ml.
sible for the hyperpigmentation events such as melasma,
freckles, ephelides, and senile lentigines. Consequently,
the suppression of skin-related enzymes (elastase,
peroxide scavenging assays (Table 2). DPPH is a rela- collagenase, hyaluronidase, and tyrosinase) which
tively stable free radical compound which allows a short predominantly integrate the degradation of those
and simple method of screening anti-radical or hydrogen ECM components including the melanin overproduc-
donor capacity of the suspicious unknown (Kim et al. tion is believed to be a key strategy in providing
2016; Lee et al. 2004). Meanwhile, hydrogen peroxide is good skin integrity and youthful and magnificent
an unstable substance which can be provoked from al- skin. In this present study, the inhibitory effects of
most all of the various oxidative stressors. It is also asso- photoaging, like anti-collagenase, anti-elastase, anti-
ciated to form hydroxyl and singlet oxygen radicals hyaluronidase, and anti-tyrosinase, of 22 HPs were
causing lipid peroxidation, cell damage, as well as cell explored at the final concentration of 1 mg/ml as
senescence (Ko et al. 2015; Heo et al. 2005). Both DPPH summarized in Tables 3 and 4.
and hydrogen peroxide scavenging assays are the Our screening found that the strongest anti-
commonly used methods to evaluate the antioxidant hyaluronidase (72.70 ± 1.24%) along with anti-elastase
activity of natural extracts and their compounds (Kim (74.88 ± 4.84%) activities among evaluated HPs were oc-
et al. 2010). cupied by S. europaea extract. The results presumably
In our results, 5 of 22 HPs (S. komarovii, A. princeps, recommended that S. europaea may contribute an excel-
I. coreana, C. soldanella, and R. rugosa, extracts) showed lent anti-wrinkle activity by interrupting the degradation
comparatively strong antioxidant activities between of elastin and HA underneath the skin. S. europaea is
82.90 ± 0.23% and 93.32 ± 0.46% inhibition measured by one of the succulent euhalophyte plants that is used as a
DPPH assay. Among them, I. coreana extract possessed folk remedy for obesity, diabetes, and cancer (Aghaleh
the highest DPPH free radical scavenging effect up to et al. 2010; Won et al. 2017). Recently, some investiga-
93.32 ± 0.46% at the concentration of 0.1 mg/ml (Table tion revealed its health benefits as anti-inflammatory,
2). Likewise, the previous study of Kim et.al. (2010) anti-bacterial, and anticoagulating agents (Kim et al.
Jiratchayamaethasakul et al. Fisheries and Aquatic Sciences (2020) 23:0 Page 7 of 9

2016; Lee et al. 2018a, 2018b). However, the inhibitory DPPH and hydrogen peroxide scavenging, anti-elastase,
effect screening of the Jeju plant by Moon et al. (2010) anti-collagenase, anti-hyaluronidase, and anti-tyrosinase
unlikely found a slight inhibition of elastase, while no in- assays. From our findings, the highest scavenging effects
hibitory effect of tyrosinase was detected at the concen- of I. coreana and A. gmelinii extracts would be intro-
tration of 500 μg/ml. duced as antioxidant agents against DPPH free radical
Aside from a good DPPH scavenging effect as de- and hydrogen peroxide, respectively. On the other hand,
scribed above, R. rugosa also presented the highest the strongest anti-wrinkle potentials of examined HPs
anti-collagenase (90.31 ± 0.05%) and high anti-elastase are provided by S. europaea (anti-elastase and anti-
activities (60.76 ± 3.58%) as well as the moderate inhi- hyaluronidase effects) and R. rugosa (anti-collagenase ef-
bitions against hyaluronidase and tyrosinase as fect), whereas S. anglica extract may be an available
48.67 ± 1.51% and 38.07 ± 5.5%, respectively. There source of tyrosinase inhibition. In conclusion, all of these
were several previous studies mentioned about the results may recommend the guidance of conceivable
splendid antioxidant activities by R. rugosa and the HPs in terms of antioxidant, anti-wrinkle, and whitening
other results by Olech et al. that revealed the effective agent for cosmeceutical development. It is noteworthy
hyaluronidase inhibition of R. rugosa extracts is prob- that the supplementary experiments of examined HPs
ably due to its the richness in polyphenol content such as cell-based experiments and the adverse effects
(Ng et al. 2004; Olech et al. 2017). Nonetheless, an- should be more investigated.
other finding suggested that the ethanol extraction of
Rosa hybrida, a plant species belonging to the same Abbreviations
family with R. rugosa, provided the anti-elastase and AAAPN: N-Succinyl-Ala-Ala-Ala-p-nitroanilide; ABTS: 2,2-Azino-bis(3-
anti-tyrosinase activity as a candidate for the improve- ethylbenzthiazoline)-6-sulfonic acid; DMAB: 4-(dimethyl amino)benzaldehyde;
DMSO: Dimethyl sulfoxide; DPPH: 1,1-Diphenyl-2-pricrylhydrazyl;
ment of skin aging (Choi et al. 2015). ECM: Extracellular matrix; HA: Hyaluronic acid; HP: Halophyte plant;
Among the 22 HP extracts, S. anglica revealed the best PBS: phosphate buffered saline; IC50: 50% Inhibitory concentration;
tyrosinase inhibition activity (58.62 ± 6.08%) together ROS: Reactive oxygen species

with a comparably high elastase inhibitory effect

(71.63 ± 2.21%) at 1 mg/ml. S. anglica was firstly discov- Acknowledgements
This work was supported by the National Research Foundation of Korea
ered in 1963; it provides various ecological and eco- (NRF) grant funded by the Korea government (MSIT; no. 2019R1A5A8083404)
nomic benefits; for example, accretion for reclamation, and was supported by the Soonchunhyang University Research Fund.
amelioration of saline soils, animal fodder, and espe-
cially, seashore stabilization (Qin et al. 1998). However, Authors’ contributions
our present study gives the first evidence for the screen- CJ, YD, JML, HSK, SCK, and SHL constituted and designed the experimental
ing of anti-aging potential in S. anglica. plan. CJ, OH, STI, YJ, and SWM conducted the experimental work. CJ wrote
the manuscript. YD and SHL edited the paper. All authors discussed and
Additionally, A. sibirica, Siberian sea rosemary, offered approved the final manuscript.
an acceptable inhibitory effect of both ECM-deteriorated
enzymes (62.24 ± 0.55% of collagenase and 74.47 ± 0.18%
Funding
of elastase). According to this, we implied that A. sibirica Not applicable
may have potential as an anti-wrinkle candidate. Never-
theless, A. sibirica extract indeed manifested high anti-
Availability of data and material
tyrosinase detected by mushroom tyrosinase assay Not applicable
(58.16 ± 7.24%) compared with other HPs (Table 4). To
the authors’ knowledge, its effect on anti-aging potential Ethics approval and consent to participate
has not yet been clarified. Not applicable
It is vital to note that our screening results are the pre-
liminary demonstration of antioxidant and anti-aging Consent for publication
activities. However, to introduce it as a candidate in Not applicable
cosmeceutical application, a future study is needed. Fur-
ther investigations including 50% inhibitory concentration Competing interests
(IC50) and in vitro cell-based experiment, chemically, bio- The authors declare that they have no competing interests.
logically, and pharmaceutically are required.
Author details
1
Department of Pharmaceutical Engineering, Soonchunhyang University,
Conclusion Asan 31538, Republic of Korea. 2Department of Applied Research, National
Our present study revealed the anti-aging activity explo- Marine Biodiversity Institute of Korea, Seochun, Chungcheongnam 33662,
Republic of Korea. 3National Marine Bio-Resources and Information Center,
rations of 70% ethanol extracts of 22 HPs collected along National Marine Biodiversity Institute of Korea, Seochun, Chungcheongnam
the coast of South Korea. All HPs were assessed by 33662, Republic of Korea.
Jiratchayamaethasakul et al. Fisheries and Aquatic Sciences (2020) 23:0 Page 8 of 9

Received: 19 August 2019 Accepted: 6 February 2020 Lee HJ, Pan CH, Kim ES, Kim CY. Online high performance liquid chromatography
(HPLC)-ABTS+ based assay and HPLC-electrospray ionization mass
spectrometry analysis of antioxidant phenolic compounds in Salsola
komarovii. J Korean Soc Appl Biol Chem. 2012;55(2):317–21.
References Lee JH, Renita M, Fioritto RJ, St. Martin SK, Schwartz SJ, Vodovotz Y. Isoflavone
Aghaleh M, Niknam V, Ebrahimzadeh H, Razavi K. Effect of salt stress on characterization and antioxidant activity of Ohio soybeans. J Agric Food
physiological and antioxidative responses in two species of Salicornia (S. Chem. 2004;52(9):2647–51.
persica and S. europaea). Acta Physiol Plant. 2010;33:1261–70. Lee JM, Yim MJ, Choi G, Lee MS, Park YG, Lee DS. Antioxidant and anti-inflammatory
Asanka SKK, Park Y, Shanura FIP, Ann YS, Ko CI, Wang L, You JJ. Soft corals activity of six halophytes in Korea. Nat Prod Sci. 2018a;24(1):40–6.
collected from Jeju Island inhibits the α-MSH-induced melanogenesis in
Lee JM, Yim MJ, Lee DS, Lee MS, Park YG, Jeon JH, Choi G. Comparison of
B16F10 cells through activation of ERK. Fish Aquatic Sci. 2018;21(21).
biological activities of Korean halophytes. Nat Prod Sci. 2018b;24(4):247–52.
Bauman L. CosmoDerm/CosmoPlast (human bioengineered collagen) for the
Liyanaarachchi GD, Samarasekera JKRR, Mahanama KRR, Hemalal KDP.
aging face. Facial Plast Surg. 2004;20:125–8.
Tyrosinase, elastase, hyaluronidase, inhibitory and antioxidant activity of
Carvalho IS, Cavaco T, Brodelius M. Phenolic composition and antioxidant
Sri Lankan medicinal plants for novel cosmeceuticals. Ind Crops Prod.
capacity of six artemisia species. Ind Crops Prod. 2011;33(2):382–8.
2018;111:597–605.
Chanda SV, Moteriya P, Padaliya H, Rathod TN, Baravalia YK. Antioxidant and
Meot-Duros L, Le FG, Magné C. Radical scavenging, antioxidant and
metal chelating activities of Lagenaria siceraria (Molina) Standl peel, pulp and
antimicrobial activities of halophytic species. J Ethnopharmacol. 2008;
aerial parts in relation to their total phenol and flavonoid content. Phcog J.
116(2):258–62.
2015;7(1):64–73.
Moon JY, Yim EY, Song G, Lee NH, Hyun CG. Screening of elastase and
Chatatikun M, Chiabchalard A. Thai plants with high antioxidant levels, free
tyrosinase inhibitory activity from Jeju Island plants. EurAsia J BioSci.
radical scavenging activity, anti-tyrosinase and anti-collagenase activity. BMC
2010;4:41–53.
Complement Altern Med. 2017;17:487.
Müller HE. Detection of hydrogen peroxide produced by microorganisms on an
Chatatikun M, Yamauchi T, Yamasaki K, Aiba S, Chiabchalard A. Anti melanogenic
ABTS peroxidase medium. Zentralbl Bakteriol Mikrobiol Hyg A. 1985;259(2):
effect of Croton roxburghii and Croton sublyratus leaves in α-MSH stimulated
151–4.
B16F10 cells. J of Tradit Complement Med. 2019;9(1):66–72.
Nanjo F, Goto K, Seto R, Suzuki M, Sakai M, Hara Y. Scavenging effects of tea
Choi EK, Guo H, Choi JK, Jang SK, Shin K, Cha YS, Choi Y, Seo DW, Lee YB, Joo SS,
catechins and their derivatives on 1,1-diphenyl-2-picrylhydrazyl radical. Free
Kim YB. Extraction conditions of white rose petals for the inhibition of
Radic Biol Med. 1996;21(6):895–902.
enzymes related to skin aging. Lab Anim Res. 2015;31(3):148–52.
Ding YL, Jiratchayamaethasakul C, Kim EA, Kim J, Heo SJ, Lee SH. Hyaluronidase Ndlovu G, Fouche G, Tselanyane M, Cordier W, Steenkamp V. In vitro
inhibitory and antioxidant activities of enzymatic hydrolysate from Jeju Island determination of the anti-aging potential of four southern African medicinal
red sea cucumber (Stichopus japonicus) for novel anti-aging cosmeceuticals. J plants. BMC Complement Altern Med. 2013;13:304.
Marine Bioscience Biotechnology. 2018;10(2):62–72. Ng TB, He JS, Niu SM, Pi ZF, Shao W, Liu F, Zhao L. A gallic acid derivative and
Dudonné S, Poupard P, Coutière P, Woillez M, Richard T, Mérillon JM, Vitrac X. polysaccharides with antioxidative activity from rose (Rosa rugosa) flowers. J
Phenolic composition and antioxidant properties of poplar bud (Populus Pharm Pharmacol. 2004;56(4):20.
nigra) extract: individual antioxidant contribution of phenolics and No JK, Soung DY, Kim YJ, Shim KH, Jun YS, Rhee SH, Yokozawa T, Chung HY.
transcriptional effect on skin aging. J Agric Food Chem. 2011;59(9):4527–36. Inhibition of tyrosinase by green tea components. Life Sci. 1999;65(21):
Fulop T, Khalil A, Larbi A. The role of elastin peptides in modulating the immune PL241–6.
response in aging and age-related diseases. Pathol Biol. 2012;60(1):28–33. Olech M, Nowak R, Zaluski D, Kepusta I, Amarowicz R, Oleszek W. Hyaluronidase,
Heo SJ, Park EJ, Lee KW, Jeon YJ. Antioxidant activities of enzymatic extracts from acetylcholinesterase inhibiting potential, antioxidant activity, and LC-ESI-MS/
brown seaweeds. Bioresour Technol. 2005;96(14):1613–23. MS analysis of polyphenolics of rose (Rosa rugosa Thunb.) teas and tinctures.
Horng CT, Wu HC, Chiang NN, Lee CF, Huang YS, Wang HY, Yang JS, Chen FA. Int J Food Prop. 2017;20(S1):S16–25.
Inhibitory effect of burdock leaves on elastase and tyrosinase activity. Exp Paliwal S, Fagien S, Sun X, Holt T, Kim T, Hee CK, et al. Skin extracellular matrix
Ther Med. 2017;14(4):3247–52. stimulation following injection of a hyaluronic acid-based dermal filler in a
Jdey A, Falleh H, Ben JS, Mkadmini HK, Dauvergne X, Magné C, Krouri R. Anti- rat model. Plast Reconstr Surg. 2014;134(6):1224–33.
aging activities of extracts from Tunisian medicinal halophytes and their Palmer DM, Kitchin JS. Oxidative damage, skin aging, antioxidants and a novel
aromatic constituents. EXCLI J. 2017;16:755–69. antioxidant rating system. J Drugs Dermatol. 2010;9(1):11–5.
Jegasothy SM, Zabolotniaia V, Bielfeldt S. Efficacy of a new topical nano- Peres PS, Terra VA, Guarnier FA, Cecchini R, Cecchini AL. Photoaging and
hyaluronic acid in humans. J Clin Aesthet Dermatol. 2014;7(3):27–9. chronological aging profile: understanding oxidation of the skin. J
Kang M, Park S-H, Oh SW, Lee SE, Yoo JA, Nho YH, et al. Anti-melanogenic effects Photochem Photobiol B. 2011;103(2):93–7.
of resorcinol are mediated by suppression of cAMP signaling and activation Popoola OK, Marnewick JL, Rautenbach F, Ameer F, Iwuoha EI, Hussein AA.
of p38 MAPK signaling. Biosci Biotechnol Biochem. 2018;82(7):1188–96. Inhibition of oxidative stress and skin aging-related enzymes by prenylated
Kendall AC, Nicolaou A. Bioactive lipid mediators in skin inflammation and chalcones and other flavonoids from Helichrysum teretifolium. Molecules.
immunity. Prog Lipid Res. 2013;52(1):141–64. 2015;20(4):7143–55.
Kim JW, Um M, Lee JW. Antioxidant activities of hot water extracts from different Qin P, Xie M, Jiang Y. Spartina green food ecological engineering. Ecol Eng. 1998;
parts of Rugosa rose (Rosa rugosa thunb.). J Korean Wood Sci and Technol. 11:147–56.
2018;46(1):38–47. Sahasrabudhe A, Deodhar M. Anti-hyaluronidase, anti-elastase activity of Garcinia
Kim M, Hyun JM, Kim SS, Seong KC, Lim CK, Kang JL, et al. In vitro screening of indica. Int J Botany. 2010;6:299–303.
subtropical plants cultivated in Jeju Island for cosmetic ingredients. Orient J Siedle B, Cisielski S, Murillo R, Löser B, Castro V, Klaas CA, Hucke O, Labahn A,
Chem. 2016;32:807–15. Melzig MF, Merfort I. Sesquiterpene lactones as inhibitors of human
Kim MJ, Park MH, Jeong MK, Yeo JD, Cho WI, Chang PS, Chung JH, Lee JH. neutrophil elastase. Bioorg Med Chem. 2002;10(9):2855–61.
Radical scavenging activity and anti-obesity effects in 3T3-L1 preadipocyte Thring TS, Hili P, Naughton DP. Anti-collagenase, anti-elastase and anti-
differentiation of Ssuk (Artemisia princeps Pamp.) extract. Food Sci Biotechnol. oxidant activities of extracts from 21 plants. BMC Complement Altern
2010;19(2):535–40. Med. 2009;9:27.
Ko EY, Kim DK, Roh SW, Yoon WJ, Jeon YJ, Ahn G. Kim NK Evaluation on Tu PTB, Tawata S. Anti-oxidant, anti-aging, and anti-melanogenic properties of
antioxidant properties of sixteen plant species from Jeju Island in Korea. the essential oils from two varieties of Alpinia zerumbet. Molecules. 2015;20:
EXCLI J. 2015;14:133–45. 16723–40.
Küpeli E, Kartal M, Aslan S, Yesilada E. Comparative evaluation of the anti- Varani J, Fisher GJ, Kang S, Voorhees JJ. Molecular mechanisms of intrinsic skin
inflammatory and antinociceptive activity of Turkish Eryngium species. J aging and retinoid-induced repair and reversal. J Investig Dermatol Symp
Ethnopharmacol. 2006;107(1):32–7. Proc. 1998;3(1):57–60.
Leach JB, Kathryn AB, Charles WPJ, Christine ES. Photocrosslinked hyaluronic acid Wang L, Lee W, Oh JY, Cui YR, Ryu B, Jeon YJ. Protective effect of sulfated
hydrogels: natural, biodegradable tissue engineering scaffolds. Biotechnol polysaccharides from Celluclast-assisted extract of Hizikia fusiforme against
Bioeng. 2003;82(5):578–89. ultraviolet B-induced skin damage by regulating NF-κB, AP-1, and MAPKs
Jiratchayamaethasakul et al. Fisheries and Aquatic Sciences (2020) 23:0 Page 9 of 9

signaling pathways in vitro in human dermal fibroblasts. Mar Drugs. 2018;

16(7):239.
Wittenauer J, Mäckle S, Sußmann D, Schweiggert-Weisz U, Carle R. Inhibitory
effects of polyphenols from grape pomace extract on collagenase and
elastase activity. Fitoterapia. 2015;101:179–87.
Won KJ, Lee KP, Baek S, Cui L, Kweon M-H, Jung SH, Ryu YK, Hong JM, Cho EA,
Shin HS, Kim B. Desalted Salicornia europaea extract attenuated vascular
neointima formation by inhibiting the MAPK pathway-mediated migration
and proliferation in vascular smooth muscle cells. Biomed Pharmacother.
2017;94:430–8.
Yang EJ, Hyun J, Lee NH, Hyun CG. In vitro screening of Korean halophytes for
cosmeceutical ingredients. Int J Chemtech Res. 2016;9(8):541–7.
Youwei Z, Yonghong P. Changes in antioxidant activity in Rosa rugosa flowers at
different stages of development. N Z J Crop Hortic Sci. 2007;35(4):397–401.
Zheng J, Yu X, Maninder M, Xu B. Total phenolics and antioxidants profiles of
commonly consumed edible flowers in China. Int J Food Prop. 2018;21(1):
1524–40.

Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.