Res. on Crops 22 (4) : 948-952 (2021) DOI : 10.31830/2348-7542.2021.

155
With four figures
Printed in India
Effectiveness of pickling on the phytochemical composition and
antioxidant ability in kelakai (Stenochlaena palustris)
creeping fern
NGUYEN PHUOC MINH*
Institute of Applied Technology
Thu Dau Mot University, Binh Duong Province, Vietnam
*(e-mail : nguyenphuocminh@tdmu.edu.vn)

(Received : June 21, 2021/Accepted : September 09, 2021)

ABSTRACT

Kelakai (Stenochlaena palustris) is an available natural herb with an excellent

source of dietary fiber and antioxidant. It’s commonly used as vegetable in daily cuisine.
In order to increase its economic value, it can be converted into pickle via lactic
fermentation. This research was conducted during 2020 at STNanotech laboratory, Vietnam
to evaluate the impact of pickling of Kelakai creeping fern on the phenolic composition
and antioxidant ability of the pickled product. The total phenolic content was determined
by the Folin-Cicalteau method. Aluminum chloride colorimetric method was used for
quantification of total flavonoid content. DPPH free radical scavenging activity and ABTS
radical scavenging activity was examined by using 2,2-diphenyl-2-picrylhydrazyl and 2,2'-
azinobis (3-ethylbenzothiazoline-6-sulfonic acid) assays. Results showed that pickling
at ambient temperature for 7 days in solution consisting of 10% salt, 4% sugar, 0.1 % L.
plantarum culture (6 log CFU/mL) greatly improved the total phenolic content (450.3±0.9
mg GAE/100 g), flavonoid content (305.1±0.4 mg QE/100 g), DPPH free radical scavenging
(149.1±0.9 µg/mL), ABTS radical scavenging activity (2.05±0.01 mM TE/g) of the pickled in
kelakai (Stenochlaena palustris) creeping fern compared to the total phenolic content
(367.5±1.6 mg GAE/100 g), flavonoid content (194.2±0.7 mg QE/100 g), DPPH free radical
scavenging (201.2±0.5 µg/mL), ABTS radical scavenging activity (1.75±0.02 mM TE/g) of
the raw material. This research demonstrated that lactic fermentation strongly enhanced
the phytochemical composition and antioxidant property of kelakai (Stenochlaena palustris)
creeping fern. Utilization of pickled kelakai (Stenochlaena palustris) creeping fern would
be more beneficial than the fresh one.

Key words : ABTS, DPPH, flavonoid, pickling, Stenochlaena palustris, total phenolic

INTRODUCTION et al., 2012; Sathiyaraj et al., 2015; Jang et al.,

Kelakai (Stenochlaena palustris Burm.
fil.) is a popular edible creeping fern containing
a great amount of nutrients and bioactive
constituents beneficial for human health.
Young leaf of this plant was utilized as
vegetables (Suksri et al., 2005). Stenochlaena
palustris leaf includes much more phenolic and
flavonoid content with remaining parts of this
plant (Ndanusa et al., 2020). Fern has been
widely consumed as a food and pharmaceutical
due to its phytochemicals such as flavonoids,
phenolics, terpenoids, and alkaloids (Lee et al.,
2014; Abdul Wahab et al., 2015). These
bioactive constituents are responsible for
numerous pharmacological functions against
ailments such as burn, ulcer and cancer (Yadav
2017; Sangeetha et al., 2017). S. palustris
manifested antibacterial activity towards P.
aeruginosa (Toji et al., 2019). Cytotoxicity of
kelakai extract against breast cancer cell was
noticed at IC50 value of 493.57 µg/mL
(Harlyanti and Itma, 2020).
The pickling is commonly converted
sugar to organic acid by lactic acid bacteria
(Nurul and Asmah, 2012). Salt is essential to
create osmotic pressure to extract nutrients
from plant. These extracted nutrients become
substrates for lactic acid bacteria growth. Sugar
is transformed into lactic acid to retard the
prol iferatio n o f spoi lage an d path og en
microorganisms. From pickling process, plant
will have a better stability, good looking, firm
texture and specific flavour with great healthful
 Effect of pickling on phenolic acids and antioxidants ability of pickled kelakai creeping fern
benefits (Choi et al., 2013; Sayin and Alkan,
2015). Moreover, pickling also creates balanced
nutrition, generate vitamins, minerals,
phytochemicals and antioxidants (Kaur and
Kapoor, 2001; Yamano et al., 2006).
Pi ckli ng affects th e ph en ol ic
composition and antioxidant properties due to
numerous factors like material, bacteria,
du rati on , te mperatu re and pH du ri ng
fermentation. Therefore, objective of our study
focused on the influence of pickling to the
phenolic composition and antioxidant ability
of the pickled kelakai creeping fern.
MATERIALS AND METHODS
Material
Kelakai a creeping fern was naturally
collected in Dong Nai province, Vietnam. It was
thoroughly washed under clean water before
experiment. L. plantarum culture was supplied
from Rainbow Co. Ltd. Chemical reagents
purchased from Merck were all analytical
grades.
Researching Method
Ke lakai (Stenochlae na palustris)
creeping fern was blanched in hot water at
100ºC within 15 seconds and quickly cooled in
ice slurry. The blanched kelakai creeping fern
was drained in 20 minutes to remove excess
water. Then it was submered in the pickling
medium including of 10% salt, 4% sugar, 0.1
% L. plantarum culture (6 log CFU/mL). The
fermentation process was conducted at
ambient temperature for 7 days. The pickled
product was finally evaluated total phenolic
content (TPC, mg GAE/ 100 g), total flavonoid
content (TF, mg QE/ 100 g), DPPH free radical
scavenging (IC 50 , µg/mL) , ABTS radi cal
scavenging activity (mM TE/g). The total
phenolic content (mg GAE/ 100 g) was
determined by the Folin-Cicalteau method
(Abdullakasim et al., 2007). Aluminum chloride
colorimetric method was applied to determine
total flavonoid content (mg QE/100) (Mandal et
al., 2013). DPPH free radical scavenging activity
(µg/mL) was evaluated following method
described Andriana et al. (2019). ABTS radical
scavenging activity (mM TE/g) was evaluated
following method described by Sowndhararajan
and Kang (2013).
949
Statistical Analysis
The experiments were run in triplicate
with different groups of samples. The data were
presented as mean ± standard deviation.
Statistical analysis was performed by the
Statgraphics Centurion version XVI.
RESULTS AND DISCUSSION
Blanching conducted at accelerated
temperature to inactivate internal enzymes
existing in the tissue as well as spoilage
mi croo rg an ism ready for th e offi ci al
fe rmen tati on ( Emmanu el et al. , 2017).
Blanching dimmed the negative impact of the
latter processing steps (Pierre et al., 2017).
Adequate salt concentration is very important
for an effective fermentation by favoring lactic
acid bacteria, inhibiting salt-sensitive spoilage
bacteria, improving the preservative and
organoleptic attribute (Brady, 2002; Arghya et
al., 2017; Erin and Suzanne, 2018). Lactic acid
bacteria proliferation was highly affected by
sugar concentration as substrate for lactic
fe rmen tatio n (Ng uy en et al. , 2020).
Lactobacillus plantarum was an effective
bacterium ideal for human gastrointestinal
health (Nguyen et al., 2020). Pickled product
included a great source of phytochemicals and
antioxidants (Isabelle et al., 2010; Takebayashi
et al., 2013). Pickling process was a alternative
approach for the retention of phenolics and
antioxidants in vegetables (Sayin and Alkan,
2015). Fig. 1 presented the impact of pickling
to the total phenolic content of the kelakai
(Stenochlaena palustris) creeping fern. The total
phenolic contents of the kelakai in raw,
blanched and pickled products were recorded
at 367.5±1.6 mg GAE/100 g, 241.1±2.2 mg GAE/
100 g an d 450.3±0.9 mg GA E/100 g,
respectively. Fig. 2 revealed the changes of total
flavonoid content during lactic fermentation
of the kelakai. Total flavonoid contents in raw,
blanched and the final products were noticed
at 194.2±0.7 mg QE/100 g, 132.1±1.4 mg QE/
100 g and 305.1±0.4 mg QE/100 g, respectively.
From Figs. 1 and 2, we clearly realized that
blanching significantly decreased the phenolic
content meanwhile pickling greatly improved
the phenolic composition in kelakai. Fig. 3
reflected the degradation of DPPH free radical
scavenging. DPPH values in raw, blanched and
finished products were defined at IC 50 of
950
Fig. 1. Effect of pickling to total phenolic content of
kelakai (Stenochlaena palustris) creeping
fern.
Fig. 2. Effect of pickling to total flanonoid content
of kelakai (Stenochlaena palustris) creeping
fern.
201.2±0.5 µg/mL, 346.3±0.7 µg/mL and
149.1±0.9 µg/mL. The lower DPPH value
indicated the higher antioxidant capacity.
Meanwhile, ABTS radical scavenging activity
(mM TE/g) in raw, blanched and pickled
products were presented at 1.75±0.02 (mM TE/
g), 1.29±0.03 mM TE/g and 2.05±0.01 mM TE/
g, respectively.
From Figs. 3 and 4, we found that
blanching degraded the antioxidant capacity
meanwhile pickling process significantly
improved the antioxidant power of kelakai
(Sten ochlaena palustris) creeping fern.
According to Ndanusa et al. (2020), the total
phenolic content of S. palustris leaf extract was
380 mg GAE/100 g. Young S. palustris leaf
extract included higher total phenolic content
than root extract (Kusmardiyani et al., 2016).
Both young and old leaves consisted of high
total phenolic content (Chear et al., 2016).
DPPH free radical scavenging activity (µg/mL)
was used to evaluate the IC50 values of the
extract. A lower IC50 value indicated the
Minh
Fig. 3. Effect of pickling to DPPH free radical
scave nging of ke lakai (Ste nochlae na
palustris) creeping fern.
Fig. 4. Effect of pickling to ABTS radical scavenging
activity of kelakai (Stenochlaena palustris)
creeping fern.
accelerated free radical scavenging power. S.
palustris leaf extract was noticed at 323.40 µg/
mL. S. palustris leaf extract showed ABTS
radical scavenging activity (0.61 mM TE/g)
(Ndanusa et al., 2020).
CONCLUSION
Ke lakai (Stenochlae na palustris)
creeping fern is naturally originated as an
edible creeping fern without any fertilizer and
pesticide. With the purpose of accelerating the
functional properties of this plant, pickling
process was conducted. Kelakai (Stenochlaena
palustris) creeping fern was blanched in hot
water at 100ºC in 15 seconds before pickling
at ambient temperature for 7 days in solution
consisting of 10% salt, 4% sugar, 0.1 % L.
plantarum culture (6 log CFU/mL). The pickled
product has long shelf life, good appearance,
firm structure and mild-acidic flavour. Daily
consumption of this pickled plant may have
better health benefits.
 Effect of pickling on phenolic acids and antioxidants ability of pickled kelakai creeping fern
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