Philippine Journal of Science

152 (3): 1053–1063, June 2023

ISSN 0031 - 7683
Date Received: 08 Feb 2023

Evaluation of Phytochemicals, Total Phenolic

Content, Antioxidant, and Antimicrobial Activities
of Philippine Forest Woody Vines

Kim Wilmer M. Balagot*, Kimberly M. Delica, Rebecca B. Lapuz,

Rowena E. Ramos, and Grace Rowena B. Bisana

Chemistry and Biotechnology Section–Material Science Division (CBS-MSD),

Forest Products Research and Development Institute,
Department of Science and Technology,
College, Laguna, Philippines

Woody vines are herbaceous plants that use different clinging structures to find vertical support
from nearby trees. They are commonly used for handicraft purposes but there were reports
that they were used for medicinal applications. With this, the stems of eight forest woody vine
species were evaluated for their phytochemical components, total phenolic content, antioxidant,
and antimicrobial activities. These were Strychnos minor Denrst. (“balakbakin”), Arcangelisia
flava (L.) Merr (“albotra”), Merremia peltata (L.) Merr. (“bulakan”), Hypserpa nitida Miers ex
Benth (“lalapau”), Symphorema luzonicum (Blanco) F.-Vill. (“mulawing-baging”), Dicranopteris
linearis (Burm. f.) (“tilob”), Connarus semidecandrus (Zoll.) Leenh. (“kamagsa brown”), and
Entada phaseoloides (L.) Merr (“gugo”), which are all native to the Philippines. Results showed
that S. minor and S. luzonicum had the highest extraction yield of 6.28 ± 0.39% and 6.26 ± 0.77%,
respectively. Phytochemical screening revealed that the crude extracts contained different
phytochemicals such as alkaloids, steroids, terpenoids, and tannins. For total phenolic content,
ethanolic extracts of woody vines ranged from 9.51–640.24 mg GAE/g dried extract with C.
semidecandrus and S. luzonicum were found to have higher amounts among forest woody vines
with values of 640.24 ± 7.13 and 398.94 ± 20.08 mg GAE/g dried extract, respectively. Moreover,
DPPH results showed that D. linearis, S. luzonicum, E. phaseoloides, and C. semidecandrus had
strong antioxidant activities, which ranged from 5.99–17.88 μg/mL IC50 and were higher than the
butylated hydroxytoluene (BHT) standard (28.47 μg/mL IC50). On the other hand, M. peltata, A.
flava, H. nitida, and S. minor antioxidant activity ranged from 40.61–78.65 μg/mL IC50. Similar
results of FRAP assay showed that S. luzonicum had the highest antioxidant activity with 2.68 ±
0.07 µmol TE/g dried extract, whereas S. minor and H. nitida had the lowest antioxidant activity
with 0.43 µmol TE/g dried extract ± 0.03 and 0.41 ± 0.02 µmol TE/g dried extract, respectively.
Among the tested microorganisms for antimicrobial assay (well diffusion method), the growth
of S. aureus was susceptible to the ethanolic extracts of S. luzonicum and C. Semidecandrus with
inhibition zones of 21.3 ± 1.4 and 20.1 ± 2.5 mm, respectively. Overall, results show that woody
vines contain phytochemicals with promising antioxidant and antimicrobial properties. These
results could be used as a baseline in the purification of woody vines’ bioactive components.

Keywords: antimicrobial assay, antioxidant activity, forest woody vines, phytochemicals

*Corresponding author: kimwilmer.balagot@fprdi.dost.gov.ph

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INTRODUCTION
Woody vines or “lianas” are herbaceous plants with roots
attached to the ground and as they grow longer, find
vertical support from nearby trees using various clinging
structures such as tendrils, trellises, and hooks (Escobin
and Banaticla 2005). Woody vines are usually found in
forests of the tropics to boreal zones of the northern and
southern hemispheres and are most diverse in areas close
to the equator (Gentry 1991). In the Philippines, there are
over 75 forest vines species already identified by Escobin
and Banaticla (2005). They have reported that forest vines
are used as raw materials for handicrafts such as baskets,
wall décors, trays, vases, and plant stand. However, the
economic potential of this natural resource has not yet
been fully maximized. Also, there are only a few studies in
the Philippines that have reported their possible utilization
in the pharmaceutical industry.
Since 2002, the chemical compositions of 19 forest woody
vine species collected in the Philippines have been studied
at the Department of Science and Technology–Forest
Products Research and Development Institute (DOST-
FPRDI). With these studies, there is now available data
on their chemical properties such as moisture, ash, lignin,
cellulose, starch content, different solvent solubilities, and
some phytochemical contents.
Phytochemicals are secondary metabolites of plants
that act as their immune system and are responsible for
protecting them from parasites or defense mechanisms
in stressful conditions (Mendoza and Silva 2018).
Phytochemicals can be extracted from different parts of
the plants such as leaves, barks, seed, seed coat, flowers,
roots, and pulps (Ingle et al. 2016). These plant chemicals
include alkaloids, flavonoids, tannins, and phenolic
compounds (Edeoga et al. 2005). The presence of phenolic
compounds in certain wood species is responsible for
preventing the activity of termites, bacteria, and fungi
(Taylor et al. 2006; Maranhão 2013).
More than a thousand known phytochemicals have
been classified, including alkaloids, tannins, saponins,
and flavonoids. Dionglay et al. (2017) screened the
phytochemicals present in various local forest vines
collected in Quezon province and found that alkaloid
is the major phytochemical found in Poikilospermum
acuminatum (“hanopol”), as evidenced by the heavy
precipitation with Dragendorff’s reagent. On the other
hand, A. flava and E. phaseoloides contain flavonoids,
whereas tannin is present in all the vines studied except
Rhaphidophora pinnata. A. flava has the highest
phenolic content (0.42 mg GAE/g) extract). Overall,
Poikilospermum acuminatum, Arcangelisia flava, Entada
phaseoloides, Merremia peltata, Strychnos sp., Strychnos
minor, and Symphorema luzonicum were found to contain
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Balagot et al.: Evaluation of Philippine Forest Woody Vines
alkaloids, flavonoids, tannins, and saponins. In order to
effectively exploit these forest woody vines in medicinal
applications, further studies must be done on their
biological activity.
Studies have been conducted in different countries such
as India and Thailand regarding the bioactivities of forest
vines. For instance, the leaves of Connarus semidecandrus
were reported to have antioxidant activity (Machana et
al. 2017), whereas those of Dicrannopteris linearis were
studied for different activities such as antioxidant (Rajesh
et al. 2016), anti-inflammatory, and anti-helminthic
(Vasudeva 1999) activities. Hyspserpa nitida was once
studied for antiviral activity (Cheng et al. 2007) and
Arcanglesia flava, being a well-known forest vine, was
extensively studied for its numerous activities (Cheng et al.
2021). However, the bioactivities of Strychnos luzonensis
and Symphorema luzonicum remained unexplored. In
Cote d’Ivoire, West Africa, 114 forest woody vine species
were reported to be extensively used by the local people.
Of these, 83 are for medicinal purposes (Abbiw 1990).
One of the important vines is the Cameroonian liana
(Ancistocladus korupensis), which is said to prevent HIV
activity due to the presence of alkaloids (Thomas 1994;
Foster and Sork 1997). Indigenous people also use it in
preparing many kinds of traditional medicine.
Despite the fact that the above species have already been
evaluated in various studies abroad, local analysis needs
to be conducted because secondary metabolites vary
depending on geographical location and environmental
conditions (Yang et al. 2018; Labarrere et al. 2019;
Recuenco et al. 2020). Thus, the aim of this study is to
screen the phytochemicals from forest woody vine extracts
collected from Pagbilao, Quezon, Philippines, as well as
evaluate their total phenolic content (TPC), antioxidant
activities, and efficacy against microorganisms.
MATERIALS AND METHODS
Plant Collection and Sample Preparation
Stem samples were collected from Pagbilao, Quezon and
their identities were authenticated at the DOST-FPRDI’s
Herbarium and Xylarium facility. Samples were air-
dried for about two weeks and manually chopped before
grinding into a coarse powder. Following the sample
preparation described in the TAPPI T257 method, samples
were ground up to 60 mesh.
Extraction of Phytochemicals
One hundred fifty grams (150 g) each of ground woody
vines were soaked in 1500 mL of 80% ethanol for 5
d. After soaking, the mixtures were filtered, and crude
Philippine Journal of Science Balagot et al.: Evaluation of Philippine Forest Woody Vines
Vol. 152 No. 3, June 2023

Figure 1. Forest woody vines: C. semidecandrus (left); H. nitida (middle); E. phaseoloides (right).

Table 1. Forest woody vines collected from Pagbilao, Quezon. Saponins (Frothing Test)
Extract (0.5 mL) was added to 5 mL of distilled water
Scientific name Local name
in a test tube. The solution was shaken vigorously and
Strychnos minor Denrst. “Balakbakin” observed for a stable, persistent froth. The frothing was
Symphorema luzonicum (Blanco) F.-Vill.) “Mulawing-baging” mixed with three drops of olive oil and shaken vigorously.
Hypserpa nitida Mierrs. “Lalapau” The appearance of a creamy mass of small bubbles
Connarus semidecandrus Jack “Kamagsa brown”
indicated the presence of saponins [modified procedure
Abiodun et al. (2011)].
Merremia peltata (L.) Merr. “Bulakan”
Arcangelisia flava (L.) Merr. “Albotra”
Tannins
Entada phaseoloides (L.) Merr. “Gugo”
Extract (0.5 mL) was boiled in 10 mL of water in a test
Dicranopteris linearis (Burm. f.) “Tilob” tube and then filtered. A few drops of 0.1% ferric chloride
were added and observed for brownish-green or blue-black
coloration, indicating the presence of tannins [modified
extracts were recovered from the filtrate using a rotary
procedure from Abiodun et al. (2011)].
evaporator. Concentrated crude extracts were subjected
to a vacuum oven at 40 °C for further drying. After which,
dried extracts of woody vines were weighed and stored in Steroids
a refrigerator (2–4 °C). Acetic anhydride (2 mL) was added to 0.5 mL of ethanol
extract of each sample with 2 mL H2SO4. The color
The extraction yield was calculated using this formula: change from violet to blue or green in some samples
indicates the presence of steroids [modified procedure
weight of dried extract from Mujeeb et al. (2014)].
Extraction yield = weight of woody vines × 100 (1)

Terpenoids (Salkowski Method)

Phytochemical Screening
The ethanolic extracts of forest woody vines were
subjected to the following phytochemical screening
procedures:
Alkaloids
Extract (0.5 mL) was diluted with 10 mL of acid alcohol,
boiled, and filtered. To 5 mL of the filtrate, 2 mL of dilute
ammonia was added. Five milliliters (5 mL) of chloroform
were added and shaken gently to extract the alkaloidal
base. The chloroform layer was extracted with 10 mL
of acetic acid, which resulted in two portions. Mayer’s
reagent was added to one portion and Draggendoff’s
reagent to the other. The formation of cream (with Mayer’s
reagent) or reddish brown precipitate (with Draggendoff’s
reagent) was regarded as positive for the presence of
alkaloids (Abiodun et al. 2011).
Extract (0.5 mL) was added to 2 mL of chloroform.
Concentrated H2SO4 (3 mL) was carefully added to
form a layer. A reddish-brown coloration of the interface
indicated the presence of terpenoids [modified procedure
from Kumar et al. (2010)].
Cardiac Glycosides (Keller-Killiani Test)
Extract (0.5 mL) was diluted to 5 mL in H2O and added
with 2 mL of glacial CH3COOH containing one drop
of FeCl3 solution. This was underplayed with 1 mL
of concentrated H2SO4. A brown ring at the interface
indicated the presence of deoxysugar characteristic of
cardenolides. A violet ring may appear below the brown
ring, whereas in the acetic acid layer, a greenish ring
may form just above the brown ring and gradually spread
throughout this layer [modified procedure from Abiodun
et al. (2011)].

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Determination of Total Phenolic Content (TPC)
The TPC was determined with Folin-Ciocalteu reagent,
according to a procedure described by Singleton and Rossi
(1965) with modifications. Forest woody vines (20 mg)
dried extracts were weighed and dissolved in 100-mL
ethanol. An aliquot of 0.50 mL (sample/standard) was
added with 2.5 mL of 0.2 mol/L Folin-Ciocalteu reagent,
followed by 2-mL saturated sodium carbonate solution
(about 75 g/L). The absorbance readings were taken using
UV-1700 SHIMADZU–UV Vis Spectrophotometer at
765 nm after incubation at 45 °C for 15 min. Gallic acid
was used as a reference standard, and the results were
expressed as milligram gallic acid equivalent (mg GAE/g
dried extract). The calibration curve ranged from 0.0–1000
ug/mL with an R2 of 0.998. The data were presented as
mean values ± standard deviation (n = 3).
DPPH (2,2-diphenyl-1-picrylhydrazyl) Radical
Scavenging Activity
In vitro antioxidant activities of the extracts were
determined using the DPPH free radical scavenging
assay described by Nithianantham et al. (2011) with
modifications.
Preparation of extract solutions. A stock solution of
different extracts of 200 ug/mL was prepared by dissolving
20 mg to 100-mL ethanol. From the sample stock solution,
12.5, 25, 50, and 100 μg/mL solutions of each extract
were prepared.
Evaluation of antioxidant potential. 0.03 mM DPPH
(3mL) ethanolic solution was added to the 1-mL
sample solution. The mixture was vigorously shaken
and left to stand at room temperature in the dark for
30 min. The decrease in solution absorbance due to the
components’ proton-donating activity was determined at
517 nm. Butylated hydroxytoluene (BHT) was used as
the standard. The fifty (50%) inhibitory concentrations
(IC50 values) of the extracts were calculated from the
graph as concentration versus percentage inhibition.
Radical scavenging activity was expressed as the
percentage of inhibition. IC50 value is the concentration
of sample required to scavenge 50% of DPPH free radical.
Measurements were taken in triplicate. IC50 of the extracts
indicates the corresponding concentration, in which the
radical scavenging potential is 50%. The IC50 of the
extract and standards were determined graphically.
% radical
Absorbanceblank − Absorvancesample
scaventing = 𝑥 100 (2)
Absorbanceblank
activity
The radical scavenging activities of the extracts are
expressed in terms of their IC50 values. The data were
presented as mean values ± standard deviation (n = 3).
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Balagot et al.: Evaluation of Philippine Forest Woody Vines
FRAP assay method [modified method from Lisanti et
al. (2016)]. The FRAP reagent stock solution (Fe 3+-
TPTZ mixture) was prepared by mixing a portion of
an aqueous 10 mmol/L solution of TPTZ reagent in
40 mmol/L HCl with the same volume of 20 mmol/L
FeCl3◾6 H2O and 10 times higher volume of acetate
buffer of pH 3.6. The mixture was incubated at 37 °C
for 5 min. 1.5 mL of the Fe3+-TPTZ mixture and 50
µL of ethanolic extracts of forest woody vines sample
were mixed and leave in the dark for 5 min, and
the absorbance at 593 nm was read using UV-1700
SHIMADZU–UV Vis Spectrophotometer. Trolox (0.5
µmol/g) was used for calibration and the results were
expressed as µmol eltat equivalent (TE)/ g dried extract.
The calibration curve ranged from 0.0–0.5 mmol/mg
with an R2 of 0.997.
Antimicrobial Assay (Well Diffusion Method)
Ethanolic extracts of woody vines were tested against
Gram-negative bacteria that include Pseudomonas
aeruginosa 1335 and Escherichia coli 1634, Gram-
positive bacteria that include Bacillus subtilis 1679
and Staphylococcus aureus 1582, and a fungus that is
Candida tropicalis. The strains used were obtained from
the UPLB-BIOTECH (University of the Philippines
Los Baños–National Institute of Molecular Biology
and Biotechnology. The stock culture was incubated at
30 °C until it achieved the turbidity of 0.5 MacFarland
standard (prepared by adding 0.5 mL of 0.048 M BaCl2
to 99.5 mL of 0.36 NH2SO4; commercially available).
The turbidity of the test bacterial suspension was
compared with that of 0.5 Mac Farland (vigorously
shaken before use) against a white background with
a contrasting black line under adequate light. The
turbidity was reduced by adding a sterile broth. Forty
microliters (40 µL) of the culture was introduced to
soft agar overlay. Around 6 mm wells were bored into
the agar overlays where the ethanolic extracts were
introduced. Fifty microliters (50 µL) of ethanolic
extracts were introduced and made to diffuse, and
the plates were incubated for 18–24 h at 35 ± 2 °C.
The diameter of the zones of inhibition was measured
and recorded to the nearest millimeter using a ruler
(Guevara 2005).
Statistical Analysis
A one-way analysis of variance was done to determine
significant differences in the group means. Statistical
significance was set at P < 0.05. Pearson correlation
analysis (P < 0.05) was done to determine the strengths
of the relationships between TPC and antioxidant
activities.
Philippine Journal of Science Balagot et al.: Evaluation of Philippine Forest Woody Vines
Vol. 152 No. 3, June 2023

Table 2. Diameter of zone of inhibition and their corresponding
inferences.
Inferences Zone of inhibition (mm)
Resistant 10 or less
Intermediate 11–15
Susceptible 16 or more
RESULTS AND DISCUSSION
Extraction Yield
Eight forest woody vine samples were subjected to
extraction with 80% ethanol as the solvent. While other
solvents can be used to obtain phytochemicals, ethanol
has been known as a good solvent for polyphenols (Dai
and Mumper 2010). An aqueous mixture of ethanol (10:1
solvent to sample ratio) was used as a solvent because
aside from being safe for human consumption (Do et al.
2014), it can easily penetrate the cellular membrane to
extract the intercellular components such as polyphenols,
tannins, flavonoids, terpenoids, steroids, and alkaloids in
the vines. On the other hand, the maceration method was
used as it is the cheapest and safest way of extraction.
Table 3 shows that the extraction yield ranged from
1.51–6.28%, in which D. linearis had the lowest yield,
whereas S. minor and S. luzonicum had the highest
extraction yield among the woody vines with 6.28 and
6.26%, respectively. Extraction yields are influenced by
several factors such as solvent used, types of extraction,
length of the extraction period, particle size, and solvent-
to-sample ratio (Stalikas 2007).
Forest woody vine extracts were screened for the presence
of phytochemicals. The three major phytochemicals class
includes terpenoids, phenolic metabolites, and alkaloids
(Do et al. 2014). These phytochemicals are known to have
medical importance exemplified by alkaloids, which are
known to have anti-inflammatory (Augusto et al. 2011),
antimalarial (Dua et al. 2013), antimicrobial (Benbott et
al. 2012), cytotoxic, antispasmodic, and pharmacological
effects (Ameyaw and Duker-Eshun 2009; Thite et al.
2013). Likewise, terpenoids also have antimicrobial,
antifungal, antiviral, and anti-inflammatory properties
(Kumar et al. 2010). These phytochemicals are widely
explored due to their potential application as natural
antioxidants, functional foods, and neutraceuticals (Do
et al. 2014).
Based on the phytochemical tests on the forest
woody vine extracts (Table 4), all samples contained
tannins, steroids, terpenoids, and cardiac glycosides.
However, alkaloids were only detected in M. peltata,
C.semidecandrus, E. phaseoloides, and D. linearis
extracts. Alkaloids are nitrogenous compounds with
promising properties as therapeutic reagents due to their
potent biological activities (Perez et al. 2015). They also
have been reported to be major antioxidants in natural
products (Gan et al. 2017). Flavonoids were also detected
in the extracts, except for S. minor and D. linearis.
These phytochemicals have been found in vitro to have
antimicrobial properties (Cushnie and Lamb 2005).
Flavonoids are also considered antioxidants, and most
of them are extensively degraded into different phenolic
acids inside the human body, giving off their free radical
scavenging capacity (Pietta 2000). Tannins were detected
in all ethanolic extracts of forest woody vines. These
phytochemicals are widely known to have antibacterial
properties (Kaczmarek 2020). With these present results,
the antioxidant and antimicrobial activities of extracts
of forest woody vines were studied.

Table 3. Extraction yield, total phenolic content, and antioxidant activities of woody vines.
Total phenolic content FRAP assay
Extraction yield DPPH assay IC50 (µg/
(mg GAE/ g dried ex- (µmol TE/g dried
Species (%)* mL)**
tract)* extract)*
Strychnos minor Denrst. 6.28 ± 0.39a 35.07 ± 1.10f 78.65 ± 3.24e 0.43 ± 0.03d
Symphorema luzonicum (Blanco) 6.26 ± 0.77a 398.94 ± 20.08b 5.99 ± 1.46a 2.68 ± 0.07a
F.-Vill.)
Hypserpa nitida Mierrs. 3.64 ± 0.38b 208.06 ± 8.23d 72.73 ± 4.27e 0.41 ± 0.02d
Connarus semidecandrus Jack 3.59 ± 0.19b 640.24 ± 7.13a 15.00 ± 0.87ab –
Merremia peltata (L.) Merr. 3.20 ± 0.31b 95.65 ± 1.54e 40.61 ± 0.94c 1.01 ± 0.02b
Arcangelisia flava (L.) Merr. 2.79 ± 0.91b 9.51 ± 1.04g 58.62 ± 1.84d 0.87 ± 0.07c

Entada phaseoloides (L.) Merr. 1.86 ± 0.31c 310.03 ± 8.26c 17.88 ± 2.91b –
Dicranopteris linearis (Burm. f.) 1.51 ± 0.12c 303.64 ± 5.06c 15.64 ± 2.72b –
*Means with the same letter are not significantly different. **DPPH assay: IC > 250 µg/mL, inactive; > 100–250 µg/mL, weakly active; > 50–100 µg/mL, moderately
50
active; 10–50 µg/mL, strongly active; < 10 µg/mL, very strongly active (Phongpaichit et al. 2007).

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Vol. 152 No. 3, June 2023

Total Phenolic Content (TPC)
Phenolic compounds are one of the most numerous plant
secondary metabolites with hydroxyl groups that facilitate
free radical scavenging and redox properties responsible
for antioxidant activities (Soobrattee et al. 2005). Due to
the polarity of ethanol, phenolic compounds can be easily
extracted from the plant and can be quantified. The TPC of
the woody vine extracts, as shown in Table 3, ranged from
9.51–640.24 mg GAE/g. The TPC of C. semidecandrus
was the highest with 640.24 ± 7.13 mg GAE among
ethanolic extracts, whereas the lowest phenolic content
was A. flava at 9.51 ± 1.04 mg GAE/g.
In a study by Machana et al. (2017), the methanolic leaf
extract of C. semidecandrus collected in the mangrove
forest of Thailand had a TPC of 39.7 ± 0.07 mg GAE/g
dried extract. This was lower than the TPC of bark extract
in the current study from which the plant sample was
collected in the Philippines. The high difference in TPC
of stem and leaves mainly depends on their chemical
components. On the other hand, there is no reported study
yet for the TPC of S. luzonicum. Moreover, the TPC and
total flavonoid content (TFC) and antioxidant activities
of hexane, ethyl acetate, and methanolic extracts of A.
flava leaves were evaluated by Wahyudi et al. (2016). It
was found that the TPC and TFC of methanolic extracts
are higher than the other extracts with 135.25 and 280.61
mg QE/g, respectively. Additionally, the methanolic
extract of A. flava leaves showed the highest scavenging
activity on superoxide and hydroxyl radicals compared
to other extracts. The TPC and TFC of methanolic and
ethanolic extracts of S. minor leaves were measured in
the study of John et al. (2015). Results show that the
TPC of methanolic and ethanolic extract contains 12.12
and 10.21 mg GAE/g, respectively, whereas the TFC
of methanolic and ethanolic extracts contains 4.00 and
1.72 mg naringin equivalent/g, respectively. The present
results show that woody vines could also be a rich source
of phenolic compounds.

Table 4. Phytochemical screening of forest woody vine extracts.

Species Alkaloids Saponins Tannins Flavonoids Steroids Terpenoids Cardiac glycosides
A. flava – + + + + + +
S. minor – – + + + + +
M. peltata + – + – + + +
H. nitida – – + + + + +
S. luzonicum – – + + + + +
C.semidecandrus + – + + + + +
E. phaseoloides + + + + + + +
D. linearis + + + – + + +
*[–] negative; [+] positive

Figure 2. % radical scavenging activity of the forest woody vine extracts.

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Antioxidant Activity
The antioxidant activities of the forest vine extracts were
evaluated using DPPH and FRAP assay. DPPH assay
measures the reduction of DPPH free radicals, whose
absorbance will decrease upon its reaction with an
antioxidant. An antioxidant compound acts by donating
hydrogen atoms to obtain radicals with stable molecular
structures that will stop the chain reaction (Santos-Sánchez
et al. 2019). The DPPH radical scavenging activity of
forest woody vines and the BHT standard is presented
in Figure 2. Scavenging activity increased with the
concentration of woody vine extracts and was comparable
to the standard.
The IC50 value is a widely used parameter in measuring
the antioxidant activity of plant samples. It is calculated
as the concentration of antioxidants needed to decrease
the initial DPPH concentration by 50% (Sánchez-Moreno
1998). Thus, the lower IC50, the higher the antioxidant
activity. As presented in Table 3, S. luzonicum possessed
very strong and the highest antioxidant activity with
IC50 of 5.99 µg/mL, followed by C. semidecandrus, D.
linearis, E. phaseoloides, and M. peltata – which was
shown to be strongly active – and A. flava, H. nitida,
and S. minor – which had moderate antioxidant activity.
This is the first time that S. luzonicum is reported to
have antioxidant activity which is due to the presence
of different phytochemicals. While the methanolic leaf
extract of C. semidecandrus has low antioxidant activity as
reported by Machana et al. (2017), the current study shows
otherwise. The species appears to be an ideal source of
antioxidants, as indicated by its strong antioxidant activity.
In addition, the finding on the antioxidant potential of D.
linearis agrees with the results reported by Rajesh et al.
(2016), in which the methanolic extract of the whole of
the D. linearis plant has high TPC and strong antioxidant
activity.
On the other hand, FRAP assay measures the reducing
power of a compound. This compound reduces the ferric
ion (Fe3+) to the ferrous ion (Fe2+), which forms a blue
complex – Fe2+/TPTZ (Fernandes et al. 2016). Among the
forest woody vines analyzed, S. luzonicum also had the
highest antioxidant activity with 2.68 ± 0.07 µmol TE/g
dried extract, whereas S. minor and H. nitida also had the
lowest antioxidant activity with 0.43 ± 0.03 µmol TE/g
dried extract and 0.41 ± 0.02 µmol TE/g dried extract.
To the best of our knowledge, this is the first time to
report the reducing power ability of ethanolic extracts
of S. luzonicum, S. minor, and H. nitida. Meanwhile, the
water extract from A. flava stems and ethyl acetate extract
from M. peltata stem is reported with FRAP values of
91.30 ± 4.91 mg TE/g (Panchakul et al. 2022) and 164.53
μmol/g (Muthi’atul Af-Idah et al. 2021). The difference
in antioxidant values could be due to differences in the
Balagot et al.: Evaluation of Philippine Forest Woody Vines
geographical location and environmental conditions of
forest woody vines and experimental conditions (Yang
et al. 2018; Labarrere et al. 2019; Recuenco et al. 2020).
Correlation between Total Phenolic Content (TPC)
and Antioxidant Activities
To correlate the TPC with antioxidant activities, linear
correlation coefficients of TPC (mg GAE/g) to DPPH
(IC50) and FRAP assays were calculated. A strong and
statistically significant inverse correlation (r= –0.75820,
P < 0.05) was observed between TPC and DPPH radical
scavenging activity. Likewise, a strong and statistically
significant positive correlation (r = 0.78815, P < 0.05)
was observed between TPC and reducing power abilities.
These results suggest that phenolic compounds could
contribute to the antioxidant activities of woody vines
extracts. However, it is not limited to phenolics since
other secondary metabolites could also contribute to the
antioxidant activities (Javanmardi et al. 2003; Wongsa
et al. 2012).
Antimicrobial Assay
For the antimicrobial activity of the forest woody vine
extracts, the zone of inhibition against different microbial
strains was used as indicator. The new ISO 20776-1
standard of susceptibility testing (Rodloff et al. 2008)
was adapted to determine the likelihood a sample to have
antimicrobial properties.
As presented in Table 5, the growth of S. aureus was
susceptible to the ethanolic extracts of S. luzonicum and C.
semidecandrus with inhibition zones of 21.3 ± 1.4 and 20.1
± 2.5 mm, respectively, whereas the growth of B. subtilis
was intermediately susceptible to the extract H. nitida
with 13.8 ± 4.4 mm inhibition zone. On the other hand,
growth of S. aureus and C. tropicalis was intermediately
susceptible to the A. flava extract with inhibition zones
of 13.4 ± 1.1 and 13.3 ± 0.3 mm, respectively. As per the
study of Setyowati et al. (2014), A. flava water extract
inhibited the growth of C. albicans with 16.65 ± 4.52 mm.
C. albicans and C. tropicalis are known to be the most
pathogenic Candida species to cause oral cavity infection
(da Costa et al. 2009). M. peltata extract showed inhibition
zone of 7.9 ± 1.8 mm against P. aeruginosa. Based on the
study of Perez et al. (2015), different doses of M. peltata
ethanolic leaf extract showed 3–5-mm inhibition zones
against P. aeruginosa.
To the best of our knowledge, there are only a few reports
on the antimicrobial properties on the investigated forest
woody vines. But there are other forest woody vines studied
in the Philippines such as Bauhinia integrifolia Roxb. and
Strongylodon paucinervis Merr that possesses antibacterial
and antifungal activity against the two Gram positive strains,
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Vol. 152 No. 3, June 2023

Table 5. Inhibition zones of the forest woody vine extracts.

Inhibition zones against indicator strains (mm)
Species
P. aeruginosa E. coli B. subtilis S. aureus C. tropicalis
A. flava 3.5 ± 3.7a 0.9 ± 1.5a 8.3 ± 0.8a 13.4 ± 1.1b 13.3 ± 0.3b
S. minor 3.0 ± 2.7a 0a 8.8 ± 1.9a 9.0 ± 1.6 a 0a
M. peltata 7.9 ± 1.8a 0a 0a 0a 0a
H. nitida 1.3 ± 4.7a 0a 13.8 ± 4.4b ND 0a
S. luzonicum 10.0 ± 4.3a 9.4±4.2a 10.4 ± 1.8a 21.3 ± 1.4c 0a
E. phaseoloides 0a 0a 0a 12.4 ± 1.1b 0a
C. semidecandrus 0a 0a 0a 20.1 ± 2.5c 0a
D. linearis 0a 0a 0a 0a 0a
Legend: [a] resistant; [b] intermediate; [c] susceptible

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These present results and other related studies suggest that
forest woody vines could possess antimicrobial properties.
Therefore, they must be continuously explored for their
potential. The extraction, isolation, and characterization
of bioactive compounds must be further investigated
to determine whether specific chemical components
or a synergistic interaction among phytochemicals are
responsible for their bioactivity.
CONCLUSION
The present results suggest that forest woody vines
contain phytochemicals that have the potential to possess
antioxidant and antimicrobial properties. Future studies
on extraction, isolation, and characterization of bioactive
compounds must be explored to fully utilize forest woody
vines other than on handicraft industry.
ACKNOWLEDGMENTS
The authors wish to thank the DOST-FPRDI management
for providing funds for this study, Mr. John Jefferson A.
Collera and Mr. Audel V. Mosteiro for sample preparation,
Ms. Mariluz S.P. Dionglay and the Chemistry and
Biotechnology Section for the support, Mr. Fernando
C. Pitargue Jr. and Mr. Mario D.R. Ramos for the
identification and collection of woody vines samples,
and Ms. Rizalina K. Araral for the language editing of
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