FICUS-NOTA-123-EDITED-1 NotaMi)

QUIRINO STATE UNIVERSITY
DIFFUN CAMPUS
Diffun, 3401 Quirino
COLLEGE OF TEACHER EDUCATION
.... promoting pedagogical excellence
Chapter I
THE PROBLEM AND ITS BACKGROUND
I. Rationale
Philippines is one of the most biodiverse countries in the world that
is rich in indigenous plants and trees (Mapatac, 2019). Ficus nota (Blancoi)
Merr. also known as “Tibbeg” in Ilocano, is an endemic plant that is often
found in Luzon and Visayas where it originated (Mancia et.al 2019; Ragasa
et.al, 2014; Mapatac, 2019; Mancia, et.al, 2019). It belongs to family
Moraceae comprises of about 850 species of woody trees, shrubs vines and
epiphytes worldwide (Gandhi et.al, 2019). It is usually found in thickets and
forest at low and medium altitudes. It is an erect, spreading, dioecious tree
of about 8 meters (m) in height, with fruits around 2 to 3.5 cm in diameter,
fleshy, green and becoming yellowish white at the base. The fruit can be
eaten raw when ripe, while the young leaves are cooked as a vegetable
(Philippine Alternative Medicine, 2011).
Traditional uses of Ficus nota include drinking its water extracted
from the standing tree for fever, and muscle pain relief and a decoction of
the roots and bark for treatment of the urinary tract infection, hypertension,
and diabetes (Philippine Alternative Medicine, 2011; Ragasa et.al, 2014).
Various studies were conducted from the different parts of the Ficus
nota. Based on the study of Arquion et. al. (2015) which focuses on the stem
extracts of Ficus nota, it exhibited cytotoxic activity against the brine shrimp
Artemia salina. According to the study conducted by Latayada & Uy (2016),
they showed that the ethanol extract from the leaf extract of Ficus nota has
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no activity against the A. niger, A. flavus, C. albicans, and S. cerevisea.
However, there are no reports yet on the antimicrobial activities and
potential cytotocixxities of the fruit extracts of F. nota.
While the Philippine biodiversity and rich cultural traditions of plant
use, scientific understanding of medicinal plants remains largely unexplored
and pharmacological investigation of the Philippine flora only grown recently.
The importance of a country’s diverse medicinal plants lies not only in their
chemotherapeutic value in traditional medicine but also in their potential
sources of new chemical entities for drug discovery. A study discovered the
therapeutic ability of some plants found in the Philippines with an end goal
of providing cheaper nature-based alternative medicine to the public in the
midst of high-priced medicine produced by pharmaceutical companies. As
such, the study aimed to determine the antibacterial and cytotoxicity of
Ficus nota (Blanco) Merr Fruits. The study provides scientific evidence on
the possible use of Ficus nota (Blanco) Merr fruits to produce potential
source of antibiotics (Mapatac, 2019).
This study was anchored in the updated Harmonized National
Research and Development Agenda (HNRDA) of Department of Science and
Technology (DOST) stipulated Drug Discovery and Development (Tuklas
Lunas) as one of the priority program areas within the purview of the
Philippine Council for Health and Development (PCHRD). Under Section 2 of
HNRDA, the Health Research and Development Agenda for the year 2023-
2028 stated the development of herbal drugs and discovery of new drugs
from local terrestrial and marine sources for development up to the clinical
stage. Drugs would be developed for infectious diseases such as Anti-biotic
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resistant bacterial infections, viral diseases, and fungal infections; and non-
communicable diseases such as cancer diabetes mellitus, cardiovascular
diseases, and other lifestyle-related diseases.
In addition, priorities for 2023-2025 will be the development of herbal
drugs by means of screening for bioactive extracts from natural sources
using validated protocols of the program; formulation of priority bioactive
extracts into standardized herbal drugs; scale-up studies for formulated
standardized bioactive herbal product; pre-clinical/In-vivo studies of priority
bioactive candidates; phase 1/2 clinical studies of candidate herbal drugs;
and cultural management/propagation of priority organisms. The drug track
would took place through isolation of bioactive compounds from natural
sources using validated protocols of the program and lead optimization of
bioactive compounds to come up with new chemical entities (NCEs). Then an
advanced step priorities for 2026-2028 similar to previous years was to be
conducted (DOST, 2022).
Hence, this study was conducted accordance with the said agenda set
by the DOST. The biochemical profiling of the alcoholic crude extracted from
Ficus nota (Tibbeg) will be useful to explore its potential applications in
health spectrum to be part of this endeavor.
The vital efforts of this study focused on the biochemical profile of
Ficus nota in order to provide new avenue for is future and potential
applications. The crux of this study centered on the bioactive components of
the alcoholic crude extract from the Ficus nota.
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Statement of the Problem
Generally, the study seeks answer to the following questions:
1. What is the percent yield of alcoholic extract from the fruit of Ficus nota?
2. What is the biochemical profile of the alcoholic fruit extract of Ficus nota
in terms of its:
a) phytochemical constituents; and
b) phenolic content equivalent?
2. What is the extent of microbial activity of the alcoholic fruit extract of
Ficus nota?
3. What are the potential application of the sample based on the results of
its biochemical profile?
Scope and Delimitation of the Study
This study was conducted in Quirino State University, particularly,
the screening processes took place at the Food Laboratory located in College
of Health Sciences.
The biochemical profiling was limited only to phytochemical
screening, total phenolics content and antimicrobial activity as basis for the
potential applications.
Significance of the Study
The Health Sector. This study may provide a potential biochemical product
that may cure certain diseases. Medicines that may be created may help in
curing patients. Also, biochemical products may provide important
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information about the outcomes of treatment, especially in antioxidant and
antimicrobial activities.
The Farmers. This study may pave another knowledge driven facts to the
field of agriculture. There are abundance of trees growing in the locality
which would then they will be able to know the value of these fruit more. The
potential agricultural and economic value that is driven by this initiative to
unravel more information about Ficus nota. Thus, an opportunity for them
to flourish these values that could possibly give them profit someday.
The Investors. By coming up with the unraveling the potential commercial
value of the essential oil extracted from Ficus nota, this study (would) may
be able to pitch new investment project that the researchers foresee in the
market. The intricate information that will be derive from this study is
evidence to attract investors to make a deal with those business enthusiasts
or the researchers themselves. Additionally, the outcome of this study would
pave another innovative product that may imbued by the alcoholic extract.
The Administrators. The study would employ new factual details regarding
the essential oil of Ficus nota, which the local government and other leaders
would craft new policy to implement. They could draw some funds to extend
in further development of identified commercial value of the oil in this study.
Also, provide the stakeholders to take care this kind of resources available
around the tropical areas. Furthermore, this study is significant to lobby the
administrators to include such value of Ficus nota and devise a plan that
offers growing of the tree in a specific land area, especially, to the concerned
agencies.
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Future Studies. The recommendations of this study would open new
continuation of further studies. It could be examining the efficacy of the
identified commercial value of the alcoholic extract of Ficus nota.
Definition of Terms
Alkaloids - any of a class of naturally occurring organic nitrogen-containing
bases. Alkaloids have diverse and important physiological effects on humans
and other animals (Britannica, 2021).
Amino acid – molecules that combine to form proteins. Amino acids and
proteins are the building blocks of life (Reddy, 2022).
Anthraquinones- are bioactive natural products, some of which are active
components in medicinal medicines, especially Chinese medicines. These
compounds exert actions including purgation, anti-inflammation,
immunoregulation, antihyperlipidemia, and anticancer effects (Wang D.,
Wang X.H., et. al., 2021).
Bioactive components - compounds in plants include alkaloids, terpenoids,
coumarins, flavonoids, nitrogen-containing compounds, organosulfur
compounds, phenolics, etc. A wide spectrum of bioactivities is exhibited by
these compounds such as anti-inflammatory, immunostimulatory,
anticancer, antioxidant, antimicrobial, etc. (Stephanie F. et al., 2021).
Biochemical profile – analysis of the bioactive component and or
metabolites from the bio-extract.
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Blanching- to scald or parboil in water or steam in order to remove the skin
from, whiten, or stop enzymatic action in such as food for freezing (Merriam-
Webster).
Chromatography- technique for separating the components, or solutes, of a
mixture on the basis of the relative amounts of each solute distributed
between a moving fluid stream, called the mobile phase, and a contiguous
stationary phase (Giddings & Keller, 2023).
Drying oven - The moisture removed from the oven chamber helps sample
drying to take place (Haque,2022).
Extract- to withdraw (something, such as a juice or a constituent element)
by physical or chemical process (Merriam-Webster).
Fatty acid - important component of lipids in plants, animals, and
microorganisms. Generally, a fatty acid consists of a straight chain of an
even number of carbon atoms, with hydrogen atoms along the length of the
chain (Britannica, 2023).
Flavonoid - also called flavone, any of a class of no nitrogenous biological
pigments extensively represented in plants. Flavonoids are water-soluble
phenolic compounds (having a –OH group attached to an aromatic ring) and
are found in the vacuoles of plant cells (Britannica, 2021).
Microbial assays - a technique to assess a compound’s potency or
concentration. This is carried out by exposing microorganisms to the
compound and determining its effect on them (Davey, n.d.).
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Phenolic content – important plant constituents with redox properties
responsible for antioxidant activity. The hydroxyl groups in plant extracts
are responsible for facilitating free radical scavenging (Wade, 2018).
Phytochemical analysis – devoted to the publication of original articles
concerning the development, improvement, validation and/or extension of
application of analytical methodology in the plant sciences (Cao X., Liao W.,
and Wang S., 2020).
Regeant- an integral part of any chemical reaction. A reagent is a substance
or compound that can facilitate a reaction, and they are used in most widely
used tests (Ryding, 2021).
Triterpenes - constitutes a large structurally diverse group of natural
compounds biogenetically derived from active isoprene. Two C15 units build
squalene or related acyclic 30-carbon precursors. As the result of their
cyclization and oxidation, various structures are formed (Nazaruk J.,
Borzym-Kluczyk., M., 2014).
Tannins – It is also called tannic acid, any of a group of phenolic compounds
in woody flowering plants that are important deterrents to herbivores and
have a number of industrial applications. As secondary metabolites, tannins
are sequestered in vacuoles within the plant cell, which protects the other
cell components (Britannica, 2021).
Thin Layer Chromotography – It is a chromatographic technique used to
separate the components of a mixture using a thin stationary phase
supported by an inert backing. (Libre Text Chemistry)
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Conceptual Framework of the Study
Commercial
Application
Possible
Biochemical commercial
Profiling application of the
Phytochemical extracted
Preliminary Screening, Total biochemical of
Treatment Phenolic Contents, Ficus nota
Washing and and Microbial
Blanching and Assay
Alcoholic Extraction
Figure 1. Conceptual framework of the study
Preliminary treatments like washing, blanching and alcoholic will be
employed. The biochemical profiling was limited only to phytochemical
screening, total phenolics content, and microbial assay. Future and potential
commercial applications of the fruit of Ficus nota were considered in the
study.
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Chapter II
LITERATURE REVIEW
This section presents the digested review of related literature and
studies that provide scaffolding support to this study.
A. Related Literature
Ficus nota (Blanco) Merr. or “tibig”, from the Moraceae family, is a
plant endemic to the Philippines (Santiago et. al., 2017). It is widely
distributed from Luzon to Visayas where it originated from primary forests at
low to medium elevations. The fruits are traditionally eaten with sugar and
the leaves are consumed as vegetable. Different parts of the plant are
ethnomedically used for the treatment of diabetes, hypertension, fever,
urinary tract infection, and muscle pain (Arquion, Nuñez, & Uy, 2015).
Botanical Description
Moraceae is a family of flowering plants comprising about 40 genera
and over 1,000 species that are mostly widespread in tropical and
subtropical regions and a medicinally important family which comprises
both wild and cultivated species (Guian, 2020). The synapomorphy of
Moraceae is the presence of laticifers and milky sapin its tissues, it has
generally useful field characters that can be observed it includes two carpels
sometimes with one reduced, compound inconspicuous flowers, and
compound fruit (Mahbubur, Rahman, & Khanom, 2013). Among the genera
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found in the family Moraceae is the Ficus nota which constitutes an
important group of trees with immense medicinal value (Guian, 2020).
The genus Ficus which comprises many different species of plants,
trees and has an immense medical value, it is also considered in many
religious groups to be sacred (Mapatac, 2019). Almost all Ficus species
exude latex as a defensive mechanism (Lasky et al., 2014). Ficus nota
(subgenus Sycomorus) occurs in Malaysia Sabah, Sarawak and the
Philippines. Ficus septica (subgenus Sycomorus) is found naturally mainly
on the islands of Eastern Asia (Indonesia, Japan, Malaysia, Papua New
Guinea, Philippines, Solomon Islands, Taiwan) and neighboring Pacific areas
(Australia, Vanuatu). Ficus ulmifolia (subgenus Sycidium) occurs naturally
in the Philippines and is a vulnerable species on the Red List. Some Ficus
species are native to or have been introduced in parts of the Unites States
and the Caribbean area (Knothe et.al, 2018).
In accordance, Ficus nota (Blanco) Merr, also known as “Tibbeg” in
Iloco, is a plant native endemic to the Philippines (Santiago et.al, 2017;
Mapatac, 2019; Guian, 2020; Arquion, Nuñez, & Uy, 2015) and they are also
native in parts of northern Borneo in Malaysia. It is a member of the Ficus
genus making it a close relation of the European fig, the tropical fig (Ficus
septica), and the wild Punjabi fig (Guian, 2020). It is widely distributed from
Luzon to Visayas where it originated from primary forests at low to medium
elevations. The tree can grow up to 10 meters in height with prominent
protuberances on the trunk. Leaves are simple, alternate, obovate,
pubescent, and are approximately 24-28 x 12-14 cm in size (Santiago et.al,
2017).
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These trees are erect, up to 9 m high, or shrubby with crooked stems.
In Florida, the trees are low and spreading, with trunks branching near the
ground, probably because of having suffered occasional frost injury. No
aerial roots were observed (Santiago et.al, 2017). The twigs are hollow when
young, about 0.7 cm thick, and densely silky pubescent. Older twigs are
chocolate-brown, with leaf scars large and prominent. The terminal buds are
2.5 cm long, 0.7 cm thick at the base, pubescent, somewhat flattened or
angular, and green to tawny. The alternative leaves are large, up to 25 cm
long and 15 cm broad, ovate, the apex bluntly acute or acuminate, and the
margins coarsely serrate to almost entire. The blades are variable as to the
base, which is sometimes obovate and narrowed to cordate with narrow
sinus, the lobes often overlapping and sometimes asymmetrical. Some
blades are distinctly narrowed from the middle or above toward the rounded
base (Santiago et.al, 2017).
The texture is chartaceous. Hydathodes are present and densely
scattered over the surface, which is pubescent only on veins above but silky
pubescent below. Venation is prominent, with 3 to 5 pairs of basal veins and
7 to 9 laterals flanking the midrib, these well elevated, widely spaced, and
branched toward the margin. The petioles are up to 5 cm long, stout, silky-
pubescent, rusty-scurfy, and reddish to chocolate-brown, abruptly changing
to green at the base of the midrib. This species is cauliflorous, with the figs
borne mostly in fascicles from the stem and branches, on pubescent
peduncles up to 1.1 cm long. The sterile figs are 3.5 cm in diameter, globular
to oblate-spherical, with the surface pubescent but glossy, and decorated
with prominent, white flecks which are thickly scattered, the larger ones
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with a corky spot in the center. The figs are green when young but change to
scarlet before dropping. The umbilicus is large and somewhat depressed
(Mapatac, 2019; Arquion, Nuñez, & Uy, 2015).
Ficus Nota is a useful tree. The fruit grows on short stems from the
branches and stem of the tree. Ripe fruits are edible and maybe eaten with
sugar and cream. Young leaves are eaten as vegetable. A drinkable water can
be obtained from a freshly cut stem. The tree is also used for firewood and
for making charcoal and is also used medicinally here in the Philippines. The
fruits of most species of Ficus nota are edible and are usually has a local
economic importance or eaten as bush food (Guian, 2020).
Ficus Nota can release thousands of liters of water per year. Tibig is a
nurse tree and a water collector. Its roots are sponge-like that absorbs water
during the rainy season and releases water during summer. It grows
abundantly in farm. They say that the presence of a tibig tree is an indicator
of the presence of water in the vicinity (Guian, 2020).
Traditional use
The fruits are traditionally eaten with sugar and the leaves are
consumed as vegetable. Different parts of the plant are ethnomedically used
for the treatment of diabetes, hypertension, fever, urinary tract infection,
and muscle pain (Santiago et.al, 2017).
In some parts provinces of the Philippines “tibig” has many kinds of
traditional medicinal practices and there are small, related research about it
that has proven its medical property from the leaves, fruit, bark and roots.
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Aribal L., Bruno A.G., & Jumawid, E. (2016) revealed that Particular
Ficus species have therapeutic components. One of the most significant
plant groups in the tropical rainforest ecosystems, fig trees are primarily
thought of as a major food source for frugivores. The range of advantages
received from fig trees led to the identification of this remarkable species of
trees. In the Philippines, the 104 species of Ficus make it the fifth most
specious genus.
When the fruit (Ficus Nota) is ripe, it can be consumed raw, while the
young leaves are cooked and eaten as a vegetable. The water extracted from
the tree is useful for the treatment of fever and to ease muscle discomfort.
Urinary tract infections, hypertension, and diabetes can all be treated with a
decoction of the roots and bark. (Ragasa C., Alimboyoguen A., & Shen C-C.
2014).
In the Philippines, the Mansaka tribe in Compostela Valley used the
decoction of the bark of this plant to treat asthma, cough, and other
respiratory condition. The Ayta people of Porac, Pampanga province in the
Philippines used the stem of this plant species as repellent against
hematophagous insects.
Plants have long been recognized as a potent source of biologically
pharmacologic active compounds, held true with various studies (Al Bayati &
Al-Mola, 2008).
Medicinal plants are considered as rich sources of ingredients which
can be used in drug development and synthesis. They play a critical role in
the development of human cultures around the whole world 1 and have been
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used for centuries throughout the world by majority of the world’s
population which until today still rely on them as traditional and alternative
medicine for primary health care needs (World Health Organization, 2015).
B. Related Studies
Aribal L., Bruno A.G., & Jumawid, E. (2016) revealed that Particular
Ficus species have therapeutic components. One of the most significant
plant groups in the tropical rainforest ecosystems, fig trees are primarily
thought of as a major food source for frugivores. The range of advantages
received from fig trees led to the identification of this remarkable species of
trees. In the Philippines, the 104 species of Ficus make it the fifth most
specious genus.
When the fruit (Ficus Nota) is ripe, it can be consumed raw, while the
young leaves are cooked and eaten as a vegetable. The water extracted from
the tree is useful for the treatment of fever and to ease muscle discomfort.
Urinary tract infections, hypertension, and diabetes can all be treated with a
decoction of the roots and bark. (Ragasa C., Alimboyoguen A., & Shen C-C.
2014).
According to (Levitah M. 2018), Escherichia coli ATTC 25922,
Staphylococcus aureus ATTC 25923, Klebsiella pneumonia UPCC 1360, and
Salmonella typhimurium UPCC 1368 were used to test the fruit (Ficus nota)
extract's antimicrobial activity against pathogenic strains. The qualitative
phytochemical tests revealed the presence of common bioactive compounds
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such as alkaloids, tannins, flavonoids, saponins, flavonoids, and
anthraquinon. As compared to the common chemotherapeutic drug
chloramphenicol, the aqueous extract has demonstrated considerable
excellent activity while the ethanolic extract has demonstrated high activity,
it was concluded that Ficus nota are rich in bioactive components and
exhibits antimicrobial activity against pathogens. To add to that, according
to Latayada F., & Uy M. (2016), their results of the study demonstrate that
the bioactive components of Ficus nota in the different leaf extracts also
have antibacterial, antifungal, and possible cytotoxic effects. These results
provide more evidence in favor of Ficus nota's use in traditional medicine.
Results revealed that the plant extract possess both antioxidant and
prooxidant properties depending on the concentration by which they are
used. The dual nature exhibited by F. nota is deemed beneficial for the
treatment and prevention of cardiovascular disease, diabetes, cancer, and
aging. It can also aid in the maintenance of cellular homeostasis, host
defense mechanisms, and various signaling processes in the body (Santiago
et.al, 2017).
Findings showed that depending on the plant extract's dosage,
prooxidant and antioxidant characteristics depending on how intensely they
are utilized. The contradictory behavior of Ficus Nota is thought to be helpful
in the treatment and prevention of cancer, diabetes, cardiovascular disease,
and aging. Moreover, it can support the body's signaling systems, host
defense mechanisms, and cellular homeostasis. (Santiago, L.A., Saguinsin,
S.G.C., Reyes, A.M.L., Guerrero, R.P., et. al. 2017)
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Moreover, Studies revealed by (Ragasa, C., Alimboyoguen A., & Shen
C-C. 2014) that Ficus nota unripe fruit extract was subjected to chemical
analysis, yielding 4-(2-hydroxyethyl)-2-methoxyphenol, a combination of
meso-2,3-butanediol, (2R,3R)-2,3-butanediol, and (2S,3S)-2,3-butanediol, as
well as -sitosterol. With the use of comprehensive 1D and 2D NMR
spectroscopy, the structure of 1 was clarified. Structural moieties of
methoxyphenols and sitosterol are known to exhibit antioxidant, prooxidant,
anti-inflammatory, antibacterial, hypocholesterolemic, anti-diabetic,
cytotoxic, and immunomodulatory effects (Subramaniam, S., Keerthiraja, M.,
& Sivasubramanian, A., 2014)
Furthermore, Findings showed according to (Banawa N.G., &
Bernasol J. D, et. al n.d) that due to its lowest IC50 value among other plant
extracts, the ethanolic crude extract of Ficus nota leaves exhibited the most
substantial alphaglucosidase inhibitory activity. The ethanolic crude extract
of Ficus nota leaves inhibited mammalian alpha-glucosidase and may have
hypoglycemic properties because nota leaves demonstrated the presence of
carbohydrates, flavonoids, saponins, tannins, and glycosides, all of which
were found to inhibit alpha-glucosidase and have antihyperglycemic activity.
The dual property of the F. nota ethanolic leaf extract was assessed to
establish its effectiveness as an antioxidant and/or prooxidant.
Phytochemical analysis indicated the presence of alkaloids, flavonoids,
phenols, terpenoids, saponins, and tannins that may have significantly
contributed to its purported bioactivity. Results from general and specific
antioxidant assays revealed that the plant extract acts as an antioxidant at
low concentrations and exhibits prooxidant effects at high concentrations. In
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this regard, F. nota can be considered as an agent with dual potential in
biological and pharmaceutical applications that is deemed beneficial for the
treatment and prevention of cardiovascular disease, diabetes, cancer, and
aging. Additionally, the two-edge nature exhibited by the plant extract can
also aid in the maintenance of cellular homeostasis, host defense
mechanisms, and various signaling processes in the body. Whereas the gap
directs to the amino acid constituent’s discovery of the essential oil other
than the fruit extract that was being studied (Santiago et. al., 2017).
Moreover, the antimicrobial activity of the fruit extract against
pathogenic strains evaluated based on the zone of inhibition using paper
disc diffusion method with tested bacteria of Escherichia coli ATTC 25922,
Staphylococcus aureus ATTC 25923, Klebsiella pneumonia UPCC 1360, and
Salmonella typhimurium UPCC 1368. The qualitative phytochemical tests
exhibited the presence of common bioactive compounds including alkaloids,
tannins, flavonoids, saponins, flavonoids and anthraquinones. It was further
suggested that analysis of other compounds on the fruit of Ficus nota must
be obtained (Mapatac, 2019).
Meanwhile, the methanolic leaf extract of Ficus nota upon
quantitative analysis, researchers determined that it has a positive result for
flavonoids. As such, standard calibration curve of Quercetin for Total
Flavonoid Content and Gallic acid for Total Phenolic Content proves that the
extracts contain flavonoid and phenolic compounds. Moreover, the Thin-
layer chromatography result supports the presence of flavonoids in the leaf
extracts of Ficus nota. The results obtained from IC50, and percent
inhibition revealed that Ficus nota possesses XO inhibitory activity. The
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study has not yet conducted in-vivo study and the isolation and
identification of active compounds found in the leaves but also the fruit
extract of Ficus nota must also be conducted to identify a potential chemical
entity for clinical use in the prevention and treatment of gout (Mancia et. al.,
2019).
A study was conducted that focuses on the stem extracts of Ficus
nota which exhibited cytotoxic activity against the brine shrimp Artemia
salina. The ethanolic extract showed greater cytotoxic activity compared to
the decoction extract. These further warrants phytochemical screening of the
ethanolic extracts of F. nota for the determination of its active cytotoxic
components. The gap was found pointing towards bioactive components of
the fruit extracts of Ficus nota (Arquion et. al., 2015).
Based on the study of Latayada and Uy (2016), the antimicrobial
activities and cytotoxic potentials of the ethanol, hexane, chloroform and
aqueous extracts of the Philippine medicinal plant F. nota was evaluated
using disc diffusion assay and brine shrimp lethality assay, respectively. The
aqueous extract (FnA) showed active to very active levels of inhibition against
the three bacterial test organisms B. subtilis, S. aureus, and E. coli, with
zones of inhibitions ranging from 18.7±1.15 mm to 20.0±0.00 mm. The
extract FnA further exhibited activity against the two fungi S. cerevisiae
(11.7±1.15 mm, partially active) and C. albicans (14.0±1.00 mm, active). This
significant antimicrobial result of the aqueous extract was followed by the
chloroform extract, FnC (17.0±5.2 mm, active against B. subtilis and
22.3±4.62 mm, very active against S. aureus). Meanwhile, the toxicity test
showed that the ethanol extract FnE scored significant toxicities against the
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test organism Artemia salina with LC50 values of 79.43 g/ml and 206.5
g/ml, respectively. This study shows that the bioactive components of F.
nota with antibacterial, antifungal, and potential cytotoxic properties were
successfully and effectively extracted and concentrated in the various leaf
extracts. These findings further support the ethno-medicinal uses of F. nota.
A chemical investigation on the dichloromethane extract of the unripe
fruits of F. nota afforded the following chemical constituents: 4-(2-
hydroxyethyl)-2-methoxyphenol, a mixture of meso-2,3-butanediol, (2R,3R)-
2,3-butanediol and (2S,3S)-2,3- [9] butanediol and β-sitosterol. Cytotoxicity
study using in vivo brined shrimp lethality of the decoction and ethanol
extracts of the stem of F. nota were active against the brine shrimp Artemia
salina with LC values of 991.00 ppm and 852.22 ppm, respectively (Arquion
et. al., 2015). The decoction and ethanol extracts of the leaves of F. nota has
been recently reported to possess considerable antioxidant properties
(Latayada, 2016:5). However, there are no reports yet on the antimicrobial
activities and potential cytotoxicities of the leaf extracts of F. nota (Latayada
& Uy, 2016).
On the clinical studies, based on the study conducted by Ares K.G. et
al. (2015), which entitled “In Vitro Assessment of The Antioxidant and
Acetylcholinesterase Inhibitory Activities Of Ficus Nota (Blanco) Merr.” where
it assesses the antioxidant activity of the crude methanolic extract of the
leaves of Ficus nota (Blanco) Merr. using scavenging assays for hydroxyl
radical, nitric oxide radical, and hydrogen peroxide, and its
acetylcholinesterase (AChE) inhibitory activity. In the study the proponents
used generic drugs which includes the ascorbic acid (Vitamin C) or ascorbate
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which is a dietary supplement and donepezil (Aricept) which is a medication
used to treat Alzheimer's disease (Ares K.G. et al., 2015).
Chapter III
MATERIALS AND METHODS
1. Preliminary Processing of the sample
The preliminary treatment of the Ficus nota fruit in the present study
included washing, blanching, and then extraction via alcohol. Aqueous
washing solutions using 10% ethanol would inhibit any microbial
contamination. Blanching was done in water at 95 – 98 OC. In this process,
blanching took only three (3) minutes. Water was heated first. Just before it
boils, fruits of the sample were dropped. The blanching process was
extended to three (3) minutes with occasional stirring after which, the fruits
were to be drained and washed immediately with cold water. The fruits were
dried via oven, pre-weighed to determine the yield in every kilogram of the
sample as alcoholic extract.
2. Biochemical Profiling
a. Phytochemical Screening
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The study primarily aimed to carry out preliminary phytochemical
screenings, to detect the major class of compounds present in the extracted
essential oil and to perform TLC profiling via Thin Layer Chromatography
(TLC). Results of this procedure were revealed the secondary metabolites
present in the fruit sample. These metabolites were different classes of
organic compounds that provide unique properties of the alcoholic extract
from the sample.
Table 1. Test outcomes for constituents tested from the given spray reagents.
Constituents Tested Spray Reagent Observable Result for
a positive test
Alkaloids Dragendorff’s reagent Brown-orange visible
spots immediately
after spraying; colors
are not stable
Phenols, Tannins, Potassium ferricyanide Blue spots
Flavonoids ferric chloride (K3Fe
(CN)6·FeCl3)
Preliminary test for H2SO4 Violet – essential oil
Essential Oils
Preliminary test for Vanillin-H2SO4 Phenols – brown
phenols, fatty acid, Fatty acid – yellow
essential oil and Essential oil – pink-
triterpenes violet
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Blue -violet -
Triterpenes
Antraquinones KOH -MeOH Yellow under UV
The phytochemical screening was based on Guevara (2005). Alcoholic
crude extracts were spotted to TLC silica gel, SGF254. The TLC plates were
developed in 5:1 ratio of CHCl3: MeOH solvent system. UV light will be used
to visualize the UV active spots corresponding to a certain metabolite. Spray
reagents will be used on the TLC plates to determine the different classes of
compounds present in the extract. Vanillin-H 2SO4 was used to test the
presence of sterols, steroids, and triterpenes. For the detection of alkaloids,
Dragendorff’s reagent was used. Potassium ferricyanide-ferric chloride
reagent was used for the presence of phenolics. For the presence of
anthrones, coumarins and anthraquinones, KOH-MeOH was used.
b. Determination of the Total Phenolic Contents
The amounts of phenolic compounds in the extract were to determine
according to the method of Ragazzi and Veronese (1973) by Folin-Ciocalteu
method with slight modification. A stock solution of 1ppm ascorbic acid was
prepared and diluted into a 2-fold serial dilution to prepare the following
concentrations, 0.5ppm, 0.25ppm, 0.125 ppm, and 0.0625 ppm of ascorbic
acid. 50 mL of sodium carbonate with a concentration of 1mg/ml was also
prepared. This was done by weighing 50 mg of Na2CO3 and dissolved with
distilled water to make a final volume of 50 mL solution. 200 µL each of the
concentrations of ascorbic acid was placed in separate plastic cuvettes,200
µL of Folin-Ciocalteu was added to each and additional 400 µL of the Na2CO3
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solution were to be added to each of the mixture. A blank was prepared by
the mixture of 200 µL distilled water, 200 µL of Folin-Ciocalteu and 400 µL
of Na2CO3 solution and placed in another plastic cuvette. Subsequently, the
mixtures were allowed to stand for few minutes and then measured at
680nm of the UV-Vis spectrophotometer (PD-303 UV Spectrophotometer)
which was run 30 minutes prior to reading. A calibration curve was
prepared after the reading. After taking the calibration curve, 200 µL of 1
ppm of the sample that showed positive result in potassium ferricyanide-
ferric chloride test in TLC was mixed with 200 µL of Folin-Ciocalteu and 400
µL of Na2CO3 solution in a cuvette and measured in the spectrophotometer.
It was being carried out in triplicates. The total phenolic content was
expressed as ascorbic acid equivalents.
Antimicrobial Activity via Diffusion Disk Assay
The procedure for the diffusion disc assay to assess the antimicrobial
activity of Ficus nota can be summarized as follows:
1. Preparation of microbial cultures: Cultures of specific
microorganisms, such as bacteria or fungi, were prepared using
appropriate growth media and conditions. These cultures served as
the test organisms to evaluate the antimicrobial properties of Ficus
nota.
2. Preparation of Ficus nota extract: The alcoholic extract of Ficus nota
was prepared using a suitable solvent, such as ethanol or methanol.
The extraction process involved grinding or blending the plant
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DIFFUN CAMPUS
material and then subjecting it to solvent extraction to obtain the
extract.
3. Sterilization of materials: All the necessary materials, including Petri
dishes, agar medium, and glass or paper discs, were sterilized to
maintain aseptic conditions and prevent contamination.
4. Preparation of agar plates: Agar plates were prepared by pouring the
appropriate agar medium into sterilized Petri dishes and allowing it to
solidify. The agar medium may contain nutrients necessary for the
growth of microorganisms.
5. Inoculation of microbial cultures: A standardized amount of microbial
culture was evenly spread on the surface of the solidified agar using a
sterile swab or loop, ensuring uniform distribution.
6. Application of Ficus nota extract: Sterile glass or paper discs were
impregnated with a known concentration of the Ficus nota extract.
The discs were placed on the surface of the inoculated agar plates,
gently pressing them to ensure proper contact.
7. Incubation: The inoculated agar plates with the Ficus nota extract
discs were incubated at an appropriate temperature and duration
suitable for the growth of the test organisms.
8. Measurement of zones of inhibition: After incubation, the plates were
examined for the presence of clear zones around the discs, indicating
inhibition of microbial growth. The diameter of these zones was
measured using a ruler or calipers.
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9. Data analysis and interpretation: The diameter of the zones of
inhibition was recorded, and the results were analyzed statistically to
assess the antimicrobial activity of the Ficus nota extract against the
tested microorganisms.
Chapter IV
RESULTS AND DISCUSSIONS
This chapter presents the results and discussion of an extensive
investigation into the biochemical profile of the alcoholic extract of Ficus
nota. The study aimed to explore the various chemical constituents present
in the extract and their potential implications for medicinal and therapeutic
purposes. The flow of the results and discussion is aligned with the research
questions posed at the outset of the study. The findings shed light on the
composition and potential bioactive compounds present in Ficus nota,
contributing to the existing body of knowledge in the field of natural
products research.
4.1 Percentage Yield
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The percentage yield is a measure of the efficiency of a chemical
reaction or extraction process. It represents the proportion of the desired
product obtained compared to the theoretical maximum yield that could be
obtained under ideal conditions.
The calculation of percentage yield involves dividing the actual yield
(the amount of product obtained in practice) by the theoretical yield (the
maximum amount of product predicted by stoichiometry or other
calculations), and then multiplying by 100 to express the result as a
percentage.
The percentage yield provides valuable information about the
effectiveness of a reaction or extraction procedure. A high percentage yield
indicates that the process is efficient, and that the majority of the starting
materials were successfully converted into the desired product. Conversely, a
low percentage yield suggests that there may be inefficiencies or losses
during the reaction or extraction process.
By comparing the actual yield to the theoretical yield, scientists can
evaluate the quality of their experimental techniques, identify sources of loss
or inefficiency, and make adjustments to improve the process. This
information is crucial for optimizing the production of desired compounds,
minimizing waste, and ensuring the cost-effectiveness of chemical processes.
In the context of this study, the yield of extraction was obtained by
determining the amount of crude alcoholic extract via gravimetric method.
The crude alcoholic extract was weighed after obtaining a syrupy
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characteristic was observed. This yield was obtained from 1 kilogram of the
fresh sample.
P ercent Yield=10 g of alcoholic crude extract ∈every 1 kilogram of the sampl e
4.2 Biochemical Profile of the Crude Alcoholic Extract of Ficus nota
4.2.1 Phytochemical Screening
A preliminary investigation was conducted to analyze the presence of
various phytoconstituents in the alcoholic extract of Ficus nota. The extract
was subjected to tests to identify alkaloids, flavonoids, tannins, phenols,
essential oils, fatty acids, triterpenes, and anthraquinones. The purpose of
this phytochemical analysis was to detect and profile the bioactive
compounds present in the fruit extract. Additionally, a TLC spot test was
performed on the alcoholic extract to further identify specific bioactive
compounds.
Figure 2. TLC Spot test results as guide in profiling bioactive compounds
present in crude alcoholic extract of Ficus nota
The figure above shows the qualitative screening on the bioactive
compounds of Ficus nota. A orange-brown, purple, and dark brown colors
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DIFFUN CAMPUS
indicate the presence of important bioactive constituents. These tests result
were obtained based from the vanillin-sulfuric acid test and preliminary
tests via TLC testing.
The table below summarizes the presence of bioactive compounds on
each sample based from the adapted phytochemical screening method.
Table 2. Bioactive components of the crude alcoholic extract of Ficus nota

Essential oils
Alkaloids
Alcoholic Extract of
phenols
Ficus nota
Profile +++ ++++ +++
Table 2 shows the bioactive components obtained based from the
method employed. The TLC detected bioactive components on the sample
and is positive to some indicated components. Phenols, alkaloids and
essential oils were all present in the dried crude alcoholic extract of Ficus
nota.
Phenols exhibit a range of applications in various fields due to their
distinctive properties. Their antioxidative capabilities make them valuable for
preserving food, cosmetics, and pharmaceutical products by protecting
against oxidative damage (Dapkevicius et al., 2015). Additionally, certain
phenols, such as carbolic acid (phenol), possess antimicrobial properties and
find use as disinfectants and antiseptics in healthcare and personal hygiene
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DIFFUN CAMPUS
products (Hamoud et al., 2017). Phenolic compounds are employed in
polymer production, particularly in the creation of phenolic resins utilized in
adhesives, coatings, and molded products (Sawpan et al., 2020).
Phenols have also been investigated for their potential pharmaceutical
applications, such as their anti-inflammatory, anticancer, and antimicrobial
properties (Li et al., 2019). Furthermore, these compounds play a role in
environmental remediation processes, including wastewater treatment, due
to their ability to degrade pollutants and eliminate toxic substances (Lu et
al., 2020). The versatility and significance of phenols across various
industries underscore their wide range of applications. It is important to
note, however, that specific applications may vary depending on the specific
type and properties of phenolic compounds.
Essential oils have a wide range of applications in various industries
and settings due to their unique chemical composition and aromatic
properties. One significant application of essential oils is in aromatherapy,
where they are used to promote relaxation, reduce stress, and enhance
overall well-being (Ali et al., 2015). These oils are also commonly used in the
fragrance and cosmetic industries, where they serve as natural and aromatic
ingredients in perfumes, lotions, soaps, and other personal care products
(Kumar et al., 2017).
In the field of medicine, essential oils have been studied for their
potential therapeutic properties, including antimicrobial, anti-inflammatory,
and antioxidant effects (Bakkali et al., 2008). They are utilized in natural
remedies, herbal medicine, and traditional healing practices for various
conditions, such as respiratory ailments, digestive disorders, and skin
conditions (Guimarães et al., 2018). Additionally, essential oils are employed
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DIFFUN CAMPUS
in the food and beverage industry as flavoring agents to enhance the taste
and aroma of products (Burt, 2004).
Furthermore, essential oils have found applications in household and
cleaning products due to their natural disinfectant and deodorizing
properties (Hussain et al., 2019). They are used in air fresheners, surface
cleaners, and laundry detergents as a safer alternative to synthetic
chemicals. In agriculture, essential oils are explored for their potential as
natural pesticides, insect repellents, and plant growth regulators (Isman,
2019).
Alkaloids are a diverse group of natural compounds that have various
applications in different fields. One prominent application of alkaloids is in
the field of medicine and pharmacology. Many alkaloids exhibit significant
biological activities and have been utilized as the basis for the development
of numerous therapeutic drugs (Mahato and Garai, 2011). For example,
alkaloids such as morphine and codeine derived from opium poppy have
been used as analgesics and pain relievers (Boyer and Shannon, 2005).
Additionally, alkaloids like quinine have been employed as antimalarial
agents (Willcox et al., 2004).
Furthermore, alkaloids have shown potential as sources of anticancer
agents. Compounds such as vinblastine and vincristine, derived from the
Madagascar periwinkle plant, have been used in chemotherapy treatments
(Kinghorn et al., 2009). Alkaloids also play a role in the field of neuroscience,
with compounds like nicotine and caffeine acting as central nervous system
stimulants (Benowitz, 2010).
In addition to their medicinal applications, alkaloids have found use
in the agrochemical industry. Certain alkaloids possess insecticidal
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DIFFUN CAMPUS
properties and have been used as natural pesticides for crop protection
(Isman, 2006). They can also function as growth regulators for plants,
contributing to enhanced crop yield and quality (Aniszewski, 2007).
It is important to note that alkaloids can have both beneficial and
toxic effects, and their applications should be carefully studied and
monitored. Ongoing research continues to explore the diverse potential of
alkaloids in various fields and their role in the development of novel
therapeutic agents.
4.2.2 Antioxidant Screening Results Via Total Phenolic Content
To quantify the phenolic compounds in terms of the ascorbic acid
equivalent (AAE), the total phenolic content was determined using the
adapted method. All experiments were performed in triplicates.
Equivalents of standards were calculated on the basis of standard
regression lines for ascorbic acid (R 2 = 0.9308) and display equation of
y=2.775x + 0.031 (See figure 4). The absorbance of the blank was used as
set intercept.
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Calibration Curve of Ascorbic Acid

3
Figure
f(x) 3. Graph ofx +the
= 2.77502414507675 calibration
0.031 curve of ascorbic acid.
2.5 R² = 0.962253723945299
The ascorbic acid equivalence of the sample was calculated by the
2
calibration curve derived from dose dependent curve of the control ascorbic
1.5
acid. A highly significant quantity of phenolic contents was found in the
alcoholic
1 extract of the fruit in terms of equivalents of ascorbic acid.
0.5
Table 3. Total Phenolic Content

0
0 0.2 0.4 0.6 0.8 1 1.2
Alcoholic Extract Weight of the Mean Abs mg AAE/ gram
samples in dry weight
grams samples
Ficus nota fruit 0.003 0.225 23.00
Figure 4. preparation of the standard calibration curve via Ascorbic Acid
Equivalent
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Result in table 3 reveals the total phenolic content of the crude alcoholic
extract of Ficus nota. Regression calculation shows that 23 mg of AAE
(Ascorbic Acid Equivalent) was present in every 1 gram of the sample.
Phenolic compounds and ascorbic acid (vitamin C) have a wide range
of applications due to their beneficial properties. Phenolic compounds,
including flavonoids, phenolic acids, and tannins, are well-known for their
antioxidant activity (Rice-Evans et al., 1996). They exhibit strong free radical
scavenging capabilities, making them valuable as natural antioxidants in
various industries. Phenolic compounds are commonly used in the food and
beverage industry as natural preservatives to prevent oxidative damage and
extend the shelf life of products (Cai et al., 2004).
Additionally, phenolic compounds possess antimicrobial properties,
showing inhibitory effects against pathogenic bacteria and fungi (Cushnie
and Lamb, 2005). This antimicrobial activity has led to the exploration of
phenolic compounds in the development of natural antimicrobial agents and
disinfectants (Cowan, 1999).
Moreover, phenolic compounds have attracted attention for their potential
health benefits. They exhibit anti-inflammatory, anticancer, and
cardioprotective properties, making them promising candidates for
therapeutic applications (Scalbert et al., 2005). Consumption of phenolic-
rich foods and beverages has been associated with a reduced risk of chronic
diseases, including cardiovascular diseases and certain types of cancer
(Pandey and Rizvi, 2009).
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Ascorbic acid, or vitamin C, is an essential nutrient with strong
antioxidant properties (Carr and Frei, 1999). The ascorbic acid equivalent is
a measure used to quantify the antioxidant capacity of phenolic compounds
based on their ability to scavenge free radicals (Cao et al., 1996). This
determination of ascorbic acid equivalent is valuable for assessing the
antioxidant potential of phenolic-rich substances such as plant extracts and
natural products.
In summary, phenolic compounds and the ascorbic acid equivalent
have diverse applications, including their use as natural antioxidants in the
food industry, potential therapeutic agents for various diseases, and their
role in promoting overall health and well-being.
4.3 Microbial Activity of the Crude Alcoholic Extract of Ficus nota via
Diffusion Disc Assay
The disc diffusion assay is a widely used method for evaluating the
antimicrobial activity of various substances. The interpretation of the results
obtained from the disc diffusion assay involves assessing the zones of
inhibition around the discs impregnated with the test substances. The size
of the zone of inhibition indicates the extent of antimicrobial activity, with
larger zones generally indicating stronger activity against the target
microorganism. The figure below shows the result on the actual zone of
inhibition of the extract against SA bacteria.
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Figure 5. Zone of Inhibition against SA
The figure above shows that the largest inhibition is about 13mm,
which can be described by the table below.
Table 4. Result of microbial assay against SA
Test Organism: ANALYSIS
Staphylococcus
aureus
Sample
Sample Average Zone of
Type
Inhibition obtained in
mm
Size of Disc : 6 cm
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Ficus Alcoholic 13 mm Intermediate (Partially
nota Extract Active)
In the context of the disc diffusion assay, the term "intermediate" is
used to describe a zone of inhibition that falls between the susceptible and
resistant categories. When interpreting the results of the assay, a zone of
intermediate size indicates that the antimicrobial activity of the test
substance is not strong enough to be classified as susceptible, but it is also
not weak enough to be classified as resistant.
The intermediate category is used when there is uncertainty or limited
clinical data available regarding the significance of the zone size in terms of
therapeutic effectiveness. It suggests that further investigation or clinical
correlation may be needed to determine the appropriate treatment approach.
The intermediate result highlights the need for caution and careful
consideration in determining the clinical significance of the antimicrobial
activity.
4.4 Potential applications based on the results of the sample
Biochemical Profile
4.4.1 Based on Phenolic Content
Due to its antioxidant property, a commercial application can be
considered such as alcoholic beverage. A wine that has phenolic rich content
can be made through fermentation of the Ficus nota fruit. Antioxidants in
alcoholic beverages have been proposed as an important contributory factor
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DIFFUN CAMPUS
to the protective effect of regular alcohol use against atherosclerotic
cardiovascular disease (Polak et al., 2020).
4.4.2 Based on Anti-Microbial Activity
A bactericidal alcohol can be made of the extract of Ficus nota. The
bactericidal activity of the substance is effective against Staphylococcus
aureus. This can be used as an alternative natural raw substance in making
alcohol.
CHAPTER V
SUMMARY, CONCLUSION, RECOMMENDATION
This experimental research aimed to study the biochemical profile of
the Ficus nota (Tibbeg) and its potential applications in commercial use. The
crude alcohol was extracted from the fruit. The researchers, then conducted
phytochemical analysis to determine certain bioactive compounds present in
the fruit. Afterwards, it was revealed that the extract contained essential oil,
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DIFFUN CAMPUS
alkaloids, and a high content phenol. Thus, the crude alcohol extract was
rich in phenolic content. The antioxidant screening via total phenolic content
which showed highly significant quantity of phenolic contents was found in
the alcoholic extract of the fruit in terms of equivalents of ascorbic acid.
Additionally, the alcoholic crude extract exhibited anti-microbial activity as
partially active against bacteria such as Staphylococcus aureus.
The researchers deduced the following based on the obtained result:
1. There are 10 g of alcoholic crude extract in every 1 kilogram of the
sample as the percentage yield.
2. The alcoholic crude extract of Ficus nota contained high phenolic
content with the identification of essential oil and alkaloids. This
means that it has antioxidant property.
3. The extract is partially active against certain bacteria, especially
Staphylococcus aureus, being validated by the presence of alkaloids
(Ferrazzano et al., 2013).
4. The production of alcoholic beverage and bactericidal alcohol are the
potential application based on the results of the study.
The current study can be interpreted as the first step in investigating
the alcoholic crude extract of Ficus nota. Agricultural entrepreneur,
investors, and administrators should be encouraged to invest and support
the production of alcoholic beverages and alcohol made of Ficus nota.
Future research could further examine the anti-microbial activity of
alcoholic crude extract to other kinds of bacteria and certain target
microbes. Although, the essential was found in the extract this may further
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DIFFUN CAMPUS
suggest different approach such as extracting the essential oil directly from
fruit. Additionally, further study on the nutraceutical profile of the essential
oil extract.
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Merr Leaf and Fruit Extracts. DOI: 10.32871/rmrj1503.02.09
Merr Leaf and Fruit Extracts. Journal 3. DOI:
10.32871/rmrj1503.02.09
Pandey, K. B., & Rizvi, S. I. (2009). Plant polyphenols as dietary
antioxidants in human health and disease. Oxidative Medicine
and Cellular Longevity, 2(5), 270-278.
Philippine Alternative Medicine. (2011). Tibig Ficus nota (Blanco) Merr.
Sacking tree. Philippine Alternative Medicine. Retrieved October 15,
2015 from http://www.stuartxchange.org/Tibig.html).
Polak J, Bartoszek M, Lowe AR, Postnikov EB, Chorążewski M. Antioxidant
Properties of Various Alcoholic Beverages: Application of a
Semiempirical Equation. Anal Chem. 2020 Jan 21;92(2):2145-2150.
Ragasa, C, Alimboyoguen A, & Shen C-C (2014) Chemical constituents of
Ficus nota. Vol. 6(4), Page 98-101, ISSN: 0975-413X
46
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Ragazzi, E., & Veronese, G. (1973). Quantitative analysis of phenolic
compounds after thin-layer chromatographic separation. Journal of
Chromatography A. Volume 77. Issue 2. Pages 369-375. ISSN
0021-9673. Rice-Evans, C. A., Miller, N. J., & Paganga, G. (1996).
Structure antioxidant activity relationships of flavonoids and phenolic
acids. Free Radical Biology and Medicine, 20(7), 933-956.
Santiago, L.A., Saguinsin, S.G.C., Reyes, A.M.L., Guerrero, R.P.,
Nuguid, A.M.N., et. al; (2017). Total phenolic and flavonoid
contents and free radical scavenging components of Ficus
nota Merr. (Moraceae) ethanolic leaf extract.
Vol. 24, Iss. 5, (2017): 2050-2058.
Sawpan, M. A., Ali, M. E., & Islam, M. A. (2020). Phenolic resin as a
versatile material for adhesives and coatings: A review. Arabian
Journal of Chemistry, 13(12), 8897-8913.
Scalbert, A., Manach, C., Morand, C., Remesy, C., & Jimenez, L. (2005).
Dietary polyphenols and the prevention of diseases. Critical Reviews
in Food Science and Nutrition, 45(4), 287-306.
Stephanie F., Jules B., Batiha G., Ali I. &Subramaniam L. (2021)
Synergistic antibacterial action of ß-sitosterol-D- glucopyranoside
isolated from Desmostachya bipinnata leaves with
antiobiotics against common human pathogens. DOI:
10.5772/intechopen.98602
TechProtocols. (2022). [online] Available at:
https://www.iitg.ac.in/biotech/BTechProtocols/Thin%20Layer
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%20Ch romatography%20Protocol.pdf> [Accessed 26
September 2022].
Willcox, M. L., Bodeker, G., & Rasoanaivo, P. (2004). Traditional medicinal
plants and malaria. CRC Press.
DOCUMENTATION
1. Washing, Blanching, and Extraction of Alcohol from Ficus nota fruit.
48
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2. Phytochemical Screening
3. Antioxidant screening via Total Phenolic Content
4. Anti-microbial activity via disc diffusion assay
49
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- Preparation of McFarland Standard
Preparation of Nutrient agar for pure culture and disc diffusion assay
Putting disc diffusion assay into pure culture of bacteria, Staphylococcus
aureus
50
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- Result of the Anti-microbial
activity in SA
Curriculum Vitae
JASSER B. RAMIREZ
Male
22
Student
Details
Andres Bonifacio, Diffun, Quirino PROFILE
Philippines
ramirezjazz458@gmail.com I am the breadwinner of my family. I am
09274353267 hardworking and competent in every work that I do. I
believe in the saying, “Consistency is a discipline that drives
success “. Small consistent progress is a progress that
Skills aggregate bigger accomplishments if I do not break my
Leadership Skills momentum. I always excel in my study and willing to take
Communication Skills
all challenges and opportunities that come around.
Graphic Design
Microsoft Skills EDUCATION
IT savvy
Time-management skills Elementary
Creative writing skills Ifugao Village Integrated School, Diffun, Quirino
Formal writing skills Valedictorian
2013-14
Habits
Watching Documentaries Senior High School
Science Content Video Quirino State University-Laboratory High School
Reading Books With High Honor (Valedictorian)
Writing 2019-20
Cooking
Tertiary
Quirino State University
Bachelor of Secondary Education
2022-present
51
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Character Reference
______________________
MARY GRACE L. SADANG, MIT
Assistant Professor II
Achievements
Regional Higher Education Press Conference
Column Writing
9th Place
2023
Curriculum Vitae
DERIQ MAORENZ G. CRUZ

Male
21
Student
Details
Mangandingay, Cabarroguis, Quirino PROFILE
Philippines
cruzderiqmaorenz@gmail.com A results-oriented and highly motivated
09286316049 professional student. Possessing a strong track record of
various achievements. Known for demonstrating excellence
in my studies. Committed to delivering exceptional
Skills solutions while maintaining a keen focus on my peer’s
Social Skills satisfaction. A collaborative student with strong
Editing Skills
communication and interpersonal skills. Seeking an
opportunity to help andEDUCATION
Microsoft Skills
contribute to school and society's
success.
Elementary
Habits
Watching Animal Planet Mangandingay Elementary School
Playing Mobile Games 9th Honor
Cooking 2013-14
Dancing
Drawing Senior High School
Quirino General High School
Character Reference With High Honor (Valedictorian)
2019-20
______________________
MARY GRACE L. SADANG, MIT Tertiary
Assistant Professor II Quirino State University
Achievements
Bachelor of Secondary Education
2022-present
CavRasuc (Socio-cultural) Hip-Hop Competition Finalist
52
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Curriculum Vitae
MARY GRACE I. GACUSAN

Female
21
Student
Details
Rizal, Diffun, Quirino PROFILE
Philippines
marygracegacusan08@gmail.com I am the breadwinner of my family. I am a
09056924061 hardworking person. I believe that all things are possible.
You just need to persevere and be optimistic in life. You
need to work hard and not be afraid in failures because that
Skills will serve as a way for you to grow and continue striving.
Can work under pressure.
Leadership Skills
EDUCATION
Time-management skills
Elementary
Habits Diffun Central School, Diffun, Quirino
Watching Youtubes With Honor
Scrolling Facebook 2013-14
Reading Books
Writing Senior High School
Cooking
Diffun National High School
Dancing
With Honor
2019-20
Character Reference
Tertiary
______________________
Quirino State University
MARY GRACE L. SADANG, MIT
Assistant Professor II Bachelor of Secondary Education
2022-present
Achievements
Regional Higher Education Press Con
Editorial Writing
9th Place
2023 53
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54
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Curriculum Vitae
ARRIANNE S. CAMELLO
Female
21
Student
Details
Aurora West, Diffun, Quirino PROFILE
Philippines
arriannecamello01@gmail.com I am the breadwinner of my family. I am a
09166967326 hardworking person. I believe in the saying, “Consistency is
a discipline that drives success “. Small consistent progress
is a progress that aggregate bigger accomplishments if I do
Skills not break my momentum. I always excel in my study and
Can work under pressure willing to take all challenges and opportunities that come
Leadership Skills
around.
Time-management skills EDUCATION
Elementary
Habits Diffun Central School, Diffun, Quirino
Watching Youtubes With Honor
Scrolling Facebook 2013-14
Reading Books
Writing
Senior High School
Cooking
Quirino State University Laboratory High School
Dancing
With Honor
2019-20
Character Reference
Tertiary
______________________
MARY GRACE L. SADANG, MIT Quirino State University
Assistant Professor II Bachelor of Secondary Education
2022-present
Achievements
Regional Higher Education Press Con
Comics Strip
3rd Place
2023
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QUIRINO STATE UNIVERSITY

THE PROBLEM AND ITS BACKGROUND

Philippines is one of the most biodiverse countries in the world that

Merr. also known as “Tibbeg” in Ilocano, is an endemic plant that is often

et.al, 2014; Mapatac, 2019; Mancia, et.al, 2019). It belongs to family

epiphytes worldwide (Gandhi et.al, 2019). It is usually found in thickets and

forest at low and medium altitudes. It is an erect, spreading, dioecious tree

of about 8 meters (m) in height, with fruits around 2 to 3.5 cm in diameter,

(Philippine Alternative Medicine, 2011).

Traditional uses of Ficus nota include drinking its water extracted

and diabetes (Philippine Alternative Medicine, 2011; Ragasa et.al, 2014).

Artemia salina. According to the study conducted by Latayada & Uy (2016),

no activity against the A. niger, A. flavus, C. albicans, and S. cerevisea.

However, there are no reports yet on the antimicrobial activities and

potential cytotocixxities of the fruit extracts of F. nota.

While the Philippine biodiversity and rich cultural traditions of plant

use, scientific understanding of medicinal plants remains largely unexplored

and pharmacological investigation of the Philippine flora only grown recently.

chemotherapeutic value in traditional medicine but also in their potential

of providing cheaper nature-based alternative medicine to the public in the

midst of high-priced medicine produced by pharmaceutical companies. As

such, the study aimed to determine the antibacterial and cytotoxicity of

source of antibiotics (Mapatac, 2019).

This study was anchored in the updated Harmonized National

Research and Development Agenda (HNRDA) of Department of Science and

Technology (DOST) stipulated Drug Discovery and Development (Tuklas

Philippine Council for Health and Development (PCHRD). Under Section 2 of

stage. Drugs would be developed for infectious diseases such as Anti-biotic

communicable diseases such as cancer diabetes mellitus, cardiovascular

diseases, and other lifestyle-related diseases.

In addition, priorities for 2023-2025 will be the development of herbal

drugs by means of screening for bioactive extracts from natural sources

using validated protocols of the program; formulation of priority bioactive

extracts into standardized herbal drugs; scale-up studies for formulated

standardized bioactive herbal product; pre-clinical/In-vivo studies of priority

bioactive candidates; phase 1/2 clinical studies of candidate herbal drugs;

and cultural management/propagation of priority organisms. The drug track

would took place through isolation of bioactive compounds from natural

sources using validated protocols of the program and lead optimization of

bioactive compounds to come up with new chemical entities (NCEs). Then an

advanced step priorities for 2026-2028 similar to previous years was to be

conducted (DOST, 2022).

Ficus nota (Tibbeg) will be useful to explore its potential applications in

health spectrum to be part of this endeavor.

The vital efforts of this study focused on the biochemical profile of

applications. The crux of this study centered on the bioactive components of

the alcoholic crude extract from the Ficus nota.

Statement of the Problem

Generally, the study seeks answer to the following questions:

a) phytochemical constituents; and

b) phenolic content equivalent?

2. What is the extent of microbial activity of the alcoholic fruit extract of

its biochemical profile?

Scope and Delimitation of the Study

This study was conducted in Quirino State University, particularly,

The biochemical profiling was limited only to phytochemical

Significance of the Study

curing patients. Also, biochemical products may provide important

information about the outcomes of treatment, especially in antioxidant and

field of agriculture. There are abundance of trees growing in the locality

potential agricultural and economic value that is driven by this initiative to

The Investors. By coming up with the unraveling the potential commercial

evidence to attract investors to make a deal with those business enthusiasts

or the researchers themselves. Additionally, the outcome of this study would