II.

REVIEW OF RELATED LITERATURE AND RELATED STUDIES

This chapter includes the ideas, finished thesis, generalization or conclusions,

methodologies, and others. Those that were included in this chapter help in familiarizing
information that is relevant and similar to the present study.

This review includes previous and related studies and literature on the chemical
composition, phytochemical screening, and antimicrobial activity of Kangkong (Ipomoea
aquatica) ethanolic leaf extract. As well as its efficacy in inhibiting and eliminating the
growth of certain microorganisms. Excerpts from research journals, internet articles, or
news reports from worldwide sources, as well as the Philippines per se. The studies
acquired are restricted to explore researchers, earlier or beyond the years 2011 and
2021, respectively.

I. Review of Related Literature

A. Kangkong (Ipomoea aquatica)
I. Aquatica is a very useful vegetable crop that has been cultivated for centuries
in tropical countries. However, because of its potential to affect natural habitats, rice
and sugarcane production, irrigation systems, navigation, and recreation, it has been
marked as a Federal Noxious Weed in the United States. Because the potential
monetary or social benefits of deregulation outweigh the potential harm and cost of
control, the listing has been maintained (Anonymous, 2020. Ipomoea aquatica
swamp morning-glory).
According to the World Crops for the Northern United States (2022), almost all
parts of the tender shoot tips and younger leaves are edible, but the tender shoot
tips and younger leaves are preferred. Water spinach is also known as: kankon, you-
sai (Japanese); ong tsoi, weng cai (Cantonese); toongsin tsai (Mandarin); kang kong
(Filipino, Malaysian); kang kung, rau muong (Vietnamese); pak bung (Thai); batata
aquatica, cancon (Portuguese). Water spinach is a frost-tolerant plant.
Temperatures between 75° and 85° F are ideal for growth, and chilling injury can
occur below 50° F. Ching Quat is grown in moist soils, frequently in beds. Direct
seeding or transplanting are both options. To ensure the viability of the seed, many
Asian growers in Massachusetts soak it until germination. Growers plant 6 – 10
seeds per foot in rows 6 – 8 inches wide in beds. Plant stems are not particularly
strong, but plants grown in beds support one another and produce longer stems with
less branching, which the market prefers. Furthermore, water spinach necessitates
significantly more water than most other vegetable crops. Because increased
irrigation can leach out readily available nutrients, it is best to use slow-release
fertilizers. The entire plant can be harvested 50 to 60 days after planting. Plants are
harvested by cutting the stem close to the ground and then applying nitrogen to
encourage regrowth.
I. Aquatica grows in moist, marshy, or inundated environments, such as shallow
pools, ditches, or wet rice fields, from sea level to 1000 m. It grows in dense masses
and can be propagated easily from cuttings. It is also grown as a vegetable in
various parts of Asia, as well as being found in the wild and as a weed (Anonymous,
2020).

B. Phytochemical Screening of Ipomoea aquatica

Water spinach or Kangkong (Ipomoea aquatica) is found in an invasive species
in many parts of the world and designated as a noxious weed, forming ragged mats
that choke and stagnate in streams and swamps. Throughout the Philippines in
stagnant streams, fresh-water swamps, and pools (Stuart, Jr., G. U. 2016).
The plant contains 1.6 percent protein, 0.2 percent fat, and vitamins C3 and B2.
Nortropane and alkaloids are two major compounds found in plant. It is also used as
a purgative, diuretic, antiepileptic, CNS depressant, anti-inflammatory,
hypolipidemic, antimicrobial, anthelmintic, and antidiabetic. Antidiabetic, antioxidant,
antiproliferative, antimicrobial, diuretic, anti-ulcer, anxiolytic, memory-enhancing,
antiepileptic, CNS depressant, hepatoprotective, nephroprotective, membrane
stabilizing, a-glucosidase inhibitory, antiurolithiatic, acetylcholinesterase enzyme
inhibiting properties have been suggested in studies (Stuart, 2016).
According to Prota4U Record Display (2022), raw kangkong has the following
nutritional composition per 100 g edible portion: water 92.5 g, energy 80 kJ (19 kcal),
protein 2.6 g, fat 0.2 g, carbohydrate 3.1 g, dietary fibre 2.1 g, Ca 77 mg, Mg 71 mg,
P 39 mg, Fe 1.7 mg, Zn 0.2 mg, vitamin A 6300 IU, thiamin 0.03 mg, riboflavin 0.10
mg (USDA, 2002). The nutritional value of leaf-blades is greater than that of petioles
and stems; however, sources do not specify whether stems and leaves or only
leaves were studied. Heavy metal accumulation in kangkong has been reported for
Asia due to the plants' frequent growth in polluted water.
The presence of various phytochemicals such as alkaloids, saponin,
carbohydrates, Steroids, and terpenoids was discovered. Which are the chemical
compositions needed and essential in the forthcoming processes to come up with an
efficient and potent disinfectant.

C. Antibacterial and Antimicrobial Activity of Ipomoea aquatica

Kang Kong has a long history of traditional medicine use, with the stems being
determined as more potent than the leaves. The plant is rich in minerals such as
calcium and iron, and contains sodium, magnesium, phosphorus, manganese,
copper and zinc. Vitamin B2, C and K are also present. Further active ingredients
include flavonoids, catechin and phenolic compounds, so there is powerful
antioxidant activity (Mudbrick Herb Cottage, 2022).
Kangkong is also used as a laxative and sedative; it is used to treat piles,
nervous disorders, headaches, and insomnia. The juice is used as an emetic in
cases of arsenical or opium poisoning in Burma. In Cambodia, it is used as a
poultice for fever with delirium, and the buds are applied to ringworm. In Ayurveda,
leaf extracts are used to treat jaundice and nervous debility. In Sri Lanka, it is used
to treat liver disease, vision problems, and constipation (Stuart, 2016).
As cited by Stuart 2016, the antimicrobial efficacy of the leaf extracts of three
herbs – A longifolia, I aquatica, and E fluctuans – on four pathogenic bacterial
strains was investigated in a study (E coli, P aeruginosa, S aureus and M luteus).
Ipomoea aquatica outperformed the other two herb extracts in terms of antimicrobial
activity against bacterial strains. Moreover, the study examined herb extracts for
antibacterial activity against Propionibacterium acnes, a gram-positive human skin
commensal involved in the pathogenesis of acne. The best antibacterial activity
against P. acnes was demonstrated by an ethanol-water spinach extract. The MIC
was 1280 g/ml, while the MBC was greater than 5120 g/ml.
Due to its antibacterial activity and antimicrobial activity, we can conclude that the
ethanolic leaf extract of Kangkong (Ipomoea aquatica) can be utilized in many
medicinal, pharmacological, and decontaminating agent purposes. Ipomoea
aquatica "Kangkong" is a very useful herb that can be used as a medicine, a
delicacy, and, most importantly, a disinfectant that we can use in our own homes.

D. Bacteria Present in Frequently Held Surfaces

Diseases are frequently spread through direct contact with other people. A
pathogen (disease-causing microorganism) can easily escape from an infected
person via sniffles, sneezes, and coughs. However, in order to make a person sick,
a pathogen must enter him in sufficient numbers to survive the initial immune system
attack and then multiply (Australian Academy of Science, 2022).
According to the Australian Academy of Science (2022), Surfaces introduce a
new layer of difficulty. A pathogen must be able to land on a surface and survive
there until you touch it. After that, it must travel from the point of contact to the area
of the body targeted by the pathogen before it can thrive. Surface contamination is a
serious problem in healthcare settings, where a large number of people carrying
various germs congregate. Elsewhere, the situation is less dire because there are
fewer pathogens in a larger area. However, science can identify the most likely risks
to avoid.
Staphylococcus aureus is a gram-positive bacterium that causes a wide range of
clinical infections. Infections caused by this pathogen are common in both
community and hospital settings. Although S. aureus does not normally cause
infection on healthy skin, if it is allowed to enter the internal tissues or bloodstream, it
can cause a variety of potentially serious infections (Taylor and Unakal, 2021).
Staphylococcus (staph) is a bacterial group, according to the National Library of
Medicine (2020). There are over 30 different kinds. The majority of infections are
caused by Staphylococcus aureus. Staph bacteria are contagious and can spread
from person to person. They can also be spread on objects like towels, clothing,
door handles, athletic equipment, and remote controls.

E. Efficacy of Kangkong Ethanolic Leaf Extract

Ipomoea aquatica (kangkong) extract has antibacterial activity and can suppress
the growth of bacteria and germs. It can also be used as an alternate treatment for
destroying bacteria and microorganisms. It's an Organic Disinfectant that helps
people stay safe by reducing and eliminating the transmission of illnesses and
contaminants.
According to the Medicinal Health Guide (2016), kangkong is widely used in
traditional Chinese medicine to treat a variety of ailments. To treat fever, kangkong
juice mixed with water is applied as a cold compress. Constipation is relieved by
drinking boiled kangkong juice. Kangkong is also used to treat worm infestations in
the intestine. Kangkong is used to treat jaundice and liver problems in Indian
Ayurvedic medicine. Furthermore, kangkong leaves are used to treat gestational
diabetes.
Kangkong greens are very low in calories and fats, according to another article
from Nutrition and You.Com (2022). Nonetheless, its succulent leaves are high in
vitamins, particularly vitamin A (6600 IU/100 grams), as well as antioxidants and
minerals. Water spinach greens, like spinach, kale, and watercress, are one of the
most abundant sources of vitamin-A. 100 grams of fresh greens contain 6300 IU, or
210 percent of the daily recommended value of this vitamin. Vitamin A is required for
mucosal integrity, hair and skin health, as well as vision. In humans, it also acts as
an anti-cancer and anti-aging factor at the cellular level. They are also high in B-
complex vitamins, such as riboflavin (8%), niacin (5.5%), vitamin B-6 (pyridoxine-
7%), folic acid (14%), and others, all of which play important roles in metabolic
functions.
The Department of Molecular Medicine at the University of Malaysia conducted a
study on the protective action of kangkong against thioacetamide-induced
hepatotoxicity in rats to determine if kangkong can indeed help treat jaundice (as
cited in Medical Health Guide, 2012). An ethanol extract of kangkong was given to
rats whose livers had been damaged by thioacetamide. When compared to a control
group, the rats demonstrated resistance to liver lesions, cell necrosis, and
inflammation. The study also suggests that kangkong extract's protective effect in
chemical-induced liver damage may be due to its modulation of detoxification
enzymes, as well as its antioxidant and free radical scavenger properties.
Furthermore, it provides scientific support for the traditional use of kangkong in the
treatment of liver disorders.
According to an article on Flora and Fauna Web (2021), a Singapore
Government Agency Website, an aqueous leaf extract of kangkong helped to reduce
blood sugar in diabetic patients.
The study's findings will aid in improving public awareness about the Relevance
and efficacy of an organic disinfectant, namely Kangkong (Ipomoea aquatica)
Ethanolic Leaf Extract as an Organic Disinfectant Concentrate. This research will
offer hospitals and clinics with valuable information the effectiveness of Kangkong
Organic Disinfectant Cleaner Concentrate in getting rid of germs because hospitals
and clinics are vulnerable to a variety of threats. The given organic disinfectant will
be particularly useful to strains of bacteria

F. Extraction of Kangkong
Whenever utilizing a plant as an organic disinfectant cleaner concentrate, the
extraction process is the most important step. Extraction is done in order to analyze
the medicinal use of a plant, whereas the desired chemical components must be
extracted from plant materials before further identification and characterization.
According to Berkem (2022), plant extraction is a process that aims to extract
specific components found in plants. It is a solid/liquid separation process in which a
solid object (the plant) comes into contact with a fluid (the solvent). The solvent is
then used to solubilize and contain the plant components of interest. The resulting
solution is the desired extract. To isolate the plant extract, the solvent will eventually
be removed. It is not necessary to separate it from the extract if it is for the food
industry. If this is not the case, a second separation operation can be used to obtain
a dry extract. The source of active plant ingredient extraction has been lost to time.
In fact, mankind discovered the benefits of plants and the first techniques for
separating out what we now call "extracts" very early on. Plant extraction is
accomplished through solid/liquid extraction, which is then followed by purification
stages. It is thus defined as the operation of solubilizing one or more constituents
(solid or liquid) contained in a solid object in order to separate them. This fluid,
commonly referred to as a solvent, can be either a liquid or a gas (water vapor or
supercritical fluids).
Finally, the extracted content can be subjected to additional testing, such as
phytochemical screening, antimicrobial assays, and the determination of the
minimum inhibitory concentration. The extraction of Kangkong will be critical in
determining the plant's efficacy and potency in eliminating microorganisms. In the
current situation, whether this study will be able to provide feasible solutions.

G. Antimicrobial Assay
Antimicrobial assays are critical tools for testing and screening the inhibitory
effects of a wide range of compounds against microorganisms before establishing
their inhibitory spectra (broad vs. narrow). Before using antimicrobial compounds in
agriculture, biotechnology, or medicine, it is critical to understand their inhibitory
spectra. There are numerous traditional and modern methods available, but their
sensitivity and efficacy vary (Bailey, T. W., 2013).
From Engineering Textiles (Second Edition) (2020), the majority of methods for
assessing the activity of food antimicrobials have been adopted, either directly or
with modifications. An in vitro or screening test is used to obtain preliminary
information about antimicrobial activity when the compound is not applied to a
product under normal use conditions. The endpoint tests provide qualitative data on
effective concentration.
Furthermore, according to Medscape (2020), antimicrobial susceptibility tests are
used to determine which antibiotics a specific bacteria or fungus is susceptible to.
Most of the time, this testing is used in conjunction with a Gram stain and culture,
the results of which are obtained much faster. Antimicrobial susceptibility tests can
help doctors decide which drugs to use and how much to give to patients with
difficult-to-treat infections. The minimal inhibitory concentration (MIC) is the lowest
concentration of drug that inhibits the growth of the organism and is commonly
reported as a result. Reports usually include a quantitative result in g/mL as well as
a qualitative interpretation.
The disc diffusion method of antimicrobial assay is the most widely used and
accepted method for determining the antimicrobial or antibacterial activity of plant
extracts. Although the disc diffusion method is counterproductive for determining the
minimum inhibitory concentration (MIC), it can be used to calculate an approximate
MIC for some microorganisms and antiseptics by comparing inhibition zones.

H. Minimum Inhibitory Concentration

From lumen, Boundless Microbiology (2022), the minimum inhibitory
concentration (MIC) of an antimicrobial (such as an antifungal, antibiotic, or
bacteriostatic) drug is the lowest concentration that will inhibit visible growth of a
microorganism after overnight incubation. After isolating a pure culture, MICs can be
determined on plates of solid growth medium (called agar, as shown in the "Kirby-
Bauer Disk Susceptibility Test" atom) or broth dilution methods (in liquid growth
media, as shown in). An MIC is generally regarded as the most basic laboratory
measurement of an antimicrobial agent's activity against an organism. Lower MIC
values indicate that less of the drug is required to inhibit organism growth, so drugs
with lower MIC scores are more effective antimicrobial agents.
Moreover, Kirby-Bauer testing determines the sensitivity of bacteria to antibiotics
by culturing bacteria on solid growth media in the presence of drug sources.
Because KB tests are conducted under controlled conditions, the minimum inhibitory
concentration for a given antibiotic can be calculated by comparing the size of the
observed zone of inhibition to known values. Zone of inhibition: a region of media in
which bacteria are unable to grow due to the presence of a drug that inhibits their
growth. The minimum inhibitory concentration of an antimicrobial drug is the lowest
 concentration that prevents visible growth of a microorganism after overnight
incubation with media.
Overall, minimum inhibitory concentration is the lowest concentration of an
antimicrobial that will inhibit visible growth of a microorganism after overnight
incubation. This will be extremely useful in determining whether the Kangkong
ethanolic leaf extract, which will act as an organic disinfectant, is effective in
inhibiting the growth of Staphylococcus aureus, which is the bacteria present on the
majority of commonly held surfaces.
II. Review of Related Studies
A. Kangkong (Ipomoea aquatica)
The I. aquatica is composed of various parts, the plant is often used medicinal
purpose in Southeastern Asia and has been reported to be useful in the treatment of
high blood pressure, as an emetic in the treatment of opium and arsenic poisoning,
and as an emetic in the treatment of opium and arsenic poisoning. The dried juice is
said to be a purgative, while the leaves and stems are cooling. Furthermore, it has
traditionally been treated for nervous and general debility, piles, worm infections,
leucoderma, leprosy, jaundice, and liver complaints (Chaudhury, Das, & Saha 2017).
Ipomoea aquatica Forssk, also known as water spinach. (Convolvulaceae) is an
edible aquatic or semi-aquatic herb. I. Aquatica has traditionally been used to treat a
variety of ailments such as diabetes, liver dysfunction, constipation, and arsenic and
heavy metal poisoning. A review of the literature revealed that I. Aquatica contains
significant amounts of phenolic compounds, flavonoids, saponins, -carotene, and
ascorbic acid (Dewanjee, Dua, Khanra, Das, Barma, Joardar, Bhattacharjee, Jaafar,
& Zia-Ul-Haq 2015).
Whereas Ipomoea aquatica (Kangkong) lives mostly in moist areas, easily
propagates, and is found almost everywhere in Asia, it can be stated that there is no
depletion problem because it is easy to propagate and can withstand a variety of
weather conditions

B. Phytochemical Screening of Ipomoea aquatica

 In the study of Kehinde, Attahlur, Addejel, and Aklnade (2018), Ipomoea aquatica
possesses phytochemical substances with medicinal benefits: the presence of a
high level of alkaloids may directly attribute to its anti-oxidative action. Polyphenolic
compounds, such as tannins, play an important role in lipid oxidation stabilization
and are linked to antioxidant activity. The colony forming unit (cfu/g) method was
used to determine which microorganisms are susceptible to plant extract.
Phytochemical studies on the ethanolic plant extract revealed the presence of
alkaloids, saponin, flavonoids, glycosides, steroids, carbohydrate, proteins, and
amino acids, as well as tannins. Plant phytochemical constituents such as tannins,
alkaloids, flavonoids, phenolic compounds, and a variety of other aromatic
compounds are secondary metabolites that act as a barrier component against
predation by a variety of microorganisms (Chaudhury, et al. 2017).
Igwenyi, Obinna, and Offor (2011) found out that the phytochemical screening of
green kangkong had high concentrations of alkaloids, reducing sugar, soluble
carbohydrate, and flavonoids, but low concentrations of steroids, phenols,
glycosides, -carotene, saponins, and tannins. The high carbohydrate, vitamin, and
mineral content of the plant material demonstrated that it can be a good source of
nutrients for both human and animal food. Aside from its nutritional value, the plant's
natural endowments of phytochemicals demonstrated that it has significant
pharmacological and therapeutic effects.

C. Antibacterial and Antimicrobial Activity of Ipomoea aquatica

Singgih, Ivan, and Fidrianny (2019), studied that water spinach (Ipomoea
aquatica Forsk.) herb extract has antibacterial activity against the bacteria
Propionibacterium acnes. The reflux method was used to extract three different
polarity solvents. A rotary evaporator was used to evaporate the extracts.
Antibacterial activities were evaluated using disc diffusion, microdilution, and
antibiotic equality. All three water spinach herb extracts were more effective against
Gram-positive bacteria than Gram-negative bacteria.
Chaudhury, et al., found out that antimicrobial compounds are abundant in
Ipomoea aquatica Plant extract. The antimicrobial activity of an ethanolic extract of
Ipomoea aquatica Forsk in vitro. The antimicrobial activity of the plant extracts was
determined using the agar-well diffusion method, as described in NCCLS, 1993.
Bacterial strains were seeded in Petri plates containing 20 mL of nutrient agar
medium. A well cutter was used to bore wells of about 10 mm in diameter. Plant
extracts were prepared in DMSO (1 mg/ml DMSO stock). Plant extracts in
concentrations of 25, 50, and 100 l were added. As positive and negative controls,
ciprofloxacin (20 l) and DMSO (100 l) were used, respectively. The plates were then
incubated for 24 hours at 37 °C. The plant extract's antimicrobials are allowed to
diffuse into the medium and interact with the test organisms in the freshly seeded
plate. After 24 hours, the diameter of the inhibition zone was measured in
millimeters. However, according to Al-Rashidi, Padma, and Rasni (2017), the
antibacterial activity was greater in concentrations of methanolic extract leaves of I.
aquatica.

D. Bacteria Present in Frequently Held Surfaces

Surfaces that do not come into direct contact with patients during care are
referred to as environmental surfaces. Medical equipment surfaces, such as knobs
or handles on hemodialysis machines, x-ray machines, and instrument carts, are
classified as medical equipment surfaces, while housekeeping surfaces, such as
floors, walls, and tabletops, are classified as housekeeping surfaces. Housekeeping
surfaces are further classified as "high-touch" or "frequent hand contact" surfaces
and "low-touch" or "minimal hand contact" surfaces. Doorknobs, bed rails, light
switches, surfaces in and around patient rooms' toilets, and the edges of privacy
curtains are examples of high-touch surfaces (Suleyman, Alangaden, & Bardossy,
2018).
These three bacteria are the most commonly encountered bacteria on a daily
basis; some are beneficial, but the majority are harmful to us. So, keep sanitized to
reduce the risk of getting bacteria, and wash your food before cooking to ensure that
no bacteria get into your food.

E. Efficacy of Kangkong Ethanolic Leaf Extract

 The development of new types of diseases has increased from year to year, and
many drugs currently circulating are no longer able to cope with these new diseases.
(Rasyid, Farida, Daud, Wiwin, Wijaya, & Tangke, 2020). Infectious diseases are
caused by microorganisms (pathogens) that invade and damage organ function.
(Rajapaksha, Singhabahu, & Rajith, 2019).
Phytochemical screening of I.aquatica revealed high concentrations of alkaloids,
reducing sugar, soluble carbohydrate, flavonoids, phenol, β-carotene, tannins etc.
Ethanol extract of the leaf had the highest flavonoid content. (Sharmin, Hossain,
Rahman, Momtaz, Sharmin, & Mossadek, 2013). Manvar and Desai (2013) found
that leaf extracts of Ipomoea aquatica showed antimicrobial activity against Gram-
positive and Gram-negative microorganisms. Which showed a zone of inhibition of
08-19 mm in agar disc diffusion method. From the above-mentioned investigation of
the plant, it is revealed that I. aquatica could be really an effective natural herb
having good nutraceutical applications as well as to cure many fatal diseases
(Malakar, et al. 2015).

F. Extraction of Kangkong
In Asia, the use of herbal medicines reflects a long history of human interactions
with the environment. (Sasidharan, Chen, Saravanan, Sundram, & Latha, 2011).
Many beneficial biological activities have been reported, including anticancer,
antimicrobial, antioxidant, antidiarrheal, analgesic, and wound healing activity. In
many cases, people claim that certain natural or herbal products are beneficial
(Sasidharan, et al. 2011). In this regard, Ipomoea Aquatica (I. aquatica)
(Convolvulaceae), a perennial herb found in India, Ceylon, Tropical Asia, Africa, and
Australia, is one of many plants being studied for their therapeutic efficacy. This
plant is grown as an aquatic plant that thrives in marshy areas. In Southeastern
Asia, various parts of the Ipomoea aquatica plant are used medicinally.
The study of medicinal plants begins with pre-extraction and extraction
procedures, which are critical steps in the extraction of bioactive constituents from
plant materials. At the Small Manufacturing Enterprise (SME) level, traditional
methods such as maceration and Soxhlet extraction are commonly used. Modern
extraction methods such as microwave-assisted (MAE), ultrasound-assisted
extraction (UAE), and supercritical fluid extraction (SFE) have made significant
advances in the processing of medicinal plants, with the goal of increasing yield at a
lower cost. Furthermore, modifications to the methods are constantly being
developed. With so many methods available, selecting the best extraction method
necessitates careful consideration (Azwanida, NN, 2015).
Extraction is the most important step in the analysis of medicinal plants whereas
the desired chemical components must be extracted from plant materials before
further separation and characterization. Pre-washing, drying of plant materials or
freeze drying, grinding to obtain a homogeneous sample, and frequently improving
the kinetics of analytic extraction as well as increasing sample surface contact with
the solvent system were all part of the basic operation. The nature of the bioactive
compound being targeted heavily influences the choice of solvent system. To extract
the bioactive compound from natural products, various solvent systems are
available. Plant extracts are also made by macerating or percolating fresh green
plants or dried powdered plant material in water and/or organic solvent systems
(Sasidharan, et al.). Fresh Kangkong leaves were weighed and thoroughly washed
with tap water to remove any adhering foreign materials or soil particles prior to
extraction. The leaves were then dried for at least 5 days in the shade and sun. The
dried leaves were then weighed again to determine the percentage of moisture
content in the plant. The dried leaves were then finely powdered with a mechanical
blender. The obtained quantity was weighed and macerated in 95 percent ethanol.
After that, it was left at room temperature for 3 to 7 days, with occasional shaking.
The filtrates were then vacuum-concentrated in a rotary evaporator to remove the
solvent and obtain solid mass. The solid mass of ethanolic extract was weighed, and
the plant's percentage yield after extraction was calculated. The extract was stored
in suitable conditions for use. (Al-Rashidi, et al. 2017).

G. Antimicrobial Assay
 As stated in the study of Balouiri, Sadiki, and Ibnsouda (2015), in past years,
there has been a surge in interest in researching and developing new antimicrobial
agents derived from a variety of sources to combat microbial resistance. As a result,
antimicrobial activity screening and evaluating methods have received increased
attention. Antimicrobial susceptibility testing could also be used for drug discovery,
epidemiology, and therapeutic outcome prediction. Plants and other natural sources
can provide a wide range of compounds that are comprehensive and structurally
diverse.
The notion that a plant extract has antimicrobial activity is intriguing, but this
preliminary data should be reliable and allow researchers to compare results,
avoiding work in which researchers use the antimicrobial activity investigation as a
supplement to a phytochemical study. (Balouiri, M., et al. 2015). Methods for
evaluating antimicrobial activity that have been widely used include disc diffusion,
well diffusion, and broth dilution. (Othman, Lon, Wiart, Khoo, Lim, & Ting, 2011).
The formal method for routine antimicrobial susceptibility testing in many clinical
microbiology laboratories is agar disk-diffusion testing, which was developed in
1940. Agar plates are inoculated with a standardized inoculum of the test
microorganism in this well-known procedure. The agar surface is then covered with
filter paper discs (about 6 mm in diameter) containing the test compound at the
desired concentration. The Petri dishes are incubated in the proper conditions. In
general, an antimicrobial agent diffuses into the agar and inhibits the test
microorganism's germination and growth, after which the diameters of inhibition
growth zones are measured. The agar disk-diffusion method, on the other hand, is
ineffective for determining the minimum inhibitory concentration (MIC) because it is
impossible to quantify the amount of antimicrobial agent diffused into the agar
medium. Nonetheless, for some microorganisms and antibiotics, an approximate
MIC can be calculated by comparing inhibition zones to stored algorithms. (Balouiri,
M., et al.).

H. Minimum Inhibitory Concentration

 Antibiotics are antimicrobial agents produced by microorganisms that inhibit or kill
the growth of other microorganisms while remaining safe for host cells. The
susceptibility of pathogens to antibiotics must be determined in order to select the
best antibiotic for treating microbial infections. Bactericidal antibiotics are those that
kill bacteria, whereas bacteriostatic antibiotics only prevent their multiplication.
However, depending on the concentration, some antibiotics can be both
bacteriostatic and bactericidal (Owuama, 2017).
In Andrew's report (as cited by Owuama, 2017), the dilution method is most
useful for determining the minimum inhibitory concentration (MIC), which is the
lowest antimicrobial agent concentration that inhibits microbial growth. Diagnostic
laboratories use MICs primarily to confirm resistance, but they are also used as a
research tool to determine the in vitro activity of new antimicrobials, and data from
such studies has been used to determine MIC breakpoints. Furthermore, Owuama,
2017 initiated that, typically, for MIC and MBC determinations, antibiotic-free agar
medium is prepared and inoculated with samples from tubes that show no turbidity
or growth. Preparing agar medium takes more time, more stress, and the use of a
Petri dish. As a result, an improved new method, the dilution tube method (DTM), is
described here, which is simpler and less expensive because it does not require
agar medium and instead uses broth medium in tubes to determine MIC and MBC.
In tubes, the MIC of Escherichia coli, Klebsiella pneumoniae, and Staphylococcus
aureus were determined by double diluting (or higher dilutions) broths containing
gentamicin concentrations that inhibit bacterial growth and incubating at 37°C for 18
to 24 hours. After incubation, the MIC tube grew and became turbid.
With relevance to our study, Van Hal, Lodise, and Paterson (2012) found out that
emerging evidence suggests that vancomycin may be less effective against serious
methicillin-resistant Staphylococcus aureus (MRSA) infections with MIC values near
the upper end of the susceptibility range. Concluding that because a high
vancomycin MIC was linked to a higher mortality rate in MRSA BSI, institutions
should consider performing Etest MICs on all MRSA BSI isolates. Although these
findings raise concerns about vancomycin, there is currently no evidence to support
improved survival rates with alternative antibiotics. Data are desperately needed to
 determine whether other agents can improve the outcomes seen with vancomycin in
MRSA infections with elevated vancomycin MIC values.

III. Synthesis
Kangkong (Ipomoea aquatica) ethanolic leaf extract can be produced as an
organic disinfectant due to its pharmacological potentiality and medicinal use, as
it contains flavonoids, which are defensive and inhibitory compounds or agents
present in the plant, as well as antimicrobial activity, which shows a significant
minimum inhibitory concentration to certain microorganisms. The above-
mentioned literature and studies are compilations of works and analyses
concerning the habitat, anatomy, physiology, and life cycle of the Kangkong. In
addition to its importance and contribution to the fields of medicine and
pharmacology.

With the previous studies provided, this recent study about the use of
Kangkong as an organic disinfectant cleaner concentrate while using the
ethanolic extract can be supported. Ethanolic extracts of Kangkong have been
shown to inhibit the growth of microorganisms such as Escherichia coli and
Staphylococcus aureus. As stated in the objectives of our study, it is well
supported by the literature and studies provided. The researcher’s specific goals
are to identify the chemical components of Kangkong that make it useful as a
disinfectant. The second step is to determine the minimum inhibitory
concentration of Staphylococcus aureus exposed to various treatment
concentrations. Finally, preliminary product costing will be performed to
determine economic feasibility.

The study, however, will only focus on developing an organic disinfectant and
will only be applied to frequently used household surfaces. The researchers will
only use the Kangkong leaf, and the extraction method will be ethanolic extract.