NAMA : ASEP SAEFULLAH

NIM: G2016 0038

Indonesia Medicus Veterinus Oktober 2016 5(5) : 464-473

pISSN : 2301-7848; eISSN : 2477-6637

(IDENTIFICATION OF CHEMICAL COMPOUNDS ETHANOL EXTRACT LEAF

MORINGA (MORINGA OLEIFERA L) IN BALI)

1 2
I Wayan Dwika Pratama Putra , Anak Agung Gde Oka Dharmayudha , Luh Made
2
Sudimartini
1
Student of veterinary profession education,
2
Laboratory Pharmacology and Veterinary Pharmacy,
Veterinary Faculty, Udayana University,
Jln. PB. Sudirman, Denpasar, Bali;
Tlp. (0361) 223791, Faks. (0361) 701808. E-mail:
jack.fkh@gmail.com

ABSTRACT
Indonesia has overflowing natural resources spreading from the west to east. One of those
natural resource is medicinal plants, which can be used for traditional medicine. Moringa
oleifera L is one of medicinal plants that has the benefit for antidiabetic and antioxidants. The
aims of this research were to find out the classificatioin of chemical compounds contained in
ethanol extracted from moringa leaves. The samples used in this study is moringa leaves
taken from the north Denpasar, Bali. The moringa leaves firstly extracted using 96% of
ethanol, next step the phytochemical test is done to detect the active compound of alkaloid,
flavonoid, saponin, fenolat, triterpenoid/steroida, and tannin. The data obtained were
analyzed using descriptive qualitative method. The researchs result showed that the ethanol
extracted from moringa leaves contain chemical compounds of alkaloid, flavonoid, saponin,
fenolat, triterpenoid/steroida, and tannin.

Key words : Moringa oleifera leaves, etanol, screening phytochemical.

PRELIMINARY

Traditional medicine has been known and used all over the world since thousands of years
ago. Traditional medicine and medicinal plants are widely used insiders. Prevention,
promotive, and rehabilitative. The use of traditional medicine as a herbal medicine has been
widespread since ancestral times and so on until preserved as a cultural heritage (Duryatmo,
2005). Raw materials of this natural medicine can come from biotic and abiotic natural
resources. Biotic resources include microorganisms, flora, and fauna as well as marine biota
while abiotic resources understand the terrestrial, sound, and space resources as well as
potential scope within them.

Indonesia has abundant natural wealth. A wide variety of medicinal plants thrive in Indonesia.
This natural wealth is very beneficial to the health of its inhabitants, even to the world.
Several studies have proved to the world that is very potential as a place to grow and develop
ingredients of medicine for the world community (Fahey, 2005). The ability of society is
limited to getting modern medicines, so making traditional medicine has a very important
meaning for society because it is easier to build without prescription (Pudjarwoto et al.,
1992). While many people assume the use of drugs or traditional medicine is relatively more
secure than synthetic drugs. Not so the effects of drugs that have no side effects are
considered when using less appropriate.
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One of the plants that can be utilized both as foodstuff and medicines is the moringa plant
(Moringa oleifera L.). Kelor belongs to the familia of Moringaceae and has many names,
such as kelor, kerol, marangghi, moltong, kelo, keloro, kawano, and ongge. Moringa plants
grow in the lowlands or highlands. This plant has a stem height of 7-11 meters. Moringa
leaves are ovoid with small size compound multiple in one stalk, can be made vegetable or
medicine. The flowers are yellowish white and green flower hoods, these flowers come out
all year long. Kelor is known to contain more than 90 types of essential nutritional vitamins,
minerals, amino acids, antiaging and anti-inflammatory. Moringa contains 539 compounds is
known in traditional African and Indian medicine and has been used in traditional medicine
for the prevention of more than 300 diseases. Various parts of the moringa plant Acting as a
cardiac and circulatory stimulant, it has antitumor, antipyretic, antiepileptic,
antiinflammatory, antiulcer, diuretic, antihypertensive, lowers cholesterol, antioxidant,
antidiabetic, antibacterial and anti-fungal (Toripah et al., 2014).

All parts of the moringa plant have been used as foodstuffs and medicines. Part of this plant
is often used as a drug is a seed, leaf, and bark, and efficacious as anti diabetic and
antioxidants (Jaiswal et al., 2009; Pari et al., 2007). The juice from the kelor plant root can be
used for the treatment of external irritation. The suspension of dry beans is known as
coagulant. Some other benefits of Moringa oleifera L. from Moringa skin as inflammatory
bowel, Moringa leaf as anti anemia (Oduro et al., 2008), leaves and stems of moringa can be
used as a high blood pressure lowering and drug Diabetes (Moringa oleifera L.) Giridhari et
al., 2011).

To be optimal, please note adequate information about the chemical classes contained in these
medicinal plants. Part of this plant is often used as a drug that is seed and leaves. Data on the
active ingredient of the kelor leaf is still very rare, some literature mentions on the leaf kelor
there is the content of flavonoids, saponins, alkaloids, tannins, and phenols (Pandey et al.,
2012). However, for the chemical content in the area of Denpasar has never existed and there
are different conditions difference there is a difference in the chemical content of the
moringa.

RESEARCH METHODS

Materials used were Moringa oleifera L. leaf, 96% ethanol, Wagner reagent, Meyer reagent,
10% NaOH reagent, Wilstater reagent, Smith-Metacalve reagent, FeCl3 1% reagent, acetic
anhydride, and anhydrous acetate. The tools used in this research are blender, measuring cup,
beaker, reaction tube, analytical balance, rotary vacuum evaporator, aluminum foil, water
heater, and spriter lamp.

Taking Moringa Leaf Extract

Moringa leaf extract was prepared by maceration as much as 50 grams of fresh moringa leaf
was destroyed by blender, 96% ethanol solvent was added, put into container, closed and left
for two days protected from sunlight. The mixture is filtered so as to obtain maserate. The
dregs are macerated with 96% ethanol using the procedure the same one. Maseration is done
until the maserat is obtained clear. The maserate was evaporated using a rotary vacuum
evaporator at a temperature of 40o C. Moringa leaf extract was further screened for
phytochemicals to detect plant compounds based on their class. Methods that have been

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developed can detect the presence of classes of alkaloids, phenolics, flavonoids, tannins,
saponins and steroids / triterpenoids.
Examination of alkaloids

Examination of alkaloids was done by one ml extract added a few drops of reagent, in this
study reagent used is wagner reagent (positive reaction if formed brown precipitate) and
meyer reagent (positive reaction if formed white precipitate).

Examination of flavonoids

The reagents used in the flavanoid examination were three reagents ie 10% NAOH reagent,
Wilsatater and Smith-Metacalve reagent. 10% NaOH reagent: 1 ml extract added a few drops
of NAOH 10%, a positive reaction if a specific color change occurs. Wilstater reagent: 1 ml
of extract added a few drops of concentrated HCl + a little magnesium powder (Mg). Positive
reaction in case of red-orange change. Smith-Metacalve reagents; 1 ml of the extract added a
few drops of concentrated HCl then heated. Positive reaction if it gives white color.

Saponin examination (foam test)

One ml of the extract is added hot water and shaken. Positive reaction if long-lasting foams
are formed.

Polyphenol examination

One ml of extract was added with 1% FeCl3 reagent. Positive reaction if blackish or dark
blue is formed.

RESULTS AND DISCUSSION

The results of phytochemical test of Moringa oleifera L. extract showed that there are
alkaloids, flavonoids, saponins, phenols, steroids / triterpenoids, tannins as in Table 1.

Tabel 1. Results of Phytochemical Test on Kelor Leaves (Moringa oleifera L.)

Test Description
Phytochemic
No. als Reaction Color Change
HCl 2N + reaction
1 Alkaloid Formed brown deposits Alkaloid (+)
Wagner
Wilstater brownish green to yellowish green
2. Flavonoid Bate Smith-Metcalfe brownish green to yellowish green Flavonoid (+)
NaOH 10% brownish green to yellowish green
Akuades,
3. Saponin heated, shake + Not formed stable foam Saponin (-)
HCl 2N
4. Fenolat FeCl3 brownish green to blue black Fenolat (+)
5. Triterpenoida Lieberman-Burchard brownish green to green purple Triterpenoid/
Steroida H2SO4 brownish green to green purple steroid (+)
FeCl3 brownish green to blackish Tannin
6. Tannin
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Gelatin Formed precipitate (+)

Description: (+) = there is a chemical content, (-) = no chemical content
The result of phytochemical test on kelor leaf (Moringa oleifera L.) showed the presence of
alkaloid compound. Alkaloid compounds are the most organic compounds found in nature.
This compound is usually found in leaves that have a bitter taste. Almost all alkaloids found
in nature have certain biological activity, some are highly toxic but some are very useful in
medicine, such as quinine, morphine, and stiknin are famous alkaloids and have physiological
and psychological effects. The function of alkaloid compounds for plants is as toxic
substances against insects or plant-eating animals and as a factor of growth influence. Other
uses of this compound in the field of pharmacology as a stimulant of the nervous system,
cough medicine, eye drops, sedatives, malaria drugs, cancer, and anti-bacterial. In addition,
the alkaloids can accelerate wound healing by increasing Transforming Growth Factor 1
(TGF-1) and Epidermal Growth Factor (EGF) (Porras-Reyee et al., 1993; Dong et al.,
2005).

Flavonoid phytochemical test of Moringa leaf extract showed positive result. In plants,
flavonoids function in the process of photosynthesis, anti microbial, anti-virus. Anti oxidation
activity is also owned by certain active flavonoid components used to inhibit bleeding and
anti scratch (Robinson, 1995). In humans flavonoids serve as antibiotics, for example in
cancer and kidney disorders. Several types of flavonoids such as slimirin and silyburn are
proven to treat liver function disorders, inhibiting prostaglandin synthesis so that it works as a
hepatoprotector. Flavonoids also work to reduce blood clots. Flavonoids on humans in small
doses are flavones, which act as heart stimulants. Hydroxylated flavones act as diuretics and
as antioxidants in fat (Tarziah, 2012).

Flavonoids provide a protective effect on endothelial function and inhibit platelet

aggregation, thereby decreasing the risk of coronary heart disease, cardiovascular disease.18
Flavonoids have a hypotensive effect by inhibiting activity mechanism Angiotensin I
Converting Enzyme (ACE), as well as diuretic (Panjaitan and Bintang, 2014). Flavonoids can
inhibit ACE. Known ACE plays a role in the formation of angiotensin II which is one cause
of hypertension. Angiotensin II causes narrowed blood vessels, which can raise blood
pressure. ACE inhibitors cause blood vessels to dilate so that more blood flows into the heart,
resulting in a decrease in blood pressure (Kane et al., 2009). In addition, flavonoids can
increase urinary and electrolyte excretion, which functions like potassium, which absorbs
fluid electrolyte ions such as sodium present in intracellular blood to the extracellular
entering the renal tubules (Nadila, 2014)

Phenolic test by using FeCl3 reagent showed positive result which showed by the change of
color from brownish green color to blackish color and contained polyphenol compound
(Harborne, 1987). Phenolate is largely an antioxidant that neutralizes oxidation reactions
from free radicals that can damage cell structure and contribute to disease and aging. The role
of several classes of phenol compounds is known, for example phenolic or polyphenolic
compounds are natural plant antioxidant compounds. The compound is multifunctional and
acts as an antioxidant because it has the ability as a reducer and free radical capture (Estiasih
and Andiyas, 2006).

Leaf Moringa ethanol extract (Moringa oleifera L.) contains steroid and triterpenoid active
compounds. Triterpenoids are compounds whose carbon skeleton is derived from six
isopropene units and biosynthesically derived from acyclic hydrocarbons, that is, squalene.

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This compound is a complicated cyclic structure, mostly in the form of alcohols, aldehydes,
or carboxylic acids. The compound is a crystalline, colorless compound, often high-melting
and optically active, which is generally difficult to characterize because there is no chemical
reactivity. Triterpenoid compounds in plants serve as a defense against insects and growth
factors (Harborne, 1987). Test is widely used Is a Lieberman-Burchard reaction (mostly
concentrated acetic anhydride-H2SO4) which is mostly triterpene and sterol in the case of
green-blue changes indicating positive steroids and if the red-purple color changes, chocolate
suggests triterpenoide (Edeoga et al., 2005).

Steroid is a group of compounds having a cyclopentanaperhydrophenantrene base framework

having four integrated rings. These compounds have certain physiological effects. This
compound has several uses for plants such as growth regulator (seskuitertenoid abisin and
giberelin), carotenoids as a dye and have a role in membentu photosynthesis process. Its use
in the pharmaceutical field is commonly used as raw material for the manufacture of drugs
(Tohir 2010). In fact, this stereoide is considered a compound found only in animals but now
more and more is also found in plants (phytosterols). Phytosterols are steroid compounds
derived from plants. Phytosterol compounds commonly found in plants are sitosterol,
stigmasterol and cholesterol (Harborne, 1987).

Tannin phytochemical test on Moringa oleifera leaf extract (Moringa oleifera L.) showed
positive results. Phytochemical tests using FeCl3 are used to determine whether the sample
contains a phenol group. The presence of phenol groups is indicated by dark green or dark
blue after being added with FeCl3, so if phytolyte with FeCl3 gives positive results it is
possible in the sample to contain phenol compounds and one of them may be tannins because
tannins are polyphenolic compounds. This is reinforced by Harborne (1987), a classic way to
detect simple phenol compounds that add extracts with 1% FeCl3 solution in water, which
gives rise to strong green, red, violet, blue or black. The formation of greenish-black or blue-
black color on the extract after being added with FeCl3 because tannin will form complex
compounds with Fe3 ions.

Complex is a compound whose formation involves the formation of covalent coordination

bonds between metal ions or metal atoms with nonmetallic atoms. In the formation of
complex compounds, metal atoms or ions are referred to as central atoms, while atoms
donating their electrons to a central atom are called donor atoms. The donor atom is present
in a neutral ion or molecule. Ions or neutral molecules that have coordinated donor atoms at
the central atom are called ligands. A molecule is said to be a ligand if tanin is a group of
active compounds that have phenolic flavor (Robinson, 1995). Tannin compounds are
widespread in many plant species, and play a role in protection from predation, and possibly
as well as pesticides, and in regulation of plant growth (Linggawati, 2002). Tannin
compounds act as antioxidants and inhibitors of tumor growth (Lenny, 2006). The tannin
compound is a polyphenol compound in plants, food and beverages (Makkar and Becker,
1998) soluble in water and organic solvents (Haslam, 1996).

The phytochemical test of saponin content in Moringa oleifera leaf extract showed negative
results due to the absence of stable foam. Saponins exist in all plants with high concentrations
in certain parts, and are affected by plant varieties and growth stages. Saponins are also used
as an anti-microbial (Robinson, 1995). Saponins can accelerate the process of regeneration
and re-epithelization as they are immunostimulators that bully the host's resistance (Nayak
and Pereira, 2006; Chokotol and Hasselt, 2005). Saponins are glycosides of steroids, alkaloid

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steroids, or steroids with a nitrogen or triterpinoid function found in plants. Charantin, a

steroidal saponin isolated from Momordicha charantia reportedly induces an activity such as
insulin, by increasing insulin release and slowing the process of glucogenesis. Beta sitosterol,
a steroid found in azadirachta indica, andrographolide, an diterpenoid lactone, isolated from
Andrographis paniculata and gymnemic saponin acid isolated from Gymnema sylvestere can
lead to potential hypoglycemic activity in animals (Prabhakar and Doble, 2008).

Based on the results of phytochemical tests conducted by using samples of Moringa leaves
taken in the area of North Denpasar, Bali, it is known that the extract of ethanol leaves kelor
(Moringa oleifera L.) contains alkaloids, flavonoids, phenolics, triterpenoids / steroids and
tannins.

CONCLUDE

Phytochemical test on Moringa leaf samples taken in North Denpasar, Bali, contains
alkaloids, flavonoids, phenolics, triterpenoids / steroids, and tannins.

SUGGESTION

Further research on moringa plants, whether isolation, purification, or biological tests of the
active compounds so that it can be applied in other fields such as medicine, agriculture,
pharmacy, and others.

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