VIII.

DISCUSSION
Chemical compounds contained in plants is the result of metabolism of the plant
itself.Many experts from the research results of chemical compounds often have the
effect of physiology and pharmacology that are beneficial to humans. The chemical
compounds known as secondary metabolites that are the result of the deviation of
the primary metabolite tumuhan. These compounds are a class of alkaloids, steroids,
terpenoids, phenols, flavonoids, and saponins.

Phytochemical tests can be performed in a preliminary examination of the active

compounds are secondary metabolites, so that the relative potency of each plant can
be measured.
Samples used in these experiments were cassava leaves or Manihot
esculenta.Manihot esculenta plants are plants that are flexible because it can grow
and produce in the lowlands to highlands, ranging from an altitude of 10-1500
m. Cassava is also suitable to be developed on marginal land, less fertile and water
shortages.
In the taxonomy of cassava can be classified as follows:
Kingdom: Plantae
Divisio: Magnoliophyta
Class: Magnoliopsida
Order: Malpighiales
Tribe: Euphorbiaceae
Subsuku: Crotonoideae
Tribe: Manihoteae
Marga: Mannihot
Species: M. esculenta

Cassava leaves have long stalks and resemble strands of palm leaves, and each
leaf stalk has about 3-8 pieces. The leaf stalks are yellow, green or red. An outline of
the leaf epidermal tissue, mesophyll and epidermis pengangkut.Sel file is located at
the outside is coated by a layer of monocot plants kutikula.Pada no differentiation of
spongy parenchyma and palisade parenchyma. In the parenchyma palisade
parenchyma cells there are variations such as oil cells, mucous cells and cell
ergastik. Long-stemmed leaves form menjari about 3-8 pieces.
Stem structure was composed of cork tissue, cortical tissue, beam transport and
monocot stem pith batang.Pada, network carriers are arranged in bundles and
spread across the surface of the rod. Carrier bundles surrounded by parenchyma
tissue.
This plant occurs on the process of photosynthesis, because plants have
photosynthetic pigments kolofil that is. Photosynthetic pigments are light-absorbing
function of red and blue, and green reflective

This experiment was conducted to examine the content of alkaloids, flavonoids,

saponins, triterpenes and steroids on cassava leaves. In phytochemical testing, to
determine the content of secondary metabolites (alkaloids, flavonoids, steroids,
triterpenoids and saponins), certain parts of the plant is crushed by using
mortar.Grinding serves to open up and expand the plant cell wall surface of the
secondary metabolites of plants so much easier out and can be extracted.
In general, alkaloid compounds in plants are not in a free state, but bound in
complex partial ionic bond with an organic acid. With the addition of chloroform
amoniakal (with a stronger basicity than the alkaloid compounds), organic acids will
be bound by the chloroform compound alkaloidnya amoniakal so freely. In the test
alkaloids, the plant has been fine-amoniakal added chloroform. This addition will
break the bond formed between the alkaloid with tannin acid, which form salts with
the alkaloids, so the alkaloid will be in its free state. Addition of chloroform-amoniakal
function is to extract the alkaloid bases are present in the tested plants. The choice
of chloroform-amoniakal as pengekstraksi because almost the same polarity of
chloroform polar alkaloid alkaloid that can dissolve in accordance with the principle
of "like dissolve like", chloroform used must amoniakal or contain ammonia because
of the plant alkaloid that is alkaline in nature usually in the form of salt, tied
with organic acids with acid called tannin.Ammonia is an alkaline will decide on ties
with the alkaloid tannin acid, tannin acid that becomes bound to the ammonia while
the alkaloid is in a free state. In the free state of the alkaloids tend to be non-polar so
it will be extracted by chloroform, which also tend to be non polar. Alkaloid
compounds found in nature, especially in some parts of the plant, is in the form of
salts, in which the alkaline alkaloid that binds with tannin acid to form salts. To
examine the content of alkaloid compounds, the first alkaloid to be released from its
salt form.
Stirring after addition of chloroform-amoniakal aims to increase the contact that
occurs between the solvent with the pulp plant. Given this, the more bonds are
broken between the alkaloid with tannin acid, so the more alakaloid compounds
extracted.
Prior to testing using reagent alkaloids, first filtered by using a clean, dry cotton to
separate the chloroform extract-amoniakal. Then the filtrate in the form of extracts
obtained by chloroform-amoniakal divided into two test tubes to be tested alkaloids.
Filtrate in a test tube first and then added to a reagent Hager pikrat acid solution is
saturated so that acid can bind to and form a salt-free alkaloid is readily soluble in
water. Then the complex formation reaction will easily happen due to both (metal
ions and alkaloids) are in a water medium. The precipitation is characterized by the
formation of turbidity and after a long yellowish precipitate is formed. An alkaloid-
containing plants tested positive for if you develop a yellowish white precipitate due
to the precipitation reaction between the alkaloid with reagents containing heavy
metals.Alkaloids contain nitrogen atoms that have unpaired atoms which can donate
electrons to the metal ion, so that the alkaloid that functions as a coordinating ligand
to form covalent bonds with the central atom. Complex formed will precipitate if the
solubility product of the alkaloid and the reactant metal ion is greater than Ksp
formation of the complex. It is obtained by making a large concentration of one
species forming a complex that is by increasing the concentration of reactants due to
the alkaloid content of plants is not known with certainty. Addition of the reagent is
made from heavy metal to make it easier to settle because it is generally salts of
heavy metals have a very small Ksp.
While the filtrate in the second tube was added 2 N sulfuric acid which is used to tie
back into salt alkaloid alkaloid that can react with the reagents that are specific to
heavy metal and form a complex alkaloid insoluble inorganic salts, which can be
separated from other secondary metabolites. After addition of 2N sulfuric acid will be
two phases due to the difference in polarity between the water level is relatively polar
with relatively less polar chloroform. Alkaloid salts will dissolve in the acid layer in the
water phase is on top. Chloroform phase was under the water phase because it has
a greater density than the density of water. Difference is due to the density of
chloroform containing halogen element that has a large density ( water = 1 g / mL;
chloroform = 1.255 g / mL at 25 C).
The upper layer containing the alkaloid salts were then divided into three reaction
tubes and tested alkaloids with Mayer's reagent, Wagner and Dragendorf.
That it contains the alkaloid compounds in the plant is characterized by the formation
of the precipitate after adding a specific reagent. Precipitation that occurs due to the
formation of insoluble complexes of heavy metals which are owned by alkaloid
reagents.Formation of these deposits can occur if the product or reactant ion
concentration product-forming complex in solution is greater than the product of
reactant solubility (Ksp) at saturated conditions. This situation is obtained when the
concentration of which is owned by the reactants is high enough. A certain amount of
metal ion complexes-soluble alkaloid samapai reach saturation. After that, the
resulting complexes in solution can not be dissolved and there is in solid form which
Alkan attracted by the force of gravity that accumulate at the bottom of the tube to
form a precipitate.
Mayer's reagent, Wagner, and Dragendorf added to the acid phase reactants such
as inorganic metals that can react with the inorganic alkaloid in solution as well,
namely in the acid phase (water phase). As for the reagent Hager (pikrat acid) or
2,4,6-trinitrofenol directly added to the chloroform extract-amoniakal because this
reagent is an organic reagent which can react in an organic solution as well.

Reaction occurs:
Mayers reagent
Alkaloids + + H2SO4 reagent complex deposition
R3N R3NH H2SO4 + + + H + + SO42-
Wagner reagent

From the experimental results is known that the leaves of cassava plants do not
contain alkaloids.
Plant cassava leaves also contain flavonoids, which after all the leaf extract was
added 2N HCl and Mg powder solution became orange.
On steroid testing, and triterpenoid saponins, after the plant is crushed, added
ethanol panas.Untuk testing triterpenoids and steroids in the sample (leaf), extract of
diethyl ether was added reagent-Buchard Lieberman (LB), which is a mixture of
acetic acid with concentrated sulfuric acid anhidrid (2:1).

Lieberman-Buchard reaction

On steroid testing, and triterpenoid saponins, after the plant is crushed, added to hot
ethanol. Ethanol solvent used for ethanol has two groups, namely the polar groups
on the alcohol and nonpolar groups in the hydrocarbon.
CH3-CH2-OH
Polar non-polar groups cluster
Steroids and triterpenoids are relatively non-polar while the saponins tend to be
polar.By using ethanol, all three compounds can be extracted. The use of hot
ethanol will increase the solubility of a compound that is expected throughout the
steroid, and triterpenoid saponins contained in the plants will be extracted into
ethanol.
Ethanol extract then evaporated in the evaporator through a heating plate with a
small fire in order to obtain concentrated extracts. Concentrated extract is then
added diethyl ether to withdraw steroids and triterpenoids that the color produced
after addition of reagents Liebermann-Buchard not too thick and easy color change
was observed.Liebermann-Buchard reagent is a mixture of acetic acid and
concentrated sulfuric anhidrid ratio of 2:1. compounds are attracted by diethyl ether
in a porcelain dish before adding reagents Liebermann-Buchard, moved into the
plate drops so that the color changes that occur can be observed easily.
Color changes that occur after the add-Buchard Liebermann reagent due to the
chromophore group in the steroid and triterpenoid so as to form a color
complex.Chromophore groups will lead to an electronic transition from n to n * or
to * absorption of light resulting in a shift towards the visible region, so it can look
like the color. Electron transitions can occur after the addition of concentrated
sulfuric acid. On this test, triterpenoid compounds will provide the color purple, pink
or red because the compound absorbs light at a wavelength of 400-424 nm for the
color purple, and 647-700 nm for red. Where triterpenoid absorbs green. While
steroids will give a green or blue because it absorbs light at a wavelength of 435-480
nm for blue, and 500-560 nm for green color. Green, blue color that is not absorbed
but complementary colors. The color orange is the color absorbed. -OH group at the
triterpenoids will experience a wavelength shift that caused different colors. So red,
purple or brown are complementary colors.
From the obtained experimental results that cassava leaves contain a steroid with
the green and did not contain triterpenoid.
The next procedure is to test the identification of saponin. To test saponin, residue
left in the vaporizer cup hot distilled water was added, and then transferred into a
test tube and shaken strongly. Presence of saponin in the plant is characterized by
the formation of stable foam for about 15 minutes. The emergence of the foam is due
to the saponin which is a surface-active compounds that have properties such as
soap, has two parts, namely the head and tail of a different nature from each
other. At the head of the hydrophilic, while the tail is hydrophobic. Section will be
attracted to the hydrophilic surface of the water and the hidrofobnya will be outside
surface of the water. Section will be attracted to the hydrophobic surface of the water
and the hidrofobnya will be outside surface of the water. Hydrophobic parts would
bond together by trapping air bubbles that will form the foam.
Presence of saponin which is a surfactant will be able to lower the surface tension of
water by breaking the hydrogen bonds on the surface by placing a bersifar
hydrophilic head and a tail on the surface is hydrophobic backs away from the water
surface. From the experiments it is known that the leaves do not contain saponin
cat's whiskers.
Lieberman-Buchard reaction based on the formation of colored compounds as a
result of reaction of steroid / triterpene with acetic acid anhydride with an acid
catalyst (H2SO4 p). According Brieskon and Herrig, the mechanism of color
formation takes place through the oxidation and dehydration of 2 pieces of
conjugated double bonds in ring B on steroids or a single bond with a methylene
group is blocked at C7. At triterpene pentasiklik, involved in the reaction is methylene
group at C11 in ring C.positive test for the presence of steroid indicated by the
formation of a green color to blue. Positive test for the presence of triterpene
compounds shown by the formation of red, brown or purple.
There are several things that can be a source of error for this phytochemical
screening process, namely:
A. Solving imperfect cell wall, causing no terekstraknya compounds are expected to
be identified in the solvent used.
2. Concentrations of compounds that are too low, leading to difficulties in
observation.

IX. CONCLUSION
9.1 Manihot esculenta leaves do not contain alkaloids based on the test Hager,
Mayer, Wagner, Dragendorff
9.2 Manihot esculenta leaves contain steroids based on the test Liebermann -
Burchard and does not contain triterpenoid and saponins.
9.3 Manihot esculenta leaves contain flavonoids with flavonoid test