Isolation of Naringin Compound from Pomelo (Citrus maxima Merr) Peel

Ni Wayan Wida Sasmining Prastiwi

Chemistry Education Study Program, Mathematics and Sciences Education Departement
Faculty of Teacher Training and Education, Mataram University
Email: [email protected]

ABSTRACT: This research aims to knowing the proper isolation method to isolate
naringin compound from pomelo peel. This research begins with a sample preparation,
extracting pomelo peel powder with methanol solvent. The technique used is maceration. The
methanol extract was concentrated and then liquid extracted with n-hexane. Purification by
crystallization using an isopropanol solvent. Analyzing the purity with Thin Layer
Chromatography (TLC) tested. Analyzing the presence of functional groups with Fourier
Transform Infrared (FT-IR) spectrophotometer test. The result of FT-IR spectrophotometer
analysis contains hydroxy group (-OH), aromatic cyclic groups (C = C), CH2, CH3, and ether
groups.

Key Words : isolation, pomelo peel, naringin, Citrus maxima Merr

INTRODUCTION used as dyes, toxins, food scents, drugs and
so forth. Secondary metabolite compounds
Indonesia is a country with abundant such as steroids, alkaloids, terpenoids,
biodiversity. The diversity of animals and phenolics, flavoinoids, polyphenols,
plants are diverse. The diversity of plants is saponins, etc. are of great benefit to
a source of secondary metabolite humans. (Raharjo, 2013).
compounds that have been known to have Naringin, dihydroflavones, are
many benefits, one of them as drugs. especially present in the peels and fruits of
Indonesian people also have long known grapefruit, pomelo and other citrus fruits. It
some plants are processed into herbs that has been shown to be highly beneficial, has
are used as a traditional medicine from various biological activities such as
generation to generation for generations. preventing cancer, inhibits microbial
Secondary metabolite compounds growth and activity of many enzymes, anti-
contained in plants can be obtained by the oxidation, lowers blood cholesterol and
implementation of isolation method triglyceride levels and maintains normal
(Raharjo, 2013). capillary permeability of the blood, thus
The biodiversity of biological demonstrating potential drug applications
resources in Indonesia is a source of (Tang et all, 2011).
chemical compounds, which are primary Pomelo (Citrus maxima Merr)
metabolite compounds such as proteins, expressed contains flavonoids (naringin) on
carbohydrates, fats used by plants for their the fruit skin and flesh. The thickness of the
growth, and secondary metabolites such as fruit skin depends on the variety. The skin
terpenoids, steroids, polyphenols, of citrus fruit is divided into three layers,
flavonoids and alkaloids which generally namely the outer skin, the middle skin and
have bioactivity and serves as a protector of the inner skin. The outer skin is green,
plants. Secondary metabolites are produced yellow or yellow. Middle fruit skin is pure
from various organisms such as plants, white and the inner skin is pink. Naringin
bacteria, and fungi. Chemical compounds isolated from the middle skin and the inner
as a result of secondary metabolites in skin of pomelo.
various types of plants have been widely
RESEARCH METHOD RESULT AND DISCUSSION
Tools and Material Table 1. Isolation steps from journal

The tools and materials used in this

research are rotary evaporator, tool sets of
infrared spectrophotometer, extraction
flask, hot plate, thin layer chromatography
plate, middle skin and inner skin (albedo)
of pomelo, methanol solvent, n-hexane
solvent, isopropanol solvent and
chloroform solvent.
Sample Preparation
Pomelo peeled, part albedo cut into
small pieces. Dried in the sun for 3 days.
Next, oven for 24 hours. After completely
dry the pieces are smoothed to produce a
powder.
Isolation Table 2. Modified isolation steps
Albedo powder 50 grams macerated
using 550 ml of methanol for 3 days. The
filtrate is separated and evaporated by a
rotary evaporator to produce a dry
methanol extract. The dry methanol extract
was then added 50 ml of water and heated
to a constant temperature of 70 ° C for 30
minutes. The solution was extracted with
20 ml of n-hexane, and was allowed to
stand for 3-4 days, extraction with n-
hexane was performed twice. The liquid
phase of the extraction was added with 25
ml of isopropanol and heated to half the
volume. Cool the refrigerator to form a
crystal.
Purification
The crystals that have been
produced are recrystallized with
isopropanol solvent, the recrystallization is
done repeatedly until the crystals are felt to Determination of isolation steps
be free from impurities. taken from a journal by modifying the
isolation steps from the journal.
Qualitative Test modification is carried out on the use of
The compounds were tested with solvents for liquid extraction and
two tests: Thin Layer Chromatography recrystallization processes. steps in the
(TLC) with 3 variations of eluent, and journal before being modified in the form
Fourier Transform Infrared (FT-IR). of maceration and liquid extraction.
whereas in the step the modification results
are added recrystallization. excess
modification steps with added
recrystallization allow the crystals
produced to be purer, because the
recrystallization process is carried out to
free crystals from impurities.
The isolation step begins with the Figure 1. The result of TLC
sample preparation, the sample is made into
a powder in order to carry out the isolation Based on purification test results
process with the maximum. The powder using TLC obtained one color stain on the
form has a wider surface so that the variation of eluent / solvent used. The color
reaction can run faster. Next is the first step stain spacing corresponds to the nature of
in isolation, ie maceration. The maseration the solvent polarity and the resulting
is carried out using methanol solvent, compound. Where the naringin compounds
because the isolated compound is polar, tested for their purity are polar.
used polar solvent to make the compound In figure (a) it can be seen that the
soluble in methanol. Evaporation with a result is a spot with a semipolar and polar
rotary evaporator to produce a dry eluent. Furthermore (b) is added with a
methanol extract, and in liquid extraction non-polar eluent, n-hexane showing a spot
with n-hexane. Non-polar solvents are used with a distance close to the starting line.
in extraction for polluting to dissolve on n- Then the last one (c) is added with the polar
hexane. Extraction was perfomed twice, for eluent ie isopropanol shows a spot of color
the impurity to be maximally dissolved in near the finish line. Difference in stain
n-hexane so that recrystallization takes spots on klt plate is influenced by eluent
place at a faster time without too much polarity. The more polar eluen the further
repetition. Then purified by the spot distance and the greater the Rf
recrystallization serves to remove value.
impurities to produce pure crystals. Then
the crystals are tested qualitatively by using Four Transform Infrared
TLC and FT-IR This test will get the result of an IR
spectrum that provides important
information about the various functional
groups possessed by the molecule.
Thin Layer Chromatography
Thin Layer Chromatography (TLC) is
conducted to determine the purity of the
resulting compound. If the color stain
produced one then there is the possibility of
the compound tested purely. To test and
reassure that the compound is truly pure,
the TLC is performed three times with
three different eluents. Results of TLC with
the following three eluents; Figure 2. The result of IR spectrum

From the result of IR spectrum

analysis giving absorption band at wave
number region 3404,91 cm-1 is the shifting
vibration of hydroxy group (OH). This is
reinforced by the vibration of the recut at the isolated compound is probably true
1264.53 cm-1 for the -OH group. The naringin compound seen from the IR
absorption band in the wavelength region spectrum showing the functional groups
of 2933.92 cm-1 is a C-H group of CH3. owned by the isolated compound similar to
While the absorption band at the wave the functional group owned naringin
number 1519,95 cm-1 shows the C = C compounds.
bond of the aromatic ring. The absorption
band in the 1410.64 cm-1 wave region is a Further research on identification of
C-H bond in the CH2 group, then the naringin compounds isolated by other
absorption band number in the wave spectrophotometer tests such as MS, NMR
number region 1371,36 cm-1 is the C-H and UV-Vis is required.
bond of the CH3 group. The absorption
bands 1076.33 cm-1 and 1057.03 cm-1 are
the C-O-C bonds of the ether. REFERENCES
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can be estimated that the tested compound Daniel et all, 1999. Comparative Analysis
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OH
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Figure 3. Naringin Structure from pomelo (Citrus grandis) peel”.
According to the naringin structure International Journal of Food
can be matched with the result of IR Science and Technology. 44: 1737–
spectrum analysis. Based on analysis of IR 1742.
spectrum results showed that the tested
compound contained a hydroxy group (- Tang, D., et all. 2011. “Extraction of
OH), aromatic cyclic groups (C = C), CH 2, naringin from pomelo peels as
CH 3, and ether groups. The functional dihydrochalcone’s precursor”.
groups are also present in the naringin Journal Sep. Sci. 34: 113-117.
structure, it is increasingly convinced that
the isolated compound is the naringin
compound.

CONCLUDING AND REMARK

From the results of the research that
has been carried out can be concluded that
the isolated compound exhibits a pure
compound as it produces one color stain
from the TLC using 3 eluent variations, and