FLAVONOIDS CONTENT DETERMINATION OF TOTAL SKIN FRUIT

EXTRACT ETHANOL Avocado (Persea americana Mill.) USING

SPECTROPHOTOMETRY UV-VIS
1
Aminah , Nurhayati Tomayahu, Zainal Abidin
Faculty of Pharmacy, University Muslim Indonesia
1
aminah.aminah@umi.ac.id

ABSTRACT
Plant Avocado (Persea americana Mill.), Belongs to Lauraceae family roomates have medicinal
properties. Avocado peel contains flavonoids, compounds that can be used to protect and reduce the skin damage
against UV rays. This study Aimed to Determine the total flavonoids content of the ethanol extract of the avocado
peel using UV-Vis spectrophotometry. The ethanol extract of avocado peel was Obtained by maceration with
ethanol. The qualitative analysis of used FeCl3by forming green complex. The determination of total flavonoids
content is conducted based on AlCl 3method with total flavonoids Expressed in QE (Quercetin equivalent) at the
maximum wavelength of 435 nm. The result Showed that the average content of total flavonoids is 4.0122 mgQE / g
extract.

Keywords: Total Flavonoids, Caucasian avocado (Persea americana Mill.), UV-Vis spectrophotometry

I. PRELIMINARY peel an avocado (Persea americana Mill.), so that the

Indonesia is a country that has a wealth of
nature with various types of plants that can be
efficacious as a traditional medicine. Traditional
medicine more and more in demand by the public for
nabatinya material easy to get, easy to mix and
affordable, so that the materials used have to be
upgraded and quality in accordance with the needs of
society.
One plant that is interesting to study is the
fruit of the family Lauraceae avocado which is a
plant that can be efficacious as a medicine, this plant
can be grown in the tropics and subtropics (Katja,
Suryanto & Wehantouw 2009, p. 58).
Avocado empirically nutritious treat diseases
such as canker sores, bladder stones, toothache, dry
face, swelling due to inflammation and diabetes (Katja,
Suryanto & Wehantouw 2009, p. 58). Leather avocado
contains flavonoid compounds that can be used to
protect the skin against UV rays or can reduce skin
damage, because these compounds work as sunscreen
active ingredients (Mokodompit, Edy and Wiyono,
2013, p. 85).
Flavonoids are one of the largest natural
phenol group compounds found in all green plants
(Markham, KR 1988). According to (Pourmorad, F
2006, p. 1143) mengemukkan that one class of
polyphenol compounds known to have properties as
catchers of free radicals, inhibiting the enzyme
hydrolysis, oxidative, and also works as an anti-
inflammatory.
Based on these descriptions, it is necessary
to more intensive study of testing a total flavonoid
concentration of ethanol extract
Indonesia Fitofarmaka Journal, Vol. 4 2
potential of this plant as a raw material medicine for the
prevention and treatment of various diseases can be
developed to the maximum.
II. RESEARCH METHODS
A. Sample Collection and Processing
Sampling avocado (Persea americana Mill.)
Done in the morning around 10:00 pm in Malino, South
Sulawesi. Wet then sorted to remove soil and other
impurities that are still attached to the sample. Then the
avocado fruit is separated from the pulp and skin of the
fruit and in the clear. The skin of an avocado (Persea
americana Mill.) Which had been cleaned carried out
changing the form by cut into small pieces, then dried
with aerated for several days in the open air with no
direct sunlight. After drying the samples are weighed
and recorded and then powdered dry weight was
weighed again after the powder sample weight was
obtained (Dahlia & Ahmad 2016, p. 16).
B.Proses extraction of avocado skin (Persea
americana Mill.)
A total of 50 gram samples of skin of an
avocado (Persea americana Mill.) Was added to the
container maceration. Then added with 200 ml of 96%
ethanol until the entire sample is submerged, then
covered and left for 24 hours. Maserat filtered using
filter paper. The filtrate obtained by filtration with a
funnel, then re-pulp macerated with 200 mL of 96%
ethanol, so that the filtrate is nearly colorless.

All the filtrate was combined and concentrated using obtain the ethanol extract of the skin of an avocado
a rotary evaporator until no more liquid dripped to (Persea americana Mill.). Lumpy skin extracts of
 avocado (Persea americana Mill.) Obtained is used
for further analysis (Gustandy, M and Soegihardjo,
C, J 2016).
C. Quantitative Analysis of Flavonoid Content A
total of 1 mg of ethanol extract of the fruit skin
Avocado (Persea americana Mill.) was added with 2
drops of FeCl3, The formation of green or blue green
color indicates flavonoid compounds in a material
(Harborne, JB 1987).
D. Content of Flavonoids Qualitative Analysis
1.Penentuan maximum wavelength (ƛ max)
quercetin
Determination of the maximum wavelength
of quercetin was done by running a solution of
quercetin in the wavelength range 400-450 nm. The
results show the maximum wavelength running
standards kuarsetin is at a wavelength of 435 nm.
The maximum wavelength used to measure the
absorbance of the sample bark ethanol extract of
avocado (Persea americana Mill.).
2. Preparation of standard curve kuarsetin
Weighed as much as 25 mg of standard raw
quercetin and dissolved in 25 mL of ethanol. Sebayak
pipette stock solution of 1 mL and paid back the volume
to 10 mL with ethanol in order to obtain a concentration
of 100 ppm. Of standard solution of quercetin 100 ppm,
then made several concentration is 6 ppm, 8 ppm, 10
ppm, 12 ppm and 14 ppm. Of each standard solution
concentrations of quercetin pipette 1 mL. Then add 1
mL of AlCl32% and 1 mL of 120 mM potassium
acetate. Samples were incubated for one hour at room
temperature. Absorbance was determined using UV-Vis
spectrophotometry at a wavelength of 435 nm
maximum (Stanković, MS, 2011, p. 65).
3. Determination of total flavonoid content of the
ethanol extract of the skin of an avocado (Persea
americana Mill.)
Weighed 15 mg of extract, dissolved in
10 mL ethanol, to obtain 1500 ppm. From pipette 1 mL
of the solution was then added 1 mL of AlCl 32% and 1
mL of 120 mM potassium acetate. Samples were
incubated for one hour
at room temperature. Absorbance was determined
using UV-Vis spectrophotometry at a wavelength of
435 nm maximum. Samples were prepared in three
replication for each analysis and
Indonesia Fitofarmaka Journal, Vol. 4 2
The extract obtained is concentrated by rotary
evaporator to obtain the extract thick dark green. Then
calculate rendamen, in order to obtain the average
percent rendamen ie 17.28%. Rendamen determination
average values obtained absorbance (Stankovic, MS,
2011, p. 65).
III. RESULTS AND DISCUSSION
Plant avocado (Persea americana Mill.) Were
used in this study is only on the skin of the fruit. The
skin of an avocado (Persea americana Mill.) Can be
used as a sunscreen active ingredient that is to protect
the skin against UV rays or can reduce skin damage,
because it contains flavonoids (Mokodompit, Edy and
Wiyono, 2013, p. 85).
Flavonoids are virtually present in all parts of
plants, including fruits, roots, leaves, stems and outer
skin. Flavonoids are natural compounds that have the
potential as antioxidants to counteract free radicals
that contribute to the onset of degenerative diseases
through the mechanism of the destruction of the
immune system, lipid and protein oxidation (Rais, IR
2015, p. 103).
In this study, avocado used were obtained
from Malino, South Sulawesi. Dirt or insects that
attach to the avocado should be cleaned because it can
interfere with the extraction process and the results.
The skin of an avocado (Persea americana Mill.) Is
performed by means of changing the form of cut into
small pieces to speed up the drying process. The
drying process is intended to reduce the water content
contained in the sample, so as to prevent spoilage by
bacteria.
Extraction process aims to take the
chemical compounds contained in the sample. The
principle is based on the displacement future
extraction component substances dissolved into the
solvent so that the displacement in the interfacial layer
and diffuses into the solvent (Harborne, JB 1987).
Solvents used in this study was 96% ethanol as polar
solvents. In terms penyarian, ethanol has advantages
compared with water and methanol. Chemical
compounds capable disari with more ethanol than
methanol and water penyari (Azizah and Salama,
2013, p. 24).
To obtain the desired chemical compound
used extraction method is a method penyarian
nutritious substances or active substances from the
plant by using a suitable solvent (Yuliani & Satuhu,
2012).
The extraction method used in this study is
the maceration, because this method is simpler,
easier and without heating. Because if you use
heating can make the levels of flavonoids reduced.
Maceration process using three replication with 200
mL of 96% ethanol for 24 hours. The addition of
ethanol made up to 3 times the extraction process.
function to determine levels of secondary
metabolites are carried away by the solvent but can
not determine the type of compounds that are
carried by the solvent (Ahmad, Juwita and Malik 2016,
p. 6). Can be seen in Table 1.
Table 1. The results of the ethanol extract of the
skin of an avocado (Persea americana Mill.)
Qualitative analysis was conducted to
determine the chemical components in plants by
using reagent iron (III) chloride (FeCl3). Observed
changes in color formed is green (Harborne, JB
1987). The identification results showed ethanol
extract of the skin of an avocado (Persea americana
Mill.) Positive contain flavonoids that can be seen in
Table 2.
Table 2. The results of the qualitative analysis of
the ethanol extract of the skin of an avocado (Persea
americana Mill.)
Quantitative analysis of total flavonoid
compounds using UV-Vis spectrophotometry was
conducted to determine how large the total flavonoid
content is contained in the ethanol extract of the skin of
an avocado (Persea americana Mill.). analysis
flavonoid do with use
UV-Vis spectrophotometry as flavonoids containing
conjugated aromatic system that shows a strong
absorption band in the ultraviolet region of the
spectrum and the spectrum of visible light (Harborne,
JB 1987).
In this study to determine the levels of total
flavonoids in the samples used kuarsetin as a standard
solution with a concentration series of 6, 8, 10, 12 and
14 ppm. Used series of concentration because of the
method in use in determining the levels is a method of
using the curve equation
Indonesia Fitofarmaka Journal, Vol. 4 2
as controller work as pemblank (mengkali zero-kan)
compounds do not need to be analyzed (Basset,
1994).
raw, to make a standard curve first made a few rows to
get the concentration of linear equations that can be
used to calculate the percent levels. Kuarsetin used as a
standard solution for flavonol quercetin is a flavonoid
group having a keto group at C-4 and has a hydroxyl
group at atom C-3 or C-5 which neighbors of flavones
and flavonols (Azizah and Faramayuda 2014, p. 48),
Measurement of maximum absorption wavelength do
the running of the wavelength 400-450 nm. The results
show the maximum wavelength running standards
kuarsetin is at a wavelength of 435 nm. The maximum
wavelength used to measure the absorbance of the
sample bark ethanol extract of avocado (Persea
americana Mill.). Can be seen in Table 3.
Table 3. Results of quercetin standard solution
absorbance measurement at 435 nm wavelength
maximum
Picture 1. The calibration curve of quercetin at
maximum wavelength 435 nm
From these measurements, it can be
concluded that the higher concentrations used the
higher the absorbance were obtained. Results
obtained raw quercetin and absorbannya plotted
between levels, so the linear regression equation is y
= 0,0438x + 0.0177 with a value of R 2obtained at
0.9944 and r is 0.997. Quercetin calibration curve
equation can be used as a benchmark to determine
the concentrations of total flavonoid compounds in
the sample extract.
Testing of quantitative analysis by UV-Vis
spectrophotometry use of the reference solution
In the measurement of total flavonoid
compound, the sample solution was added
AlCl3which can form a complex, resulting in a
shift in wavelength toward visible (visible) which
is marked with a solution produces a more yellow
color. And the addition of potassium acetate which
aims to maintain the wavelength in the visible region
(visible) (Chang et al, 2002). Treatment incubation
for 1 hour before measurement meant that the
reaction goes perfectly, so that the intensity of the
color produced is maximum (Azizah and
Faramayuda 2014, p. 48), So that the results of this
study were obtained total flavonoid content of the
ethanol extract of the skin of an avocado (Persea
americana Mill.) Of 4.0122 mgQE / g extract which
can be seen in Table 3.
Table 4. The results of the assay of total flavonoids
% (w / w) in the bark ethanol extract of avocado
(Persea americana Mill.)
According to research conducted by
Kurniasari (2006) stated that a number of medicinal
plants containing flavonoids have been
reported to have antioxidant, antibacterial, antiviral,
anti-inflammatory, antiallergic and anticancer.
IV. CONCLUSION
Based on the research that has been done
can be concluded that the levels of total flavonoids
from the ethanol extract of the skin of an avocado
(Persea americana Mill.) Is 4.0122 mgQE / g extract.
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