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Characteristics of avocado (Persea americana) and kluwih (Artocarpus

camansi) seeds starch with different extraction methods
To cite this article: E Indarti et al 2022 IOP Conf. Ser.: Earth Environ. Sci. 951 012095

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3rd International Conference on Agriculture and Bio-industry (ICAGRI 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 951 (2022) 012095 doi:10.1088/1755-1315/951/1/012095

Characteristics of avocado (Persea americana) and kluwih

(Artocarpus camansi) seeds starch with different extraction
methods

E Indarti1,2*, Nurlaila1, M Muzaifa1, S Noviasari1, Z F Rozali1,2, E M Yusup3

1
Agricultural Product Technology Department, Faculty of Agriculture, Universitas
Syiah Kuala, Banda Aceh 23111, Indonesia
2
Master Program of Agricultural Industrial Technology, Faculty of Agriculture,
Universitas Syiah Kuala, Banda Aceh 23111, Indonesia
3
Department of Mechanics, Universiti Tun Husin Onn Malaysia, Johor 86400,
Malaysia

*
Email: [email protected]

Abstract. Seed from avocado (Persea Americana) and kluwih (Artocarpus camansi) fruit are
waste and by product from processing fruit. Due to the high starch content from both seeds, there
are many extractions process have been applied. The immersed raw material in sodium
metabisulfite solution affected to starch properties. The purpose of this study is to characterize
of starch extracted with and without immersed in sodium metabisulfite solution. The results
showed that the starch yield by immersed in alkaline solution was 21.6% for avocado seed starch
and 24.4% for kluwih seed starch, while yield without immersed in sodium metabisulfite solution
was 17.5% and 18,2% for avocado seed starch and kluwih seed starch, respectively. The starch
content with sodium metabisulfite immersion process of avocado seed and kuwih seed were
79.26% and 72.62%, respectively. Meanwhile without the immersion process resulted starch of
75.13 and 67.30%, in avocado seed starch and kluwih seed starch, respectively. Furthermore,
XRD and FTIR analysis were also carried out on starch to see the crystallinity character and
functional groups contained in the extracted starch.

1. Introduction
Starch is a natural polymer derived from plants with the constituent monomer is glucose linked through
glycosidic bonds between one unit to another. Starch in plants is produced to store excess glucose as
energy reserves and can be a source of energy for human intake [1]. Some plants that produce starch
include wheat, buckwheat, green beans [2], cassava [3], corn, potato [4]; [5], rice [6], and many other
starch- by plants. Chemically, starch is a polysaccharide that is non-allergenic, renewable compound,
can be decomposed by microorganisms [7], biocompatibility, can be easily converted into thermoplastic
[8], then this compound is also insoluble in cold water, alcohol, or other solvents. Physically starch is a
white powder, soft textured, and tasteless. The function of starch is generally used as human food, animal
food and industrial uses [9].

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3rd International Conference on Agriculture and Bio-industry (ICAGRI 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 951 (2022) 012095 doi:10.1088/1755-1315/951/1/012095

Starch is found in plant cells in the form of small, semi-crystalline granules called amyloplasts or
chloroplasts. Starch binds to compounds found in plants such as water and fat. The empirical formula
of starch is (C6H10O5)n from the unit -D-glucose (anhydroglucose). The main macromolecules that make
up starch are amylose and amylopectin with a distribution of about 25% and 75%, respectively. The
molar masses of amylose and amylopectin are about 106 Da and 109 Da [8]. Amylose is composed of
straight chain glucose molecules without branches with -(1,4)-D-glycosidic bonds and has a helical
structure consisting of 200-2000 anhydroglucose units. Amylopectin is also composed of glucose
molecules with -(1,4)-D-glycosidic bonds in straight chains, but the difference with amylose is that
amylopectin has branched chains at -(1,6)-D-glycosidic bonds and consists of 10,000 – 100,000 units of
anhydroglucose [10].
Starch can be obtained from various plants or agricultural materials such as corn, rice, cassava,
potatoes, beans and seeds. However, the availability of these materials is still very much needed to meet
food needs. So that alternative sources of starch that will be used as packaging films can be taken from
agricultural waste that has not been used so far, such as durian seed waste which has a starch content of
43.6%, jackfruit seeds with a starch content of 40-50%. [11] Avocado seeds that produce 21% starch
content [12], kluwih seeds which have starch content between 30.15-39.09% [13], and other wastes that
have not been utilized optimally.
Starch can be extracted dry or wet. However, these two methods often produce starch, there is dry
and wet method, which is still not good in terms of quality. For it is necessary to modify the method of
starch extraction, for example by using chemicals. Previous research modification of taro starch
extraction method was carried out by immersion using NaCl solution. The higher the concentration of
NaCl used, the higher the yield, the degree of whiteness, and the resulting starch content [14]. Therefore,
in this study, starch extraction will be carried out using an alkaline solution in the form of sodium
metabisulfite to obtain good starch produced from waste of avocado seeds and kluwih seeds.

2. Material and methods

2.1. Materials
The materials used in this study were avocado seeds, kluwih seeds, aquadest and sodium metabisulfite
(Na2S2O5). Then the tools used are knives, blenders, immersion containers, filter cloth, sieves, analytical
equipment in the form of scales and calculating tools.

2.2. Starch extraction

2.2.1. Without immersed in alkaline solution (physical method). Seeds are cleaned and peeled using a
knife then sliced about ±1 cm to have a large surface easily extracted. The sliced seeds are mashed using
a blender to form a pulp and the seed pulp is filtered through a filter cloth. The dregs of the seed filter
are washed 3 times with clean water, while the suspension obtained is precipitated for 6-12 hours, then
the precipitate is separated from the solution. After being separated, the precipitate was dried in an oven
at 50˚C for ±48 hours until the starch was dry. The dried starch was then crushed again and sieved using
a 100-mesh sieve.

2.2.2. Immersed in alkaline solution (chemical method). Seeds are cleaned and peeled using a knife.
Then thinly sliced (±1 cm) so that the surface area is larger so that the starch is easily extracted. Seeds
that have been reduced in size are immersed with sodium metabisulfite (Na2S2O5) for 24 hours, the ratio
of seeds and solution is 1:5 (g/ml). After immersing, the seeds then mashed using a blender to form a
slurry and the seed pulp is filtered with a filter cloth. The seed filter dregs were washed 3 times with
clean water, while the suspension obtained was precipitated for 6-12 hours, then the precipitate was
separated from the solution. The precipitate was dried in an oven at 50˚C for ±48 then crushed to a
powdered form and sieved using a 100-mesh sieve.

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3rd International Conference on Agriculture and Bio-industry (ICAGRI 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 951 (2022) 012095 doi:10.1088/1755-1315/951/1/012095

2.3. Characterization of starch

2.3.1 Yield starch. The calculation of the starch yield obtained is based on [15] and is presented
in the form of % yield:

weight of dry starch after extraction

% yield = ×100% (1)
weight of dry seeds

2.2.2. Amylose and amylopectin. Amylose content test was carried out using a UV-Vis
Spectrophotometer. The step is to weigh 50 mg of pure starch into a 100 ml volumetric flask, then add
1 ml of 95% ethanol and 9 ml of 1 N NaOH. The solution is heated in a water bath for 10 minutes and
distilled water is added to the mark. A total of 1, 2, 3, 4, and 5 ml of the standard solution were pipetted
into a 100 ml volumetric flask. After being allowed to stand for 20 minutes, the absorbance of the
intensity of the blue colour formed was measured by a spectrophotometer at a wavelength of 620 nm.
The standard curve was made as the correlation between amylose content (x-axis) and absorbance (y-
axis). Amylose content in the sample is calculated based on the following formula:

CxVxF
% amylose = x100% (2)
W
Where:
C = amylose concentration of standard curve (mg/ml)
V = sample volume (ml)
F = dilution factor
W = sample weight (mg)

The amylose content in the sample can be used to estimate the amylopectin content which is
calculated based on the difference between the total starch content and the amylose content.

2.2.3. X-ray diffraction (XRD). The obtained starch was analysed by X-RD to identify the crystalline
pattern in the material and to determine the size of the starch particles using an X-ray diffractometer (D8
Advance, Axs Bruker model) with Cu Kα radiation at a voltage of 40 kV and 30 mA. The sample is
scanned between 2θ=3 and 40o.

2.2.4. Fourier transform infrared (FTIR). Characterization of starch functional groups by FTIR
spectroscopy was carried out by irradiating starch with infrared radiation with a wave number. The result
of this irradiation is absorption of waves when the radiation wave has a number corresponding to the
vibration frequency, then the missed radiation is observed with a photoelectric detector. [16]. The
components that make up the FT-IR are the energy source of the nano chromator, sample container,
detector, and recorder. The FT-IR used in this study is IRPrestige-21 with a wavelength ranging from
500-4000 cm.

3. Result and discussion

3.1 Yield
Starch extracted with and without immersed in alkaline solution produced different yields. It can be seen
that starch with alkaline solution immersion produces higher yields than without alkaline solution
immersion. This is presumably due to the presence of ions in the alkaline solution, making the starch
contained in the seeds easily separated from the seed flesh because the solution is able to penetrate the
seed flesh until the starch comes out. The Table 1 present the yield starch data obtained in this study.

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3rd International Conference on Agriculture and Bio-industry (ICAGRI 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 951 (2022) 012095 doi:10.1088/1755-1315/951/1/012095

Table 1. Yield of avocado seed and kluwih seed starch.

No Starch with alkaline solution yield (%)
1 Avocado seed + 2,16
2 Avocado seed - 1,75
3 Kluwih seed + 2,44
4 Kluwih seed - 1,82

For the next discussion avocado seed starch which immersed in sodium metabisulfite solution was
defined ASS +, and without immersion was ASS -. Either for kluwih seed starch which immersed in
sodium metabisulfite solution was defined KSS +, and without immersion was KSS -.
3.2. Starch, amylose and amylopectin
Starch is a carbohydrate consisting of amylose and amylopectin. In this study, the starch content by
immersed in alkaline solution was 72.62% of avocado seed starch and 79.26% of kluwih seed starch.
Meanwhile, starch without immersed in alkaline solution is 67.30% avocado seed starch and 75.13%
kluwih seed starch. The starch content of avocado seeds and kluwih seeds can be seen in Table 2.

Table 2. Starch, amylose, and amylopectin content of avocado seed starch (ASS) and kluwih seeds
starch (KSS).
Starch Starch (%) Amylose (%) Amylopectin (%)
ASS(+) 72,62 32,54 40,08
ASS(-) 67,30 33,44 33,86
KSS(+) 79,26 25,63 53,63
KSS(-) 75,13 31,46 43,67

Table 2 show starch with alkaline solution immersion produces higher starch content than starch
without alkaline solution immersion. However, it also has a significant effect on the levels of amylose
and amylopectin. The amylose content of starch with alkaline solution was higher than starch without
alkaline solution. Meanwhile, amylopectin levels are the opposite. [17] extracting avocado seed starch
without immersed in alkaline solution and obtained starch content of 80.1%, namely amylose 43.3%
and amylopectin 36.8%.
3.3 . X-ray diffraction (XRD)
X-ray diffraction or XRD is an analytical method that can show the crystallinity index of a material
including starch. The wavelength used in this test is 5-70 nm. The structure of the crystalline phase is
obtained from X-rays that are refracted after being fired at starch molecules. The test equipment then
performs data interference and produces patterns with different intensities. The spectrum pattern of
avocado seed starch and kluwih seed starch can be seen in Figure 1.

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3rd International Conference on Agriculture and Bio-industry (ICAGRI 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 951 (2022) 012095 doi:10.1088/1755-1315/951/1/012095

17 18
20 22 24 ASS+

ASS-

KSS+

KSS-
count

5 10 15 20 25 30 35 40
2Ɵ degree

Figure 1. X-Ray Diffraction of avocado seed starch with immersed solution (ASS+), without immer-
sion (ASS-) and kluwih seed starch starch with immersed solution (KSS+), without immersion (KSS-).

According to the XRD pattern, starch has crystallinity types A, B and C. Type A starch has strong
peaks at 15o and 23o and imperfect diffraction at 17o and 18o. The type B starch crystal pattern shows
strong peaks in the 17o region and some weak diffraction peaks at 15o, 20o, 22o, and 24o as well as special
diffraction peaks around 5.6o. While the type C crystal pattern is a combination of types A and B, which
has strong peaks in the 17o and 23o regions and several small peaks in the 5o and 15o regions [18].
Based on the figure, it can be seen that the crystalline diffraction pattern of avocado seed starch and
kluwih seed starch is a type B pattern. This is because the strongest peaks of both starches are at 17o and
some small peaks are at 15o, 20o, 22o, and 24o. Then there is a peak on 5,6o which is a special character
of the type B pattern. The shape of the diffraction pattern of avocado seed starch is much sharper than
that of kluwih seeds, this is because the amylose content of avocado seed starch is higher than that of
kluwih seed starch so that the amount of crystallinity is higher and produces sharper peaks.

3.4 . Fourier transform infrared analysis (FTIR)

The functional group structure of avocado seed starch and kluwih seed starch can be identified by FTIR
spectroscopic testing. Each group that is formed displays a vibration at a certain wave number. The wave
numbers used in this spectroscopic test range from 500-4000 cm-1. Identification of functional groups is
carried out after obtaining a vibration graph from the photoelectric detector [19]. Light that is fired at
starch will produce a certain vibration on a functional group. FTIR spectroscopic graphs of avocado
seed starch and kluwih seed starch can be seen in Figure 2.

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3rd International Conference on Agriculture and Bio-industry (ICAGRI 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 951 (2022) 012095 doi:10.1088/1755-1315/951/1/012095

ASS- ASS+ KSS- KSS+

C=O
OH
CH
C-C
Transmittance (%)

400 800 1200 1600 2000 2400 2800 3200 3600 4000
Wave numbers (cm-1)

Figure 2. FTIR chart of avocado seed starch with immersed solution (ASS+), without immersion
(ASS-) and kluwih seed starch with immersed solution (KSS+), without immersion (KSS-).
In Figure 2 show the spectrum of starch shows the absorbance character for polysaccharides, with
glucopyranose rings such as stretching vibrations (stretching) of hydroxyl groups (O-H), alkyl groups
(C-H), carbonyl groups (C=O) and alkane groups (C-C). The results of the FTIR test on avocado seed
starch and kluwih seed starch prove that the functional groups obtained are in accordance with the bonds
between the starch-forming atoms. The functional groups in avocado seeds are the O-H group at the
wave number 3270.84 cm-1, the C-H group at the wave number 2926.21, the C=O group at 1638.83 and
the C-C group at 996.69[20]. Then the functional group analysis was also carried out by Tambunan [21]
who looked at the functional groups in avocado seeds. The results of the observations are that the O-H
group is found at the wave number 3260.23 cm-1, the C-H group at the wave number 2880.17, the C=O
group at 1631.89 and the C-C group at 996.50. The functional groups found in avocado seed starch and
kluwih seed starch with and without immersed in sodium metabisulfite solution show in Table 3.

Table 3. Functional groups in avocado seed starch and kluwih seed starch as a result of FTIR analysis.
Wave number (cm-1)
Functional groups
Avocado+ Avocado - Kluwih+ Kluwih-
O-H 3840,27 3894,26 3898,14 3988,31
3479,58 3522,02 3246,20 3246,20
3207,62 3186,40 2926,01 2926,01
C-H 2929,87 2929,87 2854,65 2854,65
2558,94 2357,01 2358,94 2372,44
2059,98 2065,06 2090.84 2090,84
C=O 1876,74 1882,52 1946,18 1972,89
- - 1741,72 1741,72
1639,49 1641,42 1649,14 1647,21
C-H 1336,67 1352,10 1546,91 1517,98
1205,51 1205,51 1338,60 1205,51
C-C 1002,98 999,13 1006,84 1006,84
707,88 707,88 709,80 709,80

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3rd International Conference on Agriculture and Bio-industry (ICAGRI 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 951 (2022) 012095 doi:10.1088/1755-1315/951/1/012095

4. Conclusion
Extraction of starch by immersion in alkaline solution resulted in a higher starch yield than starch
without immersion in alkaline solution. The starch content obtained was also very clearly different,
namely starch with alkaline solution immersion resulted in higher starch content than starch without
alkaline solution immersion. Then the levels of amylose and amylopectin were inversely proportional
between the two methods. Where the immersion method with alkaline solution resulted in higher
amylopectin content than amylose while the method without immersion in alkaline solution resulted in
higher amylose content than amylopectin. For XRD the four starches showed type B starch based on the
strongest diffraction peak. Then finally, the FTIR results show that the functional groups found in each
starch are the functional groups found in starch, namely the O-H, C-H, C=O, C-C groups.

Acknowledgements
The authors express their gratitude to Universitas Syiah Kuala, Indonesia for financial a support under
H-index research grants scheme, contract no 52/UN11.2/PP/PNBP/SP3/2021.

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