Aji Zaldya NM Karakterisasi
Kimia, FMIPA, Universitas Mulawarman
KARAKTERISASI SIFAT FISIK KIMIA METIL ESTER MINYAK BIJI WIJEN
(Sesamum indicum L.) DAN KOMPOSISINYA
CHARACTERIZATION PHYSICAL AND CHEMICAL OF SESAME SEED OIL
(Sesamum indicum L.) METHYL ESTER AND COMPOSITION
Aji Zaldya NM1, Daniel1* and Irfan Ashari Hiyahara1, Agustina R. Magdaleni2
1
Chemistry Study Program FMIPA Mulawarman University
Jl. Barong Tong No. 4 Samarinda, East Kalimantan, Indonesia
2
Mulawarman University Medical Study Program, Samarinda, East Kalimantan, Indonesia
*
Corresponding Author: daniel_trg08@yahoo.com
Submitted : 13 Oktober 2020
Accepted : 20 Mei 2023
Publish : 25 Mei 2023
ABSTRACT
The methyl ester of sesame seed oil (Sesame seeds L.) was used to make diethanolamide. This process involved
extraction, transesterification, and amidation. The research revealed that extraction by maceration yielded a sesame
seed oil yield of 36.14 percent, an acid number of 1.1531 mg KOH/g, a FFA number of 0.5796 percent, and a
saponification number of 51.287 mg KOH/g. In the process of converting sesame seed oil to methyl ester, the acid
number was 0.5126 mg KOH/g, the FFA number decreased to 0.2704%, and the saponification number was 50.049
mg KOH/g. According to methyl ester GC-MS analysis, methyl oleate made up 39.13% of the methyl ester of
sesame seed oil. The methyl ester FT-IR spectrum exhibits a specific absorption peak at 1743 cm-1 for C=O ester.
Keywords : sesame seed oil, transesterification, methyl ester
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0
International License.
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Jurnal Kimia Mulawarman Volume 20 Nomor 2 Mei 2023 P-ISSN 1693-5616
doi : 10.30872/jkm.v20i2.946 E-ISSN 2476-9258
INTRODUCTION
Sesame plants are prevalent in Indonesia, and their seeds are utilized to produce oil. Sesame plants thrive in
tropical and subtropical environments. Sesame seed oil can be used as a high-quality cooking oil because the
saturated fatty acids it contains are low and non-cholesterol, which is advantageous for health. Furthermore, sesame
seed oil is rich in vitamin E, which is also beneficial for health. The shelf life of sesame seed oil is high, so it has a
relatively extended shelf life [1].
Oils derived from vegetation (seeds) are vegetable oils. Sesame seed oil is one of the vegetable oils that can
be converted into methyl esters. Sesame seed oil is composed of 14% saturated fatty acids, 39% mono-chain
unsaturated fatty acids, and 46% polyunsaturated fatty acids. Sesame oil contains the fatty acids oleic, linoleic,
linolenic, palmitic, and stearic. These fatty acids can be converted into methyl esters through esterification and
transesterification processes, where partial esterification can be achieved by adjusting the conditions of the
reaction. because the formation of these esters depends heavily on the interaction between alcohol and fatty acids
[3].
There are three methods for extracting sesame seed oil: cold pressing, heated pressing, and roasting. The
presence of fatty acids and functional compounds in sesame seed oil can be altered by one of the processes
involving heat in sesame oil processing. In addition, cold pressing can enhance the quality of the sesame seed oil
produced [4].
On the basis of the preceding information, it is necessary to conduct research on synthesizing methyl esters
from sesame seed oil via a transesterification reaction that was previously accomplished via a maceration extraction
procedure, namely by immersing the sample in n-hexane solvent. [5] with the results of the characterization of its
physical and chemical properties, including acid number, Free Fatty Acid (FFA) value, saponification number,
Hydrophilic Lipophilic Balance (HLB) value, form, Thin Layer Chromatography (TLC), and solubility, as well as
characterization using Fourier Transform -Infra Red (FT-IR) 8201 PC and Gas Chromatography-Mass
Spectrometry (GC-MS) Agilent Technologies.
RESEARCH METHODOLOGY
Equipment
This study utilized a set of laboratory glassware, a set of reflux equipment, an analytical balance, a
separating funnel, a rotary evaporator, a chamber, a TLC plate, FT-IR and GC-MS spectrophotometers, and an
analytical balance.
Material
Sesame seeds (Sesame seeds L.), n-hexane, methanol, H2SO4 (p) 4%, ethyl acetate, KOH 0.1015 N, distilled water,
HCl 0.4901 N, PP indicator, H2C2O4 0.1 N, ethanol 95%, KOH-alcoholic 0.5 N, and Na2SO4 anhydrous were used
in this investigation.
Research procedure
Sesame Seed Oil Extraction
The sesame seeds (Sesamum indicum L.) are cleaned and washed under flowing water, heated to 105
degrees Celsius in an oven, and then crushed. After that, it was extracted by maceration with n-hexane for 72 hours
[5]. A rotary evaporator was used to purify and concentrate the maceration results.
Transesterification of Sesame Seed Oil
Seventy grams of sesame seed oil were placed in a three-neck, flat-bottom flask that was connected to a
magnetic stirrer-equipped reflux apparatus. After that, 19.2 mL of methanol was added with an oil-to-methanol
mole ratio of 1:6 while agitating with a magnetic stirrer. The flask is cooled with ice cubes, and up to 1.53 mL of
H2SO4 (p) at 4% w/w by weight of oil is steadily added as a catalyst. The mixture was then refluxed for four hours
at 60 to 70 degrees Celsius. The results of this reflux are placed in a separatory funnel until two distinct phases
form. The upper phase was extracted, rinsed with distilled water, and filtered with anhydrous Na 2SO4. Then,
evaporate using a rotary evaporator to obtain purer results [6].
Test the acid number and FFA level of sesame seed oil.
10 grams of sesame seed oil were added to a 250-mL Erlenmeyer flask, dissolved with 25 mL of ethanol, 3
drops of PP indicator were added, and the mixture was homogenized before being titrated with a standard solution
of 0.1015 N KOH that had been standardized with solution H 2C2O4 0.1 N until the color changed to violet red. The
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Aji Zaldya NM Karakterisasi
Kimia, FMIPA, Universitas Mulawarman
KOH titration volume was documented, and the acid number (FFA level) was calculated using the following
formula:
56,1 × 𝑉 × 𝑁 𝐾𝑂𝐻
𝐴𝑐𝑖𝑑 𝑁𝑢𝑚𝑏𝑒𝑟 =
𝑚
282 × 𝐴 × 𝑁
𝐹𝑟𝑒𝑒 𝐹𝑎𝑡𝑡𝑦 𝐴𝑐𝑖𝑑 𝐶𝑜𝑛𝑡𝑒𝑛𝑡 (%) = × 100%
10 × 𝑚
282 = Molecular weight of oleic acid
56.1 = Molecular weight of KOH
Saponification Number
Making Blanks
After adding 25 mL of 0.5 N KOH-alcoholic to the saponification vessel, 30 minutes of refluxing followed.
After the solution has boiled, allow it to settle. Then, three droplets of PP indicator were added, and the solution
was titrated with 0.4901 N HCl solution that had been standardized with 0.5 N KOH solution until it turned violet-
red.
Sample Testing
10 grams of sesame seed oil may be added to the pumpkin. After meticulous heating, 25 mL of a 0.5 N
KOH-alcoholic solution was added. After the solution had boiled, it was permitted to chill. Then, three droplets of
PP indicator were added, and the solution was titrated with 0.4901 N HCl until a distinct color change was
observed. The test was conducted in duplicate, the volume of HCl titration used was recorded, and the
saponification number was calculated using the following formula:
56,1 × (𝐵 − 𝐶) × 𝑁 𝐻𝐶𝑙
𝑆𝑎𝑝𝑜𝑛𝑖𝑓𝑖𝑐𝑎𝑡𝑖𝑜𝑛 𝑁𝑢𝑚𝑏𝑒𝑟 (𝐸) =
𝑚
56.1 is the molecular weight of KOH.
Hydrophilic-Lipophilic Balance (HLB) Test
The Griffin method, which is the calculation of HLB for nonionic surfactants derived from fatty acid
methyl ester, can be used to measure HLB. Theoretically, the HLB value can be calculated using the following
equation:
Mh = molecular weight of the hydrophilic component
M = overall molecular weight
Using the following formula, the HLB value can be determined based on the titration between the saponification
number and the acid number.
S = saponification number of the surfactant
A is the acid number of the surfactant.
Solubility Test
Place the methyl ester sample in a test vial using a pipette. Then, solvents including methanol, ethyl acetate,
and n-hexane were added. After shaking, each test tube was allowed to stand upright. The results were then
observed.
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Jurnal Kimia Mulawarman Volume 20 Nomor 2 Mei 2023 P-ISSN 1693-5616
doi : 10.30872/jkm.v20i2.946 E-ISSN 2476-9258
Thin-layer chromatography
In the thin-layer chromatography examination, a comparison of the eluent n-hexane:methanol (2:3) was
used, and the sample was identified using a capillary tube on the TLC plate. After inserting the TLC plate into a
chamber containing solvent, the plate is irradiated with a UV lamp emitting wavelengths of 254 and 365 nm, and
the results are observed.
RESULTS AND DISCUSSION
Sesame Seed Oil Extraction
The maceration of sesame seeds with n-hexane solvent for at least 72 hours yielded a mixture of oil and
solvent. A rotary evaporator was used to purify and concentrate the maceration results. The concentration
procedure yielded a luminous yellow oil with a yield of 36.14 percent. Before transesterification, the acid number,
FFA content, and saponification number of the produced sesame seed oil were evaluated.
Table 1. Sesame seed oil test results
No. Type of Analysis Analysis results
1. Acid Number (mg KOH/g) 1,1531
2. FFA content (%) 0,5796
Saponification Number (mg
3. 51,287
KOH/g)
Transesterification of Sesame Seed Oil
The transesterification procedure transforms triglycerides into methyl esters. At this point, sesame seed oil
triglycerides are reacted with methanol, and the addition of the catalyst H 2SO4 (p) produces methyl esters and then
glycerol as a byproduct. Figure 1 depicts the formation of methyl esters during the transesterification reaction.
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Aji Zaldya NM Karakterisasi
Kimia, FMIPA, Universitas Mulawarman
Figure 1. Transesterification reaction mechanism
At this point, 70 grams of sesame seed oil were combined with 19.2 milliliters of methanol (the ratio of oil
moles to methanol moles was 1:6). The mole ratio of 1:6 is intended to shift the reaction to the right so that the
conversion of triglycerides to methyl esters can operate optimally [7]. While stirring with a magnetic stirrer, 1.53
mL of 4% w/w catalyst H2SO4(p) sesame seed oil was steadily added. The catalyst is added prior to heating in order
to slow down the catalyst's reaction rate; if the catalyst is added while reflux is occurring, the catalyst will react
more quickly, and it is anticipated that this will cause a hydrolysis reaction [8]. Methanol serves as a methyl group
donor, with the methyl group replacing the proton group (H+) on the fatty acids in sesame seed oil to create methyl
esters by bonding with ester groups. H2SO4(p) 4% by weight of sesame seed oil, specifically as a catalyst that can
speed up the rate of the transesterification reaction because the reaction will run slowly without a catalyst, in
addition to a catalyst concentration of 4% of the sample weight because it is the optimal condition for producing up
to 90% methyl ester yields [9]. In addition, the addition of an acid catalyst to a completed reaction typically
employs a catalyst concentration of 1% to 4% by weight of the sample [10]. The mixture was then refluxed for four
hours at 60–70 oC methanol boiling point. The transesterification results were placed in a separatory receptacle and
left to stand for 24 hours, at which point two phases formed. The upper phase is an opaque, yellow methyl ester,
while the lower phase is a glycerol-based byproduct. The phase difference is caused by the polarity difference
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Jurnal Kimia Mulawarman Volume 20 Nomor 2 Mei 2023 P-ISSN 1693-5616
doi : 10.30872/jkm.v20i2.946 E-ISSN 2476-9258
between methyl ester and glycerol; methyl ester is non-polar, whereas glycerol is polar. The upper phase, methyl
ester, is repeatedly rinsed with distilled water to remove solvents and other impurities. The process is continued by
purifying the methyl ester with anhydrous Na2SO4 to remove any remaining water and by evaporating the
remaining solvent with a rotary evaporator to obtain a purer methyl ester. The evaporation of methyl ester resulted
in the production of a yellow liquid with an 80% yield. Before proceeding with the amidation procedure, the methyl
ester was evaluated for acid number, FFA content, saponification number, and HLB value, as well as analyzed by
GC-MS and FT-IR to ascertain the fatty acid components and functional groups present in the methyl ester.
Table 2. Test results on sesame seed oil methyl ester
No. Type of Analysis Analysis results
1. Acid Number (mg KOH/g) 0,5126
2. FFA content (%) 0,2704
Saponification Number (mg
3. 50,049
KOH/g)
4. HLB value 2,0573
GC-MS analyses
On the basis of their molecular weight values, a GC-MS analysis of methyl esters was performed to
ascertain the composition of the fatty acids present in the transesterified sesame seed oil methyl esters.
Figure 2. GC-MS Chromatogram of Sesame Seed Oil Methyl Esters
Table 3 displays the results of the GC-MS analysis of the methyl esters in sesame seed oil.
Methyl oleate, methyl linoleate, methyl palmitate, and methyl stearate made up 39.13%, 36.13%, 11.34%,
and 6.14% of the dominant lactation components in sesame seed oil methyl esters, according to Table 3. Based on
the GC-MS results, the compound is not only a methyl ester but also an ethyl ester; this is likely due to overheating
and an inefficient reaction, as some fatty acids are not converted to methyl esters while others remain fatty acids.
The conversion of lipids to ethyl esters The methyl ester with the highest concentration, 39.13%, was identified as
the seventh peak in the GC-MS results. The following is the mass spectrum of methyl oleate, the most abundant
methyl ester found in sesame seed oil.
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Aji Zaldya NM Karakterisasi
Kimia, FMIPA, Universitas Mulawarman
Table 3. Fatty Acid Methyl Esters from Sesame Seed Oil
Peak No Retention Time Area (%)
1 Methyl palmitate 25,828 11,34
2 (Z)-Methyl hexadec-11- 26,235 0,07
enoate
3 Metil Palmitoleat 26,393 0,16
4 Hexadecanoic acid, ethyl 26,697 0,14
ester
5 Heptadecanoic acid 28,379 0,08
6 Methyl stearate 31,640 6,14
7 Methyl Oleate 32,358 39,13
8 (E)-9-octadecenoic acid, 33,502 0,36
ethyl ester oleate
9 Methyl linoleate 33,957 36,13
10 Linoleic acid ethyl ester 35,061 0,37
11 Methyl linoleate 36,157 0,27
12 9-octadecanoic acid 37,770 0,84
13 Eicosanoic acid 39,439 0,68
14 Cis-methyl 11-eicosenoate 40,246 0,18
15 9,12-octadecadienoic acid 45,624 0,53
(Z,Z)
16 Methyl 20-methyl- 49,396 0,21
heneicosanoate
17 n-Hexadecanoic acid 52,299 0,49
18 Octaethylene glycol 60,256 0,23
monododecyl
Figure 3. Methyl Oleate Mass Spectrum
The mass spectrum that appears on peak 7 in Figure 3 indicates a molecular weight of 296 and the molecular
formula C19H36O2 with a molecular weight of 296. Based on the molecular weight and molecular structure data
acquired from the seventh peak, the spectrum is a methyl oleate compound with a 39.13% peak area, according to
the data. Methyl oleate is the most abundant component of sesame seed oil's methyl ester, as determined by
analysis.
Spectrophotometer FT-IR Analysis
Using the FT-IR spectrum, the functional groups of the obtained methyl ester were identified. An FT-IR
analysis of sesame seed oil was not performed, but the acid number and FFA content of sesame seed oil with the
resulting methyl ester were evaluated. If the value of the acid number and FFA methyl ester content is less than the
value of the acid number and FFA content of sesame seed oil, then the compound resulting from transesterification
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is a methyl ester because some of the free fatty acids in sesame seed oil were converted to methyl ester, resulting in
a decrease in the value of the acid number and content of FFA methyl ester. Figure 4 demonstrates the FT-IR
results for methyl ester.
Figure 4. Spektrum FT-IR Methyl Ester
The FT-IR spectrum analysis of sesame seed oil methyl ester revealed absorption at wavenumber 2924 cm -1
and 2854 cm-1, which is vibration stretching from the aliphatic C-H group, which is reinforced by the appearance of
wavenumber 1458 cm-1 and 1373 cm-1, which is vibration bending from the aliphatic C-H group. At wavenumber
1743 cm-1, vibration extending from the C=O ester group is amplified, whereas at wavenumber 1165 cm -1,
vibration bending from the C-O group is amplified. Then absorption appears at wave number 1651 cm-1, which is
strengthened vibration stretching from the C=C group, which also appears at wavenumber >3000 cm -1, and
absorption appears at wavenumber 725 cm-1, which is a chain of methyl ester compounds -(CH2)n-. According to
the results of the FT-IR spectrum analysis, the characterized compound is a methyl ester because it contains C=O
ester, C=C, C-O, and aliphatic C-H groups.
Hydrophilic-Lipophilic Balance (HLB)
Value determination The Hydrophilic-Lipophilic Balance (HLB) of the methyl ester is derived from
theoretical calculations, specifically the aggregate of the mixed HLB values of the hydrophilic and lipophilic
groups in the methyl ester using the following formula:
where Mh represents the relative molecular mass of the hydrophilic group and M represents the diethanolamide
surfactant's relative molecular mass. The HLB value for sesame seed oil methyl ester is 2.0573 according to the
above formula.
In addition, the HLB value of methyl ester can be calculated from the acid number and saponification number using
the following formula:
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Aji Zaldya NM Karakterisasi
Kimia, FMIPA, Universitas Mulawarman
where S is the saponification number value and A is the acid number value of diethanolamide. Because the
saponification number of sesame seed oil methyl ester is greater than the acid number, the HLB calculation of
sesame seed oil methyl ester cannot be performed in practice.
Yield Characteristics of Methyl Esters
In addition to the FT-IR spectrum produced, the characteristics of the methyl ester were determined to
acquire additional data, such as its form, solubility in various solvents, and TLC tests, in order to confirm that the
synthesis product formed was a methyl ester. Table 4 displays the outcomes of the methyl ester's form, solubility,
TLC test, and FFA value.
Table 4. Yield Characteristics of Methyl Esters
Solubility KLT (2:3)
(n-hexane FFA
Sample Phase Ethyl
N-hexane methanol : (%)
acetate methanol)
Methyl Ester Liquid Soluble Not Soluble Soluble Rf=0,48 0,2704
According to Table 4, methyl ester is non-polar, so it is completely soluble in n-hexane solvent, mildly
soluble in ethyl acetate, and insoluble in methanol. As eluents for the TLC assay, n-hexane and methanol were
used. According to the results, the ratio of n-hexane to methanol was 2:3, and methyl ester had an Rf value of 0.48
at one location. It can be concluded that the greater the Rf value, the closer the polarity of a sample is to the eluent
used, so that methyl esters with low Rf values have non-polar properties in the TLC test. The FFA methyl ester test
revealed a value of 0.2704%, which is lower than sesame seed oil's FFA content of 0.5796 percent. On the basis of
the FFA from sesame seed oil and methyl ester, it can be concluded that the FFA value has decreased; this is due to
the conversion of sesame seed oil's free fatty acids into methyl ester, so that the methyl ester contains relatively few
free fatty acids.
CONCLUSION
Transesterification can produce sesame seed oil methyl esters with the addition of methanol and a 4%
H2SO4(p) catalyst in the reaction process for four hours at 60-70 oC. According to GC-MS results, sesame seed oil
methyl ester has a predominant methyl oleate content of 39.13% and yields 80%. Sesame seed oil methyl esters are
a yellow liquid that is insoluble in methanol, faintly soluble in ethyl acetate, and soluble in n-hexane; it has an acid
number of 0.5126 mg KOH/g, an FFA content of 0.2704%, and a saponification number of 50.049 mg KOH/g. The
Rf value of sesame seed oil methyl esters in 2:3 eluents of n-hexane and methanol was 0.48, while the theoretical
HLB value was 2.0573.
ACKNOWLEDGMENT
We would like to thank the Organic Chemistry Laboratory team at the Faculty of Mathematics and Natural
Sciences, Mulawarman University, as well as the instrumentation team at the Organic Chemistry Laboratory at the
Faculty of Mathematics and Natural Sciences, Gadjah Mada University, and Central Jakarta LABKESDA, who
provided the opportunity to develop this method and contributed significantly to this research from the beginning to
the end.
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