Documente Academic
Documente Profesional
Documente Cultură
chromatography
components
of
living
cells
in
plants,
animals,
and
MATERIALS
1. Gas chromatography
6. Methanol
2. Capillary column
7. Pipette
8. Vortex mixer
9. Glass vial
Palm oil)
METHODS
A.
B.
C.
4. The mixture was leaved for 20 minutes and the present of clear
upper layer that formed was observed
5. The clear upper layer was then pipette out into a clean and dry vial
for analysis
6. 0.4 uL sample was injected into the GC injector port carefully
RESULTS
A.
TR
TR 1
TR 2
(averag
PA 1
PA 2
1.5635
1057.19
1357.60
1.4000
3715.66
3471.52
e)
Stearic Acid
Palmitic Acid
1.549
1.578
0
1.386
0
1.414
Sample
TR
TR 2
(averag
Acid
Corn Oil
Palm Oil
Palmiti
c Acid
Corn Oil
Palm Oil
1.623
1.622
0
1.583
0
1.619
0
1.419
0
1.416
0
1.371
0
1.414
1.6225
1.6010
1.4175
1.3925
5
3593.59
PA 1
PA 2
PA
(averag
e)
1287.08
2026.48
1656.78
84
5554.63
80
7928.75
82
6741.69
18
64
1189.68
41
973.178
3744.61
74
6158.25
1
4951.43
21
68
45
e)
Stearic
1207.39
PA
(average
)
3040.95
CALCULATION
Response Factor
Response Factor (Rf) = Concentration / Peak Area
1. Stearic acid methyl ester
Response Factor (Rf)
= 6.6258
2. Palmitic aid methyl ester
Response Factor (Rf)
= 2.2262
Concentration
Concentration (ppm) = Retention Factor (Rf) x Peak Area (cm2)
1. Palm Oil
a) Stearic acid methyl ester
Concentration (ppm) = 6.6258 x 6741.6941
= 44669.12 ppm
b) Palmitic acid methyl ester
Concentration (ppm) = 2.2262 x 4951.4345
= 11022.88 ppm
2. Corn Oil
a) Stearic acid methyl ester
Concentration (ppm) = 6.6258 x 1656.78821
= 10977.55 ppm
b) Palmitic acid methyl ester
Concentration (ppm) = 2.2262 x 973.1781
= 2166.49 ppm
DISCUSSION
Through this experiment, the fatty acids contained in cooking oils
which are palm oil and corn oil was determined by using gas
chromatography. The standard (palmitic acid methyl ester and stearic acid
methyl ester) was prepared first into desired concentration which is 8000
ppm before it was injected to GC injector port. The samples was then
prepared appropriately and leaved for 20 minutes until clear upper layer
was formed before it was also injected to GC injector port. The retention
time and peak area data was then recorded and the result was analysed.
From the results, the retention time of standard fatty acids was
compared with unknown samples (corn oil and palm oil) to identify types
of fatty acid present. Retention time plays big role in analysing the sample
in the gas chromatography. According to Wirasnita et. al, (2013) retention
time are differentiated according to the number of carbon, molecular
weight and the boiling point of the samples. The retention time will
increase when the molecular weight and boiling point of the compound
increased.
Based on the retention times (t R) data, the average tR for stearic
acid is 1.5635 meanwhile palmitic acids average t R is 1.4000. Meanwhile,
the average tR value of corn oil and palm oil is 1.6225 and 1.6010
respectively. This result shows the nearest tR value to the stearic acid
methyl ester standard (1.5635). In contrast, the others average t R value
for corn oil and palm oil are 1.4175 and 1.3925 respectively. This data
shows the nearest tR value to palmitic acid methyl ester standard
(1.4000). Both data shows that samples contain both standard which are
stearic acid methyl ester (C18) and palmitic acid methyl ester (C16).
Meanwhile, response factor (Rf) for each standard was calculated
which are 6.6258 for stearic acid methyl ester and 2.2262 for palmitic acid
methyl ester. The response factors were calculated by using this formula:Response Factor (Rf) = Concentration / Peak Area
The response factor for each standard was then used to calculate the
concentration of the appropriate fatty acids in the samples (corn oil and
palm oil) by using this formula:Concentration (ppm) = Retention Factor (Rf) x Peak Area (cm2)
From the results obtained, the concentration of palmitic acid in corn
oil and palm oil are 2166.49 ppm and 11022.88 ppm respectively.
Meanwhile, the concentration of stearic acid in corn oil and palm oil are
10977.55 ppm and 44669.12 ppm respectively. These results show that
palm oil and corn oil contain higher number of stearic acid than palmitic
acid. In addition, corn oil also show that it contain higher number of
stearic acid contain than palm oil. These results were not corresponding
with the actual result that was based on studies. This was maybe due to
several errors during preparation. According to Zambiaki et. al (2010),
palm oil should contain a lot of palmitic acid (C16) than stearic acid (C18)
as the peak area average data of palmitic acid in palm oil has the highest
value than in corn oil which results in higher concentration of fatty acid
through the concentrations calculation.
During the experiment, they are some sources of error that occur
which may affect the accuracy of the result. For example, there are
parallax error that happen when the eyes does not directly proportional to
Methylation
Method
Retrieved
to
Determine
April
6,
Fatty
2013
Acid
from
file:///C:/Users/user/Downloads/Modified%20Methylation%20Process
%20and%20D
etermination%20of%20Fatty%20Accid.pdf
composition.pdf
http://www.jhed.mk/filemanager/JHED
%20Vol%204/02.%20FPP/06.%20Full%20pap
%20Kostik%202.pdf
er%20-
%20Vesna