Beruflich Dokumente
Kultur Dokumente
Group: AS2453D1
Lecturer name: Dr.Rozita Binti Osman
Date of experiment: 07/10/2015
Date Submit: 17/12/2015
Introduction:
Gas chromatography (GC) separate usually volatile and chemically stable compound.
Fatty acid are not volatile enough to be inject on GC, therefore it was chemically
modified the compound to become more volatile and chemically stable compound. In this
experiment, fatty acid was changed into fatty acid methyl ester (FAME) that are more
volatile than fatty acid by undergoes esterification with metholic solution with catalyst
esterification reagent. Fatty acid methyl esters (FAME) are a type of fatty acid ester that
is derived by esterification of fats with methanol. The molecules in biodiesel are
primarily fame, usually obtained from vegetable oils by esterification.
Esterification is the name given to a reaction in which a Carboxylic acid combines
with an alcohol in the presence of concentrated sulphuric acid to form an ester. For
example: Ethanoic acid reacts with carbonates and hydrogen carbonates to give rise to a
salt, carbon dioxide and water
The aim of the experiment are to introduced derivation of non-volatile fatty acid that
normally used in fat analysis in GC converted into more volatile and chemically stable
fatty acid methyl ester(FAME) and determine the amount of fatty acid methyl
ester(FAME) in derivative sample.
The amount of FAME determine by response factor (RF) formula.
Respone factor (RF) =
amount of sample(std)
peak area
Experimental:
B. Instrument set-up
Injection port
: Split (40:1)
: 250c
Column temperature
: 30mLs-1
Detector temperature
: 250C
Amount of FAME
Response factor
Peak 2
in standard(ppm)
100
360.38354
0.2775
Peak 3
Peak 4
Peak 5
Peak 6
100
100
100
100
376.79562
3612.25244
615.52899
3081.19360
0.2654
0.0277
0.1625
0.0325
Retention time
Retention time
Retention time
for standard
for sample 1
for sample
for sample 3
Peak 2
(min)
1.046
(min)
0.842
2(min)
1.046
(min)
1.046
Peak 3
1.314
1.046
1.313
1.311
Peak 4
2.032
1.131
2.026
2.024
Peak 5
3.396
2.031
3.425
3.432
Peak 6
3.578
3.427
Sample 1
Sample 2
Sample 3
Rf corresponding
Peak area
Amount of
peak
0.2775
0.2654
0.0277
0.1625
0.0325
0.2775
0.2654
0.0277
0.1625
0.0325
0.2775
0.2654
0.0277
0.1625
0.0325
(pA*s)
31.52422
481.01089
205.28319
3009.74390
3114.29810
281.60815
120.72981
1827.53076
1930.30139
23.74359
9.48938
17.95598
175.03383
-
FAME
8.7479710
127.6602
5.686344
489.08338
101.214
78.14626
32.04169
50.62260
313.6739
6.588846
2.518481
0.497380
28.44299
Peak 2
Peak 3
Peak 4
Peak 5
Peak 6
Peak 2
Peak 3
Peak 4
Peak 5
Peak 6
Peak 2
Peak 3
Peak 4
Peak 5
Peak 6
SAMPLE CALCULATION.
DISCUSSION
The component in the sample compared with the standard components by the
retention time. From the retention time of standard and sample. It is proven that
components 5, peaks is not present in all 3 sample because of the differences of the
retention time between standard and sample are too big. Peak 5 in each sample give very
large different in the amount of FAME, this may be due to un-complete separation
process during shaking process or the discarding process. From table above presents the
experimental response factor values obtain from FAME standard mixture and the other 3
sample. All of sample, we run two times to choose either one is the best analysis from this
experiment .The retention time for first samples presence is 0.842, 1.046, 1.131, 2.031,
3.427. For the second and the third sample retention time are shown on the table above.
The recommendation for this experiment is clean the flask properly before we using to
prevents from contaminants affected and the measurements of the solution must be
accurate before adding or transfer to another container
CONCLUSION
The derivatization technique used in this experiment is esterification to convert nonvolatile fatty acid to more volatile fatty acid methyl ester (FAME). There are 5
components in the standard mixture while the 3 sample only indicate 4 components as
shown in the standard mixture by comparison of the retention time. The concentration of
each component is calculated by using the response factor of the standard.
REFERENCES
1. www.ncbi.nlm.nih.gov/pmc/articles
2. Moffat AC, Osselton MD, Widdop B. Clarkes Analysis of Drugs and Poisons.
London: Pharmaceutical Press; 2004
3. http://www.en.wikipedia.org/wiki/fatty-acidmethylester