3
Results
The analysis of a 37 component FAME reference standard was successfully carried out using a
TR-FAME GC column (Figure 1). The high polarity phase on the TRACE TR-FAME GC column
provided baseline resolution of the majority of FAME components, apart from C20:3 [
cis
-8, 11,
14], C22:1 [
cis
-13], and C20:3 [
cis
-8, 14, 17], which were partially separated. These compounds,
two of which are isomeric, are known to be difficult to separate by GC due to their structural
similarities, which results in poor resolution. All FAME components exhibited excellent
chromatographic peak shape.
A qualitative analysis was performed by comparing the FAME peaks in the fat matrices using the
two derivatization methods. The components were identified using the retention times in the FAME
reference standard in Figure 1. The results from the two methods are compared in Figures 2 and 3,
under equivalent conditions (see Table 2 for comparison).
Separation Conditions
Instrumentation:
Thermo Scientific™ TRACE™ GC Ultra Gas Chromatograph
Carrier gas:
Helium
Split flow:
10 mL/min
Split ratio:
10:1
Column flow:
1.0 mL/min, constant flow
Oven temperature:
100 °C (0.2 min), 2 °C/min, 240 °C (15 min)
Injector type:
Split/Splitless
Injector mode:
Split, constant septum purge
Injector temperature:
240 °C
Detector type:
Flame ionization detector (FID)
Detector temperature:
250 °C
Detector air flow:
350 mL/min
Detector hydrogen flow:
35 mL/min
Detector nitrogen flow:
30 mL/min
Injection Conditions
Instrumentation:
TriPlus Autosampler
Injection volume:
1 µL
Figure 1: Chromatogram of 37 components FAME mixture (reference standard) separated on a TR-FAME
100 m × 0.25 mm × 0.20 µm GC column
Response (mV)
Retention Time (min)
0
20
40
60
80
100
120
140
160
180
200
220
10
20
30
40
50
60
70
Response (mV)
RetentionTime (min)
20
40
60
80
100
120
140
160
66.8
67
67.2
67.4
67.6
67.8
1
2
3
4
5
6
7
8
9 10 11
12
13 14 15
16
18
17
19
20
22
23
24
25,26
27
28, 29, 30
35
36
21
37
31
32 33 34
28
29
30
