Restek
Home / Resource Hub / Technical Literature Library / Rt-2560 Columns Ensure Accurate, Reliable AOAC 996.06 and AOCS Ce 1j-07 FAMEs Analysis

Rt-2560 Columns Ensure Accurate, Reliable AOAC 996.06 and AOCS Ce 1j-07 FAMEs Analysis

articleBanner

Restek’s Rt-2560 GC column was originally developed for detailed analysis of FAMEs in foods and food products. Its highly polar biscyanopropyl stationary phase and long column format make it an effective tool for the difficult task of separating and accurately quantifying complex mixtures of FAMEs. Recently, Restek optimized the manufacturing process and implemented a new, application-specific, QC-testing procedure for all new Rt-2560 columns (cat.# 13198). As shown here, Rt-2560 columns produced using the optimized process exhibit excellent performance for both AOAC 996.06 and AOCS Ce 1j-07 FAMEs analysis and meet all method requirements.

AOAC 996.06

AOAC official method 996.06 describes a standard procedure for the determination of total, saturated, and unsaturated fat in foods using capillary GC-FID. The procedure involves hydrolytic extraction, methylation, and capillary GC-FID analysis of the resulting fatty acid methyl esters (FAMEs). Triundecanoin, the C11:0 triglyceride, is used as an internal standard for quantitation, and response factors are included in method AOAC 996.06.

The method recommends a 100 m x 0.25 mm x 0.20 µm column with a highly polar biscyanopropyl stationary phase, such as the Rt-2560. The column must be able to separate the adjacent C18:3 and C20:1 FAME isomers as well as C22:1, C20:3, and C20:4 with a resolution of 1.0 or better. The Rt-2560 is specifically designed for FAMEs analysis and readily achieves these required critical separations as shown by peaks 22 to 24 and 29 to 31 in Figure 1.

Figure 1: Application-specific QC testing ensures that all Rt-2560 columns meet AOAC 996.06 method requirements.

cgarm-img
GC_FF1261
PeakstR (min)Conc.
(mg/mL)
Structural Nomenclature
1.Methyl butyrate13.1640C4:0
2.Methyl caproate15.5340C6:0
3.Methyl octanoate19.4340C8:0
4.Methyl decanoate24.5040C10:0
5.Methyl undecanoate27.2020C11:0
6.Methyl dodecanoate29.8740C12:0
7.Methyl tridecanoate32.4720C13:0
8.Methyl myristate34.9740C14:0
9.Methyl myristoleate37.0120C14:1 (c9)
10.Methyl pentadecanoate37.3720C15:0
11.Methyl pentadecenoate39.3620C15:1 (c10)
12.Methyl palmitate39.6860C16:0
13.Methyl palmitoleate41.3020C16:1 (c9)
14.Methyl heptadecanoate41.8620C17:0
15.Methyl heptadecenoate43.4520C17:1 (c10)
16.Methyl stearate43.9740C18:0
17.Methyl octadecenoate44.9220C18:1 (t9)
18.Methyl oleate45.3440C18:1 (c9)
PeakstR (min)Conc.
(mg/mL)
Structural Nomenclature
19.Methyl linolelaidate46.3920C18:2 (t9,t12)
20.Methyl linoleate47.3220C18:2 (c9,c12)
21.Methyl arachidate47.9240C20:0
22.Methyl linolenate48.7720C18:3 (c6,c9,c12)
23.Methyl eicosenoate49.2020C20:1 (c11)
24.Methyl linolenate 49.5420C18:3 (c9,c12,c15)
25.Methyl heneicosanoate49.7820C21:0
26.Methyl eicosadienoate51.0920C20:2 (c11,c14)
27.Methyl behenate51.6040C22:0
28.Methyl eicosatrienoate 52.5220C20:3 (c8,c11,c14)
29.Methyl erucate52.8820C22:1 (c13)
30.Methyl eicosatrienoate 53.2820C20:3 (c11,c14,c17)
31.Methyl arachidonate53.4220C20:4 (c5,c8,c11,c14)
32.Methyl tricosanoate53.6520C23:0
33.Methyl docosadienoate54.8520C22:2 (c13,c16)
34.Methyl lignocerate55.3240C24:0
35.Methyl eicosapentaenoate56.0920C20:5 (c5,c8,c11,c14,c17)
36.Methyl nervonate56.7420C24:1 (C15)
37.Methyl docosahexaenoate62.1720C22:6 (c4,c7,c10,c13,c16,c19)
ColumnRt-2560, 100 m, 0.25 mm ID, 0.20 µm (cat.# 13198)
Standard/SampleFood industry FAME mix (cat.# 35077)
Diluent:Hexane/dichloromethane
Conc.:1,000 µg/mL
Injection
Inj. Vol.:1 µL split (split ratio 20:1)
Liner:Premium 4 mm Precision liner w/wool (cat.# 23305)
Inj. Temp.:225 °C
Oven
Oven Temp.:100 °C (hold 4 min) to 240 °C at 3 °C/min (hold 15 min)
Carrier GasHe, constant flow
Flow Rate:1.0 mL/min
DetectorFID @ 285 °C
Make-up Gas Flow Rate:45 mL/min
Make-up Gas Type:N2
Hydrogen flow:30 mL/min
Air flow:300 mL/min
Data Rate:20 Hz
InstrumentAgilent 7890A GC

Conclusion

The improved manufacturing process and application-specific quality control test both ensure that Rt-2560 columns provide a consistently high level of performance in the detailed analysis of FAMEs. The Rt-2560 column meets method requirements for standard methods of fat speciation in food, including AOAC 996.06 and AOCS Ce-1j-07. In addition, Rt-2560 columns can be used for other methods that call for a 100 m high-percentage biscyanopropyl capillary GC column.

FFAR2588-UNV