Fatty acids analysis protocol used in our laboratory:
FAME (Fatty acids methyl esters) were prepared dissolving 0,5 g of oil in 6 ml of n-Hexane, adding0,25 ml of KOH 2N in Methanol. After 10 seconds of vigorous shaking, samples were centrifuged at 3000 rpm for 10 minutes and the supernatant collected.
FAME analysis was done with a Agilent 7890A Gas Chromatograph (Agilent, Palo Alto, CA), equipped with a Flame Ionization Detector (FID). Separation was carried out with a Supelco EC-1000 capillary column (30 m, 0,25 μM film
thickness), using Helium as the carrier gas, at a flow rate of 1.2 ml/min. The GC oven temperature program began with the oven held at 185°C for 17 min, then increased to 220°C at 4°C/min, maintained at 220°C for 8 min, then to 230°C at 2°C/min, for 6 minutes. The total run time was 45 min. Detector temperature was set at 300°C, H2 flow at 30 ml/min, air flow at 400 ml/min, make up gas (N2)
flow at 25 ml/min. Sample Injection (1μL) was made in Split mode (50:1) at 60 ml/min. FAME were identified comparing retention times with
those of authentic standards (Sigma Aldrich) and expressed as area units (%) in respect to the total TIC (Total Ion Current) area.
Fatty acids analysis protocol used in our laboratory:
FAME (Fatty acids methyl esters) were prepared dissolving 0,5 g of oil in 6 ml of n-Hexane, adding0,25 ml of KOH 2N in Methanol. After 10 seconds of vigorous shaking, samples were centrifuged at 3000 rpm for 10 minutes and the supernatant collected.
FAME analysis was done with a Agilent 7890A Gas Chromatograph (Agilent, Palo Alto, CA), equipped with a Flame Ionization Detector (FID). Separation was carried out with a Supelco EC-1000 capillary column (30 m, 0,25 μM film
thickness), using Helium as the carrier gas, at a flow rate of 1.2 ml/min. The GC oven temperature program began with the oven held at 185°C for 17 min, then increased to 220°C at 4°C/min, maintained at 220°C for 8 min, then to 230°C at 2°C/min, for 6 minutes. The total run time was 45 min. Detector temperature was set at 300°C, H2 flow at 30 ml/min, air flow at 400 ml/min, make up gas (N2)
flow at 25 ml/min. Sample Injection (1μL) was made in Split mode (50:1) at 60 ml/min. FAME were identified comparing retention times with
those of authentic standards (Sigma Aldrich) and expressed as area units (%) in respect to the total TIC (Total Ion Current) area.
Dear Sonam
Marco's protocol is excellent way to get the profile. Please make sure you do not have high Free Fatty Acid content in the sample [should be less than 0.5% as you may find soap formation [minor comment]
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