1. Would you please tell me if Rtx-Wax(Cat: 12423) (30 meter, 0.25 mmID, 0.25 micrometer df) column is suitable or not?
2. Is this protocol suitable for fatty acid extraction and methylation, As paper was about biodiesel not omega-3?
{Transesterification was carried out as follows: 300 μl of the extraction buffer containing methanol (MeOH) and 2% of concentrated H2SO4 was added to 4 mg of the algae sample and the sample was incubated (2 h, 80 °C, 750 rpm).
Then, 300 μlof0.9%NaClsolutionand300 μl of hexane were added into the reaction tube respectively. Then, the sample was centrifuged (3000 g, 20 °C, 3 min). Finally, the supernatant containing the hexane and FAME was used for GC analysis}?
Restek gives an example for FAMEs on its homepage with this column, surely you figered that out already: http://www.restek.com/chromatogram/view/GC_FF00538 Sounds like you already own this column, so why won't you give it a try?
The protocol is ok, I recommend using some internal standard. Anyway, I usually first extract all lipids with dichloromethane or chloroform, and add then TMSH-solution to this extract. Methylation then occurs in the injector at 250-270 °C. See http://www.mn-net.com/tabid/10234/default.aspx
Dear Sara,
We do lipid analysis using RTX-WAX, and it works. Isomers, e.g. C18:3 might not separate well if you do a relatively fast analysis, but for DHA, ARA and EPA it is ok. I agree whith Thomas on the internal standard: quantitative extrapolation from (external) standard 37 FAME mix is a bit of a strecth (but ok for peak identification.
Interesting this on-injector-transesterification. We use direct biomass transesterification, similar to the protocol described in the question.
You can use wax column for Omega fatty analysis on GCMS, subject to FAME conversion of the proposed oil sample/ any biomass sample.
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