I am currently conducting a study about the formation of free EPA in raw oysters treated with acidic sauces. For separating the polar and nonpolar lipids, I will be using Sep-pak C18 catridges. However, I am still a student and buying a large amount of cartridges (50) is out of my budget. Can you recommend any alternatives for this method?
Any answer or literature will be much appreciated, thank you!
Silicium (though you change the elution mechanism), or LH-20, which come in bulk so you can pour your own columns.
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Hi Bhea
Not sure what do you mean with free EPA, but from the rest of your question I suppose that you want to see the content of EPA in TGs and in the total polar fraction? I don't know if it's out of your budget, but I use 20x20 cm TLC precoated plates SIL G-25.
You can do double development and have the different polar and neutral lipid classes separated, or just do the development of the neutral lipids and have the TGs isolated, at the same time you get your polar fraction in the bottom of the plate. Then you would have to carefully scrape the bands and methylate them, as you would do with total lipid, to prepare your fatty acids for analysis.
Here in UK the pack of 25 is around £190, and you can have 5 samples per plate. You would need a cuvette to do the development.
I use the same plates to purify FAMEs after methylation, so you could use the same pack for both purposes.
Other way to do it is as Catherine suggested. It would involve to buy the silica and pack your own columns. I have used this with carotenoids, but I don't know if it would be cost effective and you would need to run more tests to ensure you are not loosing any fraction, etc..
I recommend you to ask quotes for the different options (sep-pack cartridges, considering cuvettes or columns, silica, solvents) and consider time and efficiency. Sometimes it is worth looking for some extra funding and have a solution that is not going to give you questionable results.
I hope it helps!
Noemi
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