Hello,
I am willing to separate different chemical species of phosphatidylcholine by HPLC (standards). The only column I have is a Supelcosil LC-18 column (25 cm X 4.6 mm, 5µm particule size). I tried with many isocratic solvents containing isopropyl alcohol, in the highest amount, plus acetonitrile or water/methanol in smallest amounts. Even after 90 min there was no peaks. I've found a protocol using ZORBAX Eclipse XDB-C18 column (15 cm x 4 mm, 5 µm), I tried to apply it using my column and it doesn't work either. Is it a problem of column?
Thank you
Hi Nabil,
3 years ago a similar question has been answered here in this forum:
https://www.researchgate.net/post/Can_anyone_with_experience_in_phospholipid_separation_using_HPLC_with_cyanopropyl_column_give_some_advice
Regards
Markus
P.S.: I think a C18 column isn't suitable for this compound class. Phospholipids belong to the class of polar lipids, but have a long and non-polar part similar to a C18 phase. They are very strongly retarded on a C18 phase.
Hi Markus,
Thank you very much.
I can't use high amounts of hexane and isopropyl-alcohol because it causes a high pressure in the system, which my deteriorate the column and other parts. In addition, hexane aimes to decrease the retention of the phospholipides by the silica, this will decrease the RT but may make no difference between the chemical spices (difference in fatty acid composition). For now, I am trying with low flow and different solvents.
Greetings
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