Phospholipids and lipophilic compounds by HPLC

[Seb]

Hello everybody,

I am strugling to develop a methop to analyse phospholipids and lipophilic compounds by HPLC.

I could use normal phase. However, I am not allowed to use chlorinated solvents or hexane. Because of this, things are complicated.

I tried Hilic mode on a NH2 without success. C8 seems to work a bit but it is not wonderful.

Does any body had to cope with such a case ?

Thank you very much for your advice !

Seb

[Bruce Hamilton]

I'd strongly consider normal phase, just because there is so much literature, and phospholipids can be quite diverse depending on the source and extraction.

You're not allowed to use hexane ( toxicity? - note that 40-60 petroleum benzine/spirits usually has <2% of the toxin n-hexane ), but can you use other hydrocarbons, such as cyclohexane ( 81C), or n-heptane or iso-octane ( 2,2,4-trimethylpentane ), which boil at 100C?. Even a higher boiling range ( 80-100) petroleum benzene could be suitable.

If you can, then they would be my first suggestion. The chromaography will only change slightly.

Bruce Hamilton

[Mark Tracy]

I did lecithin phospholipids a while ago. I used an Acclaim PolarAdvantage column with 46.5:46.5:7.0:0.1 MeCN:MeOH:H2O:formic acid and ELSD detection. The results were pretty good.

[Seb]

Thank you Bruce,

I am going to use heptane on a silica column and I hope it will give something

[Seb]

I did lecithin phospholipids a while ago. I used an Acclaim PolarAdvantage column with 46.5:46.5:7.0:0.1 MeCN:MeOH:H2O:formic acid and ELSD detection. The results were pretty good.

I have other polar embedded columns in the labs I think. I am going to try this method !

Thanks !