Chromatography Forum

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

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

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.

Thank you Bruce,

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

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 !