Chromatography Forum

Hi

I wonder if is any chance to separate different kinds of lipids groups like phospholipids, sfingolipids and others... from vegetable/fruits oil using simple paper chromatography? If yes what the best solvent should we use? I had already ninhydrine spray what I think could be usuful? Also chromatography paper, separating chamber, and a view solvents...

If not any advise about TLC simple technic, which plates and solvent are good for such a test?

I dont need very accurate technic, rather prove we have some different kinds of oils in sample or not - good if I could say which one is which

Best regards

I don´t know about paper chromatgraphy, but there are countless methods for silica based TLC. We did something years ago, it was quite simple if one just wanted to separate the main classes of lipids. The method was taken over by another clinic, I lost track of the work.
There is a old classic book on paper chromatogr. (maybe someone has the exact citation?), for TLC an excellent background is given in Stahl´s book on TLC. I suspect you will be smothered if you google.

Hans: you're right about "smothered". A Google search on "phospholipids" + "paper chromatography" turned up 15,300 hits.

PiotrMar: Ninhydrin reacts with amines. It has been widely used in paper chromatography for things like amino acids and peptides, but would probably not be very useful for lipids.

I am glad for answears, but...

Yes, most of those google finds give you nothing and I dont have hours to loking it for - two sentance and you have to pay for more ore just nothing.

I am just new here but I see is quite difficult to get stright answear on this forum, Everybody gives books titles etc, if I would like to read I will find a book I will ask in library if I wont quick advise and simple method I go, full of expectation, to forum. Most of books give You whole packet of knowledge, which I really dont need now.. want just simple method if it is possible - that's what I am asking for... And I am working at labolatory alone nobody to give me halpfull hand or show how to do it - quite taff isnt it -special if you are in other country - for short period. I was specialist in water purifing using photokathalisis TiO2 based..., and now working with healthy oils, in half a year I do PV, AV, Fatty acids profiles of maybve 20 diffrent oils, daily extraction, Moistare and Volantires, Hexane and ethanol analysis on incradible old GC and Headspace equipment... quite a lot of work, isn't it? - that's why I am here now - anyone with stright answears???

Thank you for advise about ninhydrine yes it is I guess more usuful to separated dyes colouring...

Perhaps smth like this

Dissolve drop of sample in 150 μL diethylether.
Apply the solution to a polar TLC plate as a 3 cm band.
Apply 35 μL of your (classes) standards.
Run the TLC-plate twice for 45 min. in chloroform:acetone, 90/10. Air dry 5 min in between.
Visualize the bands with dichlorofluorescein and inspect in UV light.

We are not paid consultants, we all want to learn something while helping.
The "discussion" groups on chrom. in which too few members realized this have disappeared.

Thank you for advise about ninhydrine yes it is I guess more usuful to separated dyes colouring...

No, in fact ninydrin is useless for dyes. It is useful for amines and amino acids (which are typically colorless), because it forms a colored complex.

want just simple method if it is possible - that's what I am asking for...

It is not possible. Paper chromatography appears simple, but that apparent simplicity masks a great deal of complexity. Just to put things in perspective, Martin and Synge were awarded the Nobel Prize for inventing (and explaining) it.

Whenever you want to perform chromatographic analysis of Lipids, then you should always start with W.W.Christies " Lipid Analysis ".
The 3rd edition has TLC as the main separation of groups ( especially chapter 5 for phospholipids,), along with suitable detection sprays etc.,

He also has provided a free pdf, with examples, of Lipid TLC at his web site, which has heaps of useful information...
http://www.lipidlibrary.co.uk/topics/tlc/index.htm

Please keep having fun,

Bruce Hamilton

Thank You guys very much!

I get also some method from mylnefield center, where I was on lipid course some time ago. I had plesure to here live Frank Gunstone, William Christie, Charlie Scrimgeour... What was really helpfull.

I find also:

http://www.cyberlipid.org/phlipt/pl3a0003.htm

It looks like I have to order some standards and special TLC plates. Also Visualising Spray – like Phosphomolybdic acid. OK -hope it will work in future.

Kind regards

Piotr Markiewicz