Chromatography Forum

My company is interested in using HILIC mode for some raw material purity assessments, but I can't find any procedures/recommendations for how to begin with HILIC development, as well as the peculiarities of care/maintanance of HILIC columns (i.e. is it better to store between use with water, organic, or some combination of the two)? Unfortunately, the column I purchased did not come with anything detailed at all, so I'm kind of left wondering what to do.

We use a lot of reagents that are common to peptide synthesis (PyBOP, HBTU, HOBt, etc.) as well as a few unprotected amnio acids that are difficult to analyze by reverse phase due to poor retention.

Any online resources would be very appreciated, but books or periodicals that I could purchase would be fine too.

Look for an article by Eric Grumbach in LC/GC North America to give you plenty of ideas about the subject.

I would store a HILIC column in 90% acetonitrile and 10% water.

HILIC is a good technique for polar peptides and amino acids that are difficult to retain in RP. We have been using silica HILIC columns.

I also recommend to check for relevant items at the Waters website. There is a chapter on HILIC in my book on "HPLC Columns".

A good starting point for Method Development in HILIC, is the "Practical Guide to HILIC", a compilation provided free of charge since 2005.
It can be ordered at http://www.sequant.com/default.asp?ml=11810

There is also additional information on the website.

Below are applications on Unison UK-Amino:

Angiotensins: http://www.silvertonesciences.com/files/TI374E.pdf
Basic amino acids: http://www.silvertonesciences.com/files/TI329E.pdf
Branched-chain amino acids: http://www.silvertonesciences.com/files/TI311E.pdf

The column comes with an instruction manual (which describes storage
and cleaning conditions).

Feel free to contact me if you have any questions about the column.

I am using C18 RP silica in Flash LC for separatin of tetracycline and its oxidation products at prep scale (~200 mg) but some of the products react with the RP silica and stay their permanently as visible by the brown color...I feel the acidic compounds are responsible for that. Also TC is highly tailing compound. One more problem is that I start gradient with water(amm. formate):ACN 10:90 at which sample is not very soluble and increasing ACN to dissolve the sample causes some compound to elute at solvent front.

Can HILIC help to seperate the products better than RPLC and the stationary phase available in market for flash LC?