I have to do profiling of amino acids through HILIC chromatography, so I have Zorbax NH2 Column. So, Can we use the Zorbax NH2 column as the HILIC column for the separation of amino acids?
Use Reprospher HILIC (Arginine based) HPLC column from Dr Maisch GmbH Germany
Yes. HILIC was initially developed from NH2 columns (Zorbax was one of them) back in the 1980's. However many HILIC columns manufactured today are specialty columns and have little in common with their great grandfather used back in the 1980's.
Simply yes, but you should carefully select and optimize the mobile phases. Amino columns are prone to lose NH2 ligands at extreme pHs, especially in silica-based. Polymeric and organosilica backbones are more resistant at above-neutral conditions. Why I am indicating this is the critical pairs for AAs may need TEA or similar alkaline additives for baseline separation which prevents applying the proposed strategy on this type of column. Zwitter ionic phases are the best for AA analysis according to my previous experiences. For Schiff base formation-based interaction for AA analysis, amide HILIC is more robust than amine even if they are attached to the silica surface. Well-controlled pH, meticulously optimized HILIC gradient and convenient injection solvent composition may lead you to success indeed for amine HILIC method development.
Good luck, Emir..
Reprospher HILIC itself contains Arginine as stationary phase which itself is amino acid.Use ammonium acetate 20mM and ACN in (80:20) ratio at wavelength 205nm.
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