HILIC versus normal phase chromatography

[k.vd.wetering@nki.nl]

Hi all,

I want to separate/detect the broadest range of hydrophilic compounds in biological matrices (mainly urine and plasma) and use mass spectrometry for detection. It's ment as a kind of metabolomics approach. I am thinking of buying a Zorbax HILIC plus or a Zorbax Rx SIL (normal phase) column to achieve this. As I am relatively new in the field of separation of hydrophilic compounds, I would like to get some input about the advantages/disadvantages of either column/separation technique.

Your help is well appreciated.

best,
Koen

[tom jupille]

HILIC actually *is* a variety of normal phase chromatography, and many "HILIC" packings are, in fact, bare silica. As to the "Which column is best?" question, the answer is similar to that in reversed-phase: "That depends!". Selectivity will differ considerably among the different stationary phases, and I don't think that any one can be "best" for all samples.

[Wen Jiang]

I agree with Tom that HILIC columns have very broad surface functionalities and base materials, which indeed give the different "Selectivity". Here I would like to recommend you to read followed articles to have better insights on the column chemistry and chromatographic performance.

Stationary phases for hydrophilic interaction chromatography, their characterization and implementation into multidimensional chromatography concepts
P. Jandera, J. Sep. Sci., 31 (2008) 1421-1437
DOI: 10.1002/jssc.200800051

Separation efficiencies in hydrophilic interaction chromatography
T. Ikegami, K. Tomomatsu, H. Takubo, K. Horie, N. Tanaka, J. Chromatogr. A, 1184 (2008) 474-503
DOI: 10.1016/j.chroma.2008.01.075

Hydrophilic interaction chromatography
P. Hemstrom and K. Irgum, J. Sep. Sci. 29 (2006) 1784-1821
DOI: 10.1002/jssc.200600199

Retention behavior of small polar compounds on polar stationary phases in hydrophilic interaction chromatography
Y. Guo, S. Gaiki, J. Chromatogr. A , 1074 (2005) 71-80
DOI: 10.1016/j.chroma.2005.03.058

[Bintang]

Since you are using MS for detection I would go for HILIC rater than normal phase.
I have never used NP and MS but I hear I is not easy to interfase, while HILIC works like a charm with ESI-MS.