Hydrophilic Interaction Liquid Chromatography [Bioanalytics]

posted by Helmut Homepage – Vienna, Austria, 2021-11-17 12:43 (383 d 09:57 ago) – Posting: # 22674
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Hi Selwa,

❝ I would like to know if we are called …


Nobody calls you to use anything. You can use any stationary phase (normal or reversed phase, diol, amino, ion-exchange, …) you like. As long as the method passes validation, all is good.*

❝ … to use HPLC or UPLC MS columns in normal phase in bioanalysis?


I would avoid normal phase if ever possible, since strongly non-polar mobile phases (e.g., hexane) in analytical grade are rather expensive and environmentally problematic.

I suggest HILIC instead, i.e., an RP column and a mobile phase consisting of a moderately non-polar (e.g., acetonitrile) component modified by adding <20% water. Pretty old approach.3,4
In my CRO we used it for amitriptyline/nortriptyline, clobutinol, and doxepin.



  1. Acree TE, Barnard J, Cunningham G. A procedure for the sensory analysis of gas chromatographic effluents. Food Chem. 1984; 14(4): 273–86. doi:10.1016/0308-8146(84)90082-7.
  2. Song H, Liu J. GC-O-MS technique and its applications in food flavor analysis. Food Res Int. 2018; 114: 187–98. doi:10.1016/j.foodres.2018.07.037.
  3. Schmid RW, Wolf C. Use of unmodified silica with buffered aqueous mobile phase mixtures for selective chromatography of basic drugs. Chromatographia. 1987; 24: 713–9. doi:10.1007/BF02688574.
  4. Alpert AJ. Hydrophilic-interaction chromatography for the separation of peptides, nucleic acids and other polar compounds. J Chromatogr A. 1990; 499:177–96. doi:10.1016/s0021-9673(00)96972-3.

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