HPLC

June 19-24, 2016

San Francisco Marriott Marquis Hotel, San Francisco, CA, USA

Related Materials

Related presentations, posters, and scientific talks from this event have been posted here for your reference. Please click the associated link to download.

Title Author Link
Complex LC Method Development using Method Development System and LC Simulation SoftwareK. Zhu, M. Pursch, B. GuDownload
Evaluation of ACD/AutoChrom Software for LC Method DevelopmentG.A. Von Wald and M.T. Vagnini (The Dow Chemical Company)Download
Retention modelling in hydrophilic interaction chromatography (HILIC)P. Petersson, M. Euerby, K. Kassam, A. Van Wyk, S. Bhal, and I. OshchepkovaDownload

Meet ACD/Labs Staff

Booth # 400

Carol Preisig, Account Manager

Irina Oschepkova, Technical Support Specialist

Poster Schedule

Complex LC Method Development using Method Development System and LC Simulation Software
Koudi Zhu, Matthias Pursch, Binghe Gu

Wednesday, June 22nd, 3:00 PM–4:15 PM
Yerba Buena Ballroom
Poster Session 4
P-W-1415

Retention modelling in hydrophilic interaction chromatography (HILIC)
P. Petersson, M. Euerby, K. Kassam, A. Van Wyk, S. Bhal, and I. Oshchepkova

Thursday, June 23rd, 10:00 AM–11:15 AM
Yerba Buena Ballroom
Poster Session 5
P-Th-1703

Abstract: View Abstract

Hydrophilic Interaction Chromatography (HILIC) has seen an exponential growth in use as a complimentary separation technique to Reversed-Phase Chromatography (RPC) in recent decades. HILIC may show improved retention of polar molecules such as drug compounds and their metabolites and increased sensitivity when using electrospray ionization mass spectrometry (ESI-MS) due to the volatile nature of the acetonitrile enriched eluent relative to RPC. Sample preparation is also greatly simplified in bioanalysis where the acetonitrile enriched supernatant from protein precipitations can be directly injected onto the HILIC column without any need for evaporation and reconstitution.

HILIC uses hydrophilic stationary phase with reversed phase type eluents. The separation mode of HILIC is more complicated than RPC due to a multi-retention mechanism that may contribute to the overall retention of analytes. The aim of this work was to gain a better understanding of the retention behaviour of: a range of acidic, basic, quaternary ammonium salts; and polar neutral analytes on acidic, basic and neutral stationary phases as a function of a range of HILIC operating parameters such as MeCN content, buffer concentration, pH, and temperature.

A number of both pre-existing and newly developed HILIC retention models were assessed for their ability to predict retention as a function of the HILIC operating parameters using a commercially available retention modelling program (ACD/LC Simulator).

The applicability of these models has been shown in both two dimensional isocratic and one dimensional gradient separations for a wide range of analytes with varying physicochemical properties for each of the three stationary phases investigated. The accuracy of prediction for both retention time and peak width accuracy was observed to be comparable to standard RPC retention modelling, but unfortunately it was discovered that gradient modelling could not be used to predict HILIC isocratic conditions or vice versa. A statistical approach was followed to produce a relative ranking of the importance of HILIC operating parameters on the selectivity and retention of the models. The ranking observed was as follows: the nature of the stationary phase > mobile phase pH (i.e., pH 3-6 mainly effecting the ionization of the analyte) > buffer concentration = organic content > temperature. An understanding of these relative importances should prove valuable to chromatographers in the rational design of robust HILIC method development strategies.