Quantitative Assessment of Retention Mechanisms in Hydrophilic Interaction Chromatography

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Yong Guo, and his group at Fairleigh Dickinson University are developing a new methodology to evaluate the retention mechanisms in hydrophilic interaction chromatography. Chromatographic methods are widely used to support biomedical research, environmental monitoring, and development of new drugs and therapies. To develop highly efficient and specific methods for various applications, it is important to have a thorough understanding of the chromatographic principles behind practical methods. The retention mechanisms for separation in hydrophilic interaction chromatography (HILIC) are very complex involving multiple forces and interactions. Professor Guo and his team are developing a new methodology to quantitatively understand each force or interaction involved in HILIC separation. The new methodology will investigate the separation of medicinally relevant molecules on many columns with diverse chemistry. The research results are expected to lead to a better understanding of selectivity, faciliate method development, and provide guidance on designing new columns. The proposed research will engage both graduate and undergraduate students, particularly those from traditionally underrepresented groups, and provide advanced training in separation science and chemical analysis which can prepare the students to pursue advanced degrees or careers in biotech and pharmaceutical industries. The proposed research focuses on quantitative assessment of the retention mechanisms in hydrophilic interaction chromatography (HILIC). A major objective is to develop and validate a new methodology that can be used to quantitatively determine the contribution of each retention mechanism (i.e., hydrophilic partitioning, surface adsorption, and electrostatic interactions) to the overall retention of both non-ionized and ionized analytes. The main retention mechanism for any compounds can be unambiguously identified based on the quantitative contribution data. The quantitative information on the retention mechanisms will also provide new insights into the selectivity for various compounds on different stationary phases and help design meaningful selectivity tests. The new methodology will help create a mechanistic-based classification system for the stationary phases used in hydrophilic interaction chromatography, which will facilitate column selection in method development. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.