Hydrophilic Interaction Chromatography (HILIC) for Very Polar Analytes What is it? Should I be using it? How do I do it?
Eric Grumbach
Waters Corp.
Abstract
Hydrophilic interaction chromatography (HILIC) is a useful technique for the retention and separation of very polar basic analytes such as actives, metabolites and peptides. HILIC is also referred to as an "aqueous normal phase" or "reverse reversed-phase" mode of separation that offers complimentary selectivity and greater polar compound retention when compared to traditional reversed-phase (RP) chromatography. In HILIC, a high organic-low aqueous mobile phase is used where the analytes elute in order of increasing hydrophilicity. This seminar describes the benefits and advantages of HILIC.
Specific topics include:
- An introduction to the theory and practice of Hydrophilic Interaction Chromatography (HILIC)
- The advantages and benefits realized by using HILIC
- Important and practical considerations for successful HILIC separations
- Studies of the effect of injection solvent on chromatographic peak shape
- Examples of HILIC separations that demonstrate greatly enhanced retention and complimentary selectivity as compared to RP chromatography
- LCMS examples showing greater sensitivity when using HILIC
- Several "real world" chromatographic separations from customers who have successfully used HILIC to solve difficult separation problems
This presentation is designed for chromatographers who struggle to analyze polar compounds and wish to improve and understand the retention and separation of this challenging class of compounds.
Bio.
Eric Grumbach is a Technical Manager in the Chemistry Commercial Operations Division of Waters Corporation with expertise in the areas of HILIC, UltraPerformance Liquid Chromatography, reversed-phase method development and bioanalysis.
Determination of Non-Chromophoric Small Molecule Organic Acids and Bases using Hydrophilic Interation Chromatography Coupled with Mass Spectroscopy or Corona Charged Aerosol Detection
Zongyun Huang
Bristol-Myers Squibb Co.
Abstract
Many pharmaceutical starting materials, intermediates and reagents are polar, small molecule organic acids and bases. Quantitating these ionizable impurities is part of the quality assessment of drug substance. However, this task is particularly challenging because these compounds often lack UV chromophores and their polar nature results in weak to no retention on conventional reversed phase HPLC columns. In this presentation, we provide a methodology combining electrospray ionization (ESI) mass spectrometry (MS) with hydrophilic interaction chromatography (HILIC) to quantitate organic bases such as alkylamine, and acids such as alkylsulfonic acids, at ppm level. Applications to determine these small ionizable impurities in several drug substances using this methodology were also described.
Bio.
Zongyun Huang received his MS degree from Auburn University in Physical Chemistry in 1992. He is a research scientist in Analytical Research and Development department of Bristol-Myers-Squibb in New Brunswick. His research interests are chromatography and analytical method development for API and drug products.
Schedule:
Cost:
Professionals: $20
Students/Currently Unemployed: $5
Directions:
