The Chromatography Forum

of Delaware Valley

Oral Session

• Includes presentations from:
  • Andre Striegel - Chemical Sciences Division, National Institute of Standards & Technology (NIST)

    • Can You Really Do That Using Size-Exclusion Chromatography?

      • Over the past half-century-plus, size-exclusion chromatography (SEC, also known as gel permeation or gel filtration chromatography) has become the premier method for characterizing the molar mass averages and distributions of both natural and synthetic macromolecules. Coupling SEC to a multiplicity of physical and chemical detection methods also allows the separation technique to inform our knowledge of polymer long- and short-chain branching, of copolymer chemical heterogeneity, of dilute solution conformation and thermodynamics, etc. The purpose of this presentation is to demonstrate the more complete (though sometimes unusual) picture of an analyte that is obtained by viewing the latter through the lens of different detection methods, and to show that even the inability to view an analyte using a particular detector can, in itself, be quite informative. Examples will encompass polymers, oligomers, and monomers water-soluble and not, synthetic and natural, large and small – and even of negative size!

  • Chris Pohl - Chromatography and Mass Spectrometry Division, Thermo Fisher Scientific

    • Ion-Exchange Materials in HPLC and Ion Chromatography

      • Ion exchange materials continue to play a significant role in the separation of both small ionic molecules and large molecules, especially biomolecules. There are a number of factors that are of critical relevance to design and use of ion exchange materials. I will discuss recent development of new ion exchange materials and their application to various separation challenges. In addition, I will discuss factors important in optimizing the mobile phase for ion exchange separations including proper choice of elution species and design of buffer systems. Often inappropriate choices in mobile phase will result in a large amount of pH hysteresis associated with the stationary phase such that the mobile phase pH is dramatically different than the stationary phase pH. I will review the best options for avoiding such issues. In the case of mixed-mode ion exchange stationary phases, optimization of the method is often complicated by the wide range of experimental parameters available to the analytical chemist. A general method development strategy will be presented, describing how to best approach method development when faced with a multitude of useful experimental operating parameters. Several example optimizations will be illustrated.

  • Mark Schure - Kroungold Analytical, Inc.

    • Multidimensional Liquid Chromatography: A Tutorial on Principles, Applications and Recent Developments

      • In multidimensional chromatography, sample components are fractionated by different columns sequentially, utilizing different retention mechanisms to spread the component zone(s) across an area (2D), volume (3D) and hpervolume (≥3D). To achieve successful resolution of complex sample components, dissimilar (orthogonal) retention mechanisms are required to effectively spread the peaks throughout the available separation space. A good method will maximize the resolution between adjacent zones.

      • In this tutorial our discussion of two-dimensional liquid chromatography (2DLC) includes:
        • How 2DLC works
        • Instrumentation
        • Column selection based on sample chemistry
        • Method development
        • Using orthogonality metrics to characterize the separation
        • Applications of 2DLC to separate peptides, proteins, small and large molecule pharmaceuticals, biological extracts, industrial polymers and surfactants.
      • In addition, 3D and higher dimensional separations are discussed; examples being 2DLC/MS, 2DLC/MS/MS and 2DLC/IMS/MS which have now been realized

  • Kaitlin Grinias - GlaxoSmithKline, Department of Product Development and Supply

    • Automated Forced Degradation Screening for Genotoxic Risk Assessment of Small Molecule Pharmaceutical Candidates

      • Delivering the best possible medicine to the patient is the primary goal of pharmaceutical research and development. Solubility and stability characterization of new target therapies is required to select the most viable compounds to progress into clinical trials. Finding the best molecule and formulation early in the development cycle will reduce the time and effort necessary for that therapy to reach the patient. Due to the large number of molecules in pre-clinical development, the study of any individual molecule is constrained by labor, time, and budgetary resources. Laboratory automation increases the throughput, thus expanding the scope of solubility and stability screening. The Freeslate Core Module 3 (CM3) automates sample preparation through liquid and solid dispensing (with integrated balance) as well as heating, cooling, stirring, and vortexing in a single platform. The system is easily programmable enabling the user to customize each experimental design. The platform interfaces with offline analytical instrumentation (such as HPLC, X-ay diffraction, and Raman). Using high resolution, robust chromatography techniques and automated data processing also improve the throughput of these experiments. These laboratory automation tools facilitated the design, implementation and validation of an automated forced degradation screen that focuses on understanding the genotoxic risk of therapeutic candidates.  

  • Bill Farrell - Pfizer

    • Supercritical Fluid Chromatography (SFC) as a Tool for Pharmaceutical Analysis (Hint: it’s not just for chiral purification)

      • Faced with a separation challenge, most chemists would reach for a reversed phase HPLC column and apply a generic gradient from 100% aqueous to full organic. The reason is likely because the majority of these chromatographers have built upon their own personal experience with RP-HPLC, either in academia or industry, using this approach as a starting point. The simplicity of the setup, broad applicability and the relative ease to learn has made RP-HPLC a popular, near universal technique. With recent advanced in smaller particles and more sophisticated instrumentation, the technique has only gained in popularity. However, RP-HPLC does have some limitations, especially when retaining polar compounds or samples that contain a mix of compounds with a large span of Log D's, which make method development challenging. These types of samples have led to the resurgence in Hydrophilic Interaction Liquid Chromatography (HILIC), an aqueous based, normal phase technique, which uses polar stationary phases with highly organic mobile phases.

        But if your background includes normal phase chromatography, specifically supercritical fluid chromatography (SFC), the choice of tools to apply to a given separation widens. Not solely based on partitioning, SFC separations are characteristically different than those in RP-HPLC and the solvent systems differ than those used in HILIC. Employing the same column dimensions and particles as in reversed phase, but coated with chemistries similar to those found in HILIC, SFC is able to span a wide polarity range with an alternative solvent choices while still demonstrating efficient separations and vastly different selectivity’s.

        This presentation will explore the use of SFC as the first choice for a separation technique and demonstrate the applications for a diversity of chemical compounds.

  • James Grinias - Rowan University
    • Investigating the Packing and Performance of Capillary and Microfluidic LC Columns
      • Improving column performance remains paramount to advancing liquid chromatography (LC) technology. This performance is directly related to the particle bed structure, but until recently the only measure of bed structure quality was observed through van Deemter efficiency measurements. Confocal laser microscopy has been shown to provide sufficient image resolution to allow for the characterization of bed morphology beyond van Deemter analysis. Capillary columns of varying aspect ratios were packed with porous sub-2 μm particles and tested for efficiency. The columns were then imaged and morphological parameters were calculated from a computational reconstruction.  Trends in efficiency matched observed deviations in the packed bed structure. An additional cause of broadening due to particle-size segregation was observed in larger aspect ratio columns with lower performance. This segregation effect was found to correlate with the slurry concentration used during column preparation.
      • Findings regarding slurry concentration effects on column performance were then applied to the preparation of columns on microfluidic devices.  Because pressure limits on these devices are lower than standard columns, 4-5 μm superficially porous particles were explored as an alternative to sub-2 μm packing material. As expected, capillary columns outperformed microfluidic columns due to imperfections imposed by microfluidic channel asymmetry and the world-to-chip connection. However, superficially porous particles packed in microfluidic LC columns had flatter van Deemter curves indicating the potential for better performance at high reduced velocities.  It is in this regime of faster LC separations where microfluidic columns are likely to have the biggest impact in the future. 
  • Michael Dong - MWD Consulting
    • UHPLC Instruments: Perspectives, Innovations and Issues

      • The advent of ultra-high-pressure liquid chromatography (UHPLC) and its successful commercialization in 2004 coupled with the use of sub-2µm-particle columns has set a new performance benchmark for HPLC. It is now possible to perform very fast stability-indicating assays in a few minutes or routine analysis of complex samples with peak capacities exceeding 600. Most HPLC manufacturers have UHPLC product offerings in the range of 15,000 to 22,500 psi with reduced dispersion and improved precision/sensitivity performance. UHPLC has evolved from a research tool into a standard HPLC platform for sample analysis and quality control.This presentation focuses on the modern perspectives and innovations of UHPLC instrumentation including low-dispersion modules, precise pumps with low dwell volumes, autosamplers perform well at low sampling volumes, and small UV flow cells with standard and extended path-lengths. Potential issues of UHPLC in regulatory analysis are described (viscous heating, pump blending and method “transfer” issues) together with their mitigation strategies.

  • Hernan Cortes - HJ Consulting LLC

    • Charaterization of Macromolecules via Hyphenated and Multidimensional Technologies

      • Macromolecules of synthetic and biological origin require special attention when characterization using separation techniques is needed, since their chromatographic behaviour may or may not be akin to conventional small molecules. Detailed characterization of synthetic polymers is critical for better understanding of structure-performance relationships.  Performance properties are dependent on a number of variables, such as molecular weight distribution, molecular topology, functionality and chemical composition. Because such molecules typically lack volatility, separations in the liquid phase are predominant, although large molecules can be converted to small molecule fragments that can be analyzed in the gas phase after pyrolysis treatment, or other means. However, such sample treatments create mixtures of such complexity that single dimension separations are not sufficient to yield the desired information. The use of multidimensional chromatography and appropriate detection become critical for elucidation of chemical structure as well as understanding the implications of structure on performance, since the higher peak capacities obtained as well as the ability to utilize a variety of orthogonal separation mechanisms, supply information not attainable by other means. This presentation will outline strategies used in the characterization of large molecules using hyphenation and multidimensional techniques. Examples of the applications to problems of current interest will be presented to highlight the utility of the technologies devised.

• For more information about our speakers, see below

Poster Session

• Poster sessions are strongly encouraged from the work of students presenting their science and engineering work to vendors aiming to highlight scientific capabilities and innovation
• This symposium is an ideal platform for students to hone their presentation skills and to communicate their projects with those present in the symposium audience
• The Chromatography Forum strongly encourages the presentation of new insights into chromatographic and separation science and the poster session can be a valuable experience for both presenters and attendees
• Judges are from the Chromatography Forum of Delaware Valley Executive Committee
• To submit an abstract, email publicity@cfdv.org before March 27, 2017

Exhibition

• If you are a vendor interested in participating, contact Dave Kohler at david@esind.com
• View the latest in chromatography instrumentation, columns, and sample prep productions from vendors will include:
  • 908 Devices
  • Agilent Technologies
  • Chiral Technologies
  • ES Industries
  • Macherey-Nagel Inc
  • Mac-Mod
  • Millipore Sigma
  • Perkin Elmer
  • Regis Technologies
  • Restek
  • Shimadzu
  • Tosoh Bioscience
  • Waters Corporation
  • Wyatt
• Visit all of our vendors for a chance to win great prizes!