Improving Chromatography for Analysis of Biological and Pharmaceutical Molecules


Improving Chromatography for Analysis of Biological and Pharmaceutical Molecules: James Grinias, the 2022 winner of the Emerging Leader in Chromatography Award 

March 22, 2022

James P. Grinias is an Associate Professor in the Department of Chemistry & Biochemistry at Rowan University in Glassboro, New Jersey. His research focuses on advancing UHPLC, SFC, RPLC, high-throughput LC, portable chromatography instruments, and multi-dimensional liquid chromatography instrumentation and workflows. He is the winner of the 2022 Emerging Leader in Chromatography Award, which is presented byLCGC magazine. This annual award recognizes the achievements and aspirations of a talented young chromatographer, selected by an independent scientific committee. We recently interviewed him about his research and teaching work.

You and your coworkers have recently written a review on advances in ultra-high-pressure liquid chromatography (UHPLC) and multi-dimensional liquid chromatography (MDLC) (1). What would you say are some of the greatest developments in UHPLC and MDLC over the past five years?

We are approaching the 25-year anniversary of the first report capillary UHPLC (2), which spurred the development of the wide array of commercial UHPLC instruments that are available today. In more recent years, a few trends have emerged that could have an impact on the future of chemical separations. For unidimensional separations, one interesting aspect is the increased throughput that is now available using UHPLC methods (3). High-throughput strategies involving short columns, fast injection cycles, and minimal extra-column volumes enable routine methods in the 10s of seconds rather than the typical 10s of minutes. At the capillary scale, a better understanding of LC column preparation is aiding in the production of unprecedented separation performance for packed beds (4). Parallel to this work, the commercialization of microfabricated pillar arrays and nanoflow instruments with extended pressure limits are providing the proteomics community with new tools to enhance the separation aspects of their methods. Multidimensional separations have also advanced greatly, primarily through the use of improved modulation strategies that enable fast sampling of first dimension peaks while limiting reductions in chromatographic efficiency during the process. We are exploring the adoption of these new strategies in 2D methods involving SFC for the analysis of organic acids and in the development of capillary-scale instrumentation that can provide added orthogonality by simultaneously operating multiple columns with different stationary phases.

In a published overview, you and your students have reported on advances in low-cost strategies for chemical separations (5). How do you foresee the use of 3D printing and other technologies transforming analytical chemistry in the coming years?

3D printing is already starting to play a role in chromatography, as several groups have reported the fabrication of integrated column devices with printed stationary phases in recent years. I also think that 3D printing will play a critical role in microfluidics, as it can be used to fabricate devices designed to perform electrophoretic separations. We have applied 3D printing in the development of low-cost instrument components that can be used for portable analysis, and I think these capabilities will continue to grow as cheaper ways of designing detectors and microcontroller-based interfaces become more widely available. Eventually, I hope that printing technology advances to a point where simple instrument parts and tubing fittings can be prepared as needed rather than ordered, saving both time and money.

Here in LCGC, you have written about portable capillary liquid chromatography (6). How have you used this technology in your work?

This is an exciting time to be working in the area of compact and portable chromatographic instrumentation, as several groups are advancing the field with a number of technological developments. I have been collaborating with the team at Axcend for nearly five years and we have pursued projects in the areas of point-of-care drug screening and pharmaceutical analysis,
both using their compact capillary LC instrument. We also recently devised strategies to use the instrument during classroom lecture demonstrations, as this is much more difficult to accomplish with traditional benchtop equipment. Working in New Jersey, which is often referred to as the “Garden State”, has also provided motivation to use the instrument for on-site testing in a variety of agricultural applications.

What are the main challenges you have to teaching chromatography to undergraduate students?

I usually introduce students to analytical chromatography in our second-year Quantitative Analysis course, which they take after completing one semester of our organic chemistry sequence. This means that they have previously been introduced to liquid-liquid extractions and preparatory column chromatography, making the initial introduction to the topic a bit easier. I
tend to believe that the best way to learn about chromatography is to do it, so we try to provide as much hands-on experience during the laboratory portions of the course that we can. This was more difficult during the pandemic, so we tried to employ as many virtual substitutes as we could as a way of still meeting our learning objectives.

What were some of the key challenges you have encountered during your work? What would you consider to be the most useful contribution(s) of your work?

When research projects involve the preparation of LC columns and the development of new instrument platforms, frustration often follows when something doesn’t work the way you expect it too. The title of the well-known “Art and Science of Column Packing” paper (7) is an apt description of the artisan-like column preparation process that the Jorgenson and Tallarek groups
attempted to better elucidate while I was at UNC. It took me a long time to develop the patience and persistence that is needed to advance a research project, and this is still something I work on every day, although now more in a mentorship role. I have found that providing first-hand accounts of my struggles to my students demonstrates that research can be a difficult endeavor
that is especially rewarding once the hurdles that arise are overcome. In terms of “useful” contributions, I hope to continue to promote the principles of the Open Science Movement within the analytical community and put cutting-edge measurement techniques into the hands of as many researchers as possible. One of the most rewarding outcomes of our work in this area is
when someone contacts our group and indicates an interest in using one of our open-source devices or software programs.

What are some key aspects in a scientific career that motivate you? Would you share some of your work and organizational habits that have helped you be productive and successful professionally?

The best feeling as a researcher is overcoming a technical challenge, which can be in any number of different ways in our work: difficult chromatographic method development, issues with electronics and fluidics during instrument design, challenging sample preparation, etc. I have found in my current role that helping a student mentee overcome one of these challenges is even more rewarding and is one of the best parts of my job. I have been extremely lucky to have a number of both formal and informal mentors throughout the separation science community over the past fifteen years and I now hope to pay that forward to other young scientists in the field. I definitely struggle with physical organization in my office, but I have tried to be diligent in my “digital” organization, both in terms of scientific data and other documentation. As our lives continue to become more digital, effective data organization, backup, and curation are critical tools for scientists and non-scientists alike. Another strategy I have implemented is goal-setting: daily, weekly, monthly, annual, and 5-year goals are listed in various places that I see throughout my day. Although I rarely complete all of the things that are listed, it still provides a regular opportunity to prioritize upcoming tasks as well as give a long-term perspective to day-to-day work.

What can you share with our readers regarding your next area of interest for your research?

Many of the advances in high-throughput LC and multidimensional LC in recent years have relied upon analytical-scale column technology. We hope to develop new ways of using capillary LC in these areas while maintaining system performance, as this provides an opportunity to use the techniques on lower volume samples along with greatly reducing the mobile phase solvent required to perform the analysis.

Do you have any words of advice for young people desiring a career in science?

The first advice I give to anyone who asks me about a career in science is to visit the American Chemical Society’s ChemIDP website. Their self-assessment tool provides an opportunity to critically think about your personal interests and then suggests possible chemistry career paths that best align with them. Once someone has identified these potential paths, the next step is to talk to people who are in the roles that you someday hope to be and learn as much as you can. The digital age has made this process much easier, as a short e-mail or a message over a social media platform can be sent in minutes. Although not every person you contact may respond, I have been pleasantly surprised to see how willing many of today’s leading scientists are to help younger students and junior colleagues in the field. Finally, don’t forget that great science often includes a lot of writing and presenting, so polishing these skills is just as important as improving laboratory technique. As with many areas of life, practice makes perfect, so trying to write or talk about science each day (either formally or informally) can make a big difference over several weeks or months.

Written by: Jerome Workman, Jr.

(1) J. De Vos, D. Stoll, S. Buckenmaier, S. Eeltink, and J.P. Grinias, Advances in ultra‐high‐pressure and multi‐dimensional liquid chromatography instrumentation and workflows, Anal. Sci. Adv.2(3-4), 171–192 (2021).
(2) J.E. MacNair, K.C. Lewis, and J.W. Jorgenson, Ultrahigh-Pressure Reversed-Phase Liquid Chromatography in Packed Capillary Columns, Anal. Chem.69(6), 983-989 (1997).
(3) A.S. Kaplitz, G.A. Kresge, B. Selover, L. Horvat, E.G. Franklin, J.M. Godinho, K.M. Grinias, S.W. Foster, J.J. Davis, and J. P. Grinias, High-Throughput and Ultrafast Liquid Chromatography, Anal. Chem.92(1), 67-84 (2020).
(4) L.E. Blue, E.G. Franklin, J.M. Godinho, J.P. Grinias, K.M. Grinias, D.B. Lunn, and S.M. Moore, Recent advances in capillary ultrahigh pressure liquid chromatography, J. Chromatogr. A1523, 17-39 (2017).
(5) J.J. Davis, S.W. Foster, and J.P. Grinias, Low-cost and open-source strategies for chemical separations, J. Chromatogr. A1638, 461820 (2021).
(6) J.P. Grinias, The Potential for Portable Capillary Liquid Chromatography, LCGC Nor. Am. 38(6S), 15-24 (2020).
(7) J.J. Kirkland and J.J. DeStefano, The art and science of forming packed analytical high-performance liquid chromatography columns, J. Chromatogr. A1126(1-2), 50-57 (2006).

LCGC. Improving Chromatography for Analysis of Biological and Pharmaceutical Molecules: Accessed 13 June 2022