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HPLC Column Fundamentals: Terms & Relationships


This 4-part seminar provides an introduction to how solutes can interact with Supelco columns containing porous particles (or stationary phases) to result in a chromatography separation. The efficiency, retention and selectivity terms in the resolution equation will be explained as well as how certain particle and column characteristics can alter the terms and affect resolution. The presentation should be suitable for new HPLC users, also for those who have some experience and need a quick refresher on measurement and interpretation of column performance.

Read our summary and companion visual guide to this video, Factors Affecting Resolution in HPLC.


The seminar covers the following points:

  • The separation process on a column that is filled with porous particles - how the sample is introduced and transported through the column by a mobile phase.
  • The well-known Resolution Equation along with a further definition of how the terms are calculated.
  • Retention factor, k (or capacity factor); Selectivity (separation factor); Column Efficiency.
  • How to calculate Peak Symmetry
  • Benefits and drawbacks of porous silica particles as an HPLC column platform
  • Important HPLC Variables - particle size, bonding chemistry, choice of column phase selection
  • C18 stationary phase; modern alkyl phases
  • Polar reversed-phase columns as alternatives to C18 and C8 - embedded polar group, amide phase, phenyl phase, fluorinated aromatic phase
  • Supelco HPLC columns
  • Steps to developing an HPLC method
  • Method development considerations with polar reversed-phase columns


Richard Henry

Richard Henry

ACS Subdivision on Separations


Richard Henry received his B.S. degree in Chemistry from Juniata College in 1963 and Ph.D. in Analytical Chemistry from The Pennsylvania State University in 1966. After a postdoctoral year in separations at Purdue University with Professor L. B. Rogers, he joined DuPont in 1967 at the Experimental Station in Wilmington, DE and became one of the first employees of the DuPont Instrument Products Division in 1969. Dick worked closely with Dr. Jack Kirkland and others in the development of HPLC columns and packing materials while at DuPont.

After about 10 years with Spectra-Physics and consulting on the west coast, he joined the Penn State University chemistry faculty in 1985 where he was Director of Analytical Laboratories and taught Instrumental Analysis to chemistry majors. In 1985, he also founded Keystone Scientific, Inc. where he developed HPLC columns and related separation technology. He retired from both Penn State University and Keystone Scientific in 2002, and remains active teaching short courses on separation technology and as a consultant.

Dick has research interests in separation mechanisms and all applications of new LC column technology. He is also interested in new analytical techniques, especially the rapidly growing fields of UHPLC, chiral HPLC, LC-MS, and multidimensional separation methods. He served two terms as Chairman of the ACS Subdivision on Separations (1998-2002) and has also served for many years on its Executive Committee.

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