Non-destructive chromatographic determination of surface area in reverse phase chromatographic columns

Abstract

One of the critical but underutilized parameters that affect chromatographic separation is an adsorbent surface area (S.A.). We introduce a concept of direct determination of the total surface area of chromatographic columns by utilizing a nondestructive measurement of eluent component excess adsorption isotherm measurement and the application of the Standard Excess Adsorption Isotherm from acetonitrile-water and methanol-water binary mixtures for direct determination of the S.A. in chromatographic columns. This nondestructive technique was used to determine the surface area of five commercial columns, including three core-shell particle type columns (Kinetex-C18, Ascentis C18, and Hallo C18) and in-housemade C18 columns. The traditional low-temperature nitrogen adsorption (LTNA) technique was compared to the novel chromatographic method of nondestructive surface area determination for adsorbents modified with octadecylsilanes. The chromatographically determined surface area was in good agreement with the conventional techniques for all columns used. The difference between the three techniques is less than 10%. Columns were characterized in terms of adsorbent geometry (e.g., pore volume, column void volume, interparticle volume), ligands bonding density, and the amount of the packing material in a chromatographic column.

Graphical Abstract

Keywords:

• Adsorbent surface area
• excess adsorption isotherm
• minor disturbance

Acknowledgements

The authors gratefully acknowledge Merck Research Laboratories (Merck & Co., Inc.) for access to the instrumentation used in this study.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Correction Statement

This article has been corrected with minor changes. These changes do not impact the academic content of the article.