Quantitative description of the quality of size exclusion chromatography separations.

Autor: Fekete S; Waters Corporation, located in CMU-Rue Michel Servet 1, 1211 4 Geneva, Switzerland. Electronic address: Szabolcs_Fekete@waters.com., Imiolek M; Waters Corporation, located in CMU-Rue Michel Servet 1, 1211 4 Geneva, Switzerland.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2024 Aug 16; Vol. 1730, pp. 465051. Date of Electronic Publication: 2024 Jun 05.
DOI: 10.1016/j.chroma.2024.465051
Abstrakt: Size exclusion chromatography (SEC) is unique among chromatographic methods as it allows separation of non-retained analytes. However, such mechanism often put an analytical scientist in front of relatively poorly resolved set of peaks that may have strikingly different abundance. The description of such chromatograms needs a particular approach to accurately capture the overall quality of separation. Consequently, use of a single parameter description may not be accurate enough and therefore we introduce a dimensionless separation quality factor, which is based on five SEC specific measures (peak-to-valley, elution window width, peak widths, peak-positioning and recovery). Combining several factors allowed detailed differentiation of various simulated separations, clearly correlating column characteristics with specific contributions to separation quality whether they concern a single peak pair or entire peak landscape. The method could be further elaborated by the addition of normalized priority weighting allowing for flexible quality quantification of a relevant portion of real-life nucleic acid separation on different columns. With growing complexity of biotherapeutics to be separated, such a term is predicted to be a useful response function for purposes of factorial method optimization.
Competing Interests: Declaration of competing interest The authors are employed by Waters Corporation, a company that manufactures and sells chromatography columns.
(Copyright © 2024. Published by Elsevier B.V.)
Databáze: MEDLINE
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