Column Classification/Characterisation of Strong Cation Exchange Phases for the Liquid Chromatographic Analysis of Small
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ORIGINAL
Column Classification/Characterisation of Strong Cation Exchange Phases for the Liquid Chromatographic Analysis of Small Molecular Weight Bases Jennifer K. Field1 · Ashleigh Bell1 · Ifigeneia Christopoulou1 · Patrik Petersson2 · Paul D. Ferguson3 · Melvin R. Euerby1,4 Received: 30 April 2020 / Revised: 28 July 2020 / Accepted: 5 August 2020 © The Author(s) 2020
Abstract A simple, rapid and robust protocol for the characterisation of strong cation exchange columns for the analysis of small molecular weight bases is described. A range of ten different phases were characterised, and the resultant selectivity and retention factors analysed using Principal Component Analysis. The score plots for the first and second principal components described 83% of the variability within the dataset. Score plots highlighted the large chromatographic differences observed between the phases, the validity of which was established using a larger range of bases. All the strong cation exchange materials demonstrated a synergistic mixed mode (i.e. ion exchange and hydrophobic) retention mechanism. Principal Component Analysis also highlighted the potential difficulty in locating suitable strong cation exchange “back-up” columns for the analysis of small molecular weight bases in that the characterised columns all displayed very different selectivities. The robustness of the protocol was confirmed by a factorial design experiment. Keywords Strong cation exchange stationary phases · Column characterisation · Principal Component Analysis · Robustness testing · Factorial design · Small molecular weight bases
Introduction Over the last 20 years, considerable efforts have been made to assist the chromatographer in selecting reversed-phase LC (RP-LC) columns for various purposes such as identifying ones with differing chromatographic selectivity for method development proposes and those with very similar Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10337-020-03943-7) contains supplementary material, which is available to authorized users. * Melvin R. Euerby [email protected] 1
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, UK
2
Novo Nordisk A/S, Novo Nordisk Park, 2760 Måløv, Denmark
3
AstraZeneca, Silk Road Business Park, Macclesfield SK10 2NA, UK
4
Shimadzu UK, Milton Keynes, Buckinghamshire MK12 5RD, UK
selectivity and retention as equivalent columns for “back up” purposes. This has been accomplished by the establishment of column characterisation databases which are freely available on the internet allowing the chromatographer to search the databases using predefined selection criteria. The largest RP-LC databases are based on the Snyder’s Hydrophobic Subtraction Model [1] and the extended Tanaka protocols developed by Euerby et al. [2] which can be found in the USP PQRI and the ACD databases, respectively [3–5]. Publications have also been made to characterise stationary phases for hydrophilic i
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