Abstract
This article focuses on correlating the column classification obtained from the method created at the Katholieke Universiteit Leuven (KUL), with the chromatographic resolution attained in biomedical separation. In the KUL system, each column is described with four parameters, which enables estimation of the FKUL value characterising similarity of those parameters to the selected reference stationary phase. Thus, a ranking list based on the FKUL value can be calculated for the chosen reference column, then correlated with the results of the column performance test. In this study, the column performance test was based on analysis of moclobemide and its two metabolites in human plasma by liquid chromatography (LC), using 18 columns. The comparative study was performed using traditional correlation of the FKUL values with the retention parameters of the analytes describing the column performance test. In order to deepen the comparative assessment of both data sets, factor analysis (FA) was also used. The obtained results indicated that the stationary phase classes, closely related according to the KUL method, yielded comparable separation for the target substances. Therefore, the column ranking system based on the FKUL-values could be considered supportive in the choice of the appropriate column for biomedical analysis.
Highlights
The continuously broadened application of reversed-phase high-performance liquid chromatography (RP-HPLC, RPLC) in many various areas, including drug analysis, has expanded the demand for new generations of RPLC stationary phases packing columns of different geometry which would offer better selectivity, efficiency and chemical stability
Many papers reporting methods of characterising stationary phases have been published to date to resolve this problem, including the method proposed by Galushko [5], the chromatographic tests reported by Stella et al [6,7], the quantitative structure-retention relationships (QSRRs) delivered by the Kaliszan group [8,9,10,11], the hydrophobic-subtraction model (HSM) published by the Snyder-Dolan group [12,13], and the alternative propositions to the LC column selectivity introduced by Tanaka [14], Euerby [15,16,17,18], Visky [19], Veuthey [20,21,22] and others [23,24,25,26,27]
LC columns were classified based on the Katholieke Universiteit Leuven (KUL) method and their selectivity toward moclobemide and its two metabolites in human plasma, using 18 RPLC columns
Summary
The continuously broadened application of reversed-phase high-performance liquid chromatography (RP-HPLC, RPLC) in many various areas, including drug analysis, has expanded the demand for new generations of RPLC stationary phases packing columns of different geometry which would offer better selectivity, efficiency and chemical stability. Any column classification method needs to undergo an important test to verify whether stationary phases with similar parameters will give comparable separations in real pharmaceutical and biomedical applications. Most papers only concern comparative analyses of the FKUL parameter values determined for stationary phases against the pharmacopoeial test known as System Suitability Test (SST) or Chromatographic Response Function (CRF), conducted to evaluate the separation of such columns in real pharmaceutical applications [28,29,30,31,32,33,34,35,36,37]. The FA results were compared to the results of the principal component analysis (PCA) and hierarchical clustering analysis (HCA) previously reported in the literature [28,38,43]
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