Heart-cut achiral-chiral LC-LC method development using factorial design: application to the chiral separation of ketoprofen

Abstract

A two-dimensional achiral-chiral LC-LC method in heart-cut mode for ketoprofen and its enantiomeric fraction determination was proposed. A C column was used in the first dimension, and the chiral column was an α-acid glycoprotein. The mobile phase of the chiral system was optimized by a factorial design. The effect of temperature on retention and on enantiomeric resolution was studied. Particular attention was paid to mobile phase compatibility for the two columns and to transferring time, using ketoprofen standards. The R-(-) and S-(+)-ketoprofen retention times were 9 and 11 min, respectively; the resolution was higher than 1.1 and enantiomeric fraction close to 0.5. The method was applied to capsules and gels containing ketoprofen. Factorial design was also used to establish the best conditions for gel sample preparation. Recoveries were 84 and 105 % for capsules and gels, respectively. Graphical abstract Two-dimensional chromatogram for KPF and its enantiomers.

Determination of profens in pharmaceuticals by heart‐cut achiral‐chiral two‐dimensional HPLC

“…Based on a previous work using an α 1 ‐AGP column for KPF chiral determination , enantiomer separation of the three PROs, KPF, NAP and IBU, was tried; however, in the conditions specified there KPF and IBU enantiomers overlapped. It must be emphasized that the order of enantiomer elution in this column was NAP, KPF, IBU, that is, KPF and NAP eluted in a different order in the achiral column.…”

“…In this context, 2‐D HPLC helps by cleaning‐up the PRO racemates in a first dimensional achiral column and further separating their enantiomers in a second chiral column; in this way, the sample matrix is removed and selectivity is improved . The heart‐cut mode is most often used to transfer the racemic from the first column to the second column; it consists in transferring a portion of the peak center of the selected racemic from the first column to the second column in the 2‐D HPLC . With respect to the CSP column, one of the most used is α 1 ‐acid glycoprotein (AGP), which probably covers one of the broadest fields of application; it has been employed to separate a wide range of basic, neutral and acidic drug racemates as well as compounds with different structures ; it must be emphasized that this column may allow the simultaneous separation of several enantiomer.…”

Deducing formation routes of oxylipins by quantitative multiple heart-cutting achiral-chiral 2D-LC-MS

“…This is common for achiral-chiral (multiple) heart-cutting methods, but not applicable for the structurally diverse class of oxylipins. Other 2D methods with a chiral second dimension aim to analyze individual heart-cuts, so that 2 D runtimes from four 23 , six 25 , or seven minutes 26 up to more than 30 min 43 are used, which is not applicable for multiple heart-cutting methods.…”

“…Achiral-chiral heart-cutting 2D-LC is frequently used for the analysis of amino acids 23,24 or pharmaceuticals. 25,26 By loop-based multiple cutting, the collection of a large number of 1 D sections and subsequent 2 D analysis is possible. 27 Despite its great potential, this has hardly been exploited for comprehensive targeted metabolomics.…”

Recent Advances in Chromatography for Pharmaceutical Analysis

“…Another heart-cutting achiral-chiral LC-LC method, involving RPLC (C8 phase) in the first dimension and a chiral column (alpha-1-acid glycoprotein) in the second dimension, was used for the successful separation of ketoprofen enantiomers. 35 The experimental setup included factorial designs as a mathematical approach to allow for the easy optimization of both separation conditions, with a focus on the compatibility of the mobile phases.…”