Comparison of chiral chromatography on SFC, and normal and reverse phase Ciara O'Doherty and Joshua Freem Pharmaron, UK The relationship between chiral chromatography methods on normal-phase high performance liquid chromatography (NP-HPLC) and supercritical fluid chromatography (SFC) was investigated to aid in seamless method transfer between techniques, which have been widely assumed to yield similar enantioseparation. The orthogonal technique, reverse-phase high performance liquid chromatography (RP-HPLC) was also investigated as an alternative to the above techniques, potentially yielding unique chromatographic results. A mobile and stationary phase screen in NP-HPLC, RP-HPLC and SFC was performed on four commercially available active pharmaceutical ingredients. Common chiral assay criteria of resolution, tailing and capacity factor for the enantioseparation of these compounds was critically compared. Among the investigated conditions, success was not consistent between the three techniques, with retention mechanisms discussed to potentially provide explanation for this. The results from these simultaneous screens allows a clearer understanding of the compatibility of the three techniques, highlighting certain columns and mobile phases that may lead to a consistent method transfer and allowing the mechanisms underpinning the two techniques to be compared. The outcome of this project aids in the method development for chiral chromatography when moving from a SFC column screen to a HPLC method for use in industry.
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