Assessment and application of Marfey's reagent and analogs in enantioseparation: a decade's perspective

Of the various methods available for high-performance liquid chromatography separation of enantiomers (of e.g. amino acids and amino group containing compounds) by the pre-column derivatization approach, use of Marfey's reagent has been most successful with continued application since its introduction in 1984. The reagent is prepared from difluoro dinitro benzene by nucleophilic substitution of one of its F atoms byl-alanine amide. There is flexibility to prepare several chiral variants (by substituting the F atom with different chiral auxiliaries) and to tailor the hydrophobicity and resolution, ultimately, of the diastereomeric derivatives. The present paper assesses and reviews applications of Marfey's reagent and its chiral variants (i.e. other FDNP reagents) for enantioseparation of certain amino group containing drugs/amino acids, and to provide some case studies on enantiomeric separations that are important for the pharmaceutical industry. Various explanations for separation mechanism and elution order using FDNP reagents are included and the question of the configuration of the corresponding enantiomer using an indirect approach has also been addressed.

Automated summary

Marfey's reagent, introduced in 1984, has been the most successful method for enantioseparation of amino acids and amino group containing compounds through pre-column derivatization in high-performance liquid chromatography. By substituting the F atom with different chiral auxiliaries, multiple chiral variants can be prepared to tailor the hydrophobicity and resolution of diastereomeric derivatives. Various applications of Marfey's reagent and its chiral variants for enantioseparation of certain amino group containing drugs/amino acids have been assessed in this study.