Preparation and evaluation of new Pirkle type chiral stationary phases with long alkyl chains for the separation of amino acid enantiomers derivatized with NBD-F

In order to improve the high-performance liquid chromatographic separation of alpha-amino acids derivatized with the fluorogenic reagent 3-fluoro-7-ni tro-2,1,3-benzoxadiazole (NBD-F) on commercially available chiral stationar y phases (CSPs) such as SUMICHIRAL OA-2500(S) (CSP 1) and OA-4700 (CSP 3), the preparation of two new CSPs (CSP 2 and CSP 3) having 11-aminoundecanoic acid between the aminopropyl silica gel support and the chiral moiety in C SP 1 and CSP 3 is described. CSP 2 and CSP 4 improved both the mutual and e nantiomeric separation of NBD-amino acids compared with CSP 1 and CSP 3. Th us, 17 pairs of NBD-amino acid enantiomers and NBD-glycine were separated o n CSP 2 except for six NBD-amino acids (D-Asn, D-Ser, D-Gln, L-Pro, L-Ser a nd Gly). CSP 2 and CSP 4 also showed better enantiomeric separation of NBD- amino acid esters and amides than CSP 1 and CSP 3. It was considered that t he achiral long alkyl chains in the CSPs might form a hydrophobic space whi ch assisted the stereoselective interaction of analytes with the chiral moi ety by changing the environment around the chiral moiety. On CSP 1 and CSP 2, NBD-beta-amino acid was also enantiomerically separated.