Wenya Feng^a,c, Juan Qiao^a,b,*, Dan Li^d, Li Qi^a,b,*

Abstract: Utilizing block copolymers as coatings, a protocol of chiral ligand exchange capillary electrochromatography (CLE-CEC) protocol was designed and developed with dual ligands for D,L-amino acids enantioseparation. Four block copolymers incluing poly maleic anhydride-co-styrene-co-N-methacryloyl-L-histidine methyl ester [P(MAn-St-MAH)], poly maleic anhydride-co-styrene-co-N-methacryloyl-L-lysine methyl ester [P(MAn-St-MAL)], poly maleic anhydride-co-styrene-co-N-methacryloyl-L-phenylalanine methyl ester [P(MAn-St-MAP)] and poly maleic anhydride-co-styrene-co-N-methacryloyl-L-threonine methyl ester [P(MAn-St-MAT)] were synthesized by reversible addition fragmentation chain transfer polymerization reaction. Key factors affecting the enantioresolution were optimized, including the concentration of Zn (II) central ion, pH value of buffer solution and monomers of the block copolymers. The enantioresolution of the proposed CLE-CEC system could be enhanced dramatically by employing P(MAn-St-MAH) as the immobilized chiral ligand and by coordinating the synergistic effect of free ligand in buffer solution. The principle of improved enantioresolution of the CLE-CEC system with dual ligands was discussed. Well enantioseparation was successfully realized with 7 pairs of D,Lamino acids enantiomers baseline separation and 5 pairs part separation. For quantitative analysis of D,L-alanine, a good linearity was established in the range of 9.4 μM to 1.5 mM (r2 = 0.997) with the limits of detection (LODs) 3.7 μM of D-alanine, 2.0 μM for L-alanine, and limits of quantifcation (LOQs) 9.0 μM for D-alanine and 6.0 μM for L-alanine. The peak area and migration time reproducibility (n = 6) were 4.1% and 3.5% for Dalanine, 3.7% and 3.1% for L-alanine. Further, the enzyme kinetics study of alanine aminotransferase was investigated with the constructed CLE-CEC system.

Keywords: Block copolymer coating; Chiral ligand exchange capillary; electrochromatography; Enantioseparation; Amino Acids                                                                   

文章链接：j.talanta.2018.10.059.pdf