A teicoplanin-cyclodextrin bilayer chiral stationary phase boosts chiral separation of native amino acids

Separating enantiomers is crucial for advancing scientific research. Teicoplanin-based columns exhibit unique advantages in the chiral separation of native amino acids, and its further improvement for better separation performance is valuable. This work reports double-layer bridged columns, in which different amounts of cyclodextrin (CD) were directly bridged onto teicoplanin (TK) molecules on the chiral stationary phase (TK CSP) to prepare TK-CD-2 CSP and TK-CD-6 CSP. Additionally, a column with the two chiral selectors (TK and CD) arranged in a single layer on the silica gel surface (TK + CD CSP) was prepared, which served as a control group. The results revealed that TK-CD-6 CSP exhibited better chiral separation ability for native amino acids and a wider range of separable chiral molecules than TK, TK + CD, and TK-CD-2 CSPs. The possible mechanism is that the dual-layer structure of the TK-CD-6 CSP effectively reduces the competition between the different chiral selectors and provides more chiral recognition sites, which enhances the separation ability and achieves broad-spectrum chiral separation. This work offers valuable insights for enhancing the chiral separation ability of teicoplanin-based columns and developing liquid chromatographic strategies with separation diversity.