Chiral organic-silica hybrid monolithic HPLC column with dynamic grafting BSA for the enantiomeric separation

BSA functionalized hybrid monolithic column prepared by in-situ dynamic grafting on traditional stainless steel column for enantiomeric separation. A hybrid silica monolithic column (HMC) was prepared using a single precursor (methytrimethoxysilane) in stainless steel HPLC column (50 mm × 4.6 mm) by sol–gel method. The chiral hybrid monolithic column (CHMC) was prepared by grafting 3-aminopropyl trimethoxysilane onto HMC, and then reacting bovine serum albumin (BSA) with Glutaraldehyde. The preparation conditions were optimized, and the maximum grafting amount of BSA was 48.6 mg g−1. The CHMC was characterized and its chromatographic performance was tested. The results showed that the CHMC has a homogeneous network skeleton structure, good stability and reproducibility. The CHMC was combined with HPLC, under optimized chromatographic conditions, the baseline separation of tryptophan and pidotimod enantiomers were successfully achieved.