ABSTRACT
A novel glycosyl amino acid hydrophilic interaction chromatography (HILIC) stationary phase was prepared via click chemistry. The key intermediate N(3)-glycosyl D-phenylglycine was prepared by a three steps procedure, including selective condensation of amino glucose with N-succinimidyl ester of Boc-D-phenylglycine, deprotection and transformation of amino group to azido group. The structure of all the intermediates and functionalized silica beads were confirmed by (1)H NMR, IR, elemental analysis and (13)C CP-MAS. The chromatography test showed that this new type of separation material possessed good HILIC properties and glycopeptide enrichment characteristics. Nucleosides and bases could be separated in a simple eluent composition (only acetonitrile in combined with water), and with the same condition, these model compounds could not be separated on the commercial HILIC column (Atlantis). Click glycosyl amino acid thus prepared also showed longer retention and better separation ability in the separation of polar organic acids.
Subject(s)
Amino Acids/chemistry , Chromatography, High Pressure Liquid/instrumentation , Chromatography, High Pressure Liquid/methods , Glycopeptides/chemistry , Acetonitriles/chemistry , Glycopeptides/chemical synthesis , Glycosylation , Humans , Hydrophobic and Hydrophilic Interactions , Immunoglobulin G/analysis , Immunoglobulin G/chemistry , Immunoglobulin G/isolation & purification , Models, Chemical , Molecular Structure , Nucleosides/analysis , Nucleosides/chemistry , Nucleosides/isolation & purification , Purines/analysis , Purines/chemistry , Purines/isolation & purification , Pyrimidines/analysis , Pyrimidines/chemistry , Pyrimidines/isolation & purification , Reproducibility of ResultsABSTRACT
A novel chitooligosaccharide stationary phase for hydrophilic interaction liquid chromatography (HILIC) was developed via click chemistry and showed great HILIC characteristics on separation of polar compounds and enrichment of glycopeptides.
Subject(s)
Chromatography, Liquid/instrumentation , Oligosaccharides/chemistry , Amino Acids/chemistry , Amino Acids/isolation & purification , Carbohydrates/chemistry , Carbohydrates/isolation & purification , Catalysis , Chromatography, Liquid/methods , Copper/chemistry , Glycopeptides/chemistry , Glycopeptides/isolation & purification , Nucleosides/chemistry , Nucleosides/isolation & purification , Oligosaccharides/chemical synthesis , Silicon Dioxide/chemistryABSTRACT
2D-HPLC is an important technique for the separation of complex samples. Developing new types of stationary phases is of great interest to construct 2D-LC systems with high orthogonality. In this study, a novel stationary phase-Click dipeptide (l-phenylglycine dipeptide) was prepared by immobilization of alpha-azido l-phenylglycine dipeptide on alkyne-silica via click chemistry. In the preparation of this new material, an efficient, inexpensive and shelf-stable diazo transfer reagent (imidazole-1-sulfonyl azide hydrochloride) was utilized to transfer the amino group of l-phenylglycine to corresponding azido group under mild conditions. The Click dipeptide thus prepared was confirmed by FT-IR, solid state CP/MAS (13)C NMR and elemental analysis. The Click dipeptide packing showed high orthogonality with C18, which reached 63.5%. An off-line 2D-RP/RPLC system was developed to analyze a traditional Chinese medicine (TCM)-Rheum Palmatum L. The results showed high orthogonality between Click dipeptide and C18 as well as great separating power in the practical separation of complex samples.
