"Two-dimensional" molecularly imprinted solid-phase extraction coupled with crystallization and high performance liquid chromatography for fast semi-preparative purification of tannins from pomegranate husk extract.

In this study, "two dimensional" molecularly imprinted solid-phase extraction (2D-MIP-SPE) of semi-preparative grade was constructed to fast purify ellagitannins in pomegranate husk extract with the help of crystallization and reverse-phase liquid chromatgoraphy (RPLC). Ellagic acid and punicalagin imprinted polymers were synthesized in batch mode and two semi-preparative MIP-SPE columns were individually packed. After investigaing "functional complementation", 2D-MIP-SPE was constructed using ellagic acid MIP and punicalagin MIP-SPE as the first and second dimension, respectively. Then, pomegranate husk extract was fast divided into four fractions individually enriching in ellagic acid, granatin A, punicalagin and ellagic acid glucoside by 2D-MIP-SPE. With the aid of crystallization and RPLC, ellagic acid (13.5mg) and punicalagin (53.4mg) were fast obtained in 30min. Ellagic acid glucoside was purified to the purity near 100% with a recovery of 86.1%. Granatin A (92%) was directly obtained by 2D-MIP-SPE with the recovery of 81.8%. All above indicated that 2D-MIP-SPE was highly efficient in natural product purification. The concept of "functional complementation" was expected to be a useful tool in the construction of 2D-MIP-SPE.

Synthesis and evaluation of sulfobetaine zwitterionic polymer bonded stationary phase.

Zwitterionic polymer stationary phases have attracted increasing attention in hydrophilic interaction chromatography (HILIC). In this work, a zwitterionic sulfobetaine functionalized polyacrylamide stationary phase (named TENS) based on porous silica particles was prepared via controlled surface initiated reversible addition-fragmentation transfer (RAFT) polymerization. Instead of traditional methacrylate type sulfobetaine monomer, acrylamide type sulfobetaine monomer, which has higher chemical stability and hydrophicility, was employed in this work. The characterization of elemental analysis and solid-state 13C cross polarization/magic-angle-spinning nuclear magnetic resonance indicated the successful preparation of TENS stationary phase. Meanwhile, scanning electron microscope (SEM), nitrogen adsorption experiment and study of size exclusion performance were conducted, revealing that the surface initiated polymerization was well controlled. For better understanding of TENS material under HILIC mode, chromatographic evaluation of TENS material was performed, among which, TENS material exhibited good hydrophilicity and chemical stability. To further study the applicability of TENS material, saccharides which were considered as challenging targets in HILIC, were chosen as tested analytes. Various saccharide samples, including fructooligosaccharide, trisaccharide isomers and ginsenosides, were well separated on TENS material. Moreover, TENS material displayed good selectivity for the enrichment of glycopeptides. These results demonstrated the capability of TENS as a promising material in glycomics and glycoproteomics.

A controlled thiol-initiated surface polymerization strategy for the preparation of hydrophilic polymer stationary phases.

A controlled thiol-initiated surface polymerization strategy has been successfully developed and employed to prepare hydrophilic polymer stationary phases, which exhibited excellent chromatographic performance and protein non-fouling properties.