Study Of Hydrophilic Interaction Chromatography Stationary Phases And Its Salt Effects

In this paper,chromatography silica gel was used as the substrate,the functional monomer N,N-dimethyl-N-acryloyloxyethyl-N-ammonium propane sulfonate(DMAEAPS)and N-vinylimidazole-N-ammonium propane sulfonate(VIMPS)were grafted on the surface of Sil-Br by atom transfer radical polymerization(ATRP),the sulfobetaine zwitterionic hydrophilic interaction chromatography stationary phase Sil-DMAEAPS and Sil-VIMPS were obtained.The chromatographic properties of the polar solute were investigated by observing the retention of the polar solute on the selfmade chromatographic stationary phase.As the same time,investigated the salt effect on the four different kinds of phase,those were Silica gel column(Kinetex silica),Amino column(Luna NH2),Betaine type amphoteric ioncolumn(ZIC(?)-HILIC)and Phosphorylcholine modified silica gel column(ZIC(?)-cHILIC).It's expected to provide a reference value for the separation of chinese medicine components.The paper mainly includes the following parts:1.N,N-dimethyl-N-acryloyloxyethyl-N-ammonium propane sulfonate(DMAEAPS)zwitterionic sulfobetaine functional monomer was synthesized.Dimethyl amino ethyl acrylate(DMAEA)and 1,3-propyl sulfonic acid ester as raw materials were used to synthesis the functional monomer,the DMAEAPS functional monomer was characterized by 1H-NMR and FT-IR.2-bromoisobutyryl bromide allyl silicone(Sil-Br)as the initiator,CuBr and 2,2-bipyridyl(Bpy)as the catalytic system,the functional monomer DMAEAPS was grafted on the surface of Sil-Br by ATRP,and the sulfobetaine zwitterionic hydrophilic interaction chromatography stationary phase(Sil-DMAEAPS)was obtained.The Sil-DMAEAPS stationary phase was characterized by element analysis.Chromatographic evaluation in hydrophilic interaction chromatography(HILIC)mode showed that the materials were suitable for use as stationary phase,the column showed a dual retention mechanism.HILIC mode in acetonitrile-rich(acetonitrile/water>0.7,v/v)mobile phases and RP mode in highly aqueous(acetonitrile/water<0.2,v/v)mobile phases.The separation selectivity and retention of the five polar solutes(benzoin,pyridoxine,rutin hydrate,p-coumaric acid and caffeic acid)on the Sil-DMAEAPS stationary phase were different from commercially available ZIC(?)-HILIC.A method for the determination of rutin hydrate in rutin tablets and ferulic acid in angelica,cimicifuga were established.It was simple and accurate for the determination of polar compounds.2.In this paper,1-vinyl imidazole and 1,3-propane sultone as raw material,reaction at 0? for 20 h,Synthesised of N-vinylimidazole-N-ammonium propane sulfonate(VIMPS)functional monomer by ATRP,functional monomer grafted onto the brominated silica gel initiator agent prepared containing betaine type zwitterionic stationary phase;used 1H-NMR and FT-IR methods to characterize functional monomer;N-vinylimidazole-N-ammonium propane sulfonatezwitterionic hydrophilic interaction chromatography stationary phase(Sil-VIMPS)successfully prepared by elemental analysis.This experiment also select 8 kinds of polar solutes by control acetonitrile content,pH,salt concentration to study on this chromatographic stationary phase solute reservations,the results showed that the synthesized chromatographic stationary phases have a good selection and the separation of hydrophilic properties.3.Salt effect on the change of HILIC selectivity has an important role.A systematic study of uracil B1,uridine,adenine,adenosine,cytosine,cytidine,benzoin,penicillin,pyridoxine,ferulic acid,niacin,caffeic acid and chlorogenic acid,by adjusting the salt concentration and the type of buffer systems studied polar solute component retention behavior in hydrophilic interaction chromatography.The results showed that ammonium formate has a greater influence on solute retention,ammonium sulfate has a less influence on solute retention that salt effect of hydrophilic interaction chromatography affect solute retention and the separation.