[Preparation and evaluation of new ion-exchange chromatographic stationary phase for the use in high performance liquid chromatography].
A new method for the bonding of diethylamine(DEA) on the surface of silica to prepare novel hydrophilic packings for HPLC has been studied. After allyl glycidyl ether being synthesized, the Si-DEA anion-exchange bonded phase was prepared by the reaction of the double bond in allyl group with Si-H silica. The bonded phases obtained were characterized by elemental analysis, diffuse reflectance infrared Fourier transform(DRIFT) spectroscopy and HPLC evaluation. The methods were used for both porous silica and monodisperse non-porous silica. The contents of carbon, hydrogen and nitrogen of porous Si-DEA packing (MPS-DEA) were 3.31%, 0.95% and 1.34% respectively and those of monodisperse non-porous Si-DEA packing (NPS-DEA) were 2.55%, 0.97% and 0.96% respectively. The diethylamine absorption peak can be observed at 2970 cm-1 from the Si-DEA silica DRIFT spectrum. These data revealed that the diethylamine had been bonded on MPS-DEA and NPS-DEA packings. In HPLC tests, nucleotides and nucleosides such as cytosine, uracil, cytidine-5'-monophosphate, adenosine-5'-monophosphate, inosine-5'-monophosphate and guanosine-5'-monophosphate were satisfactorily separated on the porous anion-exchange packing (MPS-DEA), and a group of proteins (lysozyme, ribonuclease, ovalbumin, bovine serum albumin, insulin and gamma-globulin) were separated within 15 minutes successfuly. All test results indicated that the new method for preparing better anion-exchange silica packings is effective for both porous silica and monodiperse non-porous silica.
Lei
X
X
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116012, China.
Wang
J
J
Cong
R
R
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003597:Cytosine Nucleotides / Q000302:isolation & purification; D004048:Diethylamines; D004852:Epoxy Compounds; D007291:Inosine Monophosphate / Q000302:isolation & purification; D009113:Muramidase / Q000302:isolation & purification*; D012260:Ribonucleases / Q000302:isolation & purification; D012822:Silicon Dioxide; D017550:Spectroscopy, Fourier Transform Infrared
D003597:Cytosine Nucleotides; D004048:Diethylamines; D004852:Epoxy Compounds; D007291:Inosine Monophosphate; D012822:Silicon Dioxide; C034281:diethylamine; D012260:Ribonucleases; D009113:Muramidase; C054361:allyl glycidyl ether