In this paper, L (þ)-Cysteine (L-Cys) and 5-norbornene-2-carboxylic acid (5-N-2-CA) were modified on the silica
spheres by cross-linking polymerization and thiol-ene click reaction. The Cys-Si-NCA was characterized by
scanning electron microscopy, nitrogen sorption porosimetry, thermogravimetric analysis and other character-
ization methods. The results indicated that the colume had a regular spherical shape with porous surfaces and
particle sizes mainly ranging from 4.5 to 5.5 μm. The surface area of the Cys-Si-NCA microspheres was 328.68
m2 g?1, and their respective average pore size was 9.27 nm. Furthermore, the colume exhibited excellent sta-
bilities and reproducibility. Experimental results demonstrate that this new stationary phase can facilitate the
separation of sulfonamides, nucleosides and nucleobases compounds in hydrophilic chromatographic (HILIC)
conditions. In addition to being able to attenuate the peak tailing of separation, it can also be used for quanti-
tative analysis of creatinine in urine. We also give the temperature and pH ranges of separation on Cys-Si-NCA
column. The successful synthesis of the Cys-Si-NCA stationary phase provides foundation for the application of
this kind of materials in liquid chromatography (LC).