Preparation and chromatographic evaluation of a newly designed steviol glycoside modified-silica stationary phase in hydrophilic interaction liquid chromatography and reversed phase liquid chromatography.
A diterpene glycoside compound, rebaudioside A (commonly abbreviated as RA), was immobilized onto porous silica surface through "thiol-ene" click chemistry strategy. The successful immobilization of the RA on the silica support was confirmed by FT-IR and elemental analysis. Chromatographic characteristics of the new stationary phase, named Click TE-RA, were evaluated by a set of diverse analytes such as carbohydrates, nucleosides, and organic acids in hydrophilic interaction liquid chromatography (HILIC) mode. The effects of water content, buffer pH and concentration were investigated and a typical HILIC retention feature of Click TE-RA was observed at high organic modifier content. The Click TE-RA stationary phase was further studied by a series of glycoside compounds. Tunable retention mechanisms from hydrophilic to hydrophobic interactions were observed. Separation of very polar compounds including oligosaccharides, nucleic acid bases and nucleosides using Click TE-RA in HILIC mode was successfully accomplished. In addition, separation of saponins both in HILIC and reversed-phase liquid chromatography (RPLC) modes was performed, demonstrating the presence of orthogonality between two different modes on Click TE-RA column. The multiple interactions induced by polar sugar group and hydrophobic aglycone group allowed this Click TE-RA to serve as a multi-mode stationary phase in two-dimensional liquid chromatography.
Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China.
Tu
T
Liang
Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China.
Qing
Q
Fu
Key Lab of Separation Science for Analytical Chemistry, Key Lab of Natural Medicine, Liaoning Province, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, PR China.
Aijin
A
Shen
Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China.
Hui
H
Wang
Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China.
Yu
Y
Jin
Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China.
Huaxia
H
Xin
Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China.
Yanxiong
Y
Ke
Key Lab of Separation Science for Analytical Chemistry, Key Lab of Natural Medicine, Liaoning Province, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, PR China. Electronic address: guozhimou@dicp.ac.cn.
Zhimou
Z
Guo
Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China; Key Lab of Separation Science for Analytical Chemistry, Key Lab of Natural Medicine, Liaoning Province, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, PR China. Electronic address: liangxm@ecust.edu.cn.
Xinmiao
X
Liang
D002241:Carbohydrates / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D057930:Click Chemistry; D045786:Diterpenes, Kaurane / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D009930:Organic Chemicals / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared; D014867:Water / Q000737:chemistry
D002241:Carbohydrates; D045786:Diterpenes, Kaurane; D009930:Organic Chemicals; D014867:Water; D012822:Silicon Dioxide; C025474:rebaudioside A
Hydrophilic interaction liquid chromatography (HILIC); Mixed-mode; Rebaudioside A; Stationary phase; Thiol-ene click chemistry