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Preparation of Medium Cation Exchange Stationary Phase of Polymeric Matrix and Their Chromatographic Properties
Author(s) -
Chen Gang,
Gong BoLin,
Bai Quan,
Geng XinDu
Publication year - 2007
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.200790021
Subject(s) - chemistry , lysozyme , chromatography , dispersity , stationary phase , ion exchange , glycidyl methacrylate , ion exchange resin , phase (matter) , salt (chemistry) , matrix (chemical analysis) , ion , polymer , organic chemistry , polymerization , biochemistry
Based on the monodisperse poly(glycidyl methacrylate‐co‐ethylenedimethacrylate) beads (PGMA/EDMA) with macropore as a medium, a new hydrophilic medium cation exchange (MCX) stationary phase for HPLC was synthesized by a new chemically modified method. The stationary phase was evaluated with the property of ion exchange, separability, reproducibility, hydrophilicity, effect of salt concentration, salt types, column loading and pH on the separation and retention of proteins in detail. It was found that it follows ion exchange chromatographic (IEC) retention mechanism. The measured bioactivity recovery for lysozyme was (96±5)%. The dynamic protein loading capacity of the synthesized MCX packings was 21.8 mg/g. Five proteins were almost completely separated within 6.0 min at a flow rate of 4 mL/min using the synthesized MCX resin. The MCX resin was also used for the rapid separation and purification of lysozyme from egg white with only one step. The purity and specific bioactivity of the purified lysozyme was found more than 95% and 70345 U/mg, respectively.