A Novel Silica-Based Metal Chelate Stationary Phase—L-Glutamic Acid–Copper(II)

A novel silica-based metal chelate stationary phase, L-glutamic acid-copper(II) (L-Glu-Cu(II)), was synthesized. Effects of the immobilized metal ion, the pH of the solution, and competitive agents on chromatographic behavior of proteins were investigated on prepared chelate column. The coordination role of proteins on L-Glu-Cu(II) column was demonstrated. Optimal separation conditions for proteins on L-Glu-Cu(II) column were discussed. According to the established chromatographic conditions, protein mixtures were effectively separated. Separation performance of proteins and the leakage of Cu2+ on L-Glu-Cu(II) column were discussed and compared with the traditional iminodiacetic acid-copper(II) (IDA-Cu(II)) column. Separation performance of L-Glu-Cu(II) column for proteins was superior to that of the IDA-Cu(II) column. Under a proper eluting condition, a less amount of Cu2+ was leaked from the L-Glu-Cu(II) column as compared with IDA-Cu(II) column.