Application of Humic Acid Bonded‐Silica as a Hydrophilic‐interaction Chromatographic Stationary Phase in Separation of Polar Compounds

Abstract

In this paper, humic acid bonded silica (HAAS) stationary phase was prepared through the spacer of aminoalkyl silanes. The high performance liquid chromatographic behavior of several polar compounds, including four alkaloids and clenbuterol, was studied on HAAS. The effect of mobile phase variables such as organic solvent content, buffer pH, and ionic strength on their chromatographic behavior was investigated. The retention mechanism of tested compounds on the stationary phase was elucidated. The results indicate that the HAAS stationary phase behaved as hydrophilic interaction chromatographic packing, using high content organic solvent as mobile phase, and the stationary phase showed good separation selectivity for four alkaloids and clenbuterol.

Keywords:

• Polar compounds
• Separation
• Humic acid
• Hydrophilic interaction chromatography

Acknowledgments

This work was partly supported by grants from the National Science Fund for Distinguished Young Scholars (No. 20625516) and the Program for New Century Excellent Talents in University (NCET‐05‐0616).