Glyphosate is a broad spectrum herbicide used to kill weeds. It is the most used herbicide. Glyphosate is an aminophosphonic analogue of the natural amino acid glycine. Glyphosate and its intermediates are very polar ionic compounds derived from glycine. Neither of intermediates can be retained on traditional reversed-phase columns. Two methods for glyphosate intermediates were developed on Primesep B, Primesep B2 and Primesep 100 columns.