Separation of cytokinins by High Pressure Liquid Chromatography

Summary A number of cytokinin reference compounds have been successfully separated by High Pressure Liquid Chromatography using columns of pellicular strong cation exchange resin and of pellicular polyamide. On polyamide, all cytokinins were eluted within 10 min with an aqueous buffer but on the cation exchange resin some cytokinins (generally those with bulky N⁶-substituents and in addition lacking an N⁹-ribosyl substituent) had excessively long retention times with aqueous buffer eluent. However, addition of methanol to the buffer enabled these cytokinins to be separated and eluted within a reasonable time. As small an amount as 5 nanogram of cytokinin could readily be detected by the procedures described.