The separation of myo-inositol phosphates by ion pair, reverse-phase high performance liquid chromatography has been investigated. The retention of the inositol phosphates is dependent on both the polarity of the hetaeron utilized and on the pH of the solvent. A method is presented which permits the isocratic separation of multiple forms of inositol phosphates including isomers of myo-inositol trisphosphate. This method appears to be superior to the anion exchange based systems currently employed because of smaller retention volumes, the low ionic strength of the solvent employed, the absence of a requirement for reequilibration, and the ability to perform separations isocratically.