The chromatographic identification of some biologically important phosphate esters

the objective of the present work was to provide a means for separating and indentifying phosphate esters involved in glycolysis in higher plants. Paper chromatography of phosphate esters has been employed by several workers, most notably Benson et al. (1) and Hanes and Isherwood (2). Benson's procedures were not primarily designed for identification of phosphate esters and gave low Rr values for the phosphate compounds of particular interest to us. The unidimensional methods of Hanes and Isherwood do not result in adequate resolution of the complex mixtures such as are obtained from our plant materials. The present procedure is based on two-dimensional chromatography with successive development in an acid and in a basic solvent. The solvents finally selected gave the best over-all resolution of the intermediates involved in plant glycolysis. Undoubtedly the resolution of certain pairs of compounds may be improved by suitable modifications. We have in addition made certain improvements in the procedure for locating the chromatographed materials.