ABSTRACT: Partisphere SAX HPLC analysis of endogenous inositol phosphates in [3H]inositol-labelled barley aleurone tissue revealed a range of isomers, including D- and/or L-Ins3P, D- and/or L-Ins(1,4)P2, D- and/or L-Ins(1,2)P2, a third unidentified InsP2, Ins(1,2,3)P3, D- and/or L-Ins(1,2,6)P3, D-and/or L-Ins(1,2,3,4)P4, D- and/or L-Ins(1,2,5,6)P4, Ins(1,3,4,5,6)P5, D- and/or L-Ins(1,2,3,4,5)P5, Ins(1,2,3,4,6)P5, InsP6 and a molecule with the chromatographic properties of an inositol pyrophosphate. The striking match between the identities of the stereoisomers, and in some cases enantiomers, detected in vivo and those stereoisomers produced in vitro by the action of wheat-bran phytase on InsP6 [Cosgrove (1980) Inositol Phosphates: Their Chemistry, Bio-chemistry and Physiology. Elsevier, Amsterdam] strongly suggests that most of the inositol phosphates identified are products of the breakdown of InsP4 by endogenous phytase(s) with stereospecificity similar to that of the wheat-bran enzyme(s).