Detection of Inositol and Inositol Diphosphate on Paper Chromatograms

Abstract

A COLOUR reaction, known as Scherer's test, is extensively used as a specific, qualitative test for inositol. It is based on the formation of the coloured alkaline earth metal salt (usually calcium salt) of rhodizonic acid on the oxidation of inositol with concentrated nitric acid^1,2. While studying phos-pholipids containing inositol we needed to identify accurately very small amounts of inositol by paper chromatography, or by spot tests on filter paper. P. Fleury et al. ³ reported a method in which they used mercuric oxide as a catalyst of the oxidation, and barium acetate as for the alkaline earth metal. This method, however, is tedious and sometimes gives uncertain results; the smallest quantity detectable is 5–10 µgm. This may be ascribed to the formation of yellow mercuric salt at the first stage of the reaction. A more sensitive, reliable and easier method was, therefore, devised by modifying that of Fleury. This modified method can, in addition, be applied to the detection of inositol diphosphate located on paper chromatograms. The procedure is as follows.