Purification and Separation of Plant Gibberellins from Their Precursors and Glucosyl Conjugates

Abstract

A procedure using 2 small preparative columns (in sequence) of C18 reverse phase Bondapak B material with methanolic extracts of plant tissue (Pisum sativum L., Malus domestica Borkh., Pimpinella anisum L.) yields 2 fractions: (i) GA precursors, and (ii) free GA/GA methyl esters (GA-Me)/GA glycosyl conjugates. The discrete separation of (iii) free GA/GA-Me from (iv) GA glucosyl conjugates is then accomplished by a combination of differential solvent solubility and SiO2 partition chromatography. All fractions are almost pigment free, and appreciable dry weight purification was accomplished for the GA precursor and free GA/GA-Me fractions. Solvent volumes can be kept low, no buffer salts are introduced, and each fraction (i, iii, iv) can be subjected directly to preparative or analytical reverse phase C18 high performance liquid chromatography without recourse to solvent partitioning, and often without further purification.