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Articles

Purification and Separation of Plant Gibberellins from Their Precursors and Glucosyl Conjugates ¹

Department of Biology, University of Calgary, Calgary, Alberta T2N 1N4 Canada

A procedure using two 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 two fractions: (i) gibberellin (GA) precursors, and (ii) free GA/GA methyl esters (GA-Me)/GA glucosyl 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.

¹ Supported by Natural Sciences and Engineering Research Council Grants A-2585 to R. P. P., GO-633 to R. P. P. and J. N. Owens, and GO-154 to J. D. Bewley, D. M. Reid, and R. P. P.

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