Rapid Separation and Quantification of Abscisic Acid from Plant Tissues Using High Performance Liquid Chromatography

Abstract

Abscisic acid (ABA) was purified from soybean (Glycine max [L.]) seed extract using a preparative high performance liquid chromatography (HPLC) procedure. The preparative procedure was rapid (70 minutes per sample), required no prior partitioning for purification and was quantitative as demonstrated with an internal standard of [2-(14)C]ABA, of which 98.9% was recovered.Following purification by the preparative HPLC procedure, the ABA in a soybean seed extract was quantified using either GLC with an electron capture detector (GLC-EC) or by analytical HPLC with a UV detector. For soybean seed extracts, two analytical HPLC column packing materials were found adequate: muPorasil and muBondapak-NH(2) (Waters Associates). However, with complex tissue extracts, such as soybean leaf and nodule tissues, only GLC-EC had the necessary selectivity and sensitivity.