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 min/sample), required no prior partitioning for purification and was quantitative as demonstrated with an internal standard of [2-14C]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, 2 analytical HPLC column packing materials were found adequate: .mu.Porasil and .mu.Bondapak-NH2. With complex tissue extracts, such as soybean leaf and nodule tissues, only GLC-EC had the necessary selectivity and sensitivity.