Liquid Chromatographic Determination of Unbound and Acetone-Soluble Bound Gossypol in Cottonseed Meals and Mixed Feeds

Abstract

Abstract A liquid chromatographic method has been developed for the determination of unbound and acetone- soluble bound gossypol in cottonseed meals and mixed feeds at levels of 0.5 ppm. The method involves extraction with aqueous acetone in the presence of ascorbic acid, hydrolysis of the "soluble bound" forms with hydrochloric acid, partitioning into chloroform, and chromatographic separation on a 10 µm C18 column by step gradient elution using a methanol-water mobile phase acidified with phosphoric acid. With minor modifications, the method permitted discriminate determination of unbound gossypol and acetone-soluble hydrophilic and lipophilic forms of bound gossypol. The gossypol peak was characterized by on-line spectral scanning and absorbance rationing. Overall relative standard deviation was 6.7% and overall recovery was 98.1 ± 3.3%. When the method was applied to several cottonseed meal samples, results were inconsistent with those obtained by the official American Oil Chemists' Society method for "free" gossypol determination.