Gossypol analysis in cottonseed oil by HPLC

Gossypol in cottonseed oil was selectively separated by the extraction of cottonseed oil with a hexane and N,N dimethyl formamide:water (2∶1, v/v) solvent mixture. After filtration, the extract was injected into the HPLC with the elution time less than 15 min. The spectrophotometric method showed 2 to 5 times higher values of gossypol content in different types of glanded cottonseed oil than did the HPLC method. This is probably due to the gossypol derivatives and coloring interferences reacting with p-anisidine to develop color and increasing the absorbance reading, whereas gossypol was separated and detected in the HPLC method.