A reverse-phase High Performance Liquid Chromatography (HPLC) column (3.9 x 150.0 mm) was used to determine amino acid composition in palm kernel cake (PKC) in 40 min (based on Waters AccQ Tag method). The amino acid analysis was carried out in three steps i.e., substrate hydrolysis, chromatographic separation and data analysis. 18 amino acids were separated after precolumn derivatisation. The amino acids separated were glutamic acid, aspartic acid, alanine, arginine, cystine, glycine, histidine, isoleucine, leucine, lysine, methionine. phenylalnine, proline, serine, threonine, tyrosine and valine. Glutamic acid and aspartic acid were the major non-essential amino acids in palm kernel cake accounting for 3.01% and 2.08% of the total recovered amino acids respectively. Trythopan, methionine and cystein which were destroyed during hydrolysis with Hydrochloric acid (HCl) required special method for the analysis.