Determination of amino acids in Buffalo horn by reversed-phase high performance liquid chromatography with o-phthaldialdelhyde pre-colomn derivatization

A pre-column derivatization procedure is described for the determination of amino acids in Buffalo horn by reversed-phase high performance liquid chromatography, o-phthaldialde/hyde/beta-mercaptoethanol was used for the derivatization of amino acids. The separation of amino acid derivatives was accomplished on a column packed with 5 mu m ODS(octadecylsilance). An acetate buffer at pH 5.6 was used as the mobile phase A, while methanol was employed as the mobile phase B. Linear gradient elution was adopted with a fluorescent detector( E-x = 340 nm, E-m = 460 nm). Twelve amino acids in buffalo horn were separated and detected in 32 min. From the results, a good linear relationship exists between peak area and concentration of amino acids ranged from 0 similar to 0.1 g/L to 0 similar to 0. 5 g/L. The correlation coefficients are in the range of 0.9975 similar to 0.9994. The recovery of 12 amino acids is in the range of 94% similar to 104%.