Determination of 18 free amino acids in giant panda serum by high performance liquid chromatography after derivatization with 2,4-dinitrochlorobenzene

A method for the determination of 18 free amino acids in giant panda serum by pre-column derivatization-high performance liquid chromatography was developed.After removing protein in serum with HPO3,the free amino acids in supernatant were derivatizated with 2,4-dinitrochlorobenzene in alkalescent media(pH 9.0) in a water bath of 90 ℃.Then the derivatives were separated by C18 column and detected by UV detector,and the amount of amino acids was quantified with internal standard method.The correlation coefficients of regression equations for the 18 free amino acids were between 0.9951 to 0.9999;the range of RSD were 1.39%～4.67%;the range of detection limit were: 0.20～7.72 μg/mL;and the recoveries were 65.9%～118%.