高效液相色谱法测定饲料中氨基酸含量的改进

Determination of amino acid in feed by improved method of HPLC

对高效液相色谱(HPLC)测定饲料中氨基酸含量的方法进行了改进。简化了样品的水解液制备过程,采用内标法定量,色谱分析采用邻苯二甲醛和氯甲酸芴甲酯联合柱前在线衍生,用反相C18短柱梯度洗脱,二极管阵列检测器检测。此方法线性关系好(相关系数为0.9876～0.9996),线性范围为20～1000μmol/L。大部分氨基酸的加标回收率94.55%～103.35%,相对标准偏差为2.36%～4.87%,在不需预先除去干扰物质的情况下,提高了氨基酸测定的准确度。

The method for determination of amino acids in feedstuffs by high performance liquid chromatography （HPLC） was improved by simplifing sample preparation,using internal standard method performing.The online derivatization with o-phthaldialdehyde and 9-fluorenylmethyl chloroformate automatically ,and analyzing on a C18 short column using gradient elution with diode array detector detection.There was a good linearity between （correlation coefficient were 0.9876 - 0.9996）the peak area of amino acids and their concentration in the range of 20 - 1000 μmol/L.The recoveries of standard spiking of amino acids were 94.55 % - 103.35 % ,except cystine and tryptophan were destroyed completely and somewhat methionine.The relative standard deviation（RSD,n=5）were 2.36 % - 4.87 %.The accuracies of determination of amino acids were imoroved without the interference being removed before analvsis.