高效液相色谱法对鲢鱼、鳙鱼和草鱼肌肉中氨基酸含量的测定

Detection Amino Acids in Muscle of Hypophthalmichthys molitrix,Aristichthys nobilis and Ctenopharyngodon idellus by HPLC

建立了高效液相色谱法检测鲢鱼(Hypophthalmichthys molitrix)、鳙鱼(Aristichthys nobilis)和草鱼(Ctenopharyngodon idellus)肌肉中氨基酸含量的方法 ,采用专用氨基酸Eelipse-AAA色谱柱,以硼酸缓冲液为pH调节剂,以邻苯二甲醛(OPA)和9-芴甲基氯甲酸酯(FMOC)为柱前衍生化试剂,磷酸氢二钠、甲醇、乙腈和水为流动相,柱温40℃条件下分离目标化合物,同时用紫外检测器(DAD)和荧光检测器(FLD)检测同一个样品,对比两种检测器的检出限,利用两种检测器的检测结果来判定氨基酸的含量。结果表明,17种氨基酸在26 min内用紫外检测器能很好的分离,16种氨基酸在26 min内用荧光检测器也能较好的分离;鲢鱼、鳙鱼、草鱼的必需氨基酸占总氨基酸的比例分别为35.75%、34.67%、36.24%。

The experiment was established by HPLC method for detection of amino acids in the muscle of aquatic products.Amino acids in samples were separated by Eelipse-AAA column. Aboric acid buffer solution as pH regulator,with two formaldehyde phthalate（OPA） and 9-fluorene methyl chloroformate（FMOC） as a precolumn derivatization reagent,the mobile phase was two sodium hydrogen phosphate,methanol,acetonitrile and water,the column temperature was 40 ℃. At the same time using ultraviolet detector（DAD） and fluorescence detector（FLD） were detected in the same sample,comparison the LOD of two kinds detectors,and determination of the content of amino acids by the results of the two detectors. The results showed that 17 kinds of amino acids could be separated well with ultraviolet detector（DAD） in 26 min,and 16 amino acids could be separated with the fluorescence detector in 26 min. The essential amino acids of Hypophthalmichthys molitrix,Aristichthys nobilis and Ctenopharyngodon idellus accounted for the proportion of total amino acids were 35.74%,34.67%,36.24%.