采用邻苯二甲醛柱前衍生化的高效液相色谱法分析鱼肉中的游离氨基酸。
HPLC method for analysis of free amino acids in fish using o-phthaldialdehyde precolumn derivatization.
作者信息
Antoine F R, Wei C I, Littell R C, Marshall M R
机构信息
Food Science and Human Nutrition Department, P.O. Box 110720, University of Florida, Gainesville, Florida 32611-0370, USA.
出版信息
J Agric Food Chem. 1999 Dec;47(12):5100-7. doi: 10.1021/jf990032+.
Precolumn derivatization applying o-phthaldialdehyde (OPA) was used to analyze free lysine, histidine, and ornithine, precursors of the respective biogenic amines cadaverine, histamine, and putrescine, which are considered indicators of fish quality and safety. This method uses 75% methanol to eliminate the use of strong acids as the extraction solution. Each analysis took 35 min, was reproducible, and allowed separation of primary amino acids in fish samples. A binary solvent delivery system coupled with a fluorescence detector and an Ultrasphere ODS column were utilized for HPLC separation. Linearity of the calibration curves was very good (r(2) = 0.99) for the amino acids of interest. Minimum concentrations of detection were 40 pmol/mL for histidine and lysine and 70 pmol/mL for ornithine. Average recoveries were 72% for lysine, 93% for histidine, and 98% for ornithine. This method used solvent gradient elution to study the levels of these analytes in mahi-mahi, bigeye tuna, and flounder.
采用邻苯二甲醛（OPA）柱前衍生化法分析游离赖氨酸、组氨酸和鸟氨酸，它们分别是生物胺尸胺、组胺和腐胺的前体，被视为鱼类质量和安全性的指标。该方法使用75%的甲醇，避免了使用强酸作为提取溶液。每次分析耗时35分钟，具有可重复性，能够分离鱼类样品中的伯氨基酸。采用二元溶剂输送系统结合荧光检测器和超球ODS柱进行高效液相色谱分离。所关注氨基酸的校准曲线线性非常好（r(2) = 0.99）。组氨酸和赖氨酸的最低检测浓度为40 pmol/mL，鸟氨酸为70 pmol/mL。赖氨酸的平均回收率为72%，组氨酸为93%，鸟氨酸为98%。该方法采用溶剂梯度洗脱来研究鲯鳅、大眼金枪鱼和比目鱼中这些分析物的含量。
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