Simultaneous analysis of amino acids and amines as their o-phthalaldehyde-ethanethiol-9-fluorenylmethyl chloroformate derivatives in cheese by high-performance liquid chromatography.
Simultaneous analysis of amino acids and amines as their o-phthalaldehyde-ethanethiol-9-fluorenylmethyl chloroformate derivatives in cheese by high-performance liquid chromatography.
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DOI：
10.1016/j.chroma.2008.07.035
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发表时间：
2008-09
期刊：
影响因子：
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通讯作者：
A. Körös;Zsófia Varga;I. Molnár-Perl
中科院分区：
文献类型：
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作者：
A. Körös;Zsófia Varga;I. Molnár-Perl
本文介绍了一种高效液相色谱-二极管阵列-荧光检测器联用测定奶酪样品中氨基酸和胺类的方法。该提案基于酸性脱蛋白、衍生化和梯度优化研究，通过一次进样从单一溶液中鉴别和定量21种氨基酸和9种胺。优化的简单方案包括脱蛋白（1 M高氯酸），离心，过滤和随后的邻苯二甲醛-乙烷乙基-9-芴基甲基氯甲酸酯（OPA-ET-FMOC）试剂衍生化。该方法具有较宽的浓度范围（6.25- 1000 pM/进样）的线性、良好的色谱重现性（平均RSD：2.69%）和良好的回收率（平均：100.2%;平均RSD：3.84%）。该方法成功地应用于波特萨鲁特干酪、蓝纹干酪和烟熏干酪样品中氨基酸和胺含量的测定。
A high-performance liquid chromatographic method (HPLC) with combined diode array and fluorescence detection of amino acids and amines in various cheese samples is described. The proposal is based on acidic deproteinization, derivatization and gradient optimization studies, resulting in the identification and quantification of 21 amino acids and 9 amines from a single solution, by one injection. The optimized, simple protocol consists of deproteinization (1M perchloric acid), centrifugation, filtration and the subsequent derivatization with the o-phthalaldehyde-ethanethiol-9-fluorenylmethyl chloroformate (OPA-ET-FMOC) reagent. The method can be characterized with a linearity of wide concentration range (6.25–1000pM/injection), a good chromatographic reproducibility (average: 2.69% RSD) and an excellent recovery (average: 100.2%; average 3.84% RSD). The developed method was successfully applied in the determination of the amino acid and amine contents of port salut cheese, blue cheese and smoked cheese samples.