Department of Environmental Chemistry, University of Łódź, Poland.
Amino Acids. 2011 Jun;41(1):187-94. doi: 10.1007/s00726-010-0521-7. Epub 2010 Mar 4.
Homocysteine (Hcy) is incorporated into protein via a reaction of the thioester Hcy-thiolactone with ε-amino group of a protein lysine residue generating N-Hcy-protein. This reaction impairs and alters protein's function and has been implicated in atherothrombotic disease. Here, we describe new high-performance liquid chromatography assays for the determination of Hcy-thiolactone, protein N-linked Hcy, and Hcy based on an on-column derivatization with o-phthaldialdehyde and fluorescence detection. The on-column derivatization generates narrow peaks, which allows fast run times (3-5 min) and facilitates determination of N-linked Hcy directly from acid hydrolysates of plasma protein. Utility of these assays was demonstrated with human urine and plasma samples.
同型半胱氨酸（Hcy）通过 Hcy-硫内酯与蛋白质赖氨酸残基的ε-氨基之间的反应掺入蛋白质中，生成 N-Hcy-蛋白质。该反应会损害和改变蛋白质的功能，并与动脉血栓疾病有关。在这里，我们描述了新的高效液相色谱法测定 Hcy-硫内酯、蛋白质 N-连接 Hcy 和 Hcy 的方法，该方法基于柱上衍生化与邻苯二甲醛和荧光检测。柱上衍生化产生窄峰，允许快速运行时间（3-5 分钟），并有助于直接从血浆蛋白的酸水解物中测定 N-连接 Hcy。这些测定方法在人尿和血浆样品中的应用得到了验证。
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