Deng Ying-Hua, Wang Hong, Zhong Lu, Zhang Hua-Shan
Department of Chemistry, Wuhan University, Wuhan, China.
Talanta. 2009 Feb 15;77(4):1337-42. doi: 10.1016/j.talanta.2008.09.013. Epub 2008 Sep 18.
In this study, a new capillary electrophoresis (CE) method is described originally for the sensitive and selective determination of short-chain aliphatic amines in biological samples. These amines were converted into their N-hydroxysuccinimidyl fluorescein-O-acetate (SIFA) derivatives and measured by micellar electrokinetic capillary chromatography with laser-induced fluorescence detection. The derivatization conditions and separation parameters for the aliphatic amines were optimized in detail. The SIFA-labeled amines were fully separated within 8.5 min using 25 mM pH 9.6 boric acid electrolyte containing 60mM sodium dodecyl sulfate (SDS). The parameters of validation such as linearity of response, precision and detection limits were determined. The detection limits were obtained in the range from 0.02 to 0.1 nM, which was the lowest value reported by CE methods. The developed method was successfully employed to monitor aliphatic amines in serum and cells samples. After comparison of other CE methods using different fluorescent probes, the present method represents a powerful tool for the trace determination of aliphatic amines in complex biological samples.
在本研究中,最初描述了一种新的毛细管电泳(CE)方法,用于灵敏且选择性地测定生物样品中的短链脂肪胺。这些胺被转化为其N-羟基琥珀酰亚胺荧光素-O-乙酸酯(SIFA)衍生物,并通过胶束电动毛细管色谱法结合激光诱导荧光检测进行测定。详细优化了脂肪胺的衍生化条件和分离参数。使用含有60 mM十二烷基硫酸钠(SDS)的25 mM pH 9.6硼酸电解质,SIFA标记的胺在8.5分钟内完全分离。确定了响应线性、精密度和检测限等验证参数。检测限在0.02至0.1 nM范围内,这是CE方法报道的最低值。所开发的方法成功用于监测血清和细胞样品中的脂肪胺。在比较了使用不同荧光探针的其他CE方法后,本方法是用于痕量测定复杂生物样品中脂肪胺的有力工具。