Cohen S A, De Antonis K M
Millipore Corporation, Milford, MA 01757.
J Chromatogr A. 1994 Feb 11;661(1-2):25-34. doi: 10.1016/0021-9673(93)E0821-B.
Primary and secondary amines are rapidly labelled by 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate to form highly fluorescent asymmetric urea derivatives which are readily amenable to analysis by liquid chromatography. Derivatization consists of a simple, one-step procedure, and the resulting labelled amines can be analyzed without further cleanup. The adducts are extremely stable with no discernible loss in response after storage for one week at room temperature, making the reagent an ideal candidate for pre-column amino acid analysis. Chromatographic methods for protein hydrolysates have been developed for the analysis of samples containing many unusual amino acids including a number of cysteine derivatives, collagen hydrolysates containing hydroxyproline and hydroxylysine, performic acid oxidized samples and glycoprotein hydrolysates containing glucosamine and galactosamine. Samples with potentially interfering matrix components such as hydrolyzed feed grains and intravenous solutions are readily analyzed and are quantified with average per cent relative standard deviations in the 1-2% range. Comparative data on these samples are in good agreement with either ion-exchange amino acid analysis or label information.
伯胺和仲胺可被6-氨基喹啉-N-羟基琥珀酰亚胺基氨基甲酸酯快速标记,形成高荧光不对称脲衍生物,这些衍生物易于通过液相色谱进行分析。衍生化过程包括一个简单的一步程序,生成的标记胺无需进一步净化即可进行分析。加合物极其稳定,在室温下储存一周后响应无明显损失,这使得该试剂成为柱前氨基酸分析的理想选择。已经开发出用于蛋白质水解产物的色谱方法,用于分析含有许多不寻常氨基酸的样品,包括一些半胱氨酸衍生物、含有羟脯氨酸和羟赖氨酸的胶原蛋白水解产物、过甲酸氧化样品以及含有氨基葡萄糖和半乳糖胺的糖蛋白水解产物。含有潜在干扰基质成分的样品,如水解饲料谷物和静脉注射液,很容易进行分析,其定量的平均相对标准偏差在1-2%范围内。这些样品的比较数据与离子交换氨基酸分析或标记信息高度一致。