Department of Chemistry, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria.
Anal Biochem. 2011 Nov 1;418(1):24-9. doi: 10.1016/j.ab.2011.07.005. Epub 2011 Jul 14.
The protein-linked glycomes and, thereby, the range of individual monosaccharides of invertebrates differ from those of mammals due to a number of special modifications; therefore, it is necessary to adapt methods for monosaccharide analysis in order to cover these. We optimized the labeling procedure for anthranilic acid (AA) and 1-phenyl-3-methyl-5-pyrazolone (PMP) and the subsequent separation of the labeled monosaccharides on high-performance liquid chromatography (HPLC), with the result that we were able to identify 26 different monosaccharides. The detection limit for anthranilic acid derivatives obtained was 65 fmol, and a reliable quantification of samples was possible up to 200 nmol under the tested conditions. PMP derivatives showed a significantly higher detection limit but allow quantification of larger sample amounts. Applying these methods on snails, their impressive set of monosaccharide constituents, including methylated sugars, was shown.
蛋白质连接的糖缀合物，以及由此产生的无脊椎动物的单糖种类范围，由于许多特殊的修饰而与哺乳动物有所不同；因此，有必要对单糖分析方法进行适应性调整，以涵盖这些修饰。我们优化了对氨基苯甲酸（AA）和 1-苯基-3-甲基-5-吡唑啉酮（PMP）的标记程序，以及随后在高效液相色谱（HPLC）上对标记的单糖进行分离，结果我们能够鉴定出 26 种不同的单糖。所获得的对氨基苯甲酸衍生物的检测限为 65 fmol，在测试条件下，可靠的定量样品量可达 200 nmol。PMP 衍生物的检测限明显更高，但允许对更大的样品量进行定量。将这些方法应用于蜗牛，显示了它们令人印象深刻的单糖组成，包括甲基化糖。
