You Jinmao, Sheng Xiao, Ding Chenxu, Sun Zhiwei, Suo Yourui, Wang Honglun, Li Yulin
Northwest Plateau Institute of Biology, Chinese Academy of Sciences, Xining 810001, PR China.
Anal Chim Acta. 2008 Feb 18;609(1):66-75. doi: 10.1016/j.aca.2007.12.022. Epub 2007 Dec 28.
A novel labeling reagent 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP) coupled with capillary electrophoresis (CE) with DAD detection for the determination of carbohydrates has been developed. The chromophore in the 1-phenyl-3-methyl-5-pyrazolone (PMP) reagent is replaced by naphthyl functional group, which results in a reagent with very high molar absorptivity (epsilon251 nm = 5.58 x 10(4) L mol(-1) cm(-1)). This permits NMP-labeled carbohydrates to be detected with UV absorbance in standard 50-mum-i.d. fused silica capillaries by zone electrophoresis. In this mode, nanomolar concentrations of detection limits are obtained. The method for the derivatization of carbohydrates with NMP is simplified. The derivatization reaction is rapid and mild in the presence of ammonia catalyst without further transfer steps. Nine monosaccharide derivatives such as mannose, galacturonic acid, glucuronic acid, rhamnose, glucose, galactose, xylose, arabinose and fucose can successfully be detected in CE mode. Good reproducibility can be obtained with relative standard deviation (R.S.D.) values of the migration times and peak area, respectively, from 0.44 to 0.48 and from 3.2 to 4.8. Furthermore, the developed method has been successfully applied to the analysis of carbohydrates in the hydrolyzed rape bee pollen samples.
已开发出一种新型标记试剂1-(2-萘基)-3-甲基-5-吡唑啉酮(NMP),与毛细管电泳(CE)联用二极管阵列检测法用于测定碳水化合物。1-苯基-3-甲基-5-吡唑啉酮(PMP)试剂中的发色团被萘基官能团取代,这使得该试剂具有非常高的摩尔吸光系数(ε251nm = 5.58×10(4)L·mol(-1)·cm(-1))。这使得NMP标记的碳水化合物能够通过区带电泳在标准内径50μm的熔融石英毛细管中用紫外吸光度进行检测。在这种模式下,可获得纳摩尔浓度的检测限。用NMP对碳水化合物进行衍生化的方法得到了简化。衍生化反应在氨催化剂存在下快速且温和,无需进一步转移步骤。甘露糖、半乳糖醛酸、葡萄糖醛酸、鼠李糖、葡萄糖、半乳糖、木糖、阿拉伯糖和岩藻糖等9种单糖衍生物能够在CE模式下成功检测。迁移时间和峰面积的相对标准偏差(R.S.D.)值分别为0.44至0.48和3.2至4.8,可获得良好的重现性。此外,所开发的方法已成功应用于水解油菜蜂花粉样品中碳水化合物的分析。