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目的:建立一种通过在1-苯基-3-甲基-5-吡唑啉酮(PMP)和对氨基苯甲酸(PABA)衍生单糖反应中加入硼砂,以提高单糖衍生效率的方法。方法:实验考察了硼砂的加入对PMP衍生8种常见醛糖和PABA衍生酮糖(以果糖为例)衍生效率的影响,优化了硼砂溶液浓度、温度及时间等衍生条件,采用毛细管区带电泳(CZE)进行方法学考察,并将改进的方法应用于中药南沙参的单糖组成测定。结果:在衍生过程中加入硼砂均可一定程度提高PMP和PABA对单糖的衍生效率,其中以葡萄糖最为显著,可提高约1.2倍。优化的PMP衍生醛糖的条件为:硼砂溶液浓度6.86 mmol·L~(-1),衍生温度70℃,衍生时间30 min。优化的PABA衍生果糖的条件为:硼砂溶液浓度20.43 mmol·L~(-1),衍生温度70℃,衍生时间60 min。在测定南沙参单糖组成时,改进的方法使一些含量较低的、原方法未发现的单糖被检测出来。结论:硼砂的加入可改变单糖构象平衡,链状构象占比增加,从而提高了单糖衍生效率。该方法准确、灵敏、操作简便,可为改善中药糖类成分分析及构效关系研究提供支持。
OBJECTIVE: To establish a method for increasing monosaccharide derivatization efficiency by adding borax to 1-phenyl-3-methyl-5-pyrazolone (PMP) and p-aminobenzoic acid (PABA) . Methods: The effects of borax on the derivatization efficiency of 8 kinds of common aldose derived from PMP and PABA-derived ketose (fructose) were investigated experimentally, and the derivatization conditions of borax solution concentration, temperature and time were optimized. The capillary zone electrophoresis (CZE) methodological study, and the improved method applied to the Chinese medicine Nansha monosaccharide composition determination. Results: The addition of borax to the derivatization process can improve the derivatization efficiency of monosaccharides of PMP and PABA to a certain extent, of which the most obvious is glucose, which can be increased about 1.2 times. The optimum conditions of PMP-derived aldose were as follows: the concentration of borax solution was 6.86 mmol·L -1, the derivatization temperature was 70 ℃ and the derivatization time was 30 min. The optimized conditions of fructose-derived PABA were as follows: the concentration of borax solution was 20.43 mmol·L -1, the derivatization temperature was 70 ℃ and the derivatization time was 60 min. In the determination of Nansha ginseng single sugar composition, the improved method to some of the lower content of the original method was not found monosaccharides were detected. Conclusion: The addition of borax can change the conformational balance of monosaccharides and increase the proportion of chain conformations, thus improving the efficiency of monosaccharide derivatization. The method is accurate, sensitive and easy to operate, which can be used to improve the analysis of carbohydrate composition and the structure-activity relationship of traditional Chinese medicine.