Using polymer column we established reversed phase high⁃performance liquid chromatography ( RP⁃HPLC) method to determine five pyrimidine and nucleosides. The influences of different factors in mobile phase on separation and analysis were studied, and the pyrimidine and nucleoside compounds were effectively separated with potassium dihydrogen phosphate (KH2PO4)buffer(pH 4. 4) and methanol as mobile phase,flow rate at 1. 0 mL/min,detection wavelength at 251 nm and column temperature at 20℃.With column efficiency over 37 400 N/m,which is higher than common C18⁃silica column(plates:21 400 N/m) ,our results provides basis for wide range applications for the polymer filler in nucleoside compounds detection.%建立了使用高聚物型色谱柱,反相高效液相色谱法( RP HPLC)测定5种嘧啶和核苷的方法,样品经甲醇溶解,以pH为4.4的0.01 mol/L KH2 PO4缓冲盐溶液和甲醇为流动相进行梯度洗脱,流速1.0 mL/min,温度20℃,紫外检测波长251 nm。考察了流动相中不同因素对分离分析的影响,实现了5种嘧啶和核苷混合物的有效分离。柱效可达37400 N/m,优于常规的C18硅胶色谱柱(柱效为21400 N/m),为该高聚物填料在嘧啶和核苷化合物检测方面的广泛应用提供了有关依据。
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机译:6.一种化合物,其包含具有根据20的EP-B1-2563920(SEQ ID NO:80)的具有由20个连接的核苷组成的核碱基序列的修饰的寡核苷酸,其中所述修饰的寡核苷酸包含:由十个连接的脱氧核苷组成的缺口片段;和由五个连接的核苷组成的5'翼区段; 3'翼段由五个连接的核苷组成;其中所述间隙片段位于5'侧翼片段和3'侧翼片段之间,其中每个侧翼片段的每个核苷包含2'-O-甲氧基乙基糖;其中修饰的寡核苷酸的每个胞嘧啶是5-甲基胞嘧啶,并且其中修饰的寡核苷酸的每个核苷间键是硫代磷酸酯键;特别是inotersen;及其衍生物,例如由EP-B1-2563920保护的其盐,包括钠盐。