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化学学报
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化学学报 ›› 1981, Vol. 39 ›› Issue (6): 553-561. 上一篇    下一篇

论文

核酸化学研究Ⅳ.常见核糖核苷及其3’-核糖核苷酸柱层析

核酸组   

  1. 中国科学院上海有机化学研究所
  • 投稿日期:1979-11-02 发布日期:2013-06-03

STUDIES ON NUCLEIC ACID CHEMISTRY Ⅳ. THE COLUMN CHROMATOGRAPHY OF COMMON RIBONUCLEOSIDES AND RIBONUCLEOTIDES

NUCLEIC ACID GROUP   

  1. Shanghai Institute of Organic Chemistry, Academia Sinica
  • Received:1979-11-02 Published:2013-06-03

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本文介绍以涂渍12%甲基三正烷基(C8～C10)氯化铵的聚三氟氯乙烯细粉(160～200目)为交换剂的反相离子交换柱层析.用碳酸铵水溶液分级洗脱,可在三小时左右一次柱层析即能把常见核糖核苷和相应的3'-核糖核苷酸共八种组分的混合物分开,并可根据各组分的峰面积定量.各组分洗脱的次序为胞嘧啶核糖核苷(CR),腺嘌呤核糖核苷(AR),尿嘧啶核糖核苷(UR),鸟嘌呤核糖核苷(GR),3'-胞嘧啶核糖核苷酸(3'-CMP),3'-腺嘌呤核糖核苷酸(3'-AMP),3'-尿嘧啶核糖核苷酸(3'-UMP),3'-鸟嘌呤核糖核苷酸(3'-GMP).表示各峰分离情况的分离度(R)除CR和AR之间为0.77分离略差外,其它各峰之间均大于1.各个组分含量达10～20微克(0.3～0.5A260)*即可定量.定量分析标准误差σ=0.06.此法已成功地用于以化学方法合成的低聚核糖核苷酸的碱基组成的测定.

A simple method for the separation and quantitative analysis of the font common ribonuoleosides and their 3'-ribonuoleotides by reversed phase anion-ezohange chromatography is described.A column of polyohlorotrifluoroethylene(16O-200 mesh) coated with 12%(ml/g)methyl-tri-n-alkyl(C8-C10) ammonium chloride and a stepwise elution process are adopted.The volnmns and concentrations of aqueous ymmoninm carbonate eluent(pH 8.2) employed in succession are 30 ml of 0.001 M,200 ml of 0.2 M and 10 ml of 0.3 M.