食品与发酵工业

双向纸电泳-纸层析结合法测定酵母核酸水解的4种核苷酸

  • 张一平 ,
  • 华洵璐 ,
  • 孙梅 ,
  • 谢骏 ,
  • 匡群 ,
  • 何义进
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网络出版日期: 2010-11-25

Determination of Four Nucleotides from Yeast RNA Hydrolysate Combined the Methods of Paper Electrophoresis and Paper Chromatography

  • Zhang Yi-ping ,
  • Hua Xun-lu ,
  • Sun Mei ,
  • Xie Jun ,
  • Kuang Qun ,
  • He Yi-jin
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Online published: 2010-11-25

摘要

建立了酵母核酸水解率简便、准确、低成本测定方法。采用双向纸电泳-纸层析结合法同时测定酵母核酸水解的4种单体核苷酸;采用变异系数(CV)和平均回收率分析该测定方法的精密度和准确度。结果表明,10%醋酸电泳结合饱和硫酸铵∶1 mol/L醋酸铵∶异丙醇(体积比,80∶18∶2)的展层剂,能同时分离4种核苷酸;测定相关系数R均大于0.985;测定核苷酸标准品混合液的变异系数分别为AMP 1.30%、GMP 1.44%、CMP1.36%、UMP 1.31%;测定酵母RNA水解液对应的变异系数分别为2.45%、2.01%、2.75%、2.06%;回收率在94.80%~106.57%,最低检出限在0.65~0.72 mg/mL。

本文引用格式

张一平 , 华洵璐 , 孙梅 , 谢骏 , 匡群 , 何义进 . 双向纸电泳-纸层析结合法测定酵母核酸水解的4种核苷酸[J]. 食品与发酵工业, 2010 , 36(11) : 150 -155 . DOI: 10.13995/j.cnki.11-1802/ts.2010.11.001

Abstract

To establish the simple,accurate and inexpensive determination method of yeast RNA hydrolyzed yield.Method: The four single nucleotides of RNA hydrolysate were determined by the combined methods of paper electrophoresis and paper chromatography;the precision and accuracy of this method were analyzed using coefficient of cariation(CV) and average recovery.Result: Combined the 10% acetic acid buffer and the saturated(NH4)2SO4:1M ammonium acetate:isopropanol(80:18:2,V/V);developing solvent,the four single nucleotides could be separated simultaneously;the relative coefficient(R) were all more than 0.985;the CV value of AMP,GMP,CMP,UMP was 1.30%,1.44%,1.36% and 1.30% respectively in the standard nucleotides mixture sample,while the CV value was 2.45%,2.01%,2.75% and 2.06% respectively in the yeast RNA hydrolization liquid;the average ratio of recovery were between 94.80% and 106.57%;the minimum detection limit were between 0.65~0.72mg/mL.Conclusion: The determination method deployed in this paper had the advantages of meticulous precision,high degree of accuracy,lower investment of instrument and cheaper cost,so it can be promoted widely.
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