CHAPTER 6 



The Separation of Nucleic Acid Derivatives by 

 Chromatography on Ion-Exchange Columns^ 



WALDO E. COHN 



Page 



I. Ion Exchange 212 



1. Ion-Exchange Resins 212 



2. Ion-Exchange Equilibria 213 



a. Hydrophilic Character 213 



b. The Distribution Coefficient 213 



c. Nonpolar Affinity 214 



d. Rate of Reaction 214 



3. Ion-Exchange Chromatography 215 



a. Sorption and Elution 215 



b. Types of Chromatography 216 



c. Scaling Up 216 



II. The Separation of Bases and Nucleosides 217 



1. Ionic Properties 217 



2. Cation Exchange 218 



3. Anion Exchange 219 



a. General 219 



b. Bases 219 



c. Ribosides 221 



III. Separation of Nucleotides 221 



1. Mononucleotides 221 



a. Ionic Properties 221 



b. Distribution Coefficients 221 



c. Elution with Dilute HCl 223 



d. Use of Higher pH; the Separation of Inorganic Phosphate and Non- 

 nucleotide Phosphoric Acid Esters 224 



e. Isomeric Nucleotides 225 



(1) Choice of Anion 225 



(2) pK's of Isomeric Nucleotides 226 



(3) Cytidylic Acids 226 



(4) Adenylic and Inosinic Acids 228 



(5) Uridylic Acids 228 



(6) Guanylic Acids 228 



(7) Application to Digests 230 



' Manuscript prepared under Contract No. W-7405-eng-26 for the Atomic Energy 

 Commission. 



211 



