HPLC Application for Separation of Nucleotide Bases

Nucleotide bases are parts of DNA and RNA. Adenine and guanine are purine-bases; uracil, thymine and cytosine are pyrimidine-bases. In the view of chromatography these compounds are very polar and similar in properties. It is hard to obtain base line separation on traditional C18 as peaks of nucleotide bases co-elute even at low organic concentration. In this application nucleobases are well retained and separated on Primesep 200 mixed-mode column. Compounds are retained by weak reverse phase and weak ion-exchange mechanisms. Method can utilize UV, ELSD, and LC/MS detection.

Condition

Column Primesep 200, 4.6*250 mm,  5 µm, 100A
Mobile Phase MeCN/H2O - 10/90%
Buffer TFA - 0.2%
Flow Rate 0.5 ml/min, 1.0 ml/min
Detection UV, 270 nm
 

Description

Class of Compounds Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds Uracil, Thymine, Cytosine, Guanine, Adenine
 

Application Analytes:

Adenine
Cytosine
Guanine
Purines
Pyrimidines
Uracil