HPLC Application for Separation of Nucleotide Bases Uracil, Thymine, Guanine, Cytosine, Adenine on Primesep 200 Column

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 HPLC 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. This HPLC 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 Column

Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Adenine
Cytosine
Guanine
Purines
Pyrimidines
Uracil

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.