Separation of Nucleic Bases
Primesep 200 separates with baseline resolution nucleic bases (uracil, thymine, cytosine, guanine, and adenine) by a combination of cation exchange and reversed phase. Uracil typically does not retain on reversed-phase column and is often used as an unretained void volume marker for C18 and C8 columns. Primesep 200 has an embedded anionic functional group which helps retain polar compounds polar and ion-exchange mechanisms. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 270 nm.
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.
HPLC Separation of Nucleic Bases at pH 4 and 5 on Obelisc N
Nucleic bases are biological compounds found in genetic molecules (DNA, RNA). They can be separated on an Obelisc N column, which offers very polar characteristics and can be used with positively or negatively charged groups. Closely-eluted adenosine and uridine can be further separated by simply adjusting the pH of the mobile phase. Mobile phase is water and acetonitrile (MeCN, ACN) with Ammonium Acetate as buffer. UV detection at 250nm.
HPLC Separation of Cytidine and Cytosine Using the Hydrogen Bonding Method
Application Notes: Nucleosides glycosylamines consisting of nucleobase linked to ribose or deoxyribose sugar. Nucleoside are building blocks for DNA and RNA. These compounds are very polar in nature and contain groups available for hydrogen bonding interaction. A method for separation of cytosine and cytidine was developed based on the strong dependence of retention time to the mobile phase composition. The mobile phase consists of acetonitrile and methanol. Order of elution for compounds depends on the amount of acetonitrile and methanol. Our method is compatible with LC/MS and preparative chromatography, and can be used for separation of other nucleobases and nucleotides.
Application Columns: SHARC 1, 3.2x100 mm, 5 um, 100A. To learn more about SHARC 1 columns click here. To order this column click here. To see more chromatographic separations check our web site.
Application Compounds: Cytosine and Cytidine
Detection Technique: UV, LC/MS