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 Separation of Uracil, Thymine, Guanine, Cytosine, Adenine on Newcrom AH
View on hplc.cloudUracil, Thymine, Guanine, Cytosine and Adenine are the nucleobases found in RNA and DNA. The nucleobases are difficult to separate on reverse-phase columns due to their polar, hydrophilic and ionic nature. Using the Newcrom AH mixed-mode column, the nucleobases can be easily separated isocratically using low organic mobile phase (5% acetonitrile) or pure water, if organic mobile phase is undesirable, with ammonium formate buffer, making the method both UV and Mass Spec compatible.