Uridine

Application Notes: Nucleosides are glycosylamines consisting of nucleobase linked to ribose or deoxyribose sugar and are building blocks for DNA and RNA. These compounds are very polar and contain groups available for hydrogen bonding interaction. Thymidine, uridine, adenosine, guanosine and cytidine were separated using a hydrogen-bonding method. There is a strong correlation between the retention time and mobile phase composition. The strength of hydrogen-bonding interaction increases as the number of hydroxyls in the analytes increase. Additionally the order of elution for compounds depends on the ratio of the mobile phases: acetonitrile and methanol. Our method is compatible with LC/MS and preparative chromatography.

Application Columns: SHARC 1, 3.2×100 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: Thymidine, uridine, adenosine, guanosine and cytidine

Uracil and uridine are separated based on their polarity. The mechanism of retention is pure HILIC mode. Retention time is controlled by varying the amount of ACN. Buffer concentration and pH will affect ionization and polarity of the stationary phase, and thus indirectly affect retention of neutral compounds. Column is compatible with all general detection techniques.

Uracil is a naturally occurring pyrimidine derivative. Uracil is used for drug delivery and as a pharmaceutical. Uridine is a product of uracil and sugar ribose. Uridine is one of components of ribonucleic acid. Uracil and uridine are slightly basic in nature. Both compounds are hydrophilic and uracil is very often is used as a void marker in reversed-phase chromatography. Uracil and uridine are separated based on their polar properties on a Primesep N HILIC column. Uracil and uridine are monitored by UV.

Uridine and uracil are separated on three HILIC columns from SIELC. Primesep N, Primesep S, and Obelisc N columns all have different polarity and different embedded acidic groups on the surface of silica gel: Primesep N is a normal-phase HPLC column with embedded acidic groups with a pKa of about 5. Primesep S is a normal-phase HPLC column with embedded acidic groups with a pKa of about 3. The Primesep S stationary phase retains basic compounds by cation-exchange at pH > 3.

In traditional HILIC mode, a charged or neutral polar analyte interacts with a water layer on the polar stationary phase surface. On Obelisc N the charges are greatly separated and independently accessible, resulting in different selectivity compared to traditional HILIC and silica columns. Mobile phase composition changes the conformation of the long hydrophilic chain. All three columns can be used to analyze uridine and uracil by HPLC with UV, LC/MS and ELSD detection.

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.