Applications:
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.
Condition
Column |
Obelisc N, 4.6x150 mm, 5 µm, 100A |
Mobile Phase |
MeCN -90% |
Buffer |
AmAc |
Flow Rate |
1.0 ml/min |
Detection |
UV, 250 nm |
Description
Class of Compounds
|
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
Analyzing Compounds |
Uracil, Uridine, Adenosine, Guanosine, Cytidine, Cytosine |
AdenosineCytidineCytosineGuanosineUracilUridine
HPLC Separation of Thymidine, Uridine, Adenosine, Guanosine, and Cytidine Using the Hydrogen Bonding Method
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 rder 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, 3.2x100 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: Thymidine, uridine, adenosine, guanosine and cytidine
Condition
Column |
Sharc 1, 3.2x100 mm, 5 µm, 100A |
Mobile Phase |
MeCN/MeOH |
Buffer |
AmFm, Formic acid |
Flow Rate |
1.0 ml/min |
Detection |
UV, 270 nm |
Description
Class of Compounds
|
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
Analyzing Compounds |
Thymidine, Uridine, Adenosine, Guanosine, Cytidine |
AdenosineCytidineGuanosineThymidineUridine
HPLC Separation of Nucleosides and Deoxynucleosides
Condition
Column |
Sharc 1, 4.6x150 mm, 5 µm, 100A |
Mobile Phase |
MeCN/MeOH |
Buffer |
AmFm, Formic acid |
Flow Rate |
1.0 ml/min |
Detection |
UV, 270 nm |
Description
Class of Compounds
|
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
Analyzing Compounds |
Thymidine, Uridine, Deoxyadenosine, Adenosine, Deoxyguanosine, Guanosine, Deoxycytidine, Cytidine |
AdenosineCytidineDeoxyadenosineDeoxycytidineDeoxyguanosineGuanosineThymidineUridine
HPLC Separation of Guanosine, Deoxyguanosine, Adenosine, Deoxyadenosine on Newcrom AH Column
Separation type: Liquid Chromatography Mixed-mode
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Nucleosides are the building blocks for DNA and RNA as well as other roles in biomechanical processes such as signal transduction. By using a Newcrom AH mixed-mode column with a cation-exchange mechanism, nucleosides: guanosine, deoxyguanosine, adenosine, and deoxyadenosine, can be baseline separated in a short time using an isocratic method with a simple mobile phase of water, acetonitrile (MeCN, ACN), and H3PO4 as a buffer. UV detection at 210 nm.
Condition
Column |
Newcrom AH, 4.6x150 mm, 5 µm, 100A |
Mobile Phase |
MeCN/H2O - 20/80% |
Buffer |
H3PO4 - 0.5% |
Flow Rate |
1.0 ml/min |
Detection |
UV, 210 nm |
Description
Class of Compounds
|
Nucleoside, Hydrophilic, Ionizable |
Analyzing Compounds |
Guanosine, Deoxyguanosine, Adenosine, Deoxyadenosine |
AdenosineDeoxyadenosineDeoxyguanosineGuanosine