Adenosine

HPLC Method for Separation of Adenine, Deoxyadenosine and Adenosine on BIST B+ by SIELC Technologies.

Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

Description

Separation type: Liquid Chromatography Mixed-mode

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High Performance Liquid Chromatography (HPLC) Method of Adenosine and Deoxyadenosine.

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: 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 ammonium formate (AmFm) buffer. Detection can be achieved with UV 260 nm, mass spectrometry (MS), evaporative light scattering detection (ELSD) and Charged aerosol detection (,.CAD).

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.

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Due to cordycepin having a very similar structure to adenosine, it has shown to have inhibitive properties on the COVID-19 coronavirus. However, due to their similar structures, the separation of the two sugars can be challenging. Both sugars can be separated isocratically in about six minutes on the Newcrom AH mixed-mode column, which has both hydrophobic and cationic exchange properties. The mobile phase consists of acetonitrile (ACN, MeCN) and water with ammonium formate as a buffer which makes it mass-spec (MS) compatible. It can also be UV detected at 260nm.

Separation type: Liquid Chromatography Mixed-mode

 
Many compounds are difficult, if not impossible, to separate on reverse-phase columns in HPLC. Other compounds cannot be separated on ion-exchange columns. That’s where the mixed-mode columns come in. By using a stationary phase with both hydrophobic and ion-exchange properties, allows the chromatographer to have additional controls over separation conditions. Here, we demonstrate the separation of compounds that can’t be achieved on a C18 column. By using both an organic gradient and buffer gradient of ammonium formate (AmFm), we can separate structurally similar compounds that can’t be separated on a reverse-phase column alone.




 

Application Notes: Nucleosides are glycosylamines consisting of a nucleobase linked to a ribose or a deoxyribose sugar. Nucleoside are building blocks for DNA and RNA. These compounds are very polar in nature and contain groups available for hydrogen bonding interactions. Method for separation of adenine and adenosine were developed using a hydrogen-bonding method. There is a strong correlation between retention time for adenine/adenosine and the mobile phase composition, which consists of acetonitrile and methanol. Order of elution for compounds depends on the amount of acetonitrile and methanol.  Furthermore, ellution of adenine and adenosine can be reversed based on the composition of the mobile phase. Our method is compatible with LC/MS and preparative chromatography.

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, 3.2×100 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

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.