HPLC Method for Analysis of Guanosine, Cytidine, Adenosine, Uridine, Thymidine on Chromni Column by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Guanosine, Cytidine, Adenosine, Uridine, Thymidine.

Thymidine, also noted as dT and dThd and known as deoxythymidine, has the chemical formula C₁₀H₁₄N₂O₅. It is a a pyrimidine deoxynucleoside found as one of the four nucleosides in DNA. It pairs with deoxyadenosine in double sided DNA and is used to synchronize the cells in G1/early S phase. Thymidine’s primary use is in the production of the antiretroviral drug known as azidothymidine. You can find detailed UV spectra of Thymine and information about its various lambda maxima by visiting the following link.

Uridine, also noted as U and Urd, has C₉H₁₂O₆ chemical formula. It is a pyrimidine analog containing uracil. It is one of the five standard nucleosides. It is a non-essential nutrient, since the body produces it as necessary. While uridine can be found in certain foods in the form of RNA, it is not bioavailable, as it is destroyed in the liver. You can find detailed UV spectra of Uracil and information about its various lambda maxima by visiting the following link.

Adenosine is a key building block of energy-carrying molecules with the chemical formula C₁₀H₁₃N₅O₄. It has a variety of other uses, including being a inhibitory neurotransmitter which helps with sleep and acting as a blood flow regulator. Medicinally, it is used as treatment for supraventricular tachycardia (SVT). You can find detailed UV spectra of Adenosine and information about its various lambda maxima by visiting the following link.

Cytidine, also noted as C or Cyd, is a nucleoside molecule with the chemical formula C₉H₁₃N₃O₅. It is primarily found in foods with high RNA contents, such as organ meets, brewer’s yeast, and beer. During digestion, Cyd is broken down into ribosyl pyrimidines. When consumed by humans, it is converted into uridine, which is suspected to be the reason behind cytidine’s metabolic effects. You can find detailed UV spectra of Cytidine and information about its various lambda maxima by visiting the following link.

Guanosine, often denoted as G or Guo, is a purine nucleoside with the chemical formula C₁₀H₁₃N₅O₅. It can be phosphorylated into many other forms, which play vital roles in biochemical possesses like synthesis of nucleic acids, proteins, photosynthesis, and more. It is also required for RNA splicing. It is used in acyclovir, an antiviral drug, to treat herpes. It can also be found in abacavir, an anti-HIV drug, and regadenoson, an A_2A adenosine receptor agonist. You can find detailed UV spectra of Guanosine  and information about its various lambda maxima by visiting the following link.

Guanosine, Cytidine, Adenosine, Uridine, Thymidine can be retained and analyzed using the Chromni stationary phase column. The analysis utilizes a gradient method with a simple mobile phase consisting of water and acetonitrile (MeCN) and ethanol. Detection is performed using UV.

Condition

Description

If you are looking for optimized HPLC method to analyze Cytidine check our HPLC Applications library

For optimal results in HPLC analysis, it is recommended to measure absorbance at a wavelength that matches the absorption maximum of the compound(s) being analyzed. The UV spectrum shown can assist in selecting an appropriate wavelength for your analysis. Please note that certain mobile phases and buffers may block wavelengths below 230 nm, rendering absorbance measurement at these wavelengths ineffective. If detection below 230 nm is required, it is recommended to use acetonitrile and water as low UV-transparent mobile phases, with phosphoric acid and its salts, sulfuric acid, and TFA as buffers.
For some compounds, the UV-Vis Spectrum is affected by the pH of the mobile phase. The spectra presented here are measured with an acidic mobile phase that has a pH of 3 or lower.

HPLC Method for Separation of Cytosine, Deoxycytidine, Cytidine on BIST B+ by SIELC Technologies.

Cytosine, also noted as C and Cyt, has the chemical formula C₄H₅N₃O. In DNA, it pairs with Guanine to create a base pair. In RNA, it is synonymous with Uracil, being an interchangeable third base. Not only that, due to it’s instability, it can change into Uracil through spontaneous deamination.

Deoxycytidine is a deoxyribonucleoside with the chemical formula C₉H₁₃N₃O₄. It is a precursor for 5-aza-2′-cytidine, which is a treatment for myelodysplastic syndrome. It works through interfering with the methylation of the P15/INK4B gene. It can also be used as a biomarker for tumor diagnosis.

Cytidine, also noted as C or Cyd, is a nucleoside molecule with the chemical formula C₉H₁₃N₃O₅. It is primarily found in foods with high RNA contents, such as organ meets, brewer’s yeast, and beer. During digestion, Cyd is broken down into ribosyl pyrimidines.

Cytosine, Deoxycytidine, Cytidine can be retained and analyzed using the BIST B+ stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a sulfuric acid buffer. Detection is performed using UV.

Condition

Description

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, 3.2×100 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: Cytosine and Cytidine

Detection Technique: UV, LC/MS

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

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.