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Purine Base and Nucleoside
Guanine and Adenosine
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printable Application Sheet
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Notes: Separation and determination of purine bases (guanine) is of interest because of their involvement in
a variety of biochemical processes. Guanine and other purine bases are building blocks in both DNA and RNA
as they play a crucial role in protein biosynthesis and in the storage of genetic information. Adenosine is an
endogenous purine nucleoside that modulates many physiologic processes and exhibits various bioactivities.
For instance, all purines can inhibit the activity of mononamine oxidase and show anti-oxidant effect,
nucleoside analogues have been used as anti-HIV drugs.
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A purine base (guanine) and a purine nucleoside (adenosine) are two of many metabolites that can be retained on the
Diamond Hydride™ HPLC columns.
Method Conditions
| Column |
Cogent Diamond Hydride™, 4µm, 100A |
| Catalog No. |
70000-7.5P |
| Dimensions |
4.6 x 75 mm |
| Isocratic |
85% B/15% A |
| Mobile Phase |
A: DI water, 0.1% formic acid + 0.001% TFA
B: acetonitrile, 0.1% formic acid, 0.001% TFA |
| Peaks |
1. Adenosine
2. Guanine |
| Sample Matrix |
| Adenosine: |
1 mg/mL was dissolved in 80% acetonitrile/ 20% DI water + 0.1% formic acid |
| Guanine: |
1 mg/mL was dissolved in 80% DI water with 0.1% TFA /20% acetonitrile |
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| Detection |
UV 245 nm |
Discussion
Both guanine and adenosine are polar compounds which elute at the void volume on many leading (excellent) C18 columns.
Using a simple mobile phase and the Cogent Diamond Hydride™ column it is possible to retain guanine and
adenosine, as well as many other polar compounds. Mixture of the two samples were prepared: 100 µL of each
sample and 800 µL of the mobile phase.
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