| CAS Number | 73-24-5 |
|---|---|
| Molecular Formula | C5H5N5 |
| Molecular Weight | 135.131 g/mol |
| InChI Key | GFFGJBXGBJISGV-UHFFFAOYSA-N |
| LogP | -0.0900 |
| Synonyms |
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Applications:
HPLC Application for Separation of Nucleotide Bases
Nucleotide bases are parts of DNA and RNA. Adenine and guanine are purine-bases; uracil, thymine and cytosine are pyrimidine-bases. In the view of chromatography these compounds are very polar and similar in properties. It is hard to obtain base line HPLC separation on traditional C18 as peaks of nucleotide bases co-elute even at low organic concentration. In this application nucleobases are well retained and separated on Primesep 200 mixed-mode column. Compounds are retained by weak reverse phase and weak ion-exchange mechanisms. This HPLC method can utilize UV, ELSD, and LC/MS detection.
| Column | Primesep 200, 4.6*250 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O - 10/90% |
| Buffer | TFA - 0.2% |
| Flow Rate | 0.5 ml/min, 1.0 ml/min |
| Detection | UV, 270 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Hormone |
| Analyzing Compounds | Uracil, Thymine, Cytosine, Guanine, Adenine |
Application Analytes:
Adenine
Cytosine
Guanine
Purines
Pyrimidines
Uracil
HPLC Separation of Adenosine and Adenine Using the Hydrogen Bonding Method
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.
| 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 |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
| Analyzing Compounds | Adenosine, Adenine |
Application Analytes:
Adenine
Adenosine
Separation of Nucleic Bases
Primesep 200 separates with baseline resolution nucleic bases (uracil, thymine, cytosine, guanine, and adenine) by a combination of cation exchange and reversed phase. Uracil typically does not retain on reversed-phase column and is often used as an unretained void volume marker for C18 and C8 columns. Primesep 200 has an embedded anionic functional group which helps retain polar compounds polar and ion-exchange mechanisms. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 270 nm.
| Column | Primesep 200, 4.6*250 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O - 10/90% |
| Buffer | TFA - 0.2% |
| Flow Rate | 0.5 ml/min, 1.0 ml/min |
| Detection | UV, 270 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Hormone |
| Analyzing Compounds | Uracil, Thymine, Cytosine, Guanine, Adenine |
Application Analytes:
Adenine
Cytosine
Guanine
Nucleic Bases
Thymine
Uracil
HPLC Separation of Adenosine, Cordycepin and Adenine on Newcrom AH Column
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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.
| Column | Newcrom AH, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O - 10/90% |
| Buffer | AmFm pH 3.0- 20 mM |
| Flow Rate | 1.0 ml/min |
| Detection | UV 260 nm, MS-compatible mobile phase |
| Class of Compounds | Hydrophilic, Drug, Xanthine, Nucleobase |
| Analyzing Compounds | Adenosine, Cordycepin, Adenine |
Application Analytes:
Adenine
Adenosine
Cordycepin
HPLC Separation of Uracil, Thymine, Guanine, Cytosine, Adenine on Newcrom AH
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Uracil, Thymine, Guanine, Cytosine and Adenine are the nucleobases found in RNA and DNA. The nucleobases are difficult to separate on reverse-phase columns due to their polar, hydrophilic and ionic nature. Using the Newcrom AH mixed-mode column, the nucleobases can be easily separated isocratically using low organic mobile phase (5% acetonitrile) or pure water, if organic mobile phase is undesirable, with ammonium formate buffer, making the method both UV and Mass Spec compatible.
| Column | Newcrom AH, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O - 5/95% |
| Buffer | AmFm pH 3.0- 30 mM |
| Flow Rate | 1.0 ml/min |
| Detection | UV 260 nm, MS-compatible mobile phase |
| Column | Newcrom AH, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | H2O - 100% |
| Buffer | AmFm pH 3.0- 10 mM |
| Flow Rate | 1.0 ml/min |
| Detection | UV 260 nm, MS-compatible mobile phase |
| Class of Compounds | Hydrophilic, Drug, Xanthine, Nucleic Bases |
| Analyzing Compounds | Uracil, Thymine, Guanine, Cytosine, Adenine |
Application Analytes:
Adenine
Cytosine
Guanine
Thymine
Uracil