Adenine

HPLC Method for Analysis of Adenosine and Adenine on Amaze HD Column
Application description

Adenosine is a purine nucleoside composed of adenine and ribose fragments. Adenine is a purine-based nucleobase. Both compounds are polar and slightly basic, with ability to form hydrogen bonding with the stationary phase. Both compounds were separated within 2 minutes with an isocratic mobile phase. Method is compatible with mass spectrometry.

Conditions of Experiment
Column: Amaze HD
Separation Modes: HILIC
Column Dimenstions: 3.2 x 50 mm
Mobile Phase: MeCN/MeOH (95/5) with 0.5% HCOOH with 0.05% AmFm
Detection: UV 275 nm
Sample: 0.3 mg/ml
Injection: 1 uL
Flow rate: 1.0 mL/min
Analytes
Class of compounds: Aromatic base, Nucleobase, Nucleoside
Nature of compounds: Basic, Hydrophilic, Polar
Compounds: Adenosine, Adenine
Ultra-Fast Separation of Nucleobases on Coresep 100 Core-Shell HPLC Column in Reversed-Phase Mixed-Mode
Application description

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 impossible to retain and obtain base-line separation for this compounds on traditional reversed-phase core-shell column. In this application, nucleobases are well retained and separated on Coresep 100 core-shell  mixed-mode column. Compounds are retained by weak reverse phase and weak ion-exchange mechanisms, but synergy of two mechanism provides efficient separation with excellent peak shape, symmetry and efficiency. All five compounds are separated within one minute. Method can utilize UV, ELSD, and LC/MS detection and can be applied for fast and efficient method using regular HPLC system.

Conditions of Experiment
Column: Coresep 100
Separation Modes: reversed-phase and cation-exchange
Column Dimenstions: 3.2 x 100 mm, 2.7 um, 90A
Mobile Phase: 20% ACN with 0.1% TFA
Detection: 230 nm, ELSD
Sample: 0.3 mg/ml
Injection: 1 uL
Flow rate: 1 ml/min
Analytes
Class of compounds: Aromatic base, Nucleobase
Nature of compounds: Basic, Hydrophilic, Neutral, Polar
Compounds: Uracil, Thymine, Cytosine, Guanine, Adenine