Uracil

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
Fast HPLC Separation of Uracil and Uridine on Core-Shell Mixed-Mode Coresep S Column in HILIC Mode
Application description

Uracil and Uridine are nucleosides which occur as a part of RNA. Both compounds are neutral in nature and very polar. Uracil is often used as a void marker in reversed-phase HPLC. A fast HILIC-based method was developed for analysis of uracil and uridine on the core-shell HILIC and cation-exchange column. Method provides good peak shape and retention control. Column and method can be used for analysis of polar compounds in HILIC, and HILIC/mixed-mode.

Conditions of Experiment
Column: Coresep S
Separation Modes: HILIC
Column Dimenstions: 3.2 x 100 mm, 2.7 um, 90A
Mobile Phase: 98% ACN with no additives
Detection: UV 250 nm
Sample: 0.1-0.3 mg/ml
Injection: 1 uL
Flow rate: 0.5 mg/ml
Analytes
Class of compounds: Nucleobase, Nucleoside
Nature of compounds: Hydrophilic, Neutral
Compounds: Uracil, Uridine