HPLC Method for Analysis of Nucleosides and Nucleoside Derivatives on Amaze HD Column
Application description

Thymidine, uridine, deoxyadenosine, adenosine, deoxyguanosine and guanosine were separated on an Amaze HD column with LC/MS compatible method. The approach can be used to analyze nucleotides, nucleosides and their derivatives in various sample matrices. This HPLC method is robust and reproducible, and can be used for other compounds that can form hydrogen bonds with stationary phase and interact by electrostatic interaction.

Conditions of Experiment
Column: Amaze HD
Separation Modes: hydrogen-bonding and cation-exchange
Column Dimenstions: 3.2 x 100 mm
Mobile Phase: MeCN/MeOH from 90/10 to 50/50 with 0.1% HCOOH and 0.01% AmFm in 8 min
Detection: UV 275 nm
Sample: 0.3 mg/ml
Injection: 1 uL
Flow rate: 0.8 mL/min
Class of compounds: Aromatic base, Nucleoside
Nature of compounds: Basic, Hydrophilic, Neutral, Polar
Compounds: Thymidine, Uridine, 2′-Deoxyadenosine, Adenosine, Guanosine
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
Class of compounds: Nucleobase, Nucleoside
Nature of compounds: Hydrophilic, Neutral
Compounds: Uracil, Uridine