HPLC Method for Analysis of Uracil, Thymine, Hypoxanthine, Guanine on Newcrom AH Column by SIELC Technologies
Separation type: Liquid Chromatography Mixed-mode
Uracil, Thymine, Hypoxanthine, and Guanine are all nitrogenous bases, each playing distinct roles in the biochemistry of nucleic acids.
Uracil (U):
- Found in: RNA
- Pairs with: Adenine (A)
- Structure: Pyrimidine
- Role: Uracil replaces thymine in RNA. In DNA, adenine pairs with thymine, but in RNA, adenine pairs with uracil.
Thymine (T):
- Found in: DNA
- Pairs with: Adenine (A)
- Structure: Pyrimidine
- Role: Thymine is specific to DNA, distinguishing it from RNA. It’s the base that pairs with adenine through two hydrogen bonds.
Hypoxanthine:
- A naturally occurring purine derivative. It’s not directly a base in standard DNA or RNA, but it plays a significant role in the metabolism of purines.
- It is the base form of the nucleoside inosine, which can be found in certain tRNAs and plays a role in wobble base pairing.
- Hypoxanthine is also an intermediate in the purine degradation pathway, leading to the production of uric acid.
Guanine (G):
- Found in: Both DNA and RNA
- Pairs with: Cytosine (C)
- Structure: Purine
- Role: Guanine is one of the four main nucleobases in the nucleic acids DNA and RNA. It forms three hydrogen bonds with cytosine, contributing to the stability of the nucleic acid structures.
These nitrogenous bases are crucial for the structure, replication, and function of nucleic acids. Understanding their properties and interactions is fundamental to molecular biology and genetics.
Nucleotides can be retained and analyzed on a mixed-mode Newcrom AH column with a mobile phase consisting of water, Acetonitrile (MeCN), and ammonium formate. This analytical method can detect compounds with high resolution and peak symmetry using UV detection at 260 nm
High Performance Liquid Chromatography (HPLC) Method for Analyses of Uracil, Thymine, Hypoxanthine and Guanine
Condition
| Column | Newcrom AH, 4.6 x 150 mm, 5 µm, 100 A |
| Mobile Phase | MeCN – 1% |
| Buffer | Ammonium formate pH 3.0- 6 mM |
| Flow Rate | 1.0 ml/min |
| Peak Retention Time | 2.21, 2.38, 2.81, 7.92 min |
| Detection | 260 nm |
Description
| Class of Compounds | Nucleotides |
| Analyzing Compounds | Uracil, Thymine, Hypoxanthine, Guanine |
Application Column
Newcrom AH
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Hypoxanthine
Thymine
Uracil
