HPLC Method for Analysis of Biopterin, Rhamnopterin, Neopterin on Primesep 100 by SIELC Technologies
Separation type: Liquid Chromatography Mixed-mode
- Biopterin: This is a coenzyme involved in the biosynthesis of several important bioactive substances, including neurotransmitters like serotonin and dopamine. It’s also integral to the creation of nitric oxide, a potent vasodilator.
- Rhamnopterin is a derivative of biopterin, demonstrating remarkable properties in dietary studies. Research has shown that when it is administered alongside certain carcinogens, rhamnopterin can decrease the occurrence of liver tumors in rats. Additionally, it serves as an internal standard for measuring biopterin and neopterin concentrations in rat plasma.
- Neopterin: This is a catabolic product of guanosine triphosphate (GTP), a molecule involved in energy transfer. It’s often used as a marker for immune system activation, particularly in response to certain diseases, including viral infections, autoimmune diseases, and cancer.
These three pterins can be retained, separated and analyzed using a reverse-phase Primesep 100, 4.6 x 150 mm, 5 µm, 100 A column. The mobile phase for this method consists of water, acetonitrile (MeCN), and ammonium formate, which serves as a buffer. This analytical method can be monitored using UV detection at 275 nm, an Evaporative Light Scattering Detector (ELSD), or any other evaporative detection method such as Charged Aerosol Detection (CAD) or Electrospray Ionization Mass Spectrometry (ESI-MS)
LOD was determined for this combination of instrument, method, and analyte, and it can vary from one laboratory to another even when the same general type of analysis is being performed.
High Performance Liquid Chromatography (HPLC) Method for Analysis of Biopterin, Rhamnopterin, Neopterin
Condition
| Column | Primesep 100, 4.6 x 150 mm, 5 µm, 100 A |
| Mobile Phase | MeCN/H2O -5/95% |
| Buffer | Ammonium formate pH 3.0 – 20 mM |
| Flow Rate | 1.0 ml/min |
| Detection | UV 275 nm |
| Peak Retention Time | 3.28, 4.01, 4.85 min |
| Sample concentration | 0.02 mg/ml |
| Injection volume | 5 µl |
| Sample diluent | H2O + NaOH |
| LOD | 20 ppb |
Description
| Class of Compounds | Pteridines |
| Analyzing Compounds | Biopterin, Rhamnopterin, Neopterin |
Application Column
Primesep 100
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Neopterin
Rhamnopterin
