HPLC Method for Separation of Biopterin, Rhamnopterin and Neopterin on Primesep 100 Column
Separation type: Liquid Chromatography Mixed-mode
These three pterins can be retained, separated and analyzed using a reverse-phase Primesep 100, 4.6 x 150 mm, 5 µm, 100 Å, dual ended 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.
Methodenparameter
| Column | Primesep 100, 4.6 x 150 mm, 5 µm, 100 Å, dual ended |
|---|---|
| Mobile Phase | MeCN/H2O -5/95% |
| Buffer | Ammonium formate pH 3.0 – 20 mM |
| Flow Rate | 1.0 mL/min |
| Detection | UV 275 nm |
| Limit of Detection | 20 ppb |
| Injection Volume | 5 µl |