HPLC Method for Analysis of Rhamnopterin on Primesep 100 Column

https://sielc.com/hplc-separation-of-rhamnopterin
⬇ PDF Download

Chromatogram

Description

· High Performance Liquid Chromatography (HPLC) Method for Analysis of Rhamnopterin .

Rhamnopterin is a derivative of L-biopterin, D-biopterin, and the oxidized form of tetrahydro-L-biopterin (BH4) with the formula C 10 H 13 N 5 O 4 . It has exhibited significant properties in dietary studies. When administered in a 0.5% concentration alongside the carcinogen 4-dimethylaminoazobenzene, it can reduce the incidence of liver tumors induced by this carcinogen in rats. In addition to these therapeutic applications, rhamnopterin has also been utilized as an internal standard for quantifying biopterin and neopterin levels in rat plasma, as demonstrated in LC-MS-based studies. You can find detailed UV spectra of Rhamnopterin and information about its various lambda maxima by visiting the following link.

Rhamnopterin retained and analyzed using a reverse-phase Primesep 100 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.

Method Parameters

Mobile PhaseMeCN/H2O -5/95%
BufferAmmonium formate pH 3.0 – 20 mM
Flow Rate1.0 ml/min
DetectionUV 275 nm
Peak Retention Time3.28 min
Sample concentration0.02 mg/ml
Injection volume5 µl
Sample diluentH2O + NaOH
LOD20 ppb
Class of CompoundsPteridines
Analyzing CompoundsRhamnopterin

HPLC Column Used

Primesep 100, 4.6 x 150 mm, 5 µm, 100 A, dual ended