HPLC Method for Analysis of Xanthurenic acid on Newcrom BH Column by SIELC Technologies
Separation type: Liquid Chromatography Mixed-mode
Xanthurenic acid is a compound associated with tryptophan metabolism and is known to be excreted in the urine. It is a non-essential, non-proteinogenic amino acid derivative and is considered to be a metabolite of tryptophan. Here are some key points about xanthurenic acid:
Structure:
Xanthurenic acid is characterized by the presence of two hydroxy groups and a carboxylic acid group. Structurally, it is composed of a bicyclic ring system that incorporates a benzene ring and a pyridine ring (with a nitrogen atom). The benzene ring is substituted with two hydroxy groups at positions 4 and 8, and the pyridine ring carries a carboxylic acid functionality.
Physiological Role and Metabolism:
- Pathway: Xanthurenic acid is part of the kynurenine pathway, which is the major catabolic route for tryptophan.
- Enzyme Involvement: It is formed from 3-hydroxykynurenine by the action of the enzyme kynurenine 3-monooxygenase and can further be converted to 2-amino-3-carboxymuconate semialdehyde by xanthurenate 8-monooxygenase.
- B6 Dependency: Synthesis of xanthurenic acid is dependent on vitamin B6 (pyridoxal phosphate), as it is a cofactor for enzymes in the tryptophan degradation pathway.
Xanthurenic acid can be retained and analyzed on a mixed-mode Newcrom BH column with a mobile phase consisting of water, Acetonitrile (MeCN), and sulfuric acid. This analytical method can detect compounds with high resolution and peak symmetry using UV detection at 200 nm
High Performance Liquid Chromatography (HPLC) Method for Analyses of Xanthurenic acid
Condition
Column | Newcrom BH, 4.6 x 150 mm, 5 µm, 100 A |
Mobile Phase | Gradient MeCN – 10-60%, 10 min |
Buffer | H2SO4 – 0.2% |
Flow Rate | 1.0 ml/min |
Peak Retention Time | 5.58 min |
Detection | 250 nm |
Description
Class of Compounds | Acid |
Analyzing Compounds | Xanthurenic acid |
Application Column
Newcrom BH
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A