HPLC Method for Separation of a Mixture of Tryptophan and its Catabolites on Primesep 100 Column

High Performance Liquid Chromatography (HPLC) Method for Analysis of Mixture of Tryptophan and its Catabolites on Primesep 100 by SIELC Technologies

Separation type: Liquid Chromatography Mixed-mode

HPLC Method for Analysis of Mixture of Tryptophan and its Catabolites on Primesep 100 Column by SIELC Technologies

Tryptophan and its catabolites participate in several biological pathways, having roles in protein synthesis, serving as precursors to bioactive molecules, and influencing several physiological processes. Here’s an overview considering a mixture of tryptophan and its catabolites:

Tryptophan:

  • Essential Amino Acid: Tryptophan is a precursor to several important compounds, including serotonin and melatonin.
  • In Protein Synthesis: Incorporated into proteins during protein synthesis.

Catabolites:

1. Serotonin:

  • Neurotransmitter: Regulates mood, appetite, and sleep, among other functions.
  • Derivative: Melatonin, which regulates the sleep-wake cycle.

2. Kynurenine Pathway (Major catabolic pathway of tryptophan):

  • Kynurenine: An intermediate and precursor to several bioactive compounds.
  • Kynurenic Acid: An NMDA receptor antagonist, believed to have neuroprotective effects.
  • Xanthurenic Acid: Its physiological roles are still being explored, but it’s often studied for its relation to diabetes and neurological conditions.
  • 3-Hydroxykynurenine: Can generate reactive oxygen species, potentially contributing to cellular stress.
  • Quinolinic Acid: A neuroactive metabolite that can act as an NMDA receptor agonist.

3. Indoleamine 2,3-dioxygenase (IDO) Pathway:

  • Tryptophan can be degraded into several catabolites via the IDO pathway, influencing immune response and cell proliferation.

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Tryptophan and its Catabolites can be retained, separated and analyzed on a Primesep 100 mixed-mode stationary phase column using an gradient analytical method with a simple mobile phase of water, Acetonitrile (MeCN), and a sulfuric acid as a buffer. This analysis method can be detected using UV at 220 nm.

High Performance Liquid Chromatography (HPLC) Method for Analyses of Mixture of Tryptophan and its Catabolites

Condition

ColumnPrimesep 100, 4.6 x 150 mm, 5 µm, 100 A
Mobile PhaseGradient MeCN 5-50% in 10 min, with 2 min hold afterwards
BufferGradient H2SO4 from 0.1-0.2% in 10 min, with 2 min hold afterwards
Flow Rate1.0 ml/min
DetectionUV 220 nm

Description

Class of CompoundsEssential Amino Acid Tryptophan and its Catabolites
Analyzing CompoundsTryptophan, Picolinic Acid, Kynurenine, 3-Hydroxykynurenine, 3-Hydroxyanthranilic acid

Application Column

Primesep 100

Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:
3-Hydroxyanthranilic acid
3-Hydroxykynurenine
Kynurenine
Picolinic Acid
Tryptophan
Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.