Tryptophan Metabolites MRM Method


Measured Analytes

  1. Nicotinic Acid
  2. 2-Picolinic Acid
  3. Quinolinic Acid
  4. Serotonin
  5. Kynurenine
  6. 5-OH Tryptophan
  7. Tryptophan
  8. Tryptamine
  9. Indole-3-lactate
  10. Indole-3-acetate

Chromatography

Liquid chromatography tandem mass spectrometry (LC-MS) was carried out with a Shimadzu Prominence HPLC system and SCIEX 4000 mass spectrometer. An aliquot of 10 µl of each sample was injected onto an T3 Atlantis 3µm C18 100 x 2.0 mm column at 40oC for gradient separation. Mobile phases were: A, ddH2O with 0.1% Formic Acid; and B, Acetonitrile with 0.1% Formic Acid. Gradient schedule started at 0%B for 0.5 min and was linearly increased to 60%B at 6 minute; then rapidly increased to 95%B at 8 minutes. At 8.25 minutes the gradient was returned to starting conditions and allowed to re-equilibrate until 12.0 minutes. Flow rate was 400µL/min.

Extraction Protocols:

Extraction protocols for sample processing are dependent on the matrices involved. Please see extraction methods section for more specific instructions.

References

Method derived from the following

[1] A mass spectrometric method for quantification of tryptophan-derived uremic solutes in human serum
Anqi Zhang, Keshab Rijal, Seng Kah Ng, Katya Ravid, and Vipul Chitalia,
JBiol Methods. 2017; 4(3): e75.
Published online 2017 Jul 31. doi: 10.14440/jbm.2017.182
PMID: 29707597

Resources

This tab might be populated with additional information like PowerPoint slides from presentations at conferences, class presentations, grant associated links, and other components that would be directly tied to the method.