Development and reproducibility of a novel high-performance liquid-chromatography monolithic column method for the detection and quantification of trans-indolyl-3-acryloylglycine in human urine

Biomed Chromatogr. 2009 Oct;23(10):1108-15. doi: 10.1002/bmc.1231.


Elevated levels of trans-indolyl-3-acryloylglycine (IAcrGly) have been reported in the urine of people with various conditions including pervasive developmental disorders (PDDs) such as autism and Asperger syndrome. Reversed-phase high-performance liquid chromatography with ultra-violet detection using traditional particle silica-based columns subsequent to solid-phase extraction (SPE) has been the preferred assay method; requiring long analytical run times, high flow rates and high solvent usage. Recent developments in monolithic HPLC column technology facilitated the development of a novel analytical method, for the detection and quantification of urinary IAcrGly. The revised method eliminates the requirement for SPE pre-treatment, reduces sample run-time and decreases solvent volumes. Five urine samples from people diagnosed with PDD were run in quadruplicate to test the intra- and inter-day reliability of the new method based on retention time, peak area and peak height for IAcrGly. Detection was by UV with IAcrGly confirmation by MS/MS-MS. Relative standard deviations showed significant improvement with the new method for all parameters. The new method represents a major advancement in the detection and quantification of IAcrGly by reducing time and cost of analysis whilst improving detection limits and reproducibility.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Area Under Curve
  • Chromatography, High Pressure Liquid / methods*
  • Glycine / analogs & derivatives*
  • Glycine / urine
  • Humans
  • Linear Models
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Solid Phase Extraction
  • Tandem Mass Spectrometry / methods


  • indolylacryloylglycine
  • Glycine