title: 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.
abstract: 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.
authors.affiliation: Autism Research Unit, Department of Pharmacy, Health and Well-being, Faculty of Applied Sciences, University of Sunderland, Sunderland SR1 3SD, UK. kevin.carr@sunderland.ac.uk
authors.forename: Kevin
authors.initials: K
authors.lastname: Carr
authors.forename: Paul
authors.initials: P
authors.lastname: Whiteley
authors.forename: Paul
authors.initials: P
authors.lastname: Shattock
mesh_terms: D019540:Area Under Curve; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005998:Glycine / Q000031:analogs & derivatives* / Q000652:urine; D006801:Humans; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: C001807:indolylacryloylglycine; D005998:Glycine