High-performance liquid chromatographic profiling of indolylacryloylglycine and its possible precursors in body fluids

Indolylacryloylglycine (IAcrGly) is one of the physiological components of urine, although its source and its role in the human organism have not yet been unambiguously established. Changes in the IAcrGly excretion level have been found under some physiological (age dependence, seasonal variations) and pathological (photodermatoses, muscle dystrophy, liver cirrhosis) conditions. The proposed method for IAcrGly, indolylacrylic acid and its possible precursors, namely indolyllactic and indolylpropionic acids, involves deproteinization and extraction of urine on a Sep Pak C-18 cartridge. HPLC analysis was carried out using a DataApex liquid chromatograph, equipped with an LCD 2082 UV detector, signals being acquired with a CSW workstation. The chromatographic column was Spherisorb ODS, 5 mu m (125 x 4 mm I.D.), the mobile phase for isocratic elution was ethanol-1% acetic acid (27:73) and the flow-rate was 0.7 ml/min. The lower response limit is about 1 mu mol/l for all metabolites at 280 nm.