HPLC using a borate form of a strongly anion-exchange resin column and an immobilized enzyme reactor for colorimetric detection was used to quantify urinary 1,5-anhydro-D-glucitol. Urine samples were introduced into the system every 7 min without any pretreatment, and after separation of interfering substances in the column, 1,5-anhydro-D-glucitol was successively detected. Quantitative determination of urinary 1,5-anhydro-D-glucitol was possible within the 1.2-300 micromol/l range. The coefficient of variance was less than 3% and the correlation between results obtained with our system (y) and those obtained by gas chromatography-mass spectrometry (x) was y=0.983x-1.287 micromol/l (n=42, r=0.998).