Cellulase treatment of cellulose fibers needs to be monitored to give proper weight loss without significant strength loss. On-line monitoring of cellulase treatment is presented which can monitor the weight loss of cotton fabrics by measuring differential refractive index in real time. On-line monitoring was tried under the condition where the cellulase treatment of scoured 100% cotton knits would give weight loss of about 6.6% during one hour cellulase treatment. For comparison, processing solutions with treatment time of 5, 10, 20, 30, 45 and 60 minutes were taken out, and subjected to sugar analyses using total organic carbon (TOC), and dinitrosalicylic acid (DNS) method as well as HPLC analyses. HPLC analyses showed that cellobiose and glucose were the major components resulting from cellulase treatment of cotton fabric. TOC and differential refractive index measurement proved to be linear to total sugar concentration while DNS method result was dependent on the type of sugar. Various sugar concentrations of the processing solutions measured by these methods were compared with HPLC measurements and correlated with the actual weight loss of the cotton fabric processed. The on-line monitoring device using differential refractometer gave a real time signal which was independent of the ratio of cellobiose and glucose but dependent on the total sugar concentration. The device was also non-destructive.