In this article, we demonstrate the potential application of polarimetry and fluorescence spectroscopy for classifying mono and disaccharides (sugar) both qualitatively and quantitatively. A phase lock-in rotating analyzer (PLRA) polarimeter has been designed and developed for real time quantification of sugar concentration in a solution. Polarization rotation in the form of phase shift in sinusoidal photovoltages of reference and sample beams occurred when incident on the two spatially distinct photodetectors. Monosaccharide (fructose and glucose) and disaccharide (sucrose) have been quantitatively determined with sensitivities of 122.06 deg ml g−1, 272.84 deg ml g−1 and 163.41 deg ml g−1 respectively. Calibration equations have been obtained from the respective fitting functions to estimate the concentration of each individual dissolved in deionized (DI) water. In comparison to the predicted results, the absolute average errors of 1.47 %, 1.63 % and 1.71 % are calculated for the readings of sucrose, glucose and fructose, respectively. Furthermore, the performance of the PLRA polarimeter has been compared with fluorescence emission results acquired from the same set of samples. The Limit of detections (LODs) attained from both experimental setups are comparable for mono and disaccharides. A linear detection response is observed by both polarimeter and fluorescence spectrometer in a wide range 0–0.28 g/ml of sugar. These results depict that PLRA polarimeter is novel, remote, precise and cost-effective for quantitative determination of optically active ingredient in the host solution.