Classification of biomaterial using polarization of light at present having difficulty for label-free and direct optical detection. The optical properties of a sample which are profoundly explored through the absorption coefficient, scattering coefficient, anisotropy coefficient and degree of linear polarization (DoLP) are neither simple nor easy to handle. In this study, Angle of polarized light (AOP) is our biggest concern. Neither need labeling procedure nor hardly to measure. Instead of linear polarization, this study determined the angle of polarized light as a potent parameter for polarization measurement at the variation axis of transmitted polarized light. Hence, this work was mainly conducted to identify the angle of polarized light for classification of agarose sample, a three-dimensional crosslinked polymer. In this work, a photodiode acts as a polarized light sensor to read voltage changes due to variable concentrations of agarose samples. At the end of the study, relationships between the Angle of Polarized light (AOP) and concentrations of agarose sample at variation axis of transmitted light were successfully investigated. Our result demonstrated a linear correlation between measured voltage (mV) and the concentration of agarose sample (g/ml) with output polarization behavioral model (AOP, ϴ◦) at variation axis of transmitted light. This outcome concluded that the polarization property of the agarose sample in perspective of angle can be identified at variation axis of transmitted light. Therefore, our polarization measurement-setup with variation axis of light transmission is reliable to determine the polarization property of the unknown three-dimensional structure of tissue-mimicking phantom in the future.