Chitosan has excellent biodegradable, biocompatible and bio-absorbable properties and has been found increasing use in the biomedical field in recent decades. The linear birefringence (LB), linear diattenuation (LD), circular birefringence (CB), circular diattenuation (CD), and depolarization properties of chitosan hydrogel films crosslinked in citrate acid buffer solution (CBS) are extracted using an analytical Mueller matrix method. It is shown that the optical phase retardance property of the hydrogel films provides a reliable indication of both the chitosan concentration of the film and the pH value of the CBS crosslinking environment. In addition, chitosan hydrogel suspension with low-concentration crosslinked in CBS environments with various pH values are studied by the speckle contrast of the projected images obtained when illuminating the suspension with a scanned laser pico-projector (SLPP). It is found that for the samples crosslinked in an acidic environment, the speckle contrast decreases with an increasing pH value. By contrast, for the samples crosslinked in an alkaline CBS environment, the speckle contrast increases as the pH value increases. It is concluded that both the phase retardance and the speckle contrast enable the pH value of the CBS crosslinking solution to be reliably determined. However, of the two methods, the SLPP method yields improved measurement sensitivity. Overall, the results presented in this study show that the analytical Mueller matrix method and SLPP method provide an effective means of characterizing the optical properties, concentration and crosslinking environment of chitosan hydrogel films and suspensions.