The goal of the study was to investigate the influence of pH and water hardness on protein contents of muscle tissues of Cirrhinus mrigala fingerlings by Fourier transform infrared spectroscopy. FT-IR spectra revealed significant differences in absorbance intensities between control and toxic metal-treated muscle tissues, reflecting a change in protein contents due to heavy metals, nickel and chromium, influence at different pH and water hardness. Metal toxicity is also more pronounced at pH 9.0 than in hard water.