The effect of cyanide concentrations on the secondary conformation of protein in the fish gill homogenate was determined using an attenuated total reflectance (ATR)/Fourier transform infrared (FT-IR) microspectroscopy. Gills from male Tilapia zillii were isolated and homogenized in pH 8.0 Tris buffer solution and subjected to FT-IR study. The results indicate that the amide I and III bands of protein in fish gill homogenate deformed markedly with the increase of cyanide concentration. The fish gill homogenate shows a maximum peak at 1650 cm(-1) in amide I band, suggesting the predominant proportion of alpha-helical conformation. Once the KCN was added into the gill homogenate, the maximum peak shifted gradually from 1650 to 1643 cm(-1) due to the random coil structure, with the increase of cyanide concentration used. Two additional shoulders at 1657 (alpha-helix) and 1627 (beta-sheet) cm(-1) also appeared gradually, implying that the cyanide can in part induce changes in protein conformation of fish gill homogenate from alpha-helix to random coil and beta-sheet conformations.