The Raman spectra of the native ribonuclease treated with nitrobenzene in aqueous solution have been obtianed. The conformation of the main chain and the side chain in ribonuclease were studied. Among the amide Ⅰ and amide Ⅲ bands, the characters of β-sheet and random coil structures are clear. The disulfide bridges assume the gauche-gauche-gauche conformation. The parts of tyrosine residue are buried. It indicates that nitrobenzene treatment on protein aqueous solution is an efficient means for obtaining better Raman spectra. The Raman spectra of the native ribonuclease treated with nitrobenzene in aqueous solution have been obtianed. The conformation of the main chain and the side chain in ribonuclease were studied. Among the amide Ⅰ and amide III bands, the characters of β-sheet and random coil The structures of clear. The disulfide bridges assume the gauche-gauche-gauche conformation. The parts of tyrosine residue are buried. It indicates that nitrobenzene treatment on protein aqueous solution is an efficient means for better Raman spectra.