The invention discloses a specific fluorescence probe for identifying thiophenol and application of the specific fluorescence probe and belongs to the field of fine chemical industry. The specific fluorescence probe is a derivative of 2-benzothiazole-6-naphthol and is prepared by the following steps: mixing the 2-benzothiazole-6-naphthol and 2, 4-dinitrobenzene into an N, N-dimethylformamide solution according to a proportion, heating, and finally, purifying by adopting silica gel chromatography to obtain the fluorescence probe. The fluorescence probe and a corresponding thiophenol content detecting process can not be interfered by matrixes and impurities in a biological system and can be used for quantitatively determining the thiophenol content in various biological systems. The specific fluorescence probe is high in specificity, can be hydrolyzed after being acted with the thiophenol, namely an ether bond is fractured; is cheap and is easy in acquisition, can be obtained by chemical synthesis, is simple and feasible in synthetic process; is high in sensitivity, is good in fluorescence attribute. A hydrolysate can be excited by a two-photon laser with 800nm as an excitation light source, a biological sample is weak in background fluorescence, and the specific fluorescence probe is suitable for detecting the thiophenol content in cells and is capable of quantitatively determining the thiophenol by drawing a standard curve.