The invention discloses a quantitative detection method for intracellular active sulfhydryl compound of white-rot fungi under heavy metal stress. The method comprises the following steps of: adding white-rot fungi spore into a culture solution, and performing shaking culture on the fungi spore till the logarithmic phase of the white-rot fungi; subsequently, adding a heavy metal solution into the white-rot fungi culture solution, performing shaking culture on the solution continually till a set sampling time, filtering and separating the culture, and washing the culture by buffer liquid at 0-4 DEG C; furthermore, mixing obtained mycelium pellets with the buffer liquid at 0-4 DEG C, carrying out ice-water bath ultrasonication on the mixture, and carrying out refrigerated centrifugation on obtained homogenate to obtain supernate which is taken as a sample; and measuring the absorbance of the sample by a method with a 5.5'-disulfenyl-2-nitrobenzene acid solution, and calculating the content of the active sulfhydryl compound in the sample. The detection method is simple, convenient, high in crushing efficiency and capable of rapidly and reliably acquiring and quantitatively detecting the content of the intracellular active sulfhydryl compound of the white-rot fungi.