In the present study, the reliability of Jung-Biggs-Moorhead method for cholesterol determination has been tested in the presence of another steroid, ergosterol. It was observed that presence of ergosterol interferes with the absorbance measurement of cholesterol. When both steroids are present in the sample maximum absorption wavelength are not shifted by the mutual interaction but absorbance values are changed considerably. Therefore this method may be used only for qualitative bu not quantitative analysis for cholesterol in the presence of ergosterol.