A derivative spectrophotometric method has been developed for the determination of uranium at mg level in presence of iron in 1M HNO3 medium. Its application was extended to the determination of uranium is magnesium diuranate (Yellow Cake), an input material to the Uranium Conversion Plant. The second derivative absorbance at 408.2 nm was used for its quantification since at this wavelength the absorbance due to iron is negligibly small and that due to uranium is significant. The uranium content was found to be directly porportional to the amplitude at 408.2 nm. It was demonstrated that the impurities present in yellow cake do not interfere. The method is simple, fast, needs no prior separation and enables direct measurement of yellow cake samples which is an important requirement from the point of view of nuclear materials accounting.