A new spectrophotometric method for determination of chromium content was developed. In both polyvinyl alcohol and phosphoric acid medium, chromium (VI) could react with potassium iodide and safranine T at room temperature, leading to coloration after microwave digestion sample and oxidation of chromium (III) to chromium (VI) with potassium permanganate under alkaline conditions. The effects of the factors such as absorption spectra, reaction conditions and coexisting ions on the reaction were discussed. The results showed that under the optimum determination conditions, Beer-Lambert law was obeyed in the mass concentration range of 0.0～0.5 μg/mL for chromium with the maximum absorption peak at 460 nm. The detection limit was 14.55 μg/L. The apparent molar absorption coefficient was 6.90×10⁴ L/(mol·cm). This method had been applied to the determination of chromium content in beer yeast. The results obtained by this method agreed well with those obtained by atomic absorption spectrometry. RSD was in the range of 1.3%～2.1% (n=5), and recovery was in the range of 97.9%～101.7%. Moreover, a large number of common coexisting ions did not affect the determination.