Abstract: A UV spectrophotometer used for determination of equol was established. Using equol standard and the urine equol from the tester who ate the soybean isoflavones for samples, the ultraviolet method character of determine equol was studied through UV spectrophotometer to determine the detection limit and quantitative limit. The result was as follows:the detection limit was 5 ×10-10g/mL, the quantitative limit was 15 ×10-10g/mL.Application for equol measurement of human urine, the precision was 4.45% and the average recovery rate was 100.25%, the result was reliable. The new UV method conforms to the quantitative analysis of the requirements was, it had advantages of simple, rapid, and high sensitivity. It was suitable for soybean isoflavone and equol established measuirement application.