Abstract:The content of orange juice in fruit juice beverage was determined by high performance liquid chromatography （HPLC）.The peak area of new hesperidin，was determined by determination of one of flavonoids in Orangejuice.Quantitative standard peak of new hesperidin in pure orange juice chromatogram.The reference substances of new hesperidin with different concentrations were prepared and the standard curve was established by peak area and concentration.The content of new hesperidin in orange juice was determined and the content of orange juice was calculated.The method was simpler and faster than the national standard method.The recovery rate was 98.52 %-102.04 %.The relative standard deviation （RSD）was 0.69 %-3.08 %.The relative error was 0.67 %-3.66 %.The precision and accuracy of the method meet the requirements of orange juice content analysis.