AIM:To establish a quantitative method for determination of hesperidin in Citri reticulatae in order to evaluate their quality. METHODS:Sample was extracted with methanol by means of sonication for 30min. ODS column was used with methanol-water-acetic acid(35∶61∶2)as mobile phase. Detection wavelength was at 284nm. RESULTS:Hesperidin in sample solution was well separated. Linearity was good(r=0.9995)in range of 0.202～2.424?g. The average recovery was 98%, RSD of repeatability was 2.3%. CONCLUSION:This method is simple, accurate and reliable, which may well be used for the determination of hesperidin.