OBJECTIVE: To establish the method for the simultaneous determination of content of 6 active components as neochlorogenic acid, mulberroside A, chlorogenic acid, astragalin, sanggenon C and morusin in Morus alba, and to provide reference for improving quality control standard of M. alba. METHODS: HPLC method was adopted. The determination was performed on Agilent 5 TC-C18 with mobile phase consisted of acetonitrile-0.1% formic acid (gradient elution) at the flow rate of 1 mL/min. The detection wavelength of 280 nm. RESULTS: The mass concentration linear range of neochlorogenic acid, mulberroside A, chlorogenic acid, astragalin, sanggenon C and morusin were 0.001 06-0.042 4, 0.001 67-0.066 8, 0.007 95-0.318, 0.001 65-0.066 0, 0.005 00-0.200 and 0.001 24-0.049 6 mg/mL, respectively (all r≥0.999 6); the limits of quantitation were 0.11, 0.14, 0.81, 0.17, 0.45 and 0.12 μg/mL, respectively; the limits of detection were 0.04, 0.05, 0.41, 0.07, 0.18 and 0.04 μg/mL, respectively; RSDs of precision test were 0.26%, 0.31%, 0.24%, 0.27%, 0.36% and 0.44% (n=6), respectively; RSDs of stability test were 0.68%, 0.54%, 0.62%, 0.53%, 0.41% and 0.73%(n=6), respectively; average method recovery rates were 99.1%, 98.8%, 98.8%, 98.4%, 98.5% and 99.9%(RSDs were 0.5%-1.5%, n=9), respectively. CONCLUSIONS: The method is simple, accurate, and can be used for simultaneous determination of 6 active components in M. alba.