Objective To explore the assay method of acidity of fresh royal jelly determined by automatic potentiometric titrimeter, Method The digital operating interface was set up with LabX software, and each parameter of automatic operating interface was set. The acidity of fresh royal jelly was determined by automatic potentiometric titrimeter. Results The recoveries were from 90.0% to 96.3 % , and the RSD was 0.24 % . Conclusion The results of acidity for fresh royal jelly determined by automatic potentiometric titrimeter were no significant difference with the results obtained by GB/T 9697-2002. The digital operating interface in whole experiment process was fulfilled, The method has the characteristic of simple, fast, accuracy and efficiency.