The method was developed and validated according to ICH requirements for specificity, linearity, accuracy, precision (repeatability and intermediate precision), LOD and LOQ. Rutin was chosen as standard for the quantification of total flavonoids in order to propose a HPLC method feasible for routine analysis of the flavonoids in the passion fruit pulp. The passion fruit pulp contained 16.226±0.050 mg L−1 of isoorientin and 158.037±0.602 mg L−1 of total flavonoid, suggesting that P. edulis fruits may be comparable with other flavonoid food sources such as orange juice or sugarcane juice.