Pizotifen maleate (PZM) is an anti‐migraine drug. Effective treatment requires accurate measurement of PZM levels in patients and constant medication concentrations in tablets. This paper presents a new spectrofluorimetric approach for detecting PZM in tablets and human blood plasma (spiked with PZM). The detection mechanism depends on the inhibition of the PZM molecule's photoinduced electron transfer (PET), resulting in a bright fluorescent intensity from PZM. Acetic acid (2.0 M) acidifies the reaction environment and causes PET inhibition. This approach detects PZM at concentrations ranging from 100 to 700 ng mL−1. Furthermore, the approach has outstanding detection and quantification limits of 30.95 and 93.78 ng mL−1, respectively. Also, the approach accurately measured PZM content homogeneity in each tablet and spiked human plasma samples without any interference. This new strategy represents a new and sensitive method for accurate PZM monitoring in different samples. Furthermore, the established fluorimetric method's greenness was evaluated.