Background:Tithonia diversifolia belongs to the Asteraceae family. The leaves of T. diversifolia have been studied lately because of the presence of tagitinin C.Objective:Looking for an easy and inexpensive method to extract tagitinin C from T. diversifolia leaves, this work aims to conduct a screening to evaluate the influence of different experimental factors using the dynamic maceration and ultrasound-assisted extraction methods with 23 factorial design based on response surface methodology in enhancing this chemical marker extraction.Materials and Methods:The experimental factors were: extraction time (ET) of 30 and 60 minutes, solid: liquid ratio (SLR) of 5 and 10 grams/grams and ethanolic strength (ES) 48 and 96% (w/w). The experiments were done tripled. The content of tagitinin C in each produced extract was quantified by HPLC method.Results:The highest concentrations of tagitinin C obtained under the experimental design were 0.53 mg/mL and 0.71 mg/mL, respectively for dynamic maceration (DM) and ultrasound-assisted extraction (UAE) from Tithonia diversifolia powdered leaves. For the UAE method, the main parameter for higher contents of tagitinin C was the solid: liquid ratio, followed by the ethanolic strength, and the extraction time was not significant for this method. As for the DM method, all the parameters (SLR, ES, and ET) were significant for a higher content of tagitinin C.Conclusion:Based on the obtained results, it was revealed that the ultrasound-assisted extraction was more effective than dynamic maceration for tagitinin C extraction from T. diversifolia powdered leaves.Summary Tithonia diversifolia leaves possess tagitinin C, a sesquiterpene lactone, as an important secondary metabolite with several biological activities, such as antimalarial, gastroprotective, chemotherapeutic adjuvants, and toxic activities.Ultrasound-assisted extraction was more effective to obtain higher levels of tagitinin C when compared with dynamic maceration extraction.Factorial design can be employed as a screening tool to find the effects of factors investigated in the extraction processes. Abbreviation used: DME: dynamic maceration extraction, UAE: ultrasound-assisted extraction, ET: extraction time, ES: ethanolic strength, SLR: solid:liquid ratio, Tag C: tagitinin C, HPLC: high-performance liquid chromatography.