Supercritical fluid extraction (SFE) with carbon dioxide as extraction medium was on-line coupled to a FT-IR spectrometer equipped with a Mercury Cadmium Telluride (MCT) detector using a tailor-made high-pressure fibre optic flow cell. This method was optimised and developed for the monitoring in real time and the quantification of dynamic extractions of tagitinin C from Tithonia diversifolia leaves. In order to demonstrate the method ability to allow the direct quantification of tagitinin C in the extract medium the standard addition method was used. The area integration of curves obtained by plotting the absorbance of the highly specific C=O stretching vibration at 1668cm(-1) versus time (i.e. extractograms) was used as instrumental response. The SFE/FT-IR process was successfully validated using the accuracy profile as decision tool. On this basis, a linear regression model was chosen for the calibration curve. The relative standard deviation for repeatability and intermediate precision were between 0.8 and 3.1 %, respectively. Moreover, the method was found to be accurate as the two-sided 95% beta-expectation tolerance interval did not exceed the acceptance limits of 85 and 115% on the analytical range investigated (500-2500microg of added amount of tagitinin C). The proposed method allowed the non-destructive extraction of tagitinin C and its on-line quantitative determination in less than 25min thus facilitating the subsequent experiments or the pharmacological studies performed on this compound.