In the present study, a novel and sensitive fluorescence probe method has been established for the determination of luteolin (LTL) based on the fluorescence quenching effect of LTL on glutathione (GSH)-capped Mn-doped CdTe quantum dots (QDs). GSH-capped Mn-doped CdTe QDs with a diameter of 6–8 nm were synthesized via a facile and inexpensive method in an aqueous system. The fluorescence of doped CdTe QDs at 541 nm was quenched in the presence of LTL with an excitation wavelength at 351 nm and the fluorescence quenching efficiency was closely related to the amount of LTL added to the QDs solution; they presented a linear relationship when the concentrations of LTL vary from 6 to 138 μM. The mechanism of the interaction between doped CdTe QDs and LTL was investigated and was confirmed to be a dynamic quenching mechanism. The proposed method was successfully applied to the determination of LTL, and satisfactory results were obtained.