Near-near infrared (N-NIR) Thermal Lens Spectroscopy (TLS) was used to monitor the transesterification reaction during biodiesel production. Fifteen aliquots were extracted from the reaction during 20 min and their thermo-optical properties were measured as a function of the reaction time. A spectroscopic investigation was performed between 855 and 990 nm to identify overtone absorptions bands (markers) to monitor the chemical reaction. The results show that the OH absorption overtone (similar to 970 nm) from alcohol, which is used in the reaction, is the most appropriated marker to monitor the biodiesel production. The time dependence of the thermo-optical parameter A(dn/dQ), which is the product between the absorption coefficient (A) and the refractive index dependence with the sample deposited heat (dn/dQ), exhibits a very good correlation with the ester production measured by chromatography. Besides, the measured thermal diffusivity values as function of reaction time indicates a strong dependence with the viscosity of the aliquots. It is possible to conclude that the used TLS in the N-NIR region contributes to the optimization of biodiesel production process by indicating the best time to stop the reaction.