Process Analytical Technology means at-line collection of analytical information from the process when the reaction is in progress. Obtained information enables process engineers to better control the Critical Process Parameters and direct the reaction to desirable routs. Near-infrared spectroscopy due to its analytical features, as well as the high capability of automation, versatile sampling and spectral acquisition methods is a useful tool in process monitoring when coupled to chemometrics. The Multiple Scatter Correction preprocessing technique and Alternative Least Square method can extract spectral and concentration information of a reaction mixture simultaneously, were employed pairing with fiber optic Near-Infrared spectroscopy in 1000-2250 nm spectral region was employed to extract the analytical information from the biodiesel production process. The obtained output demonstrated appropriate results about the concentration and type of components during the process. The kinetics of the reaction was also studied using the obtained results and showed a three-stage kinetics with different rates.