This study investigates the chemical and morphological behavior of rejuvenated asphalt binders. A TFOT aged PG 58-28 binder was mixed with three different types of rejuvenators that include waste cooking oil/ untreated used cooking oil (UT), treated used oil (TR), and Hydrolene (HL) at 3%, 6% and 9% by the weight of the total binder. To understand the characteristics and the performance of these rejuvenators, three sets of characterization tests were conducted: rheological, chemical, and morphological. This paper presents a summary of results from our chemical and morphological studies. Gas Chromatography-Mass Spectroscopy (GC-MS) was conducted to identify the chemical composition of rejuvenators, while Fourier Transformed Infrared Spectroscopy (FTIR) was conducted for obtaining chemical functional group information. Furthermore, an Atomic Force Microscopy (AFM) was studied to obtain the micro-morphological properties of rejuvenated asphalt binders. The collected data showed that rejuvenation changes the chemical composition and alters the micro-structures of binders significantly, which impacts the overall performances of the binder. In fact, this experimental study showed a good correlation between chemical compositional and morphological features with the rheological performance of the binder. The research findings are expected to contribute to the performance evaluation and characterization of rejuvenated asphalt mixes.