This study evaluated the performance of two bio-based, spray-applied rejuvenators derived from soybean oil at the National Center for Asphalt Technology (NCAT) Test Track. Phase I, conducted in the north paddock parking area, provided a short-term assessment of a sub-epoxidized soybean oil (SESO) formulation and a polymer-modified variant (SESO + polymer). Performance was assessed two and four weeks after application. Both products effectively softened the asphalt binder and enhanced relaxation, as evidenced by reductions in complex modulus, lower performance grades (PGs), and higher ΔTc values. Friction, measured using the Dynamic Friction Tester (DFT), decreased immediately after application and remained relatively stable over several hours, staying lower for up to one month. Phase II extended the evaluation to a seven-year-old trafficked pavement section (W10) to assess longer-term performance at one and six months after treatment application. Both rejuvenators reduced PGs and lowered Glover-Rowe (G-R) parameter values compared with the untreated control section, indicating sustained binder softening and improved cracking resistance. In contrast to Phase I, no reduction in surface friction was observed following treatment. Field monitoring showed no adverse effects on ride quality (IRI) or surface texture (MPD), and no rutting. After approximately 3.6 million equivalent single axle loads (ESALs) of truck traffic, the treated sections exhibited visibly less surface cracking than the control. Overall, both spray-on products effectively restored oxidized surface binders, improved cracking resistance, and maintained surface safety, demonstrating strong potential as sustainable bio-based pavement preservation treatments.