An enormous surge in the pavement sector requires the evaluation of interface bonding in asphalt composite, since the assessment of bonding brings considerable cost savings. Microscopic and mechanical analyses were performed to study the status of the interface transition zone of four groups of asphalt mixtures, using thin-slice preparation to obtain asphalt mixture slices with a flat surface for microscopic analysis. The interface transition zones were characterized using good knowledge of blending or diffusion phenomena by conducting tests both at the micro and macro levels to determine mixture quality. Asphalt mixture components were observed using fluorescence microscopy imaging and evaluated by the gray value change law. Asphalt mixture groups, (virgin, recycled of 30% aged and 70% unaged, 6%, and 4% rejuvenator dosage mixtures) under the same process parameters, which are a mixing time of 270 s and a mixing temperature of 150 °C, been considered optimum for component fusion in a hot asphalt mixture were used. This study relied on the influence of morphology law, assessed through rutting tests for high temperature performance, semi-circular bending tests for low temperature performance, and pull-off tests for interface bonding strength. The relationship between interface transition zones and macro performance was studied. The self-developed pull-off method was a research innovation which can be used as an alternative to study interface transition zones in asphalt mixtures, and provides the necessary data needed with 3D surface failure mode calculations. The device measured the bonding strength of a single aggregate in distinct positions using the bitumen penetration test method. The main goals were to determine a correction factor, identify the appropriate alteration, and compute the actual fracture surface area. Using scanning electron microscopy for interface characterization and micro-morphologies of mortar transition zone, our analysis provides adequate knowledge about interface position and the components present. The applied approaches to characterize asphalt mixture interfaces proved workable and reliable, as all methods have similar trends with useful information to determine asphalt pavement quality.