Recently, high-temperature superconductor (HTS) cables have been widely studied because of their compactness and high power capacity compared to conventional copper cables. In HTS cables, AC loss is an important issue since large losses reduce the efficiency of the power line. Among HTS cables, tri-axial cable is under intensive investigation recently, since it has a smaller amount of HTS tapes, small leakage fields and small heat loss in leak when compared with the three single-phase cables. For realizing high current capacity, more than one layer is required for each phase; therefore AC loss of the multi-layer tri-axial HTS cable should be carefully examined. In the tri-axial cable, different phase currents produce the out-of-phase magnetic fields on the other phase layers. In case of multi-layer arrangement, net magnetic fields on layer surfaces may exceed the penetration field of the HTS tape. Therefore in this paper, we analyze the AC loss of a tri-axial HTS cable which is composed of two layers per phase. Here, we treat the tri-axial cable which consists of two different longitudinal segments and thus satisfies balanced phase and homogeneous current distribution condition by controlling twist pitch and length of separate segments.