In the present study, we report a novel composition based on amylose (or starch) inclusion complex with an amphiphilic material as an effective encapsulation platform technology to incorporate guests of interest. Specifically, the encapsulation of β-carotene in amylose-surfactant and amylose/starch-ascorbyl palmitate (AscP) inclusion complexes was investigated. Surfactants of different hydrophilicity/lipophilicity were selected to cover a broad range of HLB values. The formation of the inclusion complexes was characterized by X-ray diffraction and differential scanning calorimetry. The ability of amylose-surfactant system to encapsulate β-carotene was dependent on the HLB value of the surfactants, instead of their ability to induce inclusion complexation. The incorporation of β-carotene hindered amylose-surfactant inclusion complex formation, whereas no significant effect was observed on structural and thermal properties of starch-AscP inclusion complex in the presence of β-carotene. The X-ray diffraction pattern of amylose-AscP-β-carotene showed that β-carotene molecules did not crystallize into a separated phase and thus were suggested to be homogeneously immobilized within the polycrystalline amylose-AscP inclusion complexes. During a storage period of six weeks at 20 and 30 °C, the stability of β-carotene was improved by encapsulation in starch-AscP inclusion complexes compared with that in physical mixtures of the three components.