The invention relates to preparation process of amphipathic octenyl succinic short-straight chain starch nano-particles. The preparation process comprises the following steps: (1) performing enzymolysis on gelatinized starch by using pullulanase, and obtaining short straight chain starch; (2) preparing a short straight chain starch solution, performing gelatinization, adding an octenyl succinic anhydride solution which accounts for 25-100% of the weight of dry powder of short straight chain starch, and continuously stirring for 6-10 hours, so as to obtain a modified octenyl succinic short straight chain starch solution; preparing a 1-10mg / mL solution of the octenyl succinic short-straight chain starch, stirring for 6-10 hours at 37-40 DEG C, and cooling to the room temperature, thereby obtaining a amphipathic octenyl succinic short-straight chain starch nano-particle solution. The particle size of the nano-particles is within 5-100nm, the nano-particles are good in tissue adhesion property, are capable of wrapping hydrophobic active substances, are high in loading rate and low in cost, the conveying efficiency of lyophobic active components by gastrointestinal tracts can be improved, and the bioavailability of the nano-particles can be improved. The lyophobic active components can be protected, the stability of the lyophobic active components can be improved, and the release of adverse flavor can be shielded; in addition, the addition amount and the toxic and side effects of bioactive components can be effectively reduced.