To facilitate the understanding of the intrinsic mechanism responsible for the IFTs of betaine solutions, partly hydrolyzed polyacrylamide (HPAM) and hydrophobically modified polyacrylamide (HMPAM) were employed to investigate their effect on IFTs of different betaine solutions against decane using a spinning drop interfacial tensiometer. Moreover, the effects of oleic acid on the IFTs of betaine-polymer systems were also studied. The results show that the water-soluble polymers (HPAM) form the mixed adsorption film with alkyl sulfobetaine (ASB) molecules and prevent the hydrophilic groups of ASB from lying flat, which produces a minimum IFT with aging time. Besides, the hydrophobic HMPAM forms mixed micelle-like interfacial associations with ASB molecules, which results in a stronger disruption tendency for the interfacial film. The branched xylyl substituted alkyl sulfobetaine (BSB) molecules will form a tight interfacial film, therefore HPAM can’t insert into the interfacial film easily. Meanwhile, steric hindrance makes it difficult to form interfacial aggregates with HMPAM. Thus, the addition of HPAM and HMPAM polymers has little influence on the IFTs in the BSB system. The mixed adsorption of oleic acid and betaine molecules results in a significant decrease in IFTs, especially for BSB. However, in the ASB-polymer system, HPAM forms mixed adsorption film with ASB molecules and hinders the adsorption of oleic acid, accordingly IFTs change little. In contrast, the hydrophobic interaction of HMPAM promotes the adsorption of oleic acid, so the synergistic effect reduces IFTs to some extent. In the BSB-polymer system, the strong synergistic effect between oleic acid and BSB molecules is disrupted by polymers, which results in a significant increase in IFTs. The relevant results are helpful for the design of novel and efficient chemical flooding systems.