The anti-agglomerating hydrate behavior observed for some crude oils has previously been related to crude oil composition and to surface adsorption mechanisms. Petroleum acids derived from some crude oils have been found able to convert systems with initially high risk of plugging into easily flowable dispersions. In this work, acid fractions are isolated from three oils with low tendency to form hydrate plugs and from two oils associated with high risk of hydrate plugging by using an ion-exchange resin. The extracts are further separated into four sub-fractions by solid phase extraction (SPE). The chemical composition of the fractions is studied by means of HPLC, GPC, FTIR- and UV/VIS spectroscopy and elemental analysis. The distribution of chemical compound classes in the fractions differs between the non-plugging and plugging oils, and the differences are most distinctive in one of the sub-fractions. The results imply that acid sub-fractions holding a significant proportion of more weakly polar compounds, like ester functionalities, are important for how the hydrate surfaces and the oil phase interact.