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To better understand the nature of reactive adsorption of thiophene on Ni/ZnO adsorbent, the effect of ZnO textural structure on the desulfurization activity was investigated. ZnO materials were synthesized by low-temperature solid-state reaction and the corresponding Ni/ZnO adsorbents were prepared by incipient impregnation method. The analysis results showed that the crystalline sizes of ZnO as-synthesized as well as the BET surface areas varied obviously with the calcination temperature. The activity evaluations indicated that the Ni/ZnO adsorbents prepared with ZnO possessed a favorable textural structure as active component exhibited good activity of removing thiophene. The evolutions of the main crystalline phases of Ni/ZnO adsorbents before and after reaction confirmed that ZnO played a crucial role in taking up S element and converting it into ZnS in the reactive adsorption process. It was concluded that ZnO with larger surface area and smaller crystal particles resulted in better desulfurization activity, which may be the main reason for the different activities of the Ni/ZnO adsorbents prepared with ZnO calcined at different temperatures.