Bio-oil, a major product from biomass pyrolysis, is a renewable source for production of functional carbon materials such as activated carbon (AC). Bio-oil can be activated directly via mixing with an activator or be pre-carbonized followed by subsequent activation. These two processes feature with distinct reaction network and might affect pore characteristics in different ways. This was investigated herein by direct activation of bio-oil or biochar from pre-carbonization of bio-oil using K2C2O4 or KOH as the activator at 800 ºC. The results indicated that bio-oil was pre-carbonized into biochar at 500 ºC improved yields of AC (83.6 % versus 17.5 % from direct activation of bio-oil with K2C2O4) through enhanced aromatic degree and resistivity to cracking. This also diminished specific surface area of resulting AC (716.3 versus 1035.5 m2g−1 from direct activation). Much more intensive cracking reactions in direct activation of bio-oil with K2C2O4 were confirmed with in-situ IR technique. This not only formed AC of more developed pore structures, especially more mesopores/macropores, but also generated gases as dominate product (yield: 74.5 %). Similar result was observed from activation with KOH, but KOH was more effective than K2C2O4 for cracking. This, however, did not result in AC of more developed pore structures, as the K2CO3 derived from KOH tended to be wrapped with organics in direct activation of bio-oil and was difficult to be washed away, blocking pores generated.