The degradation performance of pentachlorophenol (PCP) by the microwave-activated persulfate (MW/PS) process was investigated in this study. The results indicated that degradation efficiency of PCP in the MW/PS process followed pseudo-first-order kinetics, and compared with conventional heating, microwave heating has a special effect of increasing the reaction rate and reducing the process time. A higher persulfate concentration and reaction temperature accelerated the PCP degradation rate. Meanwhile, increasing the pH value and ionic strength of the phosphate buffer slowed down the degradation rate. The addition of ethanol and tert-butyl alcohol as hydroxyl radical and sulfate radical scavengers proved that the sulfate radicals were the dominant active species in the MW/PS process. Gas chromatography-mass spectrometry (GC-MS) was employed to identify the intermediate products, and then a plausible degradation pathway involving dechlorination, hydrolysis, and mineralization was proposed. The acute toxicity of PCP, as tested with Photobacterium phosphoreum, Vibrio fischeri, and Vibrio qinghaiensis, was negated quickly during the MW/PS process, which was in agreement with the nearly complete mineralization of PCP. These results showed that the MW/PS process could achieve a high mineralization level in a short time, which provided an efficient way for PCP elimination from wastewater.