The shock impact of toxic pollutants in petrochemical wastewater on the activated sludge in biological treatment system is a key factor restricting the treatment efficiency. It is necessary to evaluate the toxicity of these pollutants by appropriate methods. In this study, four test methods were used to evaluate the toxicity of characteristic organic pollutants in petrochemical wastewater including 2,4-dichlorophenol, formaldehyde and pyridine, as well as one frequently-used reference toxicant 3,5-dichlorophenol. The sensitivity, accuracy and response time were compared among these methods: the oxygen consumption rate inhibition method (OCRIM), the dehydrogenase activity inhibition method (DAIM), the nitrification rate inhibition method (NRIM) and the growth rate inhibition method (GRIM). Principal component analysis was used to evaluate the correlation among the results of different methods. The OCRIM was comprehensively outstanding with the highest correlation between concentration and inhibition ratio (R2 values were all higher than 0.9854), good sensitivity, best accuracy (error value of the effective concentrations below 0.15 mg/l) and fastest response (<40 min). The sensitivity of the NRIM was found to be the highest in this study (10% effective inhibition concentration (EC10) value of 3,5-dichlorophenol was only 0.03 mg/l). Therefore, combined tests of OCRIM and NRIM were suggested.