Few studies have been performed up to now on the emission factors and congener profiles of persistent organic pollutants (POPs) emitted from off-road diesel engines. This investigation elucidates the emission factors and congener profiles of various POPs, namely polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polychlorinated biphenyl (PCBs), polybrominated dibenzo-p-dioxins and polybrominated dibenzofurans (PBDD/Fs) and polybrominated diphenyl ethers (PBDEs), in the exhausts of a diesel generator fueled with different waste cooking oil-based biodiesel (WCO-based biodiesel) blends. The PCDD/Fs contributed 87.2% of total dioxin-like toxicity (PCDD/Fs+PCBs+PBDD/Fs) in the exhaust, while the PCBs and PBDD/Fs only contributed 8.2% and 4.6%, respectively. Compared with petroleum diesel, B20 (20vol% WCO-based biodiesel+80vol% diesel) reduced total toxicity by 46.5% for PCDD/Fs, 47.1% for PCBs, and 24.5% for PBDD/Fs, while B40 (40vol% WCO-based biodiesel+60vol% diesel) reduced it by 89.5% for PCDD/Fs, 57.1% for PCBs, and 63.2% for PBDD/Fs in POP emission factors. The use of WCO-based biodiesel not only solves the problem of waste oil disposal, but also lowers POP emissions from diesel generators.