Production of polychlorinated biphenyls (PCBs) has been banned since 2002 but health risks from exposure persist. PCBs are metabolized by cytochrome P450 enzymes, including CYP2B6. However, the link between CYP2B6 gene polymorphisms and PCB metabolisms is poorly characterized. This study investigated the relationships among serum levels of major indicator PCBs and hydroxylated PCBs (OH-PCBs), which are PCB metabolites, and polymorphisms in CYP2B6. Blood samples (n = 129) were analyzed for single nucleotide polymorphisms in CYP2B6 (p.Q172H and p.K262R), and the corresponding haplotypes (*1, *4, *6) were determined. Concentrations of PCBs and OH-PCBs were determined using gas chromatography and mass spectrometer. Congener-specific variations in PCB metabolism were associated with different CYP2B6 genotypes, particularly * 1/* 4 (hypothesized to increased expression) and * 6/* 6 (hypothesized to decreased expression). For certain PCBs, the * 1/* 4 genotype was linked to increased metabolite-to-parent compound ratios, while * 6/* 6 was associated with decreased ratios, as observed for PCB146 (β = 0.192, 95 % CI: [0.100, 0.283], p <
0.0002 for *1/*4
β = -0.235, 95 % CI: [-0.366, -0.105], p = 0.002 for *6/*6). However, other PCBs, such as PCB170 and PCB183, exhibited opposite or more complex patterns. Our findings indicate intricate effects of CYP2B6 gene polymorphisms on PCB metabolism and highlight the potential for genotype-specific risks in PCB-related toxicity.