Despite being one of the less-characterized human isoforms, cytochrome P450 2B6 is already known for its participation in the metabolism of many drugs and several environmental carcinogens. It has been studied in different populations, but ethnicity is a crucial variable to account for interindividual variability. ; This study aimed to investigate the genotype and allelic frequencies of CYP2B6 c.516G>T SNP in an Azerbaijani population, as the determination of SNP's prevalence will be helpful in further pharmacogenetics research and optimization of personalized drug therapy in Azerbaijan. ; Identification of CYP2B6*6 allelic and genotype frequencies in 100 volunteers was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. The obtained results were confirmed by next-generation sequencing. ; The frequency of the *6 allele was 0.275, with the *1 allele frequency being 0.725. The frequency of the CYP2B6*1/*1, CYP2B6*1/*6 and CYP2B6*6/*6 genotypes were 0.55, 0.35, 0.1, respectively. ; This is the first investigation to report the frequencies of CYP2B6*/6 alleles in the Azerbaijani population. The results of this study suggest that genetic polymorphisms in the CYP2B6 gene are abundantly present among Azerbaijani individuals.