<p> <?tk 2?>Chronic mask use in healthcare professionals might influence change in genetic composition of hypoxia inducible factor-1α (HIF-1α) gene. The study determined the genetic make of HIF-1α gene and determined the association of variants with serum HIF-1α levels and metabolic effects in individuals using masks. The study was conducted on thirty healthy health professionals using masks and twenty not using masks. Serum HIF-1α was measured using ELISA and gene sequencing done for HIF-1α gene. 28% of the study population depicted HIF-1α variants that included 23 exonic and intronic variants. Genetic variants in exposed and control groups were 30% and 25% respectively with OR 1.29 (95% CI 0.36–4.62). Only the individuals of exposed group showed presence of three different variants. 87% variations were seen in the exons. The common exonic variants in exposed group were seen in exons 6 and 3. A very novel variant in exon 14 was observed for c.2289_2290insTT (P.Asn764 LeufsTer11). HIF-1α level was significantly elevated in HIF-1α variants (<i>p</i> = 0.022), specifically in those with exon 3 and 6. The variant group depicted significantly raised pCO₂ (<i>p</i> = 0.008), low pH (<i>p</i> = 0.047) and low glucose levels (<i>p</i> = 0.02). The study depicted novel HIF-1α gene variants due to chronic usage of masks to adapt and sustain the metabolic demand of hypoxic state.</p><p> <?qj left?> <?noindent??><i>CTRI Number</i>: CTRI/2023/06/054233 [Registered on 21/06/2023].</p>