To estimate benthic denitrification in a marginal sea, we assessed the usefulness of $${\text{N}}_{2}^{*}$$ N 2 ∗ , a new tracer to measure the excess nitrogen gas (N2) using dissolved N2 and argon (Ar) with N* in the intermediate layer (26.6–27.4σ θ ) of the Okhotsk Sea. The examined parameters capable of affecting $${\text{N}}_{2}^{*}$$ N 2 ∗ are denitrification, air injection and rapid cooling. We investigated the relative proportions of these effects on $${\text{N}}_{2}^{*}$$ N 2 ∗ using multiple linear regression analysis. The best model included two examined parameters of denitrification and air injection based on the Akaike information criterion as a measure of the model fit to data. More than 80 % of $${\text{N}}_{2}^{*}$$ N 2 ∗ was derived from the denitrification, followed by air injection. Denitrification over the Okhotsk Sea shelf region was estimated to be 5.6 ± 2.4 μmol kg−1. The distribution of $${\text{N}}_{2}^{*}$$ N 2 ∗ was correlated with potential temperature (θ) between 26.6 and 27.4σ θ (r = −0.55). Therefore, we concluded that $${\text{N}}_{2}^{*}$$ N 2 ∗ and N* can act complementarily as a quasi-conservative tracer of benthic denitrification in the Okhotsk Sea. Our findings suggest that $${\text{N}}_{2}^{*}$$ N 2 ∗ in combination with N* is a useful chemical tracer to estimate benthic denitrification in a marginal sea.