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Stable isotopic analyses of soil-emitted N2O(δ15Nbulk, δ18O and δ15Nsp = 15N site preference within the linear N2O molecule) may help toquantify N2O reduction to N2, an important but rarely quantifiedprocess in the soil nitrogen cycle. The N2O residual fraction (remainingunreduced N2O, rN2O) can be theoretically calculated fromthe measured isotopic enrichment of the residual N2O. However, variousN2O-producing pathways may also influence the N2O isotopicsignatures, and hence complicate the application of this isotopicfractionation approach. Here this approach was tested based on laboratory soil incubations with twodifferent soil types, applying two reference methods for quantification ofrN2O: helium incubation with direct measurement of N2 flux and the15N gas flux method. This allowed a comparison of the measuredrN2O values with the ones calculated based on isotopic enrichment ofresidual N2O. The results indicate that the performance of the N2Oisotopic fractionation approach is related to the accompanying N2Oand N2 source processes and the most critical is the determination ofthe initial isotopic signature of N2O before reduction (δ0). We show that δ0 can be well determined experimentallyif stable in time and then successfully applied for determination ofrN2O based on δ15Nsp values. Much more problematicto deal with are temporal changes of δ0 values leading to failureof the approach based on δ15Nsp values only. For thiscase, we propose here a dual N2O isotopocule mapping approach, wherecalculations are based on the relation between δ18O and δ15Nsp values. This allows for the simultaneous estimation of theN2O-producing pathways' contribution and the rN2O value

biogeosciences

Quantifying N&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;O reduction to N&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt; based on N&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;O isotopocules – validation with independent methods (helium incubation and &amp;lt;sup&amp;gt;15&amp;lt;/sup&amp;gt;N gas flux method)