A Bacterial Method for the Nitrogen Isotopic Analysis of Nitrate in Seawater and Freshwater

We report a new method for measurement of the isotopic composition of nitrate (NO3-) at the natural-abundance level in both seawater and freshwater. The method is based on the isotopic analysis of nitrous oxide (N2O) generated from nitrate by denitrifying bacteria that lack N2Oreductase activity. The isotopic composition of both nitrogen and oxygen from nitrate are accessible in this way. In this first of two companion manuscripts, we describe the basic protocol and results for the nitrogen isotopes. The precision of the method is better than 0.2” (1 SD) at concentrations of nitrate down to 1 muM, and the nitrogen isotopic differences among various standards and samples are accurately reproduced. For samples with 1 muM nitrate or more, the blank of the method is less than 10% of the signal size, and various approaches may reduce it further.