A method for the determination of nitrification rates in oligotrophic marine seawater by gas chromatography/mass spectrometry

An isotope dilution method has been developed to determine by gas chromatography/mass spectrometry (GC/MS) the rates of ammonium and nitrite oxidation in severely oligotrophic marine waters. The method is based on the formation of sudan-1 from nitrite, or from nitrate following reduction to nitrite. Samples were collected by solid phase extraction and purified by high performance liquid chromatography (HPLC). A deuterated sudan-1 internal standard was synthesized, purified by HPLC and used for quantitative analysis. Concentrations of NO2− and NO3− were generally < 2 nmol/kg and < 5 nmol/kg respectively, typical of oligotrophic surface waters, and turnover times for the inorganic N pools ranged from < 1 day to > 10 days. Significant rates of nitrification were measured in the surface oligotrophic ocean, with rates of ammonium and nitrite oxidation generally within the range of 10–500 pmol/kg/h. Consequently, a significant proportion of daily NO3− assimilation by marine phytoplankton is regenerated, and not new. In a case study of the oligotrophic gyre of the North Atlantic, the influence of NH4+ regeneration and nitrification on f-ratio values suggests that in the oligotrophic ocean, f-ratio values may be significantly, and sometimes grossly, overestimated.