Nitrate reduction in a groundwater microcosm determined by N gas chromatography-mass spectrometry

Aerobic and anaerobic groundwater continuous-flow microcosms were designed to study nitrate reduction by the indigenous bacteria in intact saturated soil cores from a sandy aquifer with a concentration of 3.8 mg of NO(3)-N liter. Traces of NO(3) were added to filter-sterilized groundwater by using a Darcy flux of 4 cm day. Both assimilatory and dissimilatory reduction rates were estimated from analyses of N(2), N(2)O, NH(4), and N-labeled protein amino acids by capillary gas chromatography-mass spectrometry. N(2) and N(2)O were separated on a megabore fused-silica column and quantified by electron impact-selected ion monitoring. NO(3) and NH(4) were analyzed as pentafluorobenzoyl amides by multiple-ion monitoring and protein amino acids as their N-heptafluorobutyryl isobutyl ester derivatives by negative ion-chemical ionization. The numbers of bacteria and their [methyl-H]thymidine incorporation rates were simultaneously measured. Nitrate was completely reduced in the microcosms at a rate of about 250 ng g day. Of this nitrate, 80 to 90% was converted by aerobic denitrification to N(2), whereas only 35% was denitrified in the anaerobic microcosm, where more than 50% of NO(3) was reduced to NH(4). Assimilatory reduction was recorded only in the aerobic microcosm, where N appeared in alanine in the cells. The nitrate reduction rates estimated for the aquifer material were low in comparison with rates in eutrophic lakes and coastal sediments but sufficiently high to remove nitrate from an uncontaminated aquifer of the kind examined in less than 1 month.