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. 1994 Jul;60(7):2467–2472. doi: 10.1128/aem.60.7.2467-2472.1994

Determination of 15N Abundance in Nanogram Pools of NO3- and NO2- by Denitrification Bioassay and Mass Spectrometry

Ole Højberg 1,*, Henrik Saaby Johansen 2, Jan Sørensen 1
PMCID: PMC201672  PMID: 16349326

Abstract

Suspensions of two strains of Pseudomonas aeruginosa (ON12 and ON12-1) were used to reduce NO3- and NO2-, respectively, to N2O. The evolved N2O was quantified by gas chromatography with electron capture detection, and the 15N abundance was determined by mass spectrometry with a special inlet system and triple-collector detection. Sample gas containing unknown N2O pools as small as 0.5 ng of N was analyzed by use of a spike technique, in which a reference gas of N2O of natural 15N abundance was added to obtain enough total N for the mass spectrometer. In NO3- or NO2- pools, the 15N abundance could be determined in samples as small as approximately 3.5 ng of N. No cross-contamination took place between the NO3- and NO2- pools. The excellent separation of NO3- and NO2- pools, small sample size required, and low contamination risk during N2O analysis offer great advantages in isotope studies of inorganic N transformations by, e.g., nitrifying or denitrifying bacteria in the environment.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Binnerup S. J., Sørensen J. Nitrate and nitrite microgradients in barley rhizosphere as detected by a highly sensitive denitrification bioassay. Appl Environ Microbiol. 1992 Aug;58(8):2375–2380. doi: 10.1128/aem.58.8.2375-2380.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blackburn T. H. Method for Measuring Rates of NH(4) Turnover in Anoxic Marine Sediments, Using a N-NH(4) Dilution Technique. Appl Environ Microbiol. 1979 Apr;37(4):760–765. doi: 10.1128/aem.37.4.760-765.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Christensen S., Tiedje J. M. Sub-Parts-Per-Billion Nitrate Method: Use of an N(2)O-Producing Denitrifier to Convert NO(3) or NO(3) to N(2)O. Appl Environ Microbiol. 1988 Jun;54(6):1409–1413. doi: 10.1128/aem.54.6.1409-1413.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Griffiths L., Cole J. A. Lack of redox control of the anaerobically-induced nirB+ gene of Escherichia coli K-12. Arch Microbiol. 1987 May;147(4):364–369. doi: 10.1007/BF00406134. [DOI] [PubMed] [Google Scholar]
  5. Koike I., Hattori A. Simultaneous determinations of nitrification and nitrate reduction in coastal sediments by a 15N dilution technique. Appl Environ Microbiol. 1978 May;35(5):853–857. doi: 10.1128/aem.35.5.853-857.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Stouthamer A. H. Biochemistry and genetics of nitrate reductase in bacteria. Adv Microb Physiol. 1976;14(11):315–375. doi: 10.1016/s0065-2911(08)60230-1. [DOI] [PubMed] [Google Scholar]

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