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Hokkaido University Collection of Scholarly and Academic Papers >
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Determination of N-15/N-14 of Ammonium, Nitrite, Nitrate, Hydroxylamine, and Hydrazine Using Colorimetric Reagents and Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS)
| Title: | Determination of N-15/N-14 of Ammonium, Nitrite, Nitrate, Hydroxylamine, and Hydrazine Using Colorimetric Reagents and Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS) |
| Authors: | Oshiki, Mamoru Browse this author →KAKEN DB | | Nagai, Komei Browse this author | | Ishii, Satoshi Browse this author | | Suzuki, Yoshiyuki Browse this author | | Saito, Nobuo Browse this author | | Yamaguchi, Takashi Browse this author | | Araki, Nobuo Browse this author | | Okabe, Satoshi Browse this author →KAKEN DB |
| Keywords: | nitrogen cycle | | nitrogen isotope | | N-15 atom% | | mass spectrometry | | colorimetric reagent | | MALDI-TOF MS |
| Issue Date: | Apr-2022 |
| Publisher: | American Society for Microbiology |
| Journal Title: | Applied and Environmental Microbiology |
| Volume: | 88 |
| Issue: | 7 |
| Start Page: | e02416-21 |
| Publisher DOI: | 10.1128/aem.02416-21 |
| Abstract: | In the nitrogen (N) cycle, nitrogenous compounds are chemically and biologically converted to various aqueous and gaseous N species. The N-15-labeling approach is a powerful culture-dependent technique to obtain insights into the complex nitrogen transformation reactions that occur in cultures. In the N-15-labeling approach, the fates of supplemented N-15- and/or unlabeled gaseous and aqueous compounds are tracked by mass spectrometry (MS) analysis, whereas MS analysis of aqueous N species requires laborious sample preparation steps and is performed using isotope-ratio mass spectrometry, which requires an expensive mass spectrometer. We developed a simple and high-throughput MS method for determining the N-15 atoms percent of NH4+, NO2-, NO3-, NH2OH, and N2H4, where liquid samples (<0.5 mL) were mixed with colorimetric reagents (naphthylethylenediamine for NO2-, indophenol for NH4+, and p-aminobenzaldehyde for N2H4), and the mass spectra of the formed N complex dyes were obtained by matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) MS. NH2OH and NO3- were chemically converted to NO2- by iodine oxidation and copper/hydrazine reduction reaction, respectively, prior to the above colorimetric reaction. The intensity of the isotope peak (M + 1 or M + 2) increased when the N complex dye was formed by coupling with a N-15-labeled compound, and a linear relationship was found between the determined N-15/N-14 peak ratio and N-15 atom% for the tested N species. The developed method was applied to bacterial cultures to examine their N-transformation reactions, enabling us to observe the occurrence of NO2- oxidation and NO3- reduction in a hypoxic Nitrobacter winogradskyi culture. IMPORTANCE N-15/N-14 analysis for aqueous N species is a powerful tool for obtaining insights into the global N cycle, but the procedure is cumbersome and laborious. The combined use of colorimetric reagents and MALDI-TOF MS, designated color MALDI-TOF MS, enabled us to determine the N-15 atom% of common aqueous N species without laborious sample preparation and chromatographic separation steps; for instance, the N-15 atom% of NO2- can be determined from >1,000 liquid samples daily at <$1 (U.S.) per 384 samples for routine analysis. This convenient MS method is a powerful tool that will advance our ability to explore the N-transformation reactions that occur in various environments and biological samples. N-15/N-14 analysis for aqueous N species is a powerful tool for obtaining insights into the global N cycle, but the procedure is cumbersome and laborious. The combined use of colorimetric reagents and MALDI-TOF MS, designated color MALDI-TOF MS, enabled us to determine the N-15 atom% of common aqueous N species without laborious sample preparation and chromatographic separation steps; for instance, the N-15 atom% of NO2- can be determined from >1,000 liquid samples daily at <$1 (U.S.) per 384 samples for routine analysis. |
| Rights: | Copyright © American Society for Microbiology |
| Type: | article |
| URI: | http://hdl.handle.net/2115/86857 |
| Appears in Collections: | 工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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