Korean Journal of Ecology and Environment (생태와환경)

The Korean Society of Limnology (한국수생태학회)
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Introduction of Denitrification Method for Nitrogen and Oxygen Stable Isotopes (δ15N-NO3 and δ18O-NO3) in Nitrate and Case Study for Tracing Nitrogen Source

탈질미생물을 이용한 질산성 질소의 산소 및 질소 동위원소 분석법 소개

  • Lim, Bo-La (Environmental Measurement & Analysis Center, NIER) ;
  • Kim, Min-Seob (Environmental Measurement & Analysis Center, NIER) ;
  • Yoon, Suk-Hee (Environmental Measurement & Analysis Center, NIER) ;
  • Park, Jaeseon (Environmental Measurement & Analysis Center, NIER) ;
  • Park, Hyunwoo (Environmental Measurement & Analysis Center, NIER) ;
  • Chung, Hyen-Mi (Fundamental Environment Research Department, NIER) ;
  • Choi, Jong-Woo (Environmental Measurement & Analysis Center, NIER)
  • 임보라 (국립환경과학원 환경측정분석센터) ;
  • 김민섭 (국립환경과학원 환경측정분석센터) ;
  • 윤숙희 (국립환경과학원 환경측정분석센터) ;
  • 박재선 (국립환경과학원 환경측정분석센터) ;
  • 박현우 (국립환경과학원 환경측정분석센터) ;
  • 정현미 (국립환경과학원 환경기반연구부) ;
  • 최종우 (국립환경과학원 환경측정분석센터)
  • Received : 2017.12.23
  • Accepted : 2017.12.31
  • Published : 2017.12.31
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Abstract

Nitrogen (N) loading from domestic, agricultural and industrial sources can lead to excessive growth of macrophytes or phytoplankton in aquatic environment. Many studies have used stable isotope ratios to identify anthropogenic nitrogen in aquatic systems as a useful method for studying nitrogen cycle. In this study to evaluate the precision and accuracy of denitrification bacteria method (Pseudomonas chlororaphis ssp. Aureofaciens ($ATCC^{(R)}$ 13985)), three reference (IAEA-NO-3 (Potassium nitrate $KNO_3$), USGS34 (Potassium nitrate $KNO_3$), USGS35 (Sodium nitrate $KNO_3$)) were analyzed 5 times repeatedly. Measured the ${\delta}^{15}N-NO_3$ and ${\delta}^{18}O-NO_3$ values of IAEA-NO-3, USGS 34 and USGS35 were ${\delta}^{15}N:4.7{\pm}0.1$‰ ${\delta}^{18}O:25.6{\pm}0.5$‰, ${\delta}^{15}N:-1.8{\pm}0.1$‰ ${\delta}^{18}O:-27.8{\pm}0.4$‰, and ${\delta}^{15}N:2.7{\pm}0.2$‰ ${\delta}^{18}O:57.5{\pm}0.7$‰, respectively, which are within recommended values of analytical uncertainties. Also, we investigated isotope values of potential nitrogen source (soil, synthetic fertilizer and organic-animal manures) and temporal patterns of ${\delta}^{15}N-NO_3$ and ${\delta}^{18}O-NO_3$ values in river samples during from May to December. ${\delta}^{15}N-NO_3$ and ${\delta}^{18}O-NO_3$ values are enriched in December suggesting that organic-animal manures should be one of the main N sources in those areas. The current study clarifies the reliability of denitrification bacteria method and the usefulness of stable isotopic techniques to trace the anthropogenic nitrogen source in freshwater ecosystem.

본 연구는 박테리아 탈질법을 이용하여 질산성 질소의 질소 및 산소의 안정동위원소 분석법을 연구하였으며, 시료 농도, 미생물 배양시간에 따른 분석값의 변화에 대하여 고찰하였다. 탈질미생물법을 이용하여 시료 내 질산염 농도가 $0.1mg\;L^{-1}$까지 질산성 질소의 산소 및 질소 동위원소 분석이 가능하였고, 0.2‰까지 정확도를 확보하였다. 환경시료(지하수) 내 질산염의 기원을 추적하기 위하여 잠재적 질소 오염원(합성비료, 축산비료)의 동위원소비를 조사한 결과, 합성비료(${\delta}^{15}N-NO_3$ -5~10‰, ${\delta}^{18}O-NO_3$ 0~15‰)는 축산비료(${\delta}^{15}N-NO_3$ 10~23‰, ${\delta}^{18}O-NO_3$ 0~20‰)와 동위원소비가 현격한 차이를 보였다. 연구지역 지하수 동위원소비와 비교한 결과, 계절별로 서로 다른 질소 오염원의 영향을 받는 것으로 여겨진다. 과거 질산염의 안정동위원소를 분석하기 위해서 다양한 방법이 시도되었다. Revesz et al. (1997)은 양이온 교환 수지를 이용한 분리법을 보고하였으며, Silva et al. (2000)와 Fukada et al. (2003)은 음이온 교환 수지를 이용한 분리법을 보고하였고, McIlvin and Altabet (2005)는 카드뮴을 이용한 화학적 변환 방법을 보고하였다. 하지만 이러한 방법에 사용되는 시약은 독성이 강하고 복잡한 전처리 과정으로 인한 긴 전처리 시간을 소요함으로 인하여 분석비용이 비싸다는 단점이 있었다. 하지만 탈질미생물법은 소량의 질산염을 이용하기 때문에 기존 방법에 비해 분석에 사용되는 시료의 부피를 1/100까지 감소시켜 과거 분석이 어려웠던 빙하, 공극수, 해수 등에 대한 분석이 가능한 장점이 있다. 수생태계로 유입되는 다양한 질소 기원을 파악하기 위하여 탈질미생물법으로 분석된 안정동위원소비를 활용한다면 효율적인 수질 관리를 위한 해석기능을 제공할 것이다. 또한, 국내 최초로 지하수 환경시료에 적용함으로써 오염 기원 추적 기법을 확립하는 데 목적을 두고 있으며, 추후 정립된 분석기법을 토대로 환경분야에서 질산성 질소의 오염 인자 판별 연구에 널리 활용되기를 기대한다.

Keywords

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