The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY (한국해양학회지:바다)

The Korean Society of Oceanography (한국해양학회)
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Stable Isotope Measurement of Ammonium Using HPLC-RTS (high performance liquid chromatography-retention time shift)

HPLC-RTS (high performance liquid chromatography-retention time shift)를 이용한 암모늄 이온의 안정동위원소 측정방법의 개선

  • An, Soonmo (Department of Oceanography, Pusan National University) ;
  • Lee, Jiyoung (Department of Oceanography, Pusan National University) ;
  • Gardner, Wayne S. (The University of Texas Marine Science Institute)
  • Received : 2013.02.04
  • Accepted : 2013.02.12
  • Published : 2013.02.28
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Abstract

Despite the usefulness of nitrogen isotope tracer experiments in nitrogen cycling studies, there are not such many measurement data mainly due to the difficulties in analytical methods. Although Gardner et al. (1996) developed a relatively simple and accurate method that can measure ammonium isotope using HPLC and used it widely in various N dynamics studies, the technique was not adopted to other laboratories. An HPLC-RTS system using updated HPLC pumps that can perform the same measurements as that of Gardner et al. (1996) was built. The result of standard sample showed linear increase of RTS with the $^{15}N$ proportions. Centroid retention times calculated with Matlab$^{(R)}$ program enhanced the linearity of the response. In a sea water incubation experiment spiked with $^{15}NH_4{^+}$, the uptake and regeneration of ammonium could be separately estimated using the temporal change of $^{15}N/^{14}N$.

생태계 질소 순환 연구에서 동위원소 추적자를 이용한 실험은 그 유용성에도 불구하고, 무기 질소의 안정동위원소 측정이 어려워 자료의 생산이 활발하지 못한 형편이다. Gardner et al. (1996)은 HPLC (high performance liquid chromatography)를 이용하여 비교적 간편하고 정밀하게 암모늄의 동위원소 비를 측정하는 방법을 개발해 질소순환 연구에 활발하게 활용하였다. 그러나 일반적이지 않은 syringe 펌프가 필요해 다른 실험실에 이 시스템이 도입되지는 못하였다. 본 연구에서는 최신 HPLC 펌프 기술을 활용하여 Dr. Gardner 그룹 이후 최초로 암모늄 동위원소비 측정 시스템을 구축하였다. 표준시료 측정 결과 RTS (retention time shift)는 안정동위원소 비와 좋은 상관관계를 보였다. 이 시스템은 측정 자료를 디지털 형태로 변환하는 장치까지 갖추어 컴퓨터 프로그램을 이용한 다양한 자료처리가 가능하다. 특히 Matlab$^{(R)}$ 이용하여 retention time 계산에 peak 면적을 이용함으로써 측정의 정확성을 높이고, 직선성을 향상할 수 있었다. 영산강 하구 해수를 가지고 실시한 $^{15}N$-암모늄 첨가 실험에서 $^{15}N/^{14}N$ 비율 변화를 HPLC-RTS법을 통해 측정하였으며, 식물성 플랑크톤에 의한 암모늄 섭취와 재광물화 과정을 각각 독립적으로 측정하는 것이 가능했다.

Keywords

References

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