Analytical and Bioanalytical Chemistry

, Volume 405, Issue 14, pp 4811–4822 | Cite as

Discovery of safety biomarkers for realgar in rat urine using UFLC-IT-TOF/MS and 1H NMR based metabolomics

  • Yin Huang
  • Yuan Tian
  • Geng Li
  • Yuanyuan Li
  • Xinjuan Yin
  • Can Peng
  • Fengguo Xu
  • Zunjian Zhang
Research Paper


As an arsenical, realgar (As4S4) is known as a poison and paradoxically as a therapeutic agent. However, a complete understanding of the precise biochemical alterations accompanying the toxicity and therapy effects of realgar is lacking. Using a combined ultrafast liquid chromatography (UFLC) coupled with ion trap time-of-flight mass spectrometry (IT-TOF/MS) and 1H NMR spectroscopy based metabolomics approach, we were able to delineate significantly altered metabolites in the urine samples of realgar-treated rats. The platform stability of the liquid chromatography LC/MS and NMR techniques was systematically investigated, and the data processing method was carefully optimized. Our results indicate significant perturbations in amino acid metabolism, citric acid cycle, choline metabolism, and porphyrin metabolism. Thirty-six metabolites were proposed as potential safety biomarkers related to disturbances caused by realgar, and glycine and serine are expected to serve as the central contacts in the metabolic pathways related to realgar-induced disturbance. The LC/MS and NMR based metabolomics approach established provided a systematic and holistic view of the biochemical effects of realgar on rats, and might be employed to investigate other drugs or xenobiotics in the future.


Pipeline of safety biomarkers discovery for realgar in rat urine by metabolomics


Realgar Metabolomics Liquid chromatography–mass spectrometry NMR Urine Biomarker 



This work was supported by the Central University Foundation for Fundamental Research of China (no. JKY2009027).

Supplementary material

216_2013_6842_MOESM1_ESM.pdf (732 kb)
ESM 1 (PDF 731 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Yin Huang
    • 1
    • 2
  • Yuan Tian
    • 1
    • 2
  • Geng Li
    • 1
    • 2
  • Yuanyuan Li
    • 1
    • 2
  • Xinjuan Yin
    • 1
    • 2
  • Can Peng
    • 1
    • 2
  • Fengguo Xu
    • 1
    • 2
  • Zunjian Zhang
    • 1
    • 2
  1. 1.Key Laboratory of Drug Quality Control and PharmacovigilanceChina Pharmaceutical University, Ministry of EducationNanjingChina
  2. 2.Department of Pharmaceutical AnalysisChina Pharmaceutical UniversityNanjingChina

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