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GC/TOFMS Analysis of Endogenous Metabolites in Mouse Fibroblast Cells and Its Application in TiO2 Nanoparticle-Induced Cytotoxicity Study

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Abstract

In this paper, we present an optimized procedure for metabolomic analysis of endogenous metabolites in mouse fibroblast (L929) cell line using gas chromatography/time-of-flight mass spectrometry with multivariate statistics. The optimization of metabolite extraction was performed using three solvents: methanol, water, and chloroform, and then followed by methoxymation and silylation. This method was subsequently validated using 29 reference standards and cell line samples. The intra- and inter-day relative standard deviations (RSDs) of the standard compounds were lower than 15.0 and 25.0 %, respectively. As for most of the tested metabolites in cell line samples, RSDs were below 20.0 % for reproducibility and stability, respectively. We applied this approach in metabolomic study of L929 cells obtained from TiO2 nanoparticle-induced cytotoxicity model samples (n = 5) and control samples (n = 5). Metabolite markers associated with TiO2 nanoparticle-induced cytotoxicity were identified and validated by statistical methods and reference standards. Our work highlights the potential of this method for cell metabolomic study.

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Acknowledgments

This work was financially supported by Shanghai Natural Science Foundation of the Science and Technology Commission of Shanghai Municipal Government (No. 09ZR1415100), two Fundamental Key Project (No. YG2010MS92; YG2011MS66) of Shanghai Jiaotong University.

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Authors and Affiliations

  1. School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 200240, China

    Yumin Liu, Yinan Zhang & Aihua Zhao

  2. Center for Instrumental Analysis, Shanghai Jiao Tong University, Shanghai, 200240, China

    Yumin Liu, Yang Bo & Chengyu Jin

  3. Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, 200240, China

    Tianlu Chen & Xiaoyan Wang

  4. Department of Chemistry, East China Normal University, Shanghai, 200240, China

    Yu Cheng

  5. Department of Nutrition, University of North Carolina at Greensboro, North Carolina Research Campus, Kannapolis, NC, 28081, USA

    Wei Jia

Authors
  1. Yumin Liu
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  2. Yu Cheng
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  3. Tianlu Chen
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  4. Yinan Zhang
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  5. Xiaoyan Wang
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  7. Wei Jia
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  9. Chengyu Jin
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Correspondence to Chengyu Jin.

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Liu, Y., Cheng, Y., Chen, T. et al. GC/TOFMS Analysis of Endogenous Metabolites in Mouse Fibroblast Cells and Its Application in TiO2 Nanoparticle-Induced Cytotoxicity Study. Chromatographia 75, 1301–1310 (2012). https://doi.org/10.1007/s10337-012-2315-4

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  • DOI: https://doi.org/10.1007/s10337-012-2315-4

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