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Dual-calibration coefficient: a more accurate protocol for simultaneous determination of superoxide and hydrogen peroxide in human HepG2 cell extracts

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  • Special Topic Analytical Sciences at the Nano-Bio Interface
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Abstract

We present a more accurate method for the quantification of superoxide anion (O2·−) and hydrogen peroxide (H2O2) simultaneously in human HepG2 cell extracts. After the xanthine/xanthine oxidase system was added into cell extract which was devoid of O2·− and H2O2, steady-state and in-situ produced O2·− and H2O2 by xanthine/xanthine oxidase system was labeled by fluorescent probes and subsequently separated by microchip electrophoresis. Based on this method, two differential equations with the calibration coefficients were established for O2·− and H2O2, respectively. Using the established dual-calibration coefficients, we obtained the calibrated concentrations of O2·− and H2O2 that produced in human HepG2 cells, which were lower (0.66±0.03 and 0.82±0.04 µmol/L for O2·− and H2O2, respectively) than that (0.85±0.03 and 0.96±0.03 µmol/L for O2·− and H2O2, respectively) obtained from statutory working curve. The proposed dual-calibration coefficient protocol takes into account both the complex matrix effect of the biological system and real time decaying of O2·− and H2O2, providing a method with higher accuracy.

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Authors and Affiliations

  1. College of Chemistry, Chemical Engineering and Materials Science; Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong; Key Laboratory of Molecular and Nano Probes, Ministry of Education; Shandong Provincial Key Laboratory of Clean Production of Fine Chemicals, Shandong Normal University, Jinan, 250014, China

    Hongmin Li, Lu Li, Xu Wang, Qingling Li & Bo Tang

  2. Department of Chemistry, Research Center of Low-Dimensional and Functional Nano-Materials, Qilu Normal University, Jinan, 250013, China

    Hongmin Li & Miao Du

Authors
  1. Hongmin Li
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  2. Lu Li
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  3. Xu Wang
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  4. Qingling Li
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  5. Miao Du
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  6. Bo Tang
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Correspondence to Bo Tang.

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Li, H., Li, L., Wang, X. et al. Dual-calibration coefficient: a more accurate protocol for simultaneous determination of superoxide and hydrogen peroxide in human HepG2 cell extracts. Sci. China Chem. 58, 825–829 (2015). https://doi.org/10.1007/s11426-015-5396-8

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  • DOI: https://doi.org/10.1007/s11426-015-5396-8

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