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A porous polyaniline nanotube sorbent for solid-phase extraction of the fluorescent reaction product of reactive oxygen species in cells, and its determination by HPLC

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Abstract

A method is described for extracting and detecting the fluorescent reaction product (2′,7′-dichlorofluorescein, DCF) that is formed by reaction of reactive oxygen species (ROS) with dichlorodihydrofluorescein diacetate (DCFH-DA). DCF is extracted by using porous polyaniline nanotubes (PPN) which have a large specific surface and pore volume which favor the adsorption capacity. Additional attractive features include an appropriate pore size distribution, hydrophobic surface, and electron-attracting groups which contribute to DCF adsorption. A variety of methods was applied to characterize the morphology of PPN. Under optimal conditions and by performing DCF in 0.08–1.0 μM concentrations, the correlation coefficient of the calibration plot is 0.999. The limits of detection for standard DCF solutions is 20 nM. Compared with commercial sorbents for solid-phase extraction (SPE) such as commercially available carbon or Welchrom® C18, the use of the new sorbent results in better retraction recovery (92%) and longer reuse times (30 times). Doxorubicin and X-ray radiation were used to externally stimulate the ROS production in HepG2 and Hela cells. ROS was stabled by DCFH-DA and quantified by DCF. Following SPE, DCF was detected by HPLC and the concentration ROS was calculated.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (21575149, 21505146 and 21575148).

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Authors and Affiliations

  1. CAS Key Laboratory of Chemistry of Northwestern Plant Resource and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou, Gansu, 730000, People’s Republic of China

    Panhong Niu, Xiaojing Liang, Xiudan Hou, Xiaofeng Lu, Xusheng Wang & Yong Guo

  2. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Panhong Niu, Feifei Li & Xiudan Hou

  3. Department of Medical Physics, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, Gansu, 730000, People’s Republic of China

    Feifei Li & Qiang Li

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  1. Panhong Niu
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  2. Feifei Li
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  3. Xiaojing Liang
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  5. Xiaofeng Lu
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Correspondence to Xiaojing Liang or Yong Guo.

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Niu, P., Li, F., Liang, X. et al. A porous polyaniline nanotube sorbent for solid-phase extraction of the fluorescent reaction product of reactive oxygen species in cells, and its determination by HPLC. Microchim Acta 185, 468 (2018). https://doi.org/10.1007/s00604-018-3000-6

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  • DOI: https://doi.org/10.1007/s00604-018-3000-6

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