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Estimation and in-situ detection of thorium in human liver cell culture by arsenazo-III based colorimetric assay

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Abstract

To understand the biological effects of Thorium-232 (Th) in human cells and animal models as well as to assess mitigation strategies for its detoxification, there is a need to develop a sensitive, specific, high-throughput and easily-implementable assay for detection and estimation of Th in biological samples. Here, we have optimized arsenazo-III dye based colorimetric assay to detect Th in biological samples. The concentration of arsenazo-III (i.e. 50 µM) was optimized, which can reliably estimate Th in the concentration range of 2.5 to 40 µM. The optimized assay can specifically detect Th without interference from other metal ions (La, Ce, U, Fe, Ca, Cu, Zn and Mn). A significant correlation (R2 = 0.999) was found between arsenazo-III-based detection of Th and total reflection X-ray fluorescence. The conditions of present assay successfully estimated Th in cell culture medium, cell harvesting (trypsin-EDTA) solution and cell lysate obtained from human liver cell culture. Moreover, for the first time, we detected Th in-situ in adherent liver cells in culture after staining with arsenazo-III. This study confirms that Th can be specifically determined in biological samples using arsenazo-III with the sensitivity, which is relevant to thorium toxicity research.

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Acknowledgements

Rakhee Yadav would like to acknowledge graduate fellowship provided by Homi Bhabha National Institute (HBNI), Department of Atomic Energy (DAE), Government of India. This work was funded by DAE, India. Authors acknowledge Dr. N. L. Mishra and Dr. S. Dhara for their help in TXRF. We also acknowledge Mr. Manjoor Ali and Mrs. Neena Shetake for critically reading the manuscript and providing their valuable inputs.

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Authors and Affiliations

  1. Radiation Biology & Health Sciences Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400 085, India

    Rakhee Yadav, Badri N. Pandey & Amit Kumar

  2. Homi Bhabha National Institute, Anushaktinagar, Mumbai, 400094, India

    Rakhee Yadav, Badri N. Pandey & Amit Kumar

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  1. Rakhee Yadav
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  2. Badri N. Pandey
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  3. Amit Kumar
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Correspondence to Amit Kumar.

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Yadav, R., Pandey, B.N. & Kumar, A. Estimation and in-situ detection of thorium in human liver cell culture by arsenazo-III based colorimetric assay. Biometals 33, 75–85 (2020). https://doi.org/10.1007/s10534-019-00231-2

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  • DOI: https://doi.org/10.1007/s10534-019-00231-2

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