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Electrocatalytic determination of reduced glutathione in human erythrocytes

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Abstract

The determination of reduced glutathione (GSH) in human erythrocytes using a simple, fast and sensitive method employing a glassy carbon electrode modified with cobalt tetrasulfonated phthalocyanine (CoTSPc) immobilized in poly(l-lysine) (PLL) film was investigated. This modified electrode showed very efficient electrocatalytic activity for anodic oxidation of GSH, decreasing substantially the anodic overpotentials for 0.2 V versus Ag/AgCl. The modified electrode presented better performance in 0.1 mol l−1 piperazine-N,N′-bis(2-ethanesulfonic acid) buffer at pH 7.4. The other experimental parameters, such as the concentration of CoTSPc and PLL in the membrane preparation, pH, type of buffer solution and applied potential, were optimized. Under optimized operational conditions, a linear response from 50 to 2,160 nmol l−1 was obtained with a high sensitivity of 1.5 nA l nmol−1 cm−2. The detection limit for GSH determination was 15 nmol l−1. The proposed sensor presented good repeatability, evaluated in terms of the relative standard deviation (1.5%) for n = 10. The modified electrode was applied for determination of GSH in erythrocyte samples and the results were in agreement with those obtained by a comparative method described in the literature The average recovery for these fortified samples was 100 ± 1)%. Applying a paired Student’s-t test to compare these methods, we could observe that, at the 95% confidence level, there was no statistical difference between the reference and the proposed methods.

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Acknowledgements

The authors acknowledge financial support from the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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Authors and Affiliations

  1. Institute of Chemistry, UNICAMP, P.O. Box 6154, 13084-971, Campinas, SP, Brazil

    Rita de Cássia Silva Luz, Flavio Santos Damos & Lauro Tatsuo Kubota

  2. Institute of Biology, UNICAMP, P.O. Box 6109, 13084-971, Campinas, SP, Brazil

    Paulo Guimarães Gandra & Denise Vaz de Macedo

  3. Departament of Chemistry, UFMA, Avenida dos Portugueses s/n, 65085-040, São Luís, MA, Brazil

    Auro Atsushi Tanaka

Authors
  1. Rita de Cássia Silva Luz
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  2. Flavio Santos Damos
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  3. Paulo Guimarães Gandra
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  4. Denise Vaz de Macedo
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  5. Auro Atsushi Tanaka
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  6. Lauro Tatsuo Kubota
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Correspondence to Lauro Tatsuo Kubota.

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de Cássia Silva Luz, R., Damos, F.S., Gandra, P.G. et al. Electrocatalytic determination of reduced glutathione in human erythrocytes. Anal Bioanal Chem 387, 1891–1897 (2007). https://doi.org/10.1007/s00216-006-1053-6

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  • DOI: https://doi.org/10.1007/s00216-006-1053-6

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