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Article

LC-MS Evaluation of the Redox Trypanothione Balance in Leishmania infantum Parasites

Pharma-Dev UMR 152, Université de Toulouse, IRD, 118 Route de Narbonne, 31062 Toulouse CEDEX 9, France
*
Author to whom correspondence should be addressed.
These authors contributed equally to the work.
Antioxidants 2025, 14(8), 977; https://doi.org/10.3390/antiox14080977
Submission received: 11 June 2025 / Revised: 21 July 2025 / Accepted: 31 July 2025 / Published: 8 August 2025
(This article belongs to the Section ROS, RNS and RSS)

Abstract

Leishmaniases are neglected tropical diseases caused by protozoan parasites of the Leishmania genus, with a significant global health burden, particularly in low-income regions. The parasites rely on a unique thiol-based redox system centered on trypanothione, which is essential for survival under oxidative stress encountered during their life cycle in both insect vectors and mammalian hosts. Given the absence of mammalian analogs, the trypanothione system represents an attractive target for antileishmanial drug development. However, accurate quantification of the reduced and oxidized forms of trypanothione has been challenging due to its instability and structural similarity between redox states. Here, we developed and validated a rapid, sensitive liquid chromatography–mass spectrometry (LC-MS) method for assessing the trypanothione redox state in Leishmania infantum. By incorporating N-ethylmaleimide as a thiol-blocking agent during sample preparation, the native redox state was preserved, enabling precise measurement of the reduced-to-oxidized ratio. Our approach demonstrated high sensitivity (nanomolar range), a rapid analysis time (5 min/sample), and robustness across various conditions. Moreover, we validated the method’s relevance in detecting oxidative stress and response to the trypanothione reductase inhibitor auranofin. This LC-MS technique provides a valuable tool for exploring Leishmania redox biology and supports the discovery of redox-targeting therapies against leishmaniasis.
Keywords: Leishmania infantum; trypanothione redox balance; LC-MS analysis; oxidative stress; N-Ethylmaleimide Leishmania infantum; trypanothione redox balance; LC-MS analysis; oxidative stress; N-Ethylmaleimide

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MDPI and ACS Style

Villarubias, T.; Royo, J.; Perio, P.; Bourgeade-Delmas, S.; Sudor, J.; Valentin, A.; Terrisse, A.-D.; Reybier, K. LC-MS Evaluation of the Redox Trypanothione Balance in Leishmania infantum Parasites. Antioxidants 2025, 14, 977. https://doi.org/10.3390/antiox14080977

AMA Style

Villarubias T, Royo J, Perio P, Bourgeade-Delmas S, Sudor J, Valentin A, Terrisse A-D, Reybier K. LC-MS Evaluation of the Redox Trypanothione Balance in Leishmania infantum Parasites. Antioxidants. 2025; 14(8):977. https://doi.org/10.3390/antiox14080977

Chicago/Turabian Style

Villarubias, Théo, Jade Royo, Pierre Perio, Sandra Bourgeade-Delmas, Jan Sudor, Alexis Valentin, Anne-Dominique Terrisse, and Karine Reybier. 2025. "LC-MS Evaluation of the Redox Trypanothione Balance in Leishmania infantum Parasites" Antioxidants 14, no. 8: 977. https://doi.org/10.3390/antiox14080977

APA Style

Villarubias, T., Royo, J., Perio, P., Bourgeade-Delmas, S., Sudor, J., Valentin, A., Terrisse, A.-D., & Reybier, K. (2025). LC-MS Evaluation of the Redox Trypanothione Balance in Leishmania infantum Parasites. Antioxidants, 14(8), 977. https://doi.org/10.3390/antiox14080977

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