In vitro study on the competitive reactions between arsenite and selenite with glutathione

Exposure to arsenic can cause various biological effects by increasing the production of reactive oxygen species (ROS). Selenium acts as a beneficial element by regulating ROS and limiting heavy metal uptake and translocation. There are studies on the interactive effects of As and Se in plants, but the antagonistic and synergistic effects of these elements based on their binding to glutathione (GSH) molecules have not been studied yet. In this study, we aimed to investigate the antagonistic or synergistic effects of As and Se on the binding mechanism of Se and As with GSH at pH 3.0, 5.0, or 6.5. The interaction of As and Se in Se(SG)₂ + As(III) or As(SG)₃ + Se(IV) binary systems and As(III) + Se(IV) + GSH ternary system were examined depending on their ratios via liquid chromatography diode array detector/electrospray mass spectrometry (LC-DAD/MS) and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). The results showed that the formation of As(GS)₃ was not detected in the As(III) + Se(SG)₂ binary system, indicating that As(III) did not affect the stability of Se(SG)₂ complex antagonistically. However, in the Se(IV) + As(SG)₃ binary system, the addition of Se(IV) to As(SG)₃ affected the stability of As(SG)₃ antagonistically. Se(IV) reacted with GSH, disrupting the As(SG)₃ complex, and consequently, Se(SG)₂ formation was measured using LC-MS/DAD. In the Se(IV) + GSH + As(III) ternary system, Se(SG)₂ formation was detected upon mixing As(III), Se(IV), and GSH. The increase in the concentration of As(III) did not influence the stability of the Se(SG)₂ complex. Additionally, Se(IV) has a higher affinity than As(III) to the GSH, regardless of the pH of the solution. In both binary and ternary systems, the formation of the by-product glutathione trisulfide (GSSSG) was detected using LC-ESI-MS/MS.