Reaction-Based Turn-on Electrochemiluminescent Sensor with a Ruthenium(II) Complex for Selective Detection of Extracellular Hydrogen Sulfide in Rat Brain

Abstract

Hydrogen sulfide (H2S) has been drawing increasing attention because it plays an important role in the nervous system and has been deemed as a third endogenous gas signal molecule besides nitric oxide (NO) and carbon monoxide (CO). In this study, using a ruthenium complex, [Ru(bpy)2(bpy-DPA)Cu](4+) (where bpy = 2,2'-bipyridine and bpy-DPA = 4-methyl-4'-[N,N-bis(2-picolyl)aminomethylene]-2,2'-bipyridine) as recognition unit, we report a new reaction-based turn-on electrochemiluminescent (ECL) sensor to selectively detect extracellular H2S in rat brain, coupled with in vivo microdialysis for dialysate sampling. To prepare the sensor for sensing endogenous H2S, [Ru(bpy)2(bpy-DPA)](2+) is first designed and synthesized, showing high ECL efficiency with tri-n-propylamine (TPA) as a coreactant and quenching after reaction with Cu(2+) (forming [Ru(bpy)2(bpy-DPA)Cu](4+)). Then a Nafion membrane is coated on the surface of glassy carbon (GC) electrode and [Ru(bpy)2(bpy-DPA)Cu](4+) is confined onto the Nafion membrane through ion exchange. The resulting [Ru(bpy)2(bpy-DPA)Cu](4+)/Nafion/GC sensor exhibits a low ECL signal. The [Ru(bpy)2(bpy-DPA)Cu](4+)/Nafion/GC sensor demonstrates enhanced ECL signal after reacting with volatile H2S due to the high-affinity binding between sulfur and Cu(2+), returning to [Ru(bpy)2(bpy-DPA)](2+)/Nafion/GC. The changes of ECL signal at the sensor depend linearly on the concentration of Na2S in the range from 0.5 to 10 μM, with a detection limit of 0.25 μM. Moreover, the sensor demonstrates high selectivity, free from interference especially by other nonvolatile thiol-containing species, such as cysteine and glutathione. The basal dialysate level of H2S in the microdialysate from the cortex of adult male Sprague-Dawley rats is determined to be 2.3 ± 0.9 μM (n = 4). This method is reliable and is envisaged to help understand the regulation of H2S in physiological and pathological events.