Stability and Free Radical Production for CO₂ and H₂ in Air Nanobubbles in Ethanol Aqueous Solution

In this study, 8% hydrogen (H₂) in argon (Ar) and carbon dioxide (CO₂) gas nanobubbles was produced at 10, 30, and 50 vol.% of ethanol aqueous solution by the high-speed agitation method with gas. They became stable for a long period (for instance, 20 days), having a high negative zeta potential (-40 to -50 mV) at alkaline near pH 9, especially for 10 vol.% of ethanol aqueous solution. The extended Derjaguin, Landau, Verwey, and Overbeek (DLVO) theory was used to evaluate the nanobubble stability. When the nanobubble in ethanol alkaline aqueous solution changed to an acidic pH of around 5, the zeta potential of nanobubbles was almost zero and the decrease in the number of nanobubbles was identified by the particle trajectory method (Nano site). The collapsed nanobubbles at zero charge were detected thanks to the presence of few free radicals using G-CYPMPO spin trap reagent in electron spin resonance (ESR) spectroscopy. The free radicals produced were superoxide anions at collapsed 8%H₂ in Ar nanobubbles and hydroxyl radicals at collapsed CO₂ nanobubbles. On the other hand, the collapse of mixed CO₂ and H₂ in Ar nanobubble showed no free radicals. The possible presence of long-term stable nanobubbles and the absence of free radicals for mixed H₂ and CO₂ nanobubble would be useful to understand the beverage quality.