Hydrogen peroxide generation from hydrated protein drink mixes

Generation of oxygen radicals upon hydration of powdered protein products was examined using luminol-enhanced chemiluminescence. Among individual proteins powders examined oxidative bursts occurred almost immediately, and then rapidly declined in the 1st 5 min. Commercially available powdered protein drink mixes behaved differently, with an initial lag phase followed by a sustained increase in luminol-enhanced luminescence, lasting for an hour or beyond. The drink mix that produced the highest level of luminol-enhanced luminescence also contained 379 nM ascorbate radical when hydrated (28 nmole/g of powdered drink mix). The entire ascorbic acid content of this drink mix was oxidized to nondetectable levels (using HPLC-diode array detection) within 60 min of being hydrated. Treatment of the hydrated drink mixes with the enzyme catalase almost completely inhibited the luminol-enhanced luminescence from the hydrated drink mix demonstrating that hydrogen peroxide generated via a chemical reaction among the drink mixes' ingredients was a primary reactive oxygen species (ROS). This is the strongest oxidative capacity demonstrated in a food product as consumed (without any manipulation to increase ROS) and the 1st time that the ascrobate radical in a food product as been quantified. Generation of hydrogen peroxide in the hydrated drink mixes from metal catalyzed reactions involving oxygen and reducing equivalents from ascorbic acid is proposed.