Mechanism Underlying Green Discolouration of Myoglobin Induced by Atmospheric Pressure Plasma

In this study, we elucidated the mechanism underlying atmospheric pressure plasma (APP)-induced green discolouration of myoglobin. Green-coloured pigments are produced upon conversion of myoglobin into sulphmyoglobin, choleglobin, verdoheme, nitrihemin, or nitrimyoglobin. We exposed myoglobin dissolved in phosphate buffer to APP for 20 min and found a decrease in a^* value (+redness/-greenness) and increase in b^* value (+yellowness/-blueness) (P < 0.05). In the ultraviolet absorption spectrum, myoglobin treated with APP for 20 min showed absorption peaks at 503 and 630 nm, a spectrum different from that of sulphmyoglobin or choleglobin. The secondary structure and molecular weight of myoglobin were unaffected by APP treatment, excluding the possibility of verdoheme or nitrihemin formation. After APP treatment, nitrite was produced in myoglobin solution that provided a positive environment for nitrimyoglobin formation. However, the addition of 0.5% sodium dithionite, a strong reducing agent, to myoglobin solution resulted in the formation of deoxymyoglobin, which was subsequently converted to nitrosomyoglobin upon APP treatment to yield a desirable red colour. Thus, APP-induced green colouration in myoglobin solution is associated with nitrimyoglobin formation. The addition of the antioxidant resulted in the production of red colour in myoglobin solution after APP treatment owing to nitrosomyoglobin formation.