Flavonoid Deactivation of Ferrylmyoglobin in Relation to Ease of Oxidation as Determined by Cyclic Voltammetry

Abstract

Fourteen flavonoid aglycones, and the flavonoid glycoside rutin, with redox potentials ranging from 0.20 (myricetin) to 0.83 V (chrysin) vs. NHE, as determined by cyclic voltammetry at 23 degrees C in aqueous 50 mM phosphate, ionic strength 0.16 (NaCl) with pH = 7.4 and compared with redox potentials determined for four cinnamic acid derivatives, were all found to reduce ferrylmyoglobin, MbFe(IV)=O, to metmyoglobin, MbFe(III). Reaction stoichiometry depends strongly on the number of hydroxyl groups in the flavonoid B-ring. All compounds with 3',4'-dihydroxy substitution reduce 2 equivalents of MbFe(IV)=O, whereas naringenin, hesperitin and kaempferol, with one hydroxyl group in the B-ring, reduce with a one-to-one stoichiometry. As studied spectrophotometrically under pseudo-first-order conditions with flavonoids in excess, rutin and apigenin react with MbFe(IV)=O with very similar and moderately high activation enthalpies of deltaH298++ = 69 +/- 1 kJ mol(-1) and deltaH298++ = 65 +/- 3 kJ mol(-1), respectively, and with positive activation entropies of deltaS298++ = 23 +/- 4 J mol(-1) K(-1) and deltaS298++ = 13 +/- 9 J mol(-1) K(-1), respectively, in agreement with outer-sphere electron transfer as rate determining. For the fifteen plant polyphenols only qualitative relations exist between redox potential and rate constants rather than a linear free energy relationship (r2 = 0.503), and especially the flavone apigenin was found more efficient as reducing agent. For the flavanones, a linear relation (r2 = 0.971) indicate that, in the absence of a 2,3 double bond, removal of the 4-carbonyl group or addition of a 3-hydroxy group only has minor effect on reactivity. The flavonols are the most efficient reducing agents, effectively reducing MbFe(IV)=O to MbFe(III) and establishing a steady state distribution between the flavonol and MbFe(III) and oxymyoglobin, MbFe(II)O2. Oxidised flavonols reduces MbFe(III) to MbFe(II)O2 very efficiently and much faster than the parent flavonol.