X-Message-Number: 33288 Date: Sat, 29 Jan 2011 09:07:11 -0800 (PST) From: Subject: bicarbonate not useful as a buffer I made a mistake in my earlier suggestion of using bicarbonate as a buffer in a glycerol/wine solution. Turns out bicarbonate destabilizes some of the polyphenols in wine. Biosci Biotechnol Biochem. 2010;74(1):63-8. Epub 2010 Jan 7. Evaluation of resveratrol oxidation in vitro and the crucial role of bicarbonate ions. Yang NC, Lee CH, Song TY. Department of Nutrition and Health Science, Chungchou Institute of Technology, Changhua, Taiwan. Abstract Polyphenols can oxidize in culture medium and produce artifacts in cell culture studies. However, the extent and mechanism of the oxidation of resveratrol, a polyphenol abundant in red wine, is unclear. We investigated the oxidation of resveratrol in vitro and the effects of various components of the culture medium on the degradation of resveratrol and the production of H(2)O(2). We found that 96% of resveratrol at a concentration of 200 microM was degraded in Base Modified Eagle Medium after 24 h of incubation at 37 degrees C, producing about 90 microM of H(2)O(2). Including sodium bicarbonate in the medium markedly stimulated resveratrol degradation and H(2)O(2) production. In sum, we found that bicarbonate ions played a crucial role in the oxidative degradation of resveratrol in vitro, and that the degradation of resveratrol can be avoided by withdrawing sodium bicarbonate from the medium. A mechanism for the oxidation of resveratrol is proposed. PMID: 20057154 Free text> http://www.jstage.jst.go.jp/article/bbb/74/1/63/_pdf J Agric Food Chem. 2004 May 19;52(10):2866-74. Phenolic antioxidants and the protection of low density lipoprotein from peroxynitrite-mediated oxidations at physiologic CO2. Ferroni F, Maccaglia A, Pietraforte D, Turco L, Minetti M. Laboratorio di Biologia Cellulare, Istituto Superiore di Sanita, 00161 Roma, Italy. Abstract Dietary phenolic antioxidants have been shown to prevent LDL modifications mediated by several physiologic oxidants including peroxynitrite. However, more recent data demonstrated that CO(2) affected the fate of peroxynitrite in biological fluids and significantly reduced peroxynitrite scavenging by polyphenols, raising doubts concerning their antioxidant activity. We found that the oxidation of LDL lipids mediated by peroxynitrite decreased in the presence of bicarbonate, while Trp oxidation and 3-nitroTyr formation increased, suggesting a redirection of peroxynitrite reactivity toward the protein moiety. We therefore evaluated the protective activity of some phenolic antioxidants (quercetin, oleuropein, resveratrol, (+)-catechin, (-)-epicatechin, tyrosol, alpha- and gamma-tocopherol, ascorbate) on peroxynitrite-mediated oxidation of LDL aromatic residues. Some of these phenols protected LDL Trp from oxidation better than ascorbate or alpha-tocopherol, although protection at 100 microM did not exceed 30-40%. However, the same phenolic antioxidants were more active in inhibiting 3-nitroTyr formation and those with a catechin structure provided significant protection (IC(50%) 40-50 microM). Red wine, a polyphenol-rich beverage, showed a protective effect comparable to that of the most active phenolic antioxidants. Direct EPR studies showed that bicarbonate significantly increased the peroxynitrite-dependent formation of O-semiquinone radicals in red wine, supporting the hypothesis that polyphenols are efficient scavengers of radicals formed by peroxynitrite/CO(2). Ascorbate was a poor inhibitor of peroxynitrite/CO(2)-induced LDL tyrosine nitration, but the simultaneous addition to the most active polyphenols halved their IC(50%). In conclusion, although cooperation with other antioxidants can further decrease the IC(50%) of polyphenolics, as demonstrated for ascorbate, their antioxidant activity appears to occur at concentrations at least 1 order of magnitude higher than their bioavailability. PMID: 15137827 Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=33288