Antioxidant activity, protective effects and absorption of polyphenolic compounds
The objectives of the studies presented in this thesis were to identify and quantify the major phenolic components of red wine and to assess the contribution of individual compounds to the total antioxidant activity. Red wines were analysed for their phenolic content and antioxidant activity using a range of complementary techniques including HPLC-tandem mass spectrometry, preparative HPLC and HPLC with an on-line antioxidant detection system. HPLC-MS2 revealed the presence of a number of flavoids and phenolic compounds of which 19 were identified, with gallic acid, the flavin-3-ols and anthocyanins being the most abundant. Preparative HPLC was used in an effort to isolate the antioxidant components in red wine and 60 aliquots were collected. Each wine fraction was analysed for total phenolics, catechins and anthocyanins, while antioxidant activity was determined by electron spin resonance spectroscopy (ESR). The preparative HPLC step did into completely separate the compounds in red wine, nonetheless increasing antioxidant activity was highly and significantly associated with total phenolics (r = 0.816, P < 0.001) and total catechins (r = 0.188, p = 0.151). HPLC with an on-line antioxidant detection system was subsequently used to separate and identify red wine phenolics. The findings from this study indicate that gallic acid, (+)-catechin, (-)-epicatechin, and procyanidin dimersB1 and B2 were the major in vitro antioxidants identified in red wine. Collectively, the flavin-3-ols contributed > 50% of the total antioxidant capacity of each wine, while gallic acid contributed between 24-44%. The flavonols and anthocyanins were minor antioxidant components in red wines. By combining HPLC, MS2 and on-line assessment of antioxidant activity, the major phenolic compounds present in red wine were identified, together with their direct contribution to the total antioxidant activity.