Veronika Abram

Phenolics in Slovenian Bilberries (Vaccinium myrtillus L.) and Blueberries (Vaccinium corymbosum L.)

Phenolics from bilberries ( Vaccinium myrtillus L.) sampled from seven different locations and highbush blueberries ( Vaccinium corymbosum L.) from one location in Slovenia were analyzed. In samples of both species 15 anthocyanins were identified by LC-MS/MS. Their contents were expressed as cyanidin 3-glucoside equivalents (C3GE); bilberries contained 1210.3 ± 111.5 mg C3GE/100 g fw and blueberries 212.4 ± 14.1 mg C3GE/100 g fw. Glycosides of delphinidin and cyanidin were predominant (488.5 vs 363.6 mg C3GE/100 g fw) in the bilberries and glycosides of malvidin (108.0 vs 100.8 mg C3GE/100 g fw) in the blueberries, whereas the contents of peonidin were lowest (74.5 vs 4.8 mg C3GE/100 g fw) in both berries. The contents of flavanols, flavonols, phenolic acids, and stilbenes were determined by LC-MS. For the first time, rutin was identified (bilberries, 0.2 ± 0.0 mg/100 g fw; blueberries, 3.1 ± 0.1 mg/100 g fw). Chlorogenic acid (as 3-caffeoylquinic acid) was the most abundant among the phenolic acids (23.1 ± 1.0 mg/100 g fw in bilberries and 70.0 ± 3.4 mg/100 g fw in blueberries). Statistical analysis shows that the content of 27 individual flavonoids, phenolic acids, and stilbenes can be used to identify the picking region of these Slovenian bilberries.

Bilberry and blueberry anthocyanins act as powerful intracellular antioxidants in mammalian cells

Berry anthocyanins have pronounced health effects, even though they have a low bioavailability. The common mechanism underlying health protection is believed to relate to antioxidant activity. Berry extracts, chemically characterised for their phenolic content, were prepared from bilberries (Vaccinium myrtillusL.) and blueberries (Vaccinium corymbosumL.); the bilberry extract was further purified to obtain the anthocyanin fraction. The antioxidant activity of each extract was examined at the cellular level. For this purpose a specific assay, known as cellular antioxidant activity assay (CAA), was implemented in different cell lines: human colon cancer (Caco-2), human hepatocarcinoma (HepG2), human endothelial (EA.hy926) and rat vascular smooth muscle (A7r5). Here we show for the first time that anthocyanins had intracellular antioxidant activity if applied at very low concentrations (<1 μg/l; nM range), thereby providing a long-sought rationale for their health protecting effects in spite of their unfavorable pharmacokinetic properties.

Effect of flavonoid structure on the fluidity of model lipid membranes.

We investigated how the structural properties of (+)-catechin, (-)-epicatechin, (-)-epigallocatechin (EGC) and (-)-epigallocatechin-3-gallate (EGCG) and butylated hydroxytoluene (BHT) correlate with structural changes of phosphatidylcholine plus sphingomyelin (2.4:1) model lipid membranes. Changes were measured by fluorescence anisotropy, electron paramagnetic resonance, and differential scanning calorimetry. Two fluorophores and two spin probes were used to monitor membrane characteristics close to water-lipid interface and in the middle of the bilayer. The data obtained were correlated to the amount of bounded compounds, the number of H-bonds, and the topological polar surface area (TPSA) of the compounds. These correlations reflect the behaviours of (+)-catechin, (-)-epicatechin, EGC, EGCG and BHT. Our results confirm that phenolics studied here are bounded to a membrane surface predominantly via hydrogen bonds, while BTH is inserted into the lipid bilayer.

Effects of industrial and home-made spread processing on bilberry phenolics.

Bilberries processed into spreads represent an important source of anthocyanins if these remain rich in the final product. The effects of thermal processing were studied with non-ground and ground bilberries processed into spreads according to industrial and home-made procedures. Samples were analysed by LC-DAD-MS/MS and LC-MS. The spreads had 28-60% less total phenolics, 4-62% less anthocyanins, and 1-fold to 2-fold more phenolic acids and total flavonols than the bilberries, but approximately equal flavanols. The home-made spread from ground bilberries had ca. 26% higher antioxidant activity. Delphinidin 3-glucoside and cyanidin 3-glucoside were taken through the two spread procedures, with their degradation to gallic acid (38-57%), protocatechuic acid (1-2%) and 2,4,6-trihydroxybenzaldehyde determined. The amounts of gallic and protocatechuic acids did not reflect well for anthocyanin degradation. The industrial spread procedure with non-ground bilberries is a more suitable procedure to maintain the final content of anthocyanins.

Induction of Polyphenol Oxidase in Sempervivum L.

Sempervivum L. plants belong to the family of Crassulaceae (Swart, 1991), the subfamily Sempervivoideae (Stevens et al., 1992). They used to grow mainly in Southern Europe all the way from Spain to Asia Minor, but nowadays they are widespread in other parts of Europe also. Sempervivum is an evergreen plant with meaty juicy leaves. Plants can survive prolonged periods of drought and they are known to have CAM metabolism (Winter, Smith, 1996).