aspalathin and orientin

The current study investigated the physiological effects of flavonoids found in daily consumed rooibos tea, aspalathin, isoorientin, and orientin on improving processes involved in mitochondrial function in C2C12 myotubes. To achieve this, C2C12 myotubes were exposed to a mitochondrial channel blocker, antimycin A (6.25 µM), for 12 h to induce mitochondrial dysfunction. Thereafter, cells were treated with aspalathin, isoorientin, and orientin (10 µM) for 4 h, while metformin (1 µM) and insulin (1 µM) were used as comparators. Relevant bioassays and real-time PCR were conducted to assess the impact of treatment compounds on some markers of mitochondrial function. Our results showed that antimycin A induced alterations in the mitochondrial respiration process and mRNA levels of genes involved in energy production. In fact, aspalathin, isoorientin, and orientin reversed such effects leading to the reduced production of intracellular reactive oxygen species. These flavonoids further enhanced the expression of genes involved in mitochondrial function, such as

Topics: Animals; Anti-Bacterial Agents; Antimycin A; Aspalathus; Cell Line; Cells, Cultured; Chalcones; Flavonoids; Glucosides; Luteolin; Mice; Mitochondria; Muscle, Skeletal; Tea

Mounting evidence of the ability of aspalathin to target underlying metabolic dysfunction relevant to the development or progression of obesity and type 2 diabetes created a market for green rooibos extract as a functional food ingredient. Aspalathin is the obvious choice as a chemical marker for extract standardisation and quality control, however, often the concentration of a single constituent of a complex mixture such as a plant extract is not directly related to its bio-capacity, i.e. the level of in vitro bioactivity effected in a cell system at a fixed concentration. Three solvents (hot water and two EtOH-water mixtures), previously shown to produce bioactive green rooibos extracts, were selected for extraction of different batches of rooibos plant material (n = 10). Bio-capacity of the extracts, tested at 10 μg ml

Topics: 3T3-L1 Cells; Animals; Aspalathus; Caco-2 Cells; Cell Line; Chalcones; Chromatography, High Pressure Liquid; Flavonoids; Functional Food; Glucosides; Hep G2 Cells; Humans; Mice; Permeability; Phenylpyruvic Acids; Plant Extracts; Quality Control

Aspalathin and nothofagin, the major dihydrochalcones in rooibos (Aspalathus linearis), are valuable bioactive compounds, but their bioactivity has not been fully elucidated. Isolation of these compounds using high-performance countercurrent chromatography (HPCCC), a gentle, support-free, up-scalable technique, offers an alternative to synthesis for obtaining sufficient amounts. An HPLC-DAD method was adapted to allow rapid (16 min from injection to injection) quantification of the four major compounds (aspalathin, nothofagin, isoorientin, orientin) during development of the isolation protocol. The traditional shake-flask method, used to determine distribution constants (K(D)) for target compounds, was also adapted to obtain higher repeatability. Green rooibos leaves with a high aspalathin and nothofagin content were selected as source material. Sample loading of the polyphenol-enriched extract was limited due to constituents with emulsifying properties, but could be increased by removing ethanol-insoluble matter. Furthermore, problems with degradation of aspalathin during HPCCC separation and further processing could be limited by acidifying the HPCCC solvent system. Aspalathin was shown to be fairly stable at pH 3 (91% remaining after 29 h) compared to pH 7 (45% remaining after 29 h). Aspalathin and nothofagin with high purities (99% and 100%, respectively) were obtained from HPCCC fractions after semi-preparative HPLC.

Topics: Aspalathus; Chalcones; Chromatography, High Pressure Liquid; Countercurrent Distribution; Flavonoids; Glucosides; Luteolin; Plant Extracts; Plant Leaves; Polyphenols; Spectrometry, Mass, Electrospray Ionization

The anti-peroxyl radical quality of two aqueous rooibos infusions and solutions of their most abundant glycosylated polyphenols was evaluated using pyrogallol red and fluorescein-based oxygen radical absorbance ratios. It was observed that the artificial infusions, prepared using only the most abundant polyphenols present in rooibos and at concentrations similar to those found in the natural infusions, showed greater antioxidant quality than the latter infusions, reaching values close to those reported for tea infusions. Additionally, the antimicrobial activity of the natural and artificial infusions was assessed against three species of bacteria: Gram (+) Staphylococus epidermidis and Staphylococcus aureus and Gram (-) Escherichia coli. When compared to the natural infusions the artificial beverages did not demonstrate any bacterostatic/cidal activity, suggesting that the antibacterial activity of rooibos is related to compounds other than the glycosylated polyphenols employed in our study.

Topics: Anti-Bacterial Agents; Apigenin; Aspalathus; Beverages; Chalcones; Escherichia coli; Flavonoids; Free Radical Scavengers; Glucosides; Microbial Sensitivity Tests; Peroxides; Plant Extracts; Polyphenols; Quercetin; Rutin; Staphylococcus aureus; Staphylococcus epidermidis

A nested analysis of variance combined with simultaneous component analysis, ASCA, was proposed to model high-dimensional chromatographic data. The data were obtained from an experiment designed to investigate the effect of production season, quality grade and post-production processing (steam pasteurization) on the phenolic content of the infusion of the popular herbal tea, rooibos, at 'cup-of-tea' strength. Specifically, a four-way analysis of variance where the experimental design involves nesting in two of the three crossed factors was considered. For the purpose of the study, batches of fermented rooibos plant material were sampled from each of four quality grades during three production seasons (2009, 2010 and 2011) and a sub-sample of each batch was steam-pasteurized. The phenolic content of each rooibos infusion was characterized by high performance liquid chromatography (HPLC)-diode array detection (DAD). In contrast to previous studies, the complete HPLC-DAD signals were used in the chemometric analysis in order to take into account the entire phenolic profile. All factors had a significant effect on the phenolic content of a 'cup-of-tea' strength rooibos infusion. In particular, infusions prepared from the grade A (highest quality) samples contained a higher content of almost all phenolic compounds than the lower quality plant material. The variations of the content of isoorientin and orientin in the different quality grade infusions over production seasons are larger than the variations in the content of aspalathin and quercetin-3-O-robinobioside. Ferulic acid can be used as an indicator of the quality of rooibos tea as its content generally decreases with increasing tea quality. Steam pasteurization decreased the content of the majority of phenolic compounds in a 'cup-of-tea' strength rooibos infusion.

Topics: Analysis of Variance; Aspalathus; Beverages; Chalcones; Chromatography, High Pressure Liquid; Coumaric Acids; Fermentation; Flavonoids; Glucosides; Pasteurization; Phenols; Plant Leaves; Principal Component Analysis; Seasons; Steam

The effects of citric and ascorbic acids on the stability of aspalathin in rooibos (Aspalathus linearis) ready-to-drink (RTD) formulations containing fermented rooibos extract (FR), aspalathin-enriched green rooibos extract (GR) and aspalathin-enriched green rooibos extract ascorbic acid solubilisate (GR-solubilisate) were investigated during storage (12 weeks at 25 °C).. Storage of iced tea formulations containing FR and GR extracts reduced their flavonoid content. The aspalathin content of FR iced tea without citric or ascorbic acid was reduced to undetectable levels by week 8 of storage. Addition of citric acid resulted in improved stability of aspalathin, but ascorbic acid did not impart additional stability. Iso-orientin and orientin were less affected than aspalathin, presumably owing to partial conversion of aspalathin to these flavones. Similar results were obtained for GR iced tea formulations. Improved stability of aspalathin was noted in iced tea containing GR-solubilisate with or without citric acid. Lower pH was shown to favour stability, especially for fermented rooibos iced teas.. Citric and ascorbic acids contribute to the stability of rooibos flavonoids during storage. Differences in stability between formulations are not due to pH differences but may be related to the matrix.

Topics: Ascorbic Acid; Aspalathus; Beverages; Chalcones; Citric Acid; Fermentation; Flavonoids; Food Storage; Glucosides; Hydrogen-Ion Concentration; Plant Extracts; Time Factors

The phenolic quality of commercial South African fermented rooibos iced teas in terms of aspalathin, iso-orientin, and orientin contents in comparison to a "cup of tea" was shown to be inferior. The role of the different manufacturing stages of powdered extract used in iced tea formulation and, more specifically, the impact of pasteurization and sterilization on the color and phenolic content of the beverage, were assessed as potential causes of its inferior phenolic quality. Aspalathin and its corresponding flavones, iso-orientin and orientin, were found to be present at all stages of the powdered extract production process. Spray-drying did not significantly (P ≥ 0.05) alter the aspalathin, iso-orientin, or orientin content of concentrates. Simulated normal-temperature sterilization (NTS at 121 °C/15 min) and high-temperature sterilization (HTS at 135 °C/4 min), but not necessarily pasteurization (93 °C/30 min), significantly (P < 0.05) reduced the aspalathin, iso-orientin, and orientin contents of different iced tea formulations. Heat-induced losses of iso-orientin and orientin were lower than those for aspalathin. Conversion of aspalathin to the flavones is implicated. The addition of ascorbic acid and/or citric acid to the base iced tea formulation containing only rooibos extract and sugar proved to be beneficial, especially for the retention of aspalathin. Browning, that is, absorbance at 420 nm, was significantly (P < 0.05) increased in the base formulation. In the case of the formulations also containing ascorbic acid and/or citric acid, absorbance remained unchanged or decreased when subjected to NTS and HTS treatments. This was attributed to removal of brown polymers from solution as the pH values of these formulations were lower than that of the base formulation.

Topics: Ascorbic Acid; Aspalathus; Beverages; Chalcones; Citric Acid; Cold Temperature; Color; Fermentation; Flavonoids; Food Handling; Glucosides; Hot Temperature; Plant Extracts