Rooibos tea waste binary oxide composite: An adsorbent for the removal of nickel ions and an efficient photocatalyst for the degradation of ciprofloxacin.

In this study, rooibos tea waste (RTW) incorporated with a binary oxide (BO; Fe2O3-SnO2) has been reported for the first time as a highly efficient adsorbent material for the elimination of Ni(II) ions. The as-synthesised rooibos tea waste-binary oxide (RWBO) composite adsorbent was characterised using miscellaneous techniques such as FTIR, XRD, SEM, EDX, TGA, BET, and XPS. The RWBO was then tested for the removal of Ni(II) in a batch adsorption experiment. The composite adsorbent showed a great removal efficiency of about 99.75% for Ni(II) ions at 45 °C, 180 min agitation time, pH 7, and dosage of 250 mg. The adsorption process was found to be endothermic and spontaneous. Also, the spent adsorbent [RWBO-Ni(II)] was found to be solar light active with a narrow band gap of 1.4 eV. It was further used as a photocatalyst for the photocatalytic abatement of 10 mg/L ciprofloxacin with an extent of degradation of 83% obtained after 150 min. In addition, the extent of mineralisation of the ciprofloxacin by the spent adsorbent as obtained from the TOC data was found to be 64%. Overall, the RWBO composite adsorbent lends itself as an efficient, eco-friendly and promising material for environmental remediation.

Rooibos tea-in the cross fire of ROS, mitochondrial dysfunction and loss of proteostasis-positioned for healthy aging.

Impaired mitochondrial function and loss of cellular proteostasis control are key hallmarks of aging and are implicated in the development of neurodegenerative diseases. A common denominator is the cell's inability to handle reactive oxygen species (ROS), leading to major downstream oxidative damage that exacerbates neuronal dysfunction. Although we have progressed in understanding the molecular defects associated with neuronal aging, many unanswered questions remain. How much ROS is required to serve cellular function before it becomes detrimental and how does the cell's oxidative status impact mitochondrial function and protein degradation through autophagy? How does ROS regulate autophagy? Aspalathus linearis, also commonly known as rooibos, is an endemic South African plant that is gaining globally acclaim for its antioxidant properties and its role as functional medicinal beverage. In this article we dissect the role of rooibos in the context of the cell's ROS handling capacity, mitochondrial function and autophagy activity. By addressing the dynamic relationship between these critical interconnected systems, and by evaluating the functional properties of rooibos, we unravel its position for preserving cell viability and promoting healthy aging.

In vitro effects of aqueous extract of unfermented rooibos on human spermatozoa.

The inability to conceive is a baleful experience for thousands of couples worldwide. Among different well-known reproductive techniques, medicinal plants have been utilized to treat male infertility. Medicinal plants, provide a therapeutic alternative, which is available and affordable for infertile couples. We investigated the direct effect of unfermented rooibos aqueous extract on human spermatozoa. Semen samples (n = 50) collected from donors and patients consulting for fertility were reassigned as normal (n = 22) and abnormal (n = 28) samples based on the outcome of the baseline semen analysis, using the World Health Organization (WHO) cut off value. Semen samples were allowed to liquefy and subsequently washed with human tubular fluid in bovine serum albumin medium. The samples were then treated with aqueous extracts of unfermented rooibos (0, 0.15, 1.5, 15, 150 µg/ml) at 37°C for 1 h and assessed thereafter. Sperm motility, vitality, DNA fragmentation, intracellular reactive oxygen species and mitochondrial membrane potential in both groups remained unchanged (p > 0.05). However, aqueous extract of unfermented rooibos (only at 1.5 µg/ml) significantly increased capacitation and acrosome reaction in the abnormal sample group (p > 0.05). Unfermented rooibos aqueous extract had no deleterious impact on human spermatozoa's function and might be attributed to its antioxidant properties.

Aspalathin and Other Rooibos Flavonoids Trapped α-Dicarbonyls and Inhibited Formation of Advanced Glycation End Products In Vitro.

The excessive dietary intake of simple sugars and abnormal metabolism in certain diseases contribute to the increased production of α-dicarbonyls (α-DCs), such as methylglyoxal (MGO) and glyoxal (GO), the main precursors of the formation of advanced glycation end products (AGEs). AGEs play a vital role, for example, in the development of cardiovascular diseases and diabetes. Aspalathus linearis (Burman f.) R. Dahlgren (known as rooibos tea) exhibits a wide range of activities beneficial for cardio-metabolic health. Thus, the present study aims to investigate unfermented and fermented rooibos extracts and their constituents for the ability to trap MGO and GO. The individual compounds identified in extracts were tested for the capability to inhibit AGEs (with MGO or GO as a glycation agent). Ultra-high-performance liquid chromatography coupled with an electrospray ionization mass spectrometer (UHPLC-ESI-MS) was used to investigate α-DCs' trapping capacities. To evaluate the antiglycation activity, fluorescence measurement was used. The extract from the unfermented rooibos showed a higher ability to capture MGO/GO and inhibit AGE formation than did the extract from fermented rooibos, and this effect was attributed to a higher content of dihydrochalcones. The compounds detected in the extracts, such as aspalathin, nothofagin, vitexin, isovitexin, and eriodictyol, as well as structurally related phloretin and phloroglucinol (formed by the biotransformation of certain flavonoids), trapped MGO, and some also trapped GO. AGE formation was inhibited the most by isovitexin. However, it was the high content of aspalathin and its higher efficiency than that of metformin that determined the antiglycation and trapping properties of green rooibos. Therefore, A. linearis, in addition to other health benefits, could potentially be used as an α-DC trapping agent and AGE inhibitor.

Optimising the Polyphenolic Content and Antioxidant Activity of Green Rooibos (Aspalathus linearis) Using Beta-Cyclodextrin Assisted Extraction.

Antioxidant activity associated with green rooibos infusions is attributed to the activity of polyphenols, particularly aspalathin and nothofagin. This study aimed to optimise ß-cyclodextrin (ß-CD)-assisted extraction of crude green rooibos (CGRE) via total polyphenolic content (TPC) and antioxidant activity assays. Response surface methodology (RSM) permitted optimisation of ß-CD concentration (0−15 mM), temperature (40−90 °C) and time (15−60 min). Optimal extraction conditions were: 15 mM ß-CD: 40 °C: 60 min with a desirability of 0.985 yielding TPC of 398.25 mg GAE·g−1, metal chelation (MTC) of 93%, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging of 1689.7 µmol TE·g−1, ferric reducing antioxidant power (FRAP) of 2097.53 µmol AAE·g−1 and oxygen radical absorbance capacity (ORAC) of 11,162.82 TE·g−1. Aspalathin, hyperoside and orientin were the major flavonoids, with quercetin, luteolin and chrysoeriol detected in trace quantities. Differences (p < 0.05) between aqueous and ß-CD assisted CGRE was only observed for aspalathin reporting the highest content of 172.25 mg·g−1 of dry matter for extracts produced at optimal extraction conditions. Positive, strong correlations between TPC and antioxidant assays were observed and exhibited regression coefficient (R2) between 0.929−0.978 at p < 0.001. These results demonstrated the capacity of ß-CD in increasing polyphenol content of green rooibos.

Green, Black and Rooibos Tea Inhibit Prostaglandin E2 Formation in Human Monocytes by Inhibiting Expression of Enzymes in the Prostaglandin E2 Pathway.

The formation of prostaglandin E2 (PGE2) is associated with adverse inflammatory effects. However, long-term treatment with nonsteroidal anti-inflammatory drugs (NSAIDs) comes with risk of severe side effects. Therefore, alternative ways to inhibit PGE2 are warranted. We have investigated the effects of tea extracts and the polyphenols epigallocatechin gallate (EGCG) and quercetin on PGE2 formation, determined by immunoassay, and protein expression, determined by immunoblotting, of cytosolic phospholipase A2 (cPLA2), cyclooxygenase 2 (COX-2) and microsomal PGE synthase-1 (mPGES-1) in human monocytes. Green and black tea extracts, and with a lower potency, Rooibos tea extract, inhibited lipopolysaccharide (LPS) and calcium ionophore-induced PGE2 formation. In addition, all tea extracts inhibited the LPS-induced expression of mPGES-1, and the green and black tea extracts also inhibited, to a lesser extent, COX-2 expression. The tea extracts only marginally reduced cPLA2 expression and had no effect on COX-1 expression. EGCG, present in green and black tea, and quercetin, present in all three teas, also inhibited PGE2 formation and expression of mPGES-1, COX-2 and cPLA2. Cell-based and cell-free assays were also performed to evaluate direct effects on the enzymatic activity of COX and PGE synthases. Mainly, the cell-free assay demonstrated partial inhibition by the tea extracts and polyphenols. However, the inhibition required higher doses compared to the effects demonstrated on protein expression. In conclusion, green and black tea, and to a lesser extent Rooibos tea, are potent inhibitors of PGE2 formation in human monocytes, and mediate their effects by inhibiting the expression of the enzymes responsible for PGE2 formation, especially mPGES-1.

Rooibos (Aspalathus linearis) influence on health and ovarian functions.

This paper reviews provenance, processing and properties of rooibos (Aspalathus linearis, Brum.f) and its numerous biologically active constituents, as well as the currently available knowledge concerning their physiological and medicinal effects and their possible extra- and intracellular mechanisms of action. Search for literature was performed in agreement with the preferred reporting items for systematic review criteria in Cochrane Library, PubMed, Web of Science and SCOPUS databases between the years 2000 and 2021. The limited number of in vitro studies suggests an influence of rooibos on basic ovarian cell functions, as well as its potential applicability to control female reproduction and prevent the effect of environmental contaminants on ovarian functions. Nevertheless, further studies are required for better understanding of the character and mechanisms of action, as well as of rooibos' application in reproductive biology and medicine.

Shelf-Life Stability of Ready-to-Use Green Rooibos Iced Tea Powder-Assessment of Physical, Chemical, and Sensory Properties.

Green rooibos extract (GRE), shown to improve hyperglycemia and HDL/LDL blood cholesterol, has potential as a nutraceutical beverage ingredient. The main bioactive compound of the extract is aspalathin, a C-glucosyl dihydrochalcone. The study aimed to determine the effect of common iced tea ingredients (citric acid, ascorbic acid, and xylitol) on the stability of GRE, microencapsulated with inulin for production of a powdered beverage. The stability of the powder mixtures stored in semi-permeable (5 months) and impermeable (12 months) single-serve packaging at 30 °C and 40 °C/65% relative humidity was assessed. More pronounced clumping and darkening of the powders, in combination with higher first order reaction rate constants for dihydrochalcone degradation, indicated the negative effect of higher storage temperature and an increase in moisture content when stored in the semi-permeable packaging. These changes were further increased by the addition of crystalline ingredients, especially citric acid monohydrate. The sensory profile of the powders (reconstituted to beverage strength iced tea solutions) changed with storage from a predominant green-vegetal aroma to a fruity-sweet aroma, especially when stored at 40 °C/65% RH in the semi-permeable packaging. The change in the sensory profile of the powder mixtures could be attributed to a decrease in volatile compounds such as 2-hexenal, (Z)-2-heptenal, (E)-2-octenal, (E)-2-nonenal, (E,Z)-2,6-nonadienal and (E)-2-decenal associated with "green-like" aromas, rather than an increase in fruity and sweet aroma-impact compounds. Green rooibos extract powders would require storage at temperatures ≤ 30 °C and protection against moisture uptake to be chemically and physically shelf-stable and maintain their sensory profiles.

Effect of Rooibos (Aspalathus linearis) extract on atorvastatin-induced toxicity in C3A liver cells.

Rooibos (Aspalathus linearis) has various health benefits. Two case studies have associated chronic Rooibos consumption with conventional prescription medications, including atorvastatin (ATV), with hepatotoxicity. Statins act by inhibiting hydroxymethylglutaryl-coenzyme A reductase, a rate-limiting enzyme in cholesterol synthesis. Although rare, statins are potentially hepatotoxic. The aim was to investigate interactions between aspalathin-rich Rooibos extract GRT™ and ATV-induced hepatotoxicity in C3A liver cells cultured with and without palmitate. Effects of co-treatment of GRT + ATV on cell viability, oxidative stress, apoptosis, mitochondrial integrity, and cellular reactive oxygen species (ROS) production were assessed. Significantly increased ROS production was observed in cells exposed to ATV and palmitate. Combination therapy of GRT + ATV also showed significant increases in ROS production. Under palmitate-treated conditions, ATV-induced significant apoptosis which was not ameliorated by GRT + ATV co-treatment. Despite studies purporting hepatoprotection from Rooibos, our study showed that GRT was unable to modulate ATV-induced hepatotoxic effects in this model.

Fumonisin B1 -induced mitochondrial toxicity and hepatoprotective potential of rooibos: An update.

Fumonisins are a family of potentially carcinogenic mycotoxins produced by Fusarium verticillioides. Several fumonisins have been identified with fumonisin B1 (FB1 ) being the most toxic. The canonical mechanism of FB1 toxicity is centered on its structural resemblance with sphinganine and consequent competitive inhibition of ceramide synthase and disruption of lipidomic profiles. Recent and emerging evidence at the molecular level has identified the disruption of mitochondria and excessive generation of toxic reactive oxygen species (ROS) as alternative/additional mechanisms of toxicity. The understanding of how these pathways contribute to FB1 toxicity can lead to the identification of novel, effective approaches to protecting vulnerable populations. Natural compounds with antioxidant properties seem to protect against the induced toxic effects of FB1 . Rooibos (Aspalathus linearis), endemic to South Africa, has traditionally been used as a medicinal herbal tea with strong scientific evidence supporting its anecdotal claims. The unique composition of phytochemicals and combination of metabolic activators, adaptogens and antioxidants make rooibos an attractive yet underappreciated intervention for FB1 toxicoses. In the search for a means to address FB1 toxicoses as a food safety problem in developing countries, phytomedicine and traditional knowledge systems must play an integral part. This review aims to summarize the growing body of evidence succinctly, which highlights mitochondrial dysfunction as a secondary toxic effect responsible for the FB1 -induced generation of ROS. We further propose the potential of rooibos to combat this induced toxicity based on its integrated bioactive properties, as a socio-economically viable strategy to prevent and/or repair cellular damage caused by FB1 .

The Rooibos Benefit Sharing Agreement-Breaking New Ground with Respect, Honesty, Fairness, and Care.

The 1992 Convention on Biological Diversity (CBD) and its 2010 Nagoya Protocol brought about a breakthrough in global policy making. They combined a concern for the environment with a commitment to resolving longstanding human injustices regarding access to, and use of biological resources. In particular, the traditional knowledge of indigenous communities was no longer going to be exploited without fair benefit sharing. Yet, for 25 years after the adoption of the CBD, there were no major benefit sharing agreements that led to significant funding streams for indigenous communities. This changed with the signing of the Rooibos Benefit Sharing Agreement in South Africa, described in this paper. As the authors report, the Rooibos Agreement is a superlative in two respects. It is the biggest benefit sharing agreement between industry and indigenous peoples to date. It is also the first industry-wide agreement to be formed in accordance with biodiversity legislation. This article is a co-production between traditional knowledge holders, the lawyer who represented their interests, the Co-Chair of the Nagoya Protocol negotiations, and an ethicist who analyzed the major challenges of this historic agreement. With no precedent in the benefit sharing world, the agreement stands as a concrete example of the 'art of the possible.' Although the rooibos case is unique in a number of aspects, the experience offers many transferable insights, including: patience; incrementalism; honesty; trust; genuine dialogue; strong legal support; a shared recognition that a fair, win-win deal is possible; government leadership; and unity amongst indigenous peoples. Such ingredients of success can apply well beyond southern Africa.

Phytochemical properties of black tea (Camellia sinensis) and rooibos tea (Aspalathus linearis); and their modulatory effects on key hyperglycaemic processes and oxidative stress.

The comparative phytochemicals, antioxidative and antidiabetic activities of Camellia sinensis (black tea) and Aspalathus linearis (rooibos tea) were studied in vitro and ex vivo. Concentrated infusions of the teas showed significant free radical scavenging activities in vitro. They significantly increased the glutathione level, superoxide dismutase and catalase enzyme activities in oxidative hepatic injury, while concomitantly depleting malondialdehyde level. The teas significantly inhibited intestinal glucose absorption and α-amylase activities, and elevated muscle glucose uptake. LCMS phytochemical profiling revealed the presence of hydroxycaffeic acid, l-threonate, caffeine, vanillic acid, n-acetylvaline, and spinacetin 3-glucoside in C. sinensis. While quinolinic acid, coumestrol, phloroglucinol, 8-hydroxyquercetagetin, umbelliferone, and ajoene were identified in A. linearis. These results portray the antioxidant and antidiabetic potencies of both teas, with A. linearis showed better activity compared to C. sinensis. These teas may thus be used as functional foods in the management of diabetes and other oxidative stress related metabolic disorders.

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK Pathways.

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced insulin resistance in C2C12 skeletal muscle and 3T3-L1 fat cells by modulating key effectors of insulin signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic insulin resistance is unknown. The effects of GRE on lipid-induced hepatic insulin resistance using palmitate-exposed C3A liver cells and obese insulin resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and lipid metabolism in treated C3A cells and improved insulin sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose metabolism, such as insulin receptor (Insr) and insulin receptor substrate 1 and 2 (Irs1 and Irs2), as well as those involved in lipid metabolism, including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic insulin resistance by improving insulin sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated pathways.

Visualization of Aspalathin in Rooibos (Aspalathus linearis) Plant and Herbal Tea Extracts Using Thin-Layer Chromatography.

Aspalathin, the main polyphenol of rooibos (Aspalathus linearis), is associated with diverse health promoting properties of the tea. During fermentation, aspalathin is oxidized and concentrations are significantly reduced. Standardized methods for quality control of rooibos products do not investigate aspalathin, since current techniques of aspalathin detection require expensive equipment and expertise. Here, we describe a simple and fast thin-layer chromatography (TLC) method that can reproducibly visualize aspalathin in rooibos herbal tea and plant extracts at a limit of detection (LOD) equal to 178.7 ng and a limit of quantification (LOQ) equal to 541.6 ng. Aspalathin is a rare compound, so far only found in A. linearis and its (rare) sister species A. pendula. Therefore, aspalathin could serve as a marker compound for authentication and quality control of rooibos products, and the described TLC method represents a cost-effective approach for high-throughput screening of plant and herbal tea extracts.

Aspalathin-Rich Green Rooibos Extract Lowers LDL-Cholesterol and Oxidative Status in High-Fat Diet-Induced Diabetic Vervet Monkeys.

Type 2 diabetic patients possess a two to four fold-increased risk for Cardiovascular Diseases (CVD). Hyperglycemia, oxidative stress associated with endothelial dysfunction and dyslipidemia are regarded as pro-atherogenic mechanisms of CVD. In this study, high-fat diet-induced diabetic and non-diabetic vervet monkeys were treated with 90 mg/kg of aspalathin-rich green rooibos extract (Afriplex GRT) for 28 days, followed by a 1-month wash-out period. Supplementation showed improvements in both the intravenous glucose tolerance test (IVGTT) glycemic area under curve (AUC) and total cholesterol (due to a decrease of the low-density lipoprotein [LDL]) values in diabetics, while non-diabetic monkeys benefited from an increase in high-density lipoprotein (HDL) levels. No variation of plasma coenzyme Q10 (CoQ10) were found, suggesting that the LDL-lowering effect of Afriplex GRT could be related to its ability to modulate the mevalonate pathway differently from statins. Concerning the plasma oxidative status, a decrease in percentage of oxidized CoQ10 and circulating oxidized LDL (ox-LDL) levels after supplementation was observed in diabetics. Finally, the direct correlation between the amount of oxidized LDL and total LDL concentration, and the inverse correlation between ox-LDL and plasma CoQ10 levels, detected in the diabetic monkeys highlighted the potential cardiovascular protective role of green rooibos extract. Taken together, these findings suggest that Afriplex GRT could counteract hyperglycemia, oxidative stress and dyslipidemia, thereby lowering fundamental cardiovascular risk factors associated with diabetes.

Rooibos agro-processing waste as herbal tea products: optimisation of soluble solids extraction from dust and application to improve sensory profile, colour and flavonoid content of stem infusions.

BACKGROUND: Rooibos represents 10% of the global herbal tea market. Shrinking production areas as a result of climate change necessitate the maximum conversion of plant biomass to product. The present study aimed to determine the potential of rooibos tea processing waste (i.e. fine dust and coarse stems) as potential flavour and herbal tea ingredients, respectively. RESULTS: Hot water extraction of soluble solids (SS) from rooibos dust was optimised and extracts from different production batches (n = 20) were prepared. Their sensory profiles were similar, although less intense than that of infusions of commercial rooibos (n = 20) when diluted to the same SS content. The turbidity and flavonoid content of the diluted extracts was mostly lower (P < 0.05) than that of commercial rooibos. An atypical and negative aroma attribute, 'planky/pencil shavings', was predominant in the stem infusions (n = 20), which contained less SS (P < 0.05) than commercial rooibos. Blends of stem infusion and extract could not effectively mask this negative aroma note (P > 0.05). CONCLUSION: Rooibos dust could be used to produce a rooibos flavour extract, whereas the prominent atypical, negative 'planky/pencil shavings' aroma note of the stems would limit their inclusion in commercial rooibos blends. © 2019 Society of Chemical Industry.

The Potential of South African Herbal Tisanes, Rooibos and Honeybush in the Management of Type 2 Diabetes Mellitus.

Diabetes mellitus is a metabolic disease that can lead to high morbidity, mortality and long-term complications. Available treatment strategies, which are mainly based on treating hyperglycemia, with insulin and other pharmacological agents are not completely efficient and can even lead to development of unwanted side effects. Scientific evidence suggests that bioactive compounds from teas and other plant-based foods, which are known source of natural antioxidants, could be an attractive strategy to preferentially treat and manage type 2 diabetes mellitus (T2DM) and thus, have significant therapeutic implications. In this review, we attempt an in-depth analysis and discussion of the current progress in our understanding of the antidiabetic potential of two commercialized South Africa herbal tisanes-Rooibos and Honeybush and their polyphenols.

A Beneficial Role of Rooibos in Diabetes Mellitus: A Systematic Review and Meta-Analysis.

In a rapid increase in cases of diabetes mellitus worldwide, there has been interested in the use of plant-derived polyphenols as nutraceuticals to prevent the onset and progression of diabetes mellitus and its associated complications. Aspalathus linearis, commonly known as rooibos, is a rich source of uncommon glycosylated plant polyphenols with various critical health-promoting properties, including the prevention and treatment of diabetes mellitus (DM). This study aimed to examine these effects by meta-analyzing the current evidence in diabetic rodent models. Peer-reviewed studies written in English from two databases, PubMed and Embase, were searched up to 28 February 2018. Studies reporting blood glucose levels in diabetic rodents with and without receiving rooibos extracts or their major phenolic compounds are included. Twelve studies enrolling 88 diabetic rodents treated with rooibos extracts or their polyphenols and 85 diabetic control males reported blood glucose levels. The pooled effect size was -0.89 (95% CI: -1.44 to -0.35) with a substantial heterogeneity (I² = 67.0%). This effect was likely to be modified by type of rooibos extracts and their polyphenols and treatment period. Blood glucose levels were significantly lower in diabetic rodent models treated with the phenolic compound rich in rooibos extracts, PPAG.

Direct effect of pholyphenol-rich plants, rooibos and ginkgo, on porcine ovarian cell functions.

The polyphenol-rich plants rooibos and ginkgo are widely used in folk medicine and in preparation of foods and drinks, but their effect on reproduction has not been properly studied yet. The aim of our in vitro experiments was to examine the possible direct effect of rooibos and ginkgo on the basic ovarian cell functions-proliferation, apoptosis and release of hormones progesterone (P4) and leptin (L). Porcine ovarian granulosa cells were cultured in the presence of rooibos or ginkgo extract (0, 1, 10 and 100 µg/ml of medium). The accumulation of markers of proliferation (PCNA and cyclin B1) and apoptosis (bax) and their mRNAs was analysed using immunocytochemistry and by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Release of P4 and L was evaluated by radioimmunoassay. It was observed that rooibos or ginkgo addition was able to inhibit proliferation (down-regulates PCNA, cyclin B1 and their mRNAs), to promote apoptosis (accumulation of bax) and to suppress both L and P4 release by ovarian cells. These data suggest a direct inhibitory (anti-proliferative, pro-apoptotic and hormone-suppressing) effect of polyphenol-containing plants rooibos and ginkgo on ovarian functions. The potential anti-reproductive effect of these medical plants is to be taken into account by their consumption.

Evaluation of capillary electrophoresis for the analysis of rooibos and honeybush tea phenolics.

Rooibos and honeybush are popular herbal teas produced from the shrubs of Aspalathus linearis and Cyclopia spp., respectively, which are indigenous to South Africa. Both herbal teas are rich in polyphenols and their consumption is associated with several health benefits, partly ascribed to their phenolic constituents. Quantification of phenolics in extracts and teas for quality control and research purposes is generally performed using HPLC, although dedicated and often species-specific methods are required. CE offers an attractive alternative to HPLC for the analysis of phenolics, with potential benefits in terms of efficiency, speed and operating costs. In this contribution, we report quantitative CZE methods for the analysis of the principal honeybush and rooibos phenolics. Optimal separation for honeybush and rooibos phenolics was achieved in 21 and 32 min, respectively, with good linearity and repeatability. Quantitative data for extracts of "unfermented" and "fermented" rooibos and two honeybush species were statistically comparable with those obtained by HPLC for the majority of compounds. The developed methods demonstrated their utility for the comparison of phenolic contents between different species and as a function of manufacturing processes, thus offering cost effective, although less sensitive and robust, alternatives to HPLC analysis.