Chemical composition and in vitro iron bioavailability of extruded and open-pan cooked germinated and ungerminated pearl whole millet "Pennisetum glaucum (L.) R. Br."

This study aimed to evaluate the effect of extrusion and of open-pan cooking on whole germinated and non-germinated grains of pearl millet (Pennisetum glaucum L. R. Br.), on its chemical-nutritional composition and in vitro iron bioavailability. The experimental design consisted of three flours: non-germination open-pan cooked millet flour (NGOPCMF), germination open-pan cooked millet flour (GOPCMF), and extrusion cooked millet flour (ECMF). The ECMF increased the carbohydrates, iron, manganese, diosmin, and cyanidin and decreased the total dietary fiber, resistant starch, lipids, and total vitamin E, in relation to NGOPCMF. The GOPCMF increased the lysine and vitamin C and decreased the phytate, lipids, total phenolic, total vitamin E, and riboflavin concentration, in relation to NGOPCMF. Furthermore, germinated cooked millet flour and extruded millet flour improved iron availability in vitro compared to non-germinated cooked millet flour. GOPCMF and ECMF generally preserved the chemical-nutritional composition of pearl millet and improved in vitro iron bioavailability; therefore, they are nutritionally equivalent and can be used to develop pearl millet-based products.

Simultaneous extraction, separation, and analysis of 5-caffeoylquinic acid and caffeine from coffee co-product by PLE-SPE × HPLC-PDA two-dimensional system.

This study proposed an integrated and automated procedure to extract, separate, and quantify bioactive compounds from a coffee co-product by pressurized liquid extraction (PLE) coupled inline with solid phase extraction (SPE) and online with HPLC-PDA (PLE-SPE × HPLC-PDA). The efficiency of the two-dimensional system in performing real-time analysis was verified by comparing HPLC-PDA results acquired by the system (online) and carried out after the extract fraction collection (offline). Different flow rates (1.5 mL/min for 336 min, 2 mL/min for 246.4 min, and 2.5 mL/min for 201.6 min) were evaluated to optimize the extraction, separation, and analysis method by PLE-SPE × HPLC-PDA. Subcritical water at 125 °C and 15 min of static time allowed the highest extraction yields of caffeine and 5-caffeoylquinic acid (5-CQA). Caffeine was retained during the aqueous extraction in the SPE adsorbent and eluted from the column by exchanging the solvent for a hydroethanolic mixture. Thus, caffeine was separated from 5-CQA and other phenolic compounds, producing extracts with different compositions. The solvent flow rate did not have a significant effect (p-value ≥ 0.05) on the extraction, separation, and analysis (by online and offline methods) of 5-CQA. However, the online quantification of retained compounds in the SPE (i.e., caffeine) can underestimate concentration compared to offline analysis. Nevertheless, the results suggest that coupling of advanced techniques can be used to efficiently extract, separate, and analyze fractions of phenolic compounds, supplying an integrated method to produce high-added value ingredients for several applications.

High-intensity ultrasound-based process strategies for obtaining edible sunflower (Helianthus annuus L.) flour with low-phenolic and high-protein content.

The sunflower Helianthus annuus L. represents the 4th largest oilseed cultivated area worldwide. Its balanced amino acid content and low content of antinutrient factors give sunflower protein a good nutritional value. However, it is underexploited as a supplement to human nutrition due to the high content of phenolic compounds that reduce the sensory quality of the product. Thus, this study aimed at obtaining a high protein and low phenolic compound sunflower flour for use in the food industry by designing separation processes with high intensity ultrasound technology. First, sunflower meal, a residue of cold-press oil extraction processing, was defatted using supercritical CO2 technology. Subsequently, sunflower meal was subjected to different conditions for ultrasound-assisted extraction of phenolic compounds. The effects of solvent composition (water: ethanol) and pH (4 to 12) were investigated using different acoustic energies and continuous and pulsed process approaches. The employed process strategies reduced the oil content of sunflower meal by up to 90% and reduced 83% of the phenolic content. Furthermore, the protein content of sunflower flour was increased up to approximately 72% with respect to sunflower meal. The acoustic cavitation-based processes using the optimized solvent composition were efficient in breaking down the cellular structure of the plant matrix and facilitated the separation of proteins and phenolic compounds, while preserving the functional groups of the product. Therefore, a new ingredient with high protein content and potential application for human food was obtained from the residue of sunflower oil processing using green technologies.

Extraction of potential bioactive compounds from industrial Tahiti lime (Citrus latifólia Tan.) by-product using pressurized liquids and ultrasound-assisted extraction.

This work evaluated two emerging techniques in extracting phenolic compounds from Tahiti lime pomace - pressurized liquid extraction (PLE) and ultrasound-assisted extraction (UAE). PLE was performed at different temperatures (60 - 110 °C) and times (5 - 40 min), and UAE was carried out varying ultrasound power (160 - 792 W), time (2 - 10 min), and solvent to feed mass ratio (20 - 40 kg solvent/kg dried pomace). Both used ethanol and water (3:1, wt.) as the solvent. The effects of these variables were evaluated on global extraction yield, polyphenols, hesperidin, narirutin yields, and antioxidant capacity. PLE was strongly affected by temperature and extraction time, and the highest temperature (110 °C) provided the best results for global yield, total phenolic, and ORAC, except for hesperidin and narirutin, which were not significative affected by temperature. UAE revealed a weak dependency on power, S/F, and time; however, the lowest power level significantly increased the yields compared to no power application. Thus, P = 480 W, t = 6 min, and S/F = 30 was chosen as the best condition in the UAE in terms of overall extraction yield, total phenolics, specific phenolics, antioxidant capacities, and solvent and energy expenditures. UAE mechanisms were investigated by comparing with heated and stirred maceration, and scanning electron microscopy suggested that total phenolic yield was influenced by mechanisms that only ultrasound can provide. Micrographics confirmed the cavitation effect on Tahiti lime pomace particles' surface. To sum up, PLE resulted in the highest yields and antioxidant capacity, followed by UAE.

Anticancer effects of root and beet leaf extracts (Beta vulgaris L.) in cervical cancer cells (HeLa).

Cervical cancer is the fourth leading cause of cancer mortality in women worldwide. Beetroot (Beta vulgaris L.) has bioactive compounds that can inhibit the progression of different types of cancer. To analyze the antiproliferative effects of beet leaf and root extracts, we performed MTT, clonogenic survival, cell cycle analysis, Annexin/PI labeling, and western blotting. Here, we report that 10 and 100 µg/ml of root and leaf extracts decreased cell viability and potentiated rapamycin and cisplatin effects while decreased the number of large colonies, especially at 10 µg/ml (293.6 of control vs. 200.0 of leaf extract, p = .0059; 138.6 of root extract, p = .0002). After 48 hr, 100 µg/ml of both extracts led to increased sub-G1 and G0/G1 populations. In accordance, 100 µg/ml of root extract induced early apoptosis (mean = 0.64 control vs. 1.56 root; p = .048) and decreased cell size (p < .0001). Both extracts decreased phosphorylation and expression of mechanistic Target of Rapamycin (mTOR) signaling, especially by inhibiting ribosomal protein S6 (S6) phosphorylation, increasing cleaved poly(ADP-ribose) polysomerase 1 (PARP1) and Bcl-2-like protein 11 (BIM), and decreasing cyclin D1 expression, which regulates cell cycle progression. Here, we demonstrate that beetroot and leaf extracts could be an efficient strategy against cervical cancer.

Beetroot and leaf extracts present protective effects against prostate cancer cells, inhibiting cell proliferation, migration, and growth signaling pathways.

Beet (Beta vulgaris L.) has high nutritional value, containing bioactive compounds such as betalains and flavonoids. Scientific evidence points to the use of these natural compounds in the treatment of several types of cancer, such as prostate cancer, one of the main causes of morbidity and mortality in men. Here, we compared beet roots and leaves extracts, and their main compounds, apigenin, and betanin, respectively, in DU-145 and PC-3 prostate cancer cell lines. Both cells presented the proliferation decreased for beetroot and beet leaves extracts. The apigenin treatment also reduced the proliferation of both cell lines. Regarding cell migration, beet leaves extract was able to decrease the scratch area in both cell lines, whereas apigenin affected only PC-3 cells' migration. In colony formation assay, both extracts were effective in reducing the number of colonies formed. Besides, the beet leaves extracts and apigenin presented strong inhibition of growth-related signaling pathways in both cell lines, and the beetroot extract and betanin presented effects only in DU-145 cells. Furthermore, the extracts and isolated compounds were able to reduce the levels of apoptotic and cell cycle proteins. This study reveals that beet extracts have important anti-cancer effects against prostate cancer cells.

Characterization of pomegranate peel extracts obtained using different solvents and their effects on cell cycle and apoptosis in leukemia cells.

Pomegranate (Punica granatum L.) has been used in traditional herbal medicine by several cultures as an anti-inflammatory, antioxidant, antihyperglycemic, and for treatment and prevention of cancer and other diseases. Different parts of the fruit, extraction methods, and solvents can define the chemical profile of the obtained extracts and their biological activities. This study aimed to characterize the chemical profile of peel extracts collected using different extraction solvents and their biological effects on the cell cycle and apoptosis of THP-1 leukemic cells. Aqueous extract presented the highest content of punicalagins (α pun = 562.26 ± 47.14 mg/L and ß pun = 1,251.13 ± 22.21 mg/L) and the lowest content of ellagic acid (66.38 ± 0.21 mg/L), and it promoted a significant impairment of the cell cycle S phase. In fact, punicalagin-enriched fraction, but not an ellagic acid-enriched fraction, caused an S phase cell cycle arrest. All extracts increased the number of apoptotic cells. Punicalagin-enriched fraction increased the percentage of cells with fragmented DNA, which was intensified by ellagic acid combination. The treatment combining punicalagin and ellagic acid fractions increased the apoptotic cleaved PARP1 protein and reduced the activation of the growth-related mTOR pathway. Thus, these results evidence that solvent choice is critical for the phenolic compounds profile of pomegranate peel extracts and their biological activities.

Extraction of Flavonoids From Natural Sources Using Modern Techniques.

Flavonoids are one of the main groups of polyphenols found in natural products. Traditional flavonoid extraction techniques are being replaced by advanced techniques to reduce energy and solvent consumption, increase efficiency and selectivity, to meet increased market demand and environmental regulations. Advanced technologies, such as microwaves, ultrasound, pressurized liquids, supercritical fluids, and electric fields, are alternatives currently being used. These modern techniques are generally faster, more environmentally friendly, and with higher automation levels compared to conventional extraction techniques. This review will discuss the different methods available for flavonoid extraction from natural sources and the main parameters involved (temperature, solvent, sample quantity, extraction time, among others). Recent trends and their industrial importance are also discussed in detail, providing insight into their potential. Thus, this paper seeks to review the innovations of compound extraction techniques, presenting in each of them their advantages and disadvantages, trying to offer a broader scope in the understanding of flavonoid extraction from different plant matrices.

Protective effects of beet (Beta vulgaris) leaves extract against oxidative stress in endothelial cells in vitro.

Beetroot is an herb used worldwide as a food product, raw material for food industry, ethanol production and source of food coloring. Beet leaves are an unconventional food with antioxidant properties, which might neutralize reactive oxygen species (ROS) induced by oxidized Low-Density Lipoprotein (LDL) present in dyslipidemias. This study aimed to elucidate the effects of beet leaves on the suppression of LDL oxidative processes. Beet leaves extract was produced, characterized, and tested for its antioxidant capacity using endothelial cells in vitro. A model of human umbilical vein endothelial cells was used in various tests, including viability assay, molecular analysis of antioxidant genes, ROS labeling, and macrophage adhesion assay. The extract improved the antioxidative protection of endothelial cells against different agents including oxidized LDL-cholesterol and H2 O2 . It acted on ROS directly due to its high content of natural antioxidants, but also due to the activation and improvement of cellular defenses such as Superoxide dismutase 1, Superoxide dismutase 2, and catalase. The inhibition of LDL-mediated oxidative effects on endothelial cells may turn this unconventional food a functional food with great potential for phytotherapy of atherosclerosis as an adjuvant for medicinal treatments.

Pomegranate Juice and Peel Extracts are Able to Inhibit Proliferation, Migration and Colony Formation of Prostate Cancer Cell Lines and Modulate the Akt/mTOR/S6K Signaling Pathway.

Pomegranate (Punica granatum) is known to contain polyphenols with many potential health benefits, including anti-tumoral, anti-inflammatory, and anti-microbial properties. It has been used in popular medicine for cancer treatment, which still represents the major cause of cancer-related deaths in men worldwide. Importantly, pomegranate peels are valuable by-products of the food industry that are rich in polyphenols. Here we report a comparison between juice and peel aqueous extracts in prostate cancer DU-145 and PC-3 cell lines. Both extracts were able to inhibit the proliferation, migration and colony formation of those cells, although peel extracts presented more robust effects compared to juice. Besides, the growth-related mTOR/S6K signaling pathway presented strong inhibition after pomegranate extracts treatment. This study presents evidence that both juice and isolated peel extracts from promegate fruit have important anti-cancer effects against prostate cancer cells, modulating the mTOR/S6K signaling pathway.

Combining pressurized liquids with ultrasound to improve the extraction of phenolic compounds from pomegranate peel (Punica granatum L.).

The combination of ultrasound and pressurized liquid extraction (UAPLE) was evaluated for the extraction of phenolic compounds from pomegranate peels (Punica granatum L.). The influence of several variables of the process on extraction yield, including solvent type (water, ethanol + water 30, 50 and 70% v:v), temperature (50-100 °C), ultrasound power (0-800 W at the generator, or 0-38.5 W at the tip of the probe), mean particle size (0.68 and 1.05 mm), and number of cycles (1-5), were analyzed according to the yield of 20 different phenolic compounds. The most suitable temperatures for the extraction of phenolic compounds using water were from 70 to 80 °C. In general, 100 °C was not adequate since the lowest extraction yields were observed. Results suggested that ultrasound had a greater impact on extraction yields using large particles and that intermediate ultrasound power (480-640 W at the generator, or 23.1-30.8 W at the tip of the probe) produced the best results. Using small particles (0.68 mm) or large particles (1.05 mm), extraction with ultrasound was 1 cycle faster. Ultrasound may have offset the negative effect of the use of large particles, however, did not increase the yield of phenolic compounds in any of the cases studied after five cycles. Additionally, the continuous clogging problems observed with small particles were avoided with the use of large particles, which combined with ultrasound allowed consistent operation with good intra and inter-day reproducibility (>95%). Using samples with large particle size, the best extraction conditions were achieved with water extraction solvent, 70 °C extraction temperature, ultrasound power at 480 W, and 3 cycles, yielding 61.72 ±â€¯7.70 mg/g. UAPLE demonstrated to be a clean, efficient and a green alternative for the extraction of phenolic compounds from pomegranate peels. These findings indicate that UAPLE has a great potential to improve the extraction of bioactive compounds from natural products.

Fast analysis of curcuminoids from turmeric (Curcuma longa L.) by high-performance liquid chromatography using a fused-core column.

The recent development of fused-core technology in HPLC columns is enabling faster and highly efficient separations. This technology was evaluated for the development of a fast method for the analysis of main curcuminoids (curcumin, demethoxycurcumin and bisdemethoxycurcumin) present in extracts of turmeric (Curcuma longa L.). A step-by-step strategy was used to optimize temperature (40-55 °C), flow rate (1.0-2.5 mL min(-1)), mobile phase composition and equilibration time (1-5 min). A gradient method was developed using acidified water and acetonitrile combined with high column temperature (55 °C) and flow rate (2.5 mL min(-1)). Optimized conditions provided a method for the separation of these three curcuminoids in approximately 1.3 min with a total analysis time (sample-to-sample) of 7 min, including the clean-up and the re-equilibration of the column. Evaluation of chromatographic performance revealed excellent intraday and interday reproducibility (>99%), resolution (>2.23), selectivity (>1.12), peak symmetry (1.24-1.42) while presenting low limits of detection (<0.40 mg L(-1)) and quantification (<1.34 mg L(-1)). The robustness of the method was calculated according to the concentration/dilution of the sample and the injection volume. Several combinations of methanol and ethanol with water as sample solvents were evaluated and the best chromatographic results and extraction rate were obtained using 100% methanol. Finally, the developed method was validated with different extracts of turmeric rhizome and products that use turmeric in their formulation.