Supercritical CO2 Extraction of Fatty Acids, Phytosterols, and Volatiles from Myrtle (Myrtus communis L.) Fruit.

BACKGROUND: Myrtle (Myrtus communis L.) is a coastal Mediterranean aromatic medicinal plant rich in essential oil components, flavonoids, and phenolic acids. Studies highlight the potential health benefits of myrtle bioactive compounds with antioxidant and antiproliferative properties. Since limited research exists on myrtle fruit's lipid fraction, the aim of this study was to apply supercritical CO2 extraction to obtain bioactive compounds from myrtle berries focusing on the fatty acids, sterols, and essential oils. METHODS: The optimization of the supercritical CO2 extraction of myrtle fruit using CO2 as solvent was carried out using the response surface methodology with Box-Behnken experimental design. The following conditions were tested: temperature (40, 50, and 60 °C), pressure (200, 300, and 400 bar), and flow rate (20, 30, and 40 g min-1) on the yield of lipid extract as well as on the yield of fatty acids, phytosterols, and volatiles present in the extract and constituting its bioactive potential. RESULTS: In the extracts examined, 36 fatty acids, 7 phytosterols, and 13 volatiles were identified. The average yield of the extract was 5.20%, the most abundant identified fatty acid was essential cis-linolenic acid (76.83%), almost 90% of the total phytosterols were ß-sitosterol (12,465 mg kg-1), while myrtenyl acetate (4297 mg kg-1) was the most represented volatile compound. The optimal process conditions obtained allow the formulation of extracts with specific compositions.

New Steroidal Saponins Isolated from the Rhizomes of Paris mairei.

The genus Paris is an excellent source of steroidal saponins that exhibit various bioactivities. Paris mairei is a unique species and has been widely used as folk medicine in Southwest China for a long time. With the help of chemical methods and modern spectra analysis, five new steroidal saponins, pamaiosides A-E (1-5), along with five known steroidal saponins 6-10, were isolated from the rhizomes of Paris mairei. The cytotoxicity of all the new saponins was evaluated against human pancreatic adenocarcinoma PANC-1 and BxPC3 cell lines.

Progress in the Medicinal Value, Bioactive Compounds, and Pharmacological Activities of Gynostemma pentaphyllum.

Gynostemma pentaphyllum (Thunb.) Makino (GP), also named Jiaogulan in Chinese, was known to people for its function in both health care and disease treatment. Initially and traditionally, GP was a kind of tea consumed by people for its pleasant taste and weight loss efficacy. With the passing of the centuries, GP became well known as more than just a tea. Until now, numbers of bioactive compounds, including saponins (also named gypenosides, GPS), polysaccharides (GPP), flavonoids, and phytosterols were isolated and identified in GP, which implied the great medicinal worth of this unusual tea. Both in vivo and in vitro tests, ranging from different cell lines to animals, indicated that GP possessed various biological activities including anti-cancer, anti-atherogenic, anti-dementia, and anti-Parkinson's diseases, and it also had lipid-regulating effects as well as neuroprotection, hepatoprotective, and hypoglycemic properties. With the further development and utilization of GP, the research on the chemical constituents and pharmacological properties of GP were deepening day by day and had made great progress. In this review, the recent research progress in the bioactive compounds, especially gypenosides, and the pharmacological activities of GP were summarized, which will be quite useful for practical applications of GP in the treatment of human diseases.

Chemical Composition and in vitro Anti-inflammatory Activity of Wheat Germ Oil Depending on the Extraction Procedure.

This study aimed to examine the chemical composition of wheat germ oil extracted by three different methods, and to evaluate its inhibitory effect on the cyclooxygenase and proteinase activities. The results showed that the contents of policosanols, tocopherols and phytosterols were affected by the extraction procedure. However, the fatty acid composition of the different oil extracts was nearly the same. Among the tested oils samples, cold pressed oil exhibited the strongest inhibitory activity against proteinase (93.4%, IC50 =195.7 µg/mL) and cyclooxygenase 1 (80.5%, IC50 =58.6 µg/mL). Furthermore, the cold pressed oil had the highest content of octacosanol, ß-sitosterol and α-linolenic acid, suggesting that those bioactive compounds could be essential for the potent ani-cyclooxygenase activity. The present data revealed that wheat germ oil contained cyclooxygenase and trypsin inhibitors, which are the promising therapeutic target for the treatment of various inflammatory diseases. Thus, wheat germ oil might be used to develop functional foods and pharmaceutic products for the human health.

Effects of Enzymatic Pretreatment of Seeds on the Physicochemical Properties, Bioactive Compounds, and Antioxidant Activity of Pomegranate Seed Oil.

Enzymatic pretreatment of seeds is a novel approach that enhances the health benefits of the extracted oil. The study investigated the influence of the enzymatic pretreatment of seeds on the quality of oil from different pomegranate cultivars. The quality of the ultrasound-assisted (and ethanol-extracted) oil was studied, with respect to the refractive index (RI), yellowness index (YI), conjugated dienes (K232), peroxide value (PV) ρ-anisidine value (AV), total oxidation value (TOTOX), total carotenoid content (TCC), total phenolic compounds (TPC), fatty acid composition, phytosterol composition, ferric reducing antioxidant power (FRAP), and 2.2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging capacity. The seeds of three different pomegranate cultivars ('Wonderful', 'Herskawitz', and 'Acco') were digested with an equal mixture of Pectinex Ultra SPL, Flavourzyme 100 L, and cellulase crude enzymes, at a concentration, pH, temperature, and time of 1.7%, 4.5, 40 °C, and 5 h, respectively. Enzymatic pretreatment of PS increased oil yield, PV, TPC, TCC, and DPPH radical scavenging capacity, but decreased the YI. The levels of K232, AV and TOTOX, fatty acids, phytosterols, RI, and FRAP, were not significantly affected by enzymatic pretreatment of PS. Principal component analysis (PCA) established that oil extracted from the 'Acco' seed after enzymatic pretreatment had higher yield, TPC, TCC, and DPPH radical scavenging capacity. Therefore, enzyme-pretreated 'Acco' pomegranate fruit seed is a source of quality seed oil with excellent antioxidant properties.

Microwave and Ultrasound Pretreatment of Moringa oleifera Lam. Seeds: Effects on Oil Expression, Oil Quality, and Bioactive Component.

This study investigates the application of green technologies (microwave and ultrasound pretreatment) in the extraction of Moringa oleifera Lam. seed oil and its effects on oil expression, oil quality, and bioactive component. Moringa seeds were pretreated with microwave (90 W, 60 s) or ultrasound (50 W, 1 h) before mechanical expression. A separate group received no pretreatment before oil extraction. Oils from these groups were then compared. Results show that oil yield increased with ultrasound pretreatment (1.24%) and significantly increased with microwave pretreatment (3.11%). For oil flow rate, the microwave and ultrasound pretreatment resulted in faster extraction (7.67 and 6.93 kg/h respectively) as compared with the control (6.51 kg/h). For physicochemical parameters, the microwave and ultrasound group had significantly less free fatty acids and significantly greater unsaponifiable matter as compared with the control. For fatty acid composition, results show that moringa seeds procured from Davao Oriental had greater oleic acid content (~77%) as compared with those reported by other literature. For phytosterol content, the predominant phytosterols found were ß-sitosterol, stigmasterol, and campesterol. Microwave and ultrasound pretreatment significantly increased total phytosterol (680.58 and 369.32 mg/kg respectively) as compared with the control (72.69 mg/kg) due to the mass transfer of the phytosterols. Microwave and ultrasound pretreatment also led to stigmastanol formation. For antioxidant activity, a comparison of both DPPH and FRAP assays depicts that the microwave group exhibited the best overall antioxidant activity. Lastly, for oil stability, a lower peroxide value was found in the microwave and ultrasound groups across time intervals, which may be attributed to their antioxidant activity. In summary, ultrasound and microwave pretreatment can improve oil expression, oil quality, and bioactive content of the mechanically expressed moringa oils.

Discovery of the signal pathways and major bioactive compounds responsible for the anti-hypoxia effect of Chinese cordyceps.

ETHNOPHARMACOLOGICAL RELEVANCE: Hypoxia will cause an increase in the rate of fatigue and aging. Chinese cordyceps, a parasitic Thitarodes insect-Ophiocordyceps sinensis fungus complex in the Qinghai-Tibet Plateau, has long been used to ameliorate human conditions associated with aging and senescence, it is principally applied to treat fatigue, night sweating and other symptoms related to aging, and it may play the anti-aging and anti-fatigue effect by improving the body's hypoxia tolerance. AIMS OF THE STUDY: The present study investigated the anti-hypoxia activity of Chinese cordyceps and explore the main corresponding signal pathways and bioactive compounds. MATERIALS AND METHODS: In this study, network pharmacology analysis, molecular docking, cell and whole pharmacodynamic experiments were hired to study the major signal pathways and the bioactive compounds of Chinese cordyceps for anti-hypoxia activity. RESULTS: 17 pathways which Chinese cordyceps acted on seemed to be related to the anti-hypoxia effect, and "VEGF signal pathway" was one of the most important pathway. Chinese cordyceps improved the survival rate and regulated the targets related VEGF signal pathway of H9C2 cells under hypoxia, and also had significant anti-hypoxia effects to mice. Chorioallantoic membrane model experiment showed that Chinese cordyceps and the main constituents of (9Z,12Z)-octadeca-9,12-dienoic acid and cerevisterol had significant angiogenic activity in hypoxia condition. CONCLUSION: Based on the results of network pharmacology and molecular docking analysis, cell and whole pharmacodynamic experiments, promoting angiogenesis by regulating VEGF signal pathway might be one of the mechanisms of anti-hypoxia effect of Chinese cordyceps, (9Z, 12Z)-octadeca-9,12-dienoic acid and cerevisterol were considered as the major anti-hypoxia bioactive compounds in Chinese cordyceps.

Novel Fast Chromatography-Tandem Mass Spectrometric Quantitative Approach for the Determination of Plant-Extracted Phytosterols and Tocopherols.

Phytosterols and tocopherols are commonly used in food and pharmaceutical industries for their health benefits. Current analysis methods rely on conventional liquid chromatography, using an analytical column, which can be tedious and time consuming. However, simple, and fast analytical methods can facilitate their qualitative and quantitative analysis. In this study, a fast chromatography-tandem mass spectrometric (FC-MS/MS) method was developed and validated for the quantitative analysis of phytosterols and tocopherols. Omitting chromatography by employing flow injection analysis-mass spectrometry (FIA-MS) failed in the quantification of target analytes due to analyte-to-analyte interferences from phytosterols. These interferences arise from their ambiguous MS fingerprints that would lead to false identification and inaccurate quantification. Therefore, a C18 guard column with a 1.9 µm particle size was employed for FC-MS/MS under isocratic elution using acetonitrile/methanol (99:1 v/v) at a flow rate of 600 µL/min. Analyte-to-analyte interferences were identified and eliminated. The false peaks could then be easily identified due to chromatographic separation. In addition, two internal standards were evaluated, namely cholestanol and deuterated cholesterol. Both internal standards contributed to the observed analyte-to-analyte interferences; however, adequate shift in the retention time for deuterated cholesterol eliminated its interferences and allowed for an accurate quantification. The method is fast (1.3 min) compared to published methods and can distinguish false peaks observed in FIA-MS. Seven analytes were quantified simultaneously, namely brassicasterol, campesterol, stigmasterol, ß-sitosterol, α-tocopherol, δ-tocopherol, and γ-tocopherol. The method was successfully applied in the quantitative analysis of phytosterols and tocopherols present in the unsaponifiable matter of canola oil deodorizer distillate (CODD). ß-sitosterol and γ-tocopherol were the most abundant phytosterols and tocopherols, respectively.

Evaluation of Antioxidant and Enzyme Inhibition Properties of Croton hirtus L'Hér. Extracts Obtained with Different Solvents.

Croton hirtus L'Hér methanol extract was studied by NMR and two different LC-DAD-MSn using electrospray (ESI) and atmospheric pressure chemical ionization (APCI) sources to obtain a quali-quantitative fingerprint. Forty different phytochemicals were identified, and twenty of them were quantified, whereas the main constituents were dihydro α ionol-O-[arabinosil(1-6) glucoside] (133 mg/g), dihydro ß ionol-O-[arabinosil(1-6) glucoside] (80 mg/g), ß-sitosterol (49 mg/g), and isorhamnetin-3-O-rutinoside (26 mg/g). C. hirtus was extracted with different solvents-namely, water, methanol, dichloromethane, and ethyl acetate-and the extracts were assayed using different in vitro tests. The methanolic extracts presented the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), and ferric reducing antioxidant power (FRAP) values. All the tested extracts exhibited inhibitory effects on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with a higher activity observed for dichloromethane (AChE: 5.03 and BChE: 16.41 mgGALAE/g), while the methanolic extract showed highest impact against tyrosinase (49.83 mgKAE/g). Taken together, these findings suggest C. hirtus as a novel source of bioactive phytochemicals with potential for commercial development.

Comparative Study on Functional Components, Physicochemical Properties and Antioxidant Activity of Amaranthus Caudatus L. Oils Obtained by Different Solvents Extraction.

Functional compositions, physicochemical properties and antioxidant activities of Amaranthus caudatus L. oils (ACO) obtained by different solvents were comparatively investigated. All the resulted ACO were enrich in 75% unsaturated fatty acid and in squalene of about 4 g/100 g. Different solvents showed varying in oil extraction, where acetone results a highest yield of 6.80 g/100 g. ACO extracted by ethanol showed a highest tocopherol (1351.26 mg/kg), polyphenols (211.28 mg/kg) and squalene (42519.13 mg/kg). However, phytosterols in ACO extracted by hexane (27571.20 mg/kg) was higher than that by acetone (19789.91 mg/kg), ethanol (22015.73 mg/kg) and petroleum ether (24763.30 mg/kg). Furthermore, antioxidant activity of ACO was also measured by DPPH, ABTS and FRAP assay. According to principal component and correlation analysis, squalene was correlated with the DPPH scavenging ability, but phytosterols and tocopherols was correlated with the ABTS and ferric reducing ability of the oils, respectively. This study provides a promising excellent source of functional oil for food industries.

Combination of temperature-assisted ternary phase homogenous liquid-liquid extraction with deep eutectic solvent-based dispersive liquid-liquid microextraction for the extraction of phytosterols from cow milk and cream samples.

A three-phase homogenous liquid-liquid extraction method followed by deep eutectic solvent-based dispersive liquid-liquid microextraction has been used for the extraction and preconcentration of some phytosterols from cow milk and cream samples. In this method, cow milk or melted cream was transferred into a glass test tube and a mixture of sodium hydroxide solution (as fatty acids saponification agent) and acetonitrile (as extraction/dispersive solvent) was added to the solution. The mixture was shaken manually and placed in a water bath. After dissolving sodium chloride and centrifugation, the obtained supernatant phase (acetonitrile) was removed and mixed with ethyl methyl ammonium chloride: phenyl acetic acid deep eutectic solvent at microliter level. Under optimum conditions, low limits of detection (1.6-4.1 µg/L) and quantification (5.3-13.1 µg/L), high enrichment factors (138-207), and extraction recoveries (55-83%), and good precision (relative standard deviations ≤ 6.8%) can be obtained. The method was done on different milk and cream samples and the results showed that total phytosterols level in all samples were in the permitted level established by the National Standard Organization Guidelines, except for three cream samples.

Chemistry, oxidative stability and bioactivity of oil extracted from Rosa rugosa (Thunb.) seeds by supercritical carbon dioxide.

Rosa rugosa Thunb. seed oil (RR) extracted by supercritical CO2 was investigated. RR chemical composition, radical scavenging effect and oxidative stability were evaluated. RR aqueous emulsions were examined for cell cytotoxicity, proliferation, redox state and migration using mouse embryonic fibroblast Balb/3T3, human dermal fibroblast NHDF cell lines, and on neoplastic cell lines: acute monocytic leukemia THP-1 and lung adenocarcinoma A549. RR total contents of phytosterols, tocopherols, carotenoids and phenolics were 10115.23, 784.16, 40.32 and 10.30 mg/kg, respectively. Rich antioxidant composition of RR was reflected in its high antioxidant activity (2.1 mM/kg Trolox equivalent) as well as oxidative stability (activation energy 105.6 kJ/mol). The RR emulsions led to marked augmentation of the total cell protein content in BALB/3T3 and NHDF cultures, inhibited cancer cell migration and reduced ROS formation. The studied RR oil proved to have a remarkable combination of bioactive compounds and to exert an antioxidative and chemopreventive effects.

Characterization of press and solvent extraction oils from new sunflower seeds with modified phytosterol compositions.

BACKGROUND: Phytosterols are plant components with health benefits. Oleaginous seed hybridization can be relevant to increase phytosterols in diet through enriched oils. Sunflower oils obtained by press (PO) and subsequent solvent extraction (SO) from three types of phytosterol-enriched seeds were characterized. One presented a phytosterol composition of common sunflower seeds, whereas the other two were rich in campesterol and Δ7-stigmasterol, respectively. Seeds from two different harvests, 2015 and 2017, were studied. RESULTS: The type of extraction did not have a significant influence on the fatty acid composition. However, considerable differences were found between harvests. The oleic-to-linoleic ratio decreased from 0.71 in 2015 to 0.47 in 2017. The phytosterol compositions of the PO were similar to their SO homologues and no substantial differences were found between harvests. However, the SO presented higher total contents of phytosterols (4849-9249 mg kg-1 ) than the PO (2839-5284 mg kg-1 ) and the oils of 2017 showed higher levels (4476-9249 mg kg-1 ) compared to 2015 (2839-5754 mg kg-1 ). Unlike phytosterols, no significant differences were found in the tocopherol contents between the PO and SO or between harvests. The PO met Codex specifications for edible oils, except for trace metals, with concentrations close or above the limits for Cu, Fe, Pb and As. CONCLUSIONS: Differences in environmental and/or cultivation conditions between harvests may result in substantial differences in the fatty acid composition and phytosterol content in oils from the new sunflower seeds. Rigorous measures and controls to avoid trace metal contamination are required so that the PO can be considered as edible virgin oils. © 2020 Society of Chemical Industry.

Application of response surface methodology (RSM) for the optimization of supercritical CO2 extraction of oil from patè olive cake: Yield, content of bioactive molecules and biological effects in vivo.

The two-phase technology for olive oil extraction generates large amounts of patè olive cake (POC), a by-product that is rich in bioactive health-promoting compounds. Here, response surface methodology (RSM) was used to maximize supercritical-CO2 oil extraction from POC, while minimizing operative temperature, pressure and time. Under the optimal parameters (40.2 °C, 43.8 MPa and time 30 min), the oil yield was 14.5 g·100 g-1 dw (~65% of the total oil content of the freeze-dried POC matrix), as predicted by RSM. Compared with freeze-dried POC, the oil contained more phytosterols (13-fold), tocopherols (6-fold) and squalene (8-fold) and was a good source of pentacyclic triterpenes. When the biological effects of POC oil intake (20-40 µL·die-1) were evaluated in the livers of BALB/c mice, no significant influence on redox homeostasis was observed. Notably, a decline in liver triglycerides alongside increased activities of NAD(P)H:Quinone Oxidoreductase 1, Carnitine Palmitoyl-CoA Transferase and mitochondrial respiratory complexes suggested a potential beneficial effect on liver fatty acid oxidation.

Effect of different processing methods on physicochemical properties, chemical compositions and in vitro antioxidant activities of Paeonia lactiflora Pall seed oils.

A research was performed to determine and compare the physicochemical properties, chemical compositions and in vitro antioxidant activities of Paeonia lactiflora Pall seed oils with ultrasonic-assisted solvent extraction, pressing and supercritical fluid extraction. Paeonia lactiflora Pall seed oil contained a high percentage of polyunsaturated fatty acids and monounsaturated fatty acids, especially oleic (31.62-32.88%) and α-linolenic acids (37.55-39.95%). The beneficial multiple dietary phytochemicals (tocopherol, phytosterols and squalene) and in vitro antioxidant activity were significantly influenced by the hull and processing method (P＜0.05). However, higher tocopherol (596.67-738.76 mg/kg) and phytosterols (5775.01-6055.62 mg/kg) contents were found in supercritical fluid extraction oils. Additionally, ten individual polyphenols were quantified, and significantly influenced by the hull and processing method (P＜0.05), with the content of benzoic acid and several individual flavonoids being the higher. According to the results, pressing might be the best process for extracting oil with a high number of polyphenols.

Optimization of the silica-gel adsorption technique for the extraction of phytosterol glycosides from soybean lecithin powder using response surface methodology and artificial neural network models.

Phytosterol glycosides (PGs), comprising both acylated steryl glycosides (ASGs) and steryl glycosides (SGs), are active ingredients with benefits for human use. Here, we aimed to optimize the silica-gel adsorption technique for the extraction of PGs from soybean lecithin powder, which contains 5 to 10% of these glycolipids. Both response surface methodology (RSM) and artificial neural networks (ANNs) were applied to optimize the PG extraction parameters (X1 = silica-gel dosage, X2 = adsorption temperature, and X3 = lecithin concentration) for high-purity phospholipid and PG production, and their prediction and optimization accuracies were compared. Although both models fitted well with the experimental data, the ANN model demonstrated better accuracy for predicting and optimizing the conditions using four interrelated dependent variables (Y1 = phospholipid yield, Y2 = ASG recovery, Y3 = SG recovery, and Y4 = PG purity) and had a higher coefficient of determination and lower root mean square error and absolute average deviation. After digitally setting the percentages of the four dependent variables for phospholipid and PG production, the ANN-optimized phospholipid product (Y1 = 88.07%, Y2 = 98.89%, Y3 = 100%, and Y4 = 49.03%) was acquired at X1 = 3.54 g/g, X2 = 26 °C, and X3 = 43 mg/mL, whereas the PG product (Y1 = 83.83%, Y2 = 97.64%, Y3 = 100%, and Y4 = 59.21%) was obtained at X1 = 2.00 g/g, X2 = 28.38 °C, and X3 = 41 mg/mL. In conclusion, the ANN method was better than RSM for the optimization of the silica-gel adsorption technique for PG extraction from soybean lecithin powder. PRACTICAL APPLICATION: This paper lays a theoretical foundation for the optimization of the industrial production of phytosterol glycosides and the comprehensive utilization of lecithin resources.

Antidiabetes constituents, cycloartenol and 24-methylenecycloartanol, from Ficus krishnae.

Ficus krishnae stem bark and leaves are used for diabetes treatment in traditional medicines. Stem bark of F. krishnae was sequentially extracted with hexane, methanol and water, and these extracts were tested for their antihyperglyceamic activity by oral glucose tolerance test (OGTT) in overnight fasted glucose loaded normal rats. Hexane extract showed significant glucose lowering activity in OGTT, and the triterpene alcohols (cycloartenol+24-methylenecycloartanol) (CA+24-MCA) were isolated together from it by activity guided isolation and characterized by NMR and mass spectroscopy. The ratio of the chemical constituents CA and 24-MCA in (CA+24-MCA) was determined as 2.27:1.00 by chemical derivatization and gas chromatographic quantification. (CA+24-MCA) in high fat diet-streptozotocin induced type II diabetic rats showed significant antidiabetes activity at 1 mg/kg and ameliorated derailed blood glucose and other serum biochemical parameters. Cytoprotective activity of (CA+24-MCA) from glucose toxicity was evaluated in cultured RIN-5F cells by MTT assay and fluorescent microscopy. (CA+24-MCA) in in vitro studies showed enhanced cell viability in RIN-5F cells and significant protection of beta cells from glucose toxicity. Both in in vivo and in vitro studies (CA+24-MCA) showed enhancement in insulin release from the beta cells. In short term toxicity studies in mice (CA+24-MCA) did not show any conspicuous toxic symptoms. The combination of the phytosterols (CA+24-MCA) obtained through activity guided isolation of the stem bark of F. krishnae showed significant activity, and therefore is a promising candidate for new generation antidiabetes drug development.

Evaluation of Antioxidant and Antibacterial Activities, Cytotoxicity of Acacia seyal Del Bark Extracts and Isolated Compounds.

Water extract of Acacia seyal bark is used traditionally by the population in Djibouti for its anti-infectious activity. The evaluation of in vitro antibacterial, antioxidant activities and cytotoxicity as well as chemical characterization of Acacia seyal bark water and methanolic extracts were presented. The water extract has a toxicity against the MRC-5 cells at 256 µg/mL while the methanolic extract has a weak toxicity at the same concentration. The methanolic extract has a strong antioxidant activity with half maximal inhibitory concentration (IC50) of 150 ± 2.2 µg/mL using 1-diphenyl-2-picrylhydrazyl (DPPH) and IC50 of 27 ± 1.3 µg/mL using 2,2'-azino-bis 3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical methods. For ferric reducing/antioxidant power (FRAP) assay, the result is 45.74 ± 5.96 µg Vitamin C Equivalent (VCE)/g of dry weight (DW). The precipitation of tannins from methanol crude extract decreases the MIC from 64 µg/mL to 32 µg/mL against Staphylococcus aureus and Corynebacterium urealyticum. However, the antioxidant activity is higher before tannins precipitation than after (IC50 = 150 µg/mL for methanolic crude extract and 250 µg/mL after tannins precipitation determined by DPPH method). By matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, the results showed that the condensed tannins consist of two types of catechin and gallocatechin-based oligomers. The fractionation led to the identification of three pure compounds: two flavanols catechin and epicatechin; one triterpene as lupeol; and a mixture of three steroids and one fatty acid: campesterol, stigmasterol, clionasterol, and oleamide.

Himalayan Nettle Girardinia diversifolia as a Candidate Ingredient for Pharmaceutical and Nutraceutical Applications-Phytochemical Analysis and In Vitro Bioassays.

Girardinia diversifolia, also known as Himalayan nettle, is a perennial herb used in Nepal to make fiber as well as in traditional medicine for the treatment of several diseases. To date, phytochemical studies and biological assays on this plant are scarce. Thus, in the present work, the G. diversifolia extracts have been evaluated for their potential pharmaceutical, cosmetic and nutraceutical uses. For this purpose, detailed phytochemical analyses were performed, evidencing the presence of phytosterols, fatty acids, carotenoids, polyphenols and saponins. The most abundant secondary metabolites were ß- and γ-sitosterol (11 and 9% dw, respectively), and trans syringin (0.5 mg/g) was the most abundant phenolic. Fatty acids with an abundant portion of unsaturated derivatives (linoleic and linolenic acid at 22.0 and 9.7 mg/g respectively), vitamin C (2.9 mg/g) and vitamin B2 (0.12 mg/g) were also present. The antioxidant activity was moderate while a significant ability to inhibit acetylcholinesterase (AChE), butyrilcholinesterase (BuChE), tyrosinase, α-amylase and α-glucosidase was observed. A cytotoxic effect was observed on human ovarian, pancreatic and hepatic cancer cell lines. The effect in hepatocarcinoma cells was associated to a downregulation of the low-density lipoprotein receptor (LDLR), a pivotal regulator of cellular cholesterol homeostasis. These data show the potential usefulness of this species for possible applications in pharmaceuticals, nutraceuticals and cosmetics.

Phytosterols extraction from hickory (Carya cathayensis Sarg.) husk with a green direct citric acid hydrolysis extraction method.

This study investigated the direct citric acid hydrolysis extraction method to optimize phytosterols extraction from hickory husk. Single factor experiments followed by a three-level three-factor Box-Behnken experiments were performed. The optimal extraction parameters were determined as: pH of 2.0, liquid-to-solid ratio of 17.12: 1 mL/g, and temperature of 55.81 °C. Practical experiments were carried out in triplicate, and subsequently yielded phytosterols of 912.452 ± 17.452 µg/g DW, in good consistence with the predicted extraction yield of 902.874 µg/g DW. The conductivity of the extract was also found to play effective role under direct citric acid hydrolysis and recorded 36.30 ± 1.08 µs/cm at optimum extraction condition. ß-Sitosterol stigmasterol, campsterol, ergosterol and lupeol were detected as main PSs and triterpenoids in hickory husk using UPLC-Triple-TOF/MS. Finally, the comparison between direct hydrolysis extraction and traditional solvent extraction showed that this new method was more effective and eco-friendlier to extract both free and conjugated phytosterols.