A novel terpenoid glycoside and other bioactive constituents from the seeds of Cichoriumintybus.

Phytochemical investigation of the seeds of Cichorium intybus L (C. intybus) led to isolate n-hexacosane (CI-1), an aliphatic higher ketone, n-nonacosan-3-one (CI-2), two aliphatic acid esters characterized as n-octacosanyl decanoate (CI-3) and n-tricosanyl hexadecanoate (CI-4), two mixed glycerides identified as as glyceryl-1-(hexadec-7- enoyl)-2- tetradecanoyl-3-hexadecanoate (Cl-5) and glyceryl-1- (eicos-9-enoyl)-2,3, bis-eicosanoate (Cl-6), and three e steroidal constituents and their structures were elucidated as as stigmast-5, 22-dien-3ß -ol-21-oic acids (Cl-7), stigmasterol-3ß-d-glucopyranoside (Cl-8) and stigmast-5, 22-dien-3-ß-ol-3-ß-d-glucuronopyranoside (Cl-9). The dry seeds powder was defatted and finally extracted with ethanol by using a maceration method. The ethanol was evaporated near to dryness and silica gel was added to the extract and a slurry with the help of methanol solvent was prepared. The slurry was loaded to the column by using petroleum ether and was eluted with a mixture of chloroform and methanol. A series of test tubes were collected and each test tube with 2 mL eluents was collected. Based on the thin layer chromatography (TLC) the content of nine test tubes were considered as pure compounds. The solvent was evaporated from the test tube at room temperature. All the nine compounds from the column were characterized by using Infrared (IR), Nuclear Magnetic Resonance (NMR) and Mass spectrometry (MS). Eight compounds were previously isolated from the plant and they showed various biological activities. A new compound was isolated for the first time from the plant kingdoms. Based on the chromatographic and spectroscopic analysis the new compound was characterized as stigmasterol carboxylic acid (CI-9). The isolated new compound could be used to treat liver and cardiac diseases.

Potential of Kalanchoe pinnata as a Cancer Treatment Adjuvant and an Epigenetic Regulator.

Cancer is a global public health problem that is related to different environmental and lifestyle factors. Although the combination of screening, prevention, and treatment of cancer has resulted in increased patient survival, conventional treatments sometimes have therapeutic limitations such as resistance to drugs or severe side effects. Oriental culture includes herbal medicine as a complementary therapy in combination with chemotherapy or radiotherapy. This study aimed to identify the bioactive ingredients in Kalanchoe pinnata, a succulent herb with ethnomedical applications for several diseases, including cancer, and reveal its anticancer mechanisms through a molecular approach. The herb contains gallic acid, caffeic acid, coumaric acid, quercetin, quercitrin, isorhamnetin, kaempferol, bersaldegenin, bryophyllin a, bryophyllin c, bryophynol, bryophyllol and bryophollone, stigmasterol, campesterol, and other elements. Its phytochemicals participate in the regulation of proliferation, apoptosis, cell migration, angiogenesis, metastasis, oxidative stress, and autophagy. They have the potential to act as epigenetic drugs by reverting the acquired epigenetic changes associated with tumor resistance to therapy-such as the promoter methylation of suppressor genes, inhibition of DNMT1 and DNMT3b activity, and HDAC regulation-through methylation, thereby regulating the expression of genes involved in the PI3K/Akt/mTOR, Nrf2/Keap1, MEK/ERK, and Wnt/ß-catenin pathways. All of the data support the use of K. pinnata as an adjuvant in cancer treatment.

A new isoquinoline and ceramide from the stem barks of Discoglypremna caloneura (Pax) Prain (Euphorbiaceae) with antiproteinase and cytotoxic activities.

Two new compounds, an isoquinoline (1) and caloneuramide (2), a ceramide were isolated from the stem bark of Discoglypremna caloneura together with seven known compounds namely aurantiamide acetate (3), acetylaleuritolic acid (4), 3α-hydroxylaleuritolic acid 2α-p-hydroxybenzoate (5), mixture of stigmasterol (6) and ß-sitosterol (7), mixture of 7-oxo-stigmasterol (8) and 7-oxo-ß-sitosterol (9). Their structures were determined based on data from literature and spectroscopic methods. Derivatization reactions on the isoquinoline led to two new compounds, the methylated (10) and acetylated (11) derivatives. Some compounds and extracts were evaluated for their cytotoxic and antiproteinase activity. Antiproteinase effect of compounds 1, 10 and 11 exhibited IC50 values of 10.77, 1.19 and 3.61 µg/mL respectively; significantly low compared to the standard drug, acetyl salicylic acid (IC50 = 20.28 µg/mL). Ethyl acetate and methanol extract exhibited moderate cytotoxicity activity on Chang liver cells with CC50 values of 167.90 ± 2.20 and 106.30 ± 2.03 µg/mL compared to the reference drug cucurmin (CC50 = 11.05 ± 1.04 µg/mL).

Yield, Characterization, and Possible Exploitation of Cannabis Sativa L. Roots Grown under Aeroponics Cultivation.

Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and psychoactive plant. Today, the nutraceutical potential of C.sativa is being increasingly reappraised; however, C. sativa roots remain poorly studied, despite citations in the scientific literature. In this direction, we identified and quantified the presence of valuable bioactives (namely, ß-sitosterol, stigmasterol, campesterol, friedelin, and epi-friedelanol) in the root extracts of C. sativa, a finding which might pave the way to the exploitation of the therapeutic potential of all parts of the C. sativa plant. To facilitate root harvesting and processing, aeroponic (AP) and aeroponic-elicited cultures (AEP) were established and compared to soil-cultivated plants (SP). Interestingly, considerably increased plant growth-particularly of the roots-and a significant increase (up to 20-fold in the case of ß-sitosterol) in the total content of the aforementioned roots' bioactive molecules were observed in AP and AEP. In conclusion, aeroponics, an easy, standardized, contaminant-free cultivation technique, facilitates the harvesting/processing of roots along with a greater production of their secondary bioactive metabolites, which could be utilized in the formulation of health-promoting and health-care products.

Characterization of seven sterols in five different types of cattle feedstuffs.

This paper provides a method for the quantification of sterols in different types of calf feedstuffs based on soy, sunflower, hay, calf feed and a mixture of all of them. The free fraction and the total sterolic fraction, after saponification and acidic hydrolysis of the samples, are extracted by solvent and the sterols are identified/quantified by reversed phase HPLC coupled to tandem mass spectrometry by atmospheric pressure chemical ionization. After the recovery evaluation, the method is validated in terms of linearity (coefficient of determination R2), repeatability (coefficient of variation RSD), limit of detection and quantification. In most of the cases, the most representative phytosterol is ß-sitosterol, followed by campesterol or stigmasterol and by other minor sterols such as fucosterol, and Δ-5-avenasterol. In addition, also cholesterol and ergosterol, if present, are evaluated in all the samples. As far as we know, very little information is available on the investigated feeds, which are commonly used on farms. The results of this survey were compared to other studies, if present in literature, showing good agreement. The proposed method resulted to be simple, fast and suitable for application to other sterols, feedstuffs and derived foods. The knowledge of the sterolic content and composition is getting more and more important, both in terms of comprehension of the vegetal biochemistry and as basis for sterolomic studies.

Isolation, characterization and quantitative HPLC-DAD analysis of components of charantin from fruits of Momordica charantia.

Charantin, a steroidal glycoside, exists as a mixture of stigmasterol glucoside (STG) and ß-sitosterol glucoside (BSG) in the fruits of Momordica charantia. Charantin has anti-diabetic activity comparable to insulin. The present work discusses a method for separation of components of charantin namely STG and BSG by simple extraction technique followed by preparative HPLC. The identity of separated components was established by chromatographic as well as spectral techniques. Also reversed phase HPLC-DAD method was developed and validated for estimation of STG and BSG present in fruits of Momordica charantia. The method used C18 column (75 mm × 4.6 mm, 3.5 µm) as stationary phase and methanol: water (98:02, v/v) as mobile phase. Retention times of STG and BSG were found to be 10.707 min and 11.870 min, respectively. The validated method was applied to evaluate content of these components in different extracts and some commercial herbal formulations containing fruits of Momordica charantia.

Nutritional Component and Chemical Characterization of Chinese Highland Barley Bran Oil.

The nutritional composition and chemical properties of the Chinese highland barley bran oil were characterized in this study. The barley bran oil extracted with solvent possessed relatively high acid value and peroxide value, indicating that the oil should be further refined before using. The fatty acid composition of the oil showed that the content of unsaturated fatty acids was 80.12 g/100 g, in which the content of polyunsaturated fatty acids was as high as 60.41 g/100 g. The overall triacylglycerol profile showed that the oil contained 27 TAGs including 21 regioisomers. Major TAGs included LLL (21.08 g/100 g), PLL (19.27 g/100 g), LLO (12.24 g/100 g), and LLLn (12.17 g/100 g). The total unsaponifiable matter of the oil reached up to 10.74 g/100 g oil. The total phytosterol content reached 7.90 g/100 g oil, in which ß-sitosterol was the most predominant, with the content of 5.69 g/100 g oil. Other important sterols included campesterol (1.32 g/100 g oil), lanosterol (0.70 g/100 g oil) and stigmasterol (0.19 g/100 g oil).

Spinasterol, 22,23-Dihydrospinasterol and Fernenol from Citrullus Colocynthis L. with Aphicidal Activity against Cabbage Aphid Brevicoryne Brassicae L.

Brevicoryne brassicae is a problematic pest in cabbage and other field crops. Synthetic pesticides are used to control this pest, but they are injurious for human health and the environment. The present study aimed to purify and identify the active compounds from Citrullus colocynthis leaves with an appraisal of their efficacy against B. brassicae. Separation and purification were performed via different chromatographic techniques. Molecular analysis and chemical structures were recognized by mass spectrum (MS) and nuclear magnetic resonance (NMR), respectively. Moreover, in vitro and in vivo aphicidal activity was assessed using various concentrations, i.e., 6.25, 12.5, 25 and 50 µg/mL at 12, 24, 48 and 72 h exposure. The outcome shows that mass spectrum analyses of the purified compounds suggested the molecular formulae are C30H50O and C29H50O, C29H48O. The compounds were characterized as fernenol and a mixture of spinasterol, 22,23-dihydrospinasterol by 1H-NMR and 13C-NMR spectrum analysis. The toxicity results showed that the mixture of spinasterol and 22,23-dihydrospinasterol showed LC50 values of 32.36, 44.49 and 37.50 µg/mL by contact, residual and greenhouse assay at 72 h exposure, respectively. In contrast, fernenol recorded LC50 values as 47.99, 57.46 and 58.67 µg/mL, respectively. On the other hand, spinasterol, 22,23-dihydrospinasterol showed the highest mortality, i.e., 66.67%, 53.33% and 60% while, 30%, 23.33% and 25% mortality was recorded by fernenol after 72 h at 50 µg/mL by contact, residual and greenhouse assay, respectively. This study suggests that spinasterol, 22,23-dihydrospinasterol are more effective against B. brassicae which may be introduced as an effective and suitable substitute of synthetic chemical pesticides.

Phytosterolaemia associated with parenteral nutrition administration in adult patients.

Vegetable lipid emulsions (LE) contain non-declared phytosterols (PS). We aimed to determine PS content depending on the brand and LE batch, and in adult hospitalised patients treated with parenteral nutrition (PN), to establish the association between plasma and administered PS. Part I was the LE study: totals and fractions of PS in three to four non-consecutive batches from six LE were analysed. Part II was the patient study: patients with at least 7 previous days of PN with 0·8 g/kg per d of an olive/soyabean (O/S) LE were randomised (day 0) 1:1 to O/S or 100 % fish oil (FO) at a dose of 0·4 g/kg per d for 7 d (day 7). Plasma PS, its fractions, total cholesterol on days 0 and 7, their clearance and their association with PS administered by LE were studied. In part I, LE study: differences were found in the total PS, their fractions and cholesterol among different LE brands and batches. Exclusive soyabean LE had the highest content of PS (422·36 (sd 130·46) µg/ml). In part II, patient study: nineteen patients were included. In the O/S group, PS levels were maintained (1·11 (sd 6·98) µg/ml) from day 0 to 7, while in the FO group, significant decreases were seen in total PS (-6·21 (sd 4·73) µg/ml) and their fractions, except for campesterol and stigmasterol. Plasma PS on day 7 were significantly associated with PS administered (R2 0·443). PS content in different LE brands had great variability. PS administered during PN resulted in accumulation and could be prevented with the exclusive administration of FO LE.

Phytosterols Content in Vegetable Oils of Brazil: Coconut, Safflower, Linseed and Evening Primrose

Abstract In the last years phytosterols, natural components of plants, have received more attention due to association of their consumption with reducing risk of cardiovascular diseases and cancer. There are several scientific studies about phytosterols in vegetable oils, but they are scarce in unconventional oils. The objective of this research was evaluating the content of phytosterols (β-sitosterol, stigmasterol and campesterol) in vegetable oils sold in São Paulo city, in Brazil. The analysis included cold alkaline saponification, derivatization with hexamethyldisilazane and trimethylchlorosilane reagents, and quantification by gas chromatography using flame ionization detection and internal standardization. The quality control parameters indicated that the method was suitable for analysis. Total sterols were between 272.3 mg kg-1 (coconut oil) to 6169.7 mg kg-1 (evening primrose oil). β-sitosterol was the component found in higher concentrations and evening primrose oil was the most representative in quantity of phytosterols.

Simultaneous determination of tocopherols, carotenoids and phytosterols in edible vegetable oil by ultrasound-assisted saponification, LLE and LC-MS/MS.

A method was developed to simultaneously determine eight bioactive compounds in edible oil based on ultrasound-assisted saponification, liquid-liquid extraction and liquid chromatography coupled with tandem mass spectrometry. Central composite design was employed to optimize ultrasonic temperature and time of saponification. Sample treatment was conducted by ultrasound-assisted saponification at temperature of 75 °C for 40 min. Limits of detection and limits of quantification ranged from 2.0 to 3.2 and from 6.1 to 10.0 ng/mL, respectively. Linear correlations were obtained (R2 > 0.99) and the recoveries at three spiked levels were between 81.7% and 112.0%. This method was employed to determine eight compounds in camellia oils and olive oils. As results, the contents of stigmasterol, δ-tocopherol, γ-tocopherol, ß-carotene and lutein in camellia oils were significantly higher than those in olive oils (p < 0.05). The proposed method can be successfully used to determination of these eight active compounds in camellia oil and other edible oils.

Insights in cyanobacteria lipidomics: A sterols characterization from Phormidium autumnale biomass in heterotrophic cultivation.

Sterol profiles were obtained from cyanobacteria Phormidium autumnale, cultivated in a heterotrophic system using three distinct sources of carbon: glucose, sucrose, and agroindustrial slaughterhouse wastewater. A simultaneous saponification-extraction ultrasound-assisted method was performed to determine sterol and other non-saponified compounds in the dry biomasses. A total of 24 compounds were observed in the biomasses, including hope-22,29-en-3-one, squalene, and 22 other sterols. Using wastewater as a carbon source, the microalgae biomass produced a diversity of sterols such as stigmasterol (455.3 µg g-1) and ß-sitosterol (279.0 µg g-1). However, with glucose it is possible to produce ergosterol (1033.3 µg g-1). Squalene was found in all the cultures, with 1440.4 µg g-1, 225.4 µg g-1, and 425.6 µg g-1 for glucose, sucrose, and slaughterhouse wastewater biomasses, respectively. Several intermediate compounds from those sterols were found. These data provide the construction of the sterol metabolism according to the literature for P. autumnale heterotrophically cultured.

Development and validation of RP-HPLC method for the determination of stigmasterol in the botanical extract of Ficus deltoidea.

A simple, rapid, accurate and precise RP-HPLC method was developed for the determination of stigmasterol in botanical extract of Ficus deltoidea. Separation was achieved with acetonitrile and acetic acid in water (75:25% v/v) in isocratic mode at 210 nm. Single sharp peak of standard stigmasterol was detected at retention time 3.17 min which overlay with the peak of plant extract at 3.14 min. The calibration curve was found to be linear in a concentration range of 2-10 µg/ml with correlation coefficient of 0.998. The LOD and LOQ were found to be 1.50 µg/ml and 4.55 µg/ml respectively. Accuracy and precision was determined with overall recovery of 99.6-100.1% for stigmasterol and RSD values in both intra-day and inter-day repeatability assay lesser than 0.340%, respectively. The robustness study also indicated that there is no influence of minor changes in detecting wavelength and flow rate of mobile phase on the response.

Phytosterols in supplements containing Serenoa repens: an example of variability of active principles in commercial plant based products.

Phytosterols are one of the bioactive components responsible for the beneficial effects of Serenoa repens in Benign Prostate Hyperplasia. The aim of this study was to verify the actual variability of the phytosterols content in supplements containing serenoa, in order to provide useful elements to check the effectiveness of these preparations. The amount of campesterol, stigmasterol and ß-sitosterol were determined by gas-chromatography in commercial raw materials and supplements containing serenoa in association or not with other botanicals. The experimental data were used to calculate amounts of phytosterols for recommended daily dose. The overall results of this study show an extreme variability in the content and also in the amounts per daily dose of phytosterols of the examined supplements (both mono/multi components). These data confirm that the characterization of serenoa based supplements is insufficient to ensure comparable effects between different products.

Analysis of functional components and radical scavenging activity of 21 algae species collected from the Japanese coast.

The functional chemical substances and the antioxidant activity of lipids in 21 marine algae along the Japanese coast were investigated. Principal component analysis was performed to detect any correlation between the chemical substances and algae phylum. Chlorophyta contained a high level of ß-carotene. Rhodophyta contained high amounts of cholesterol, ß-sitosterol, and saturated fatty acids. Phaeophyta were rich in fucosterol, α-tocopherol, fucoxanthin, and polyphenol. Phaeophyta algae also showed the highest antioxidant activity compared with other phylum. This suggests that Phaeophyta has the greatest potential to be used as a functional food. Consumption of the beneficial Phaeophyta species, such as Eisenia arborea Areschoug and Ecklonia cava Kjellman should be encouraged as not only as food products but also as nutraceuticals and dietary supplements. These beneficial ingredients should be encouraged to be studied in depth with the possibility to develop specific formulated products target to special consumer's population with added nutritional value.

Chemical profile and antihyperlipidemic effect of Portulaca oleracea L. seeds in streptozotocin-induced diabetic rats.

Hypolipidemic effect of Portulaca oleracea L. seed extract and its fractions have been studied on streptozotocin (STZ) at dose 75 mg/kg b.wt. After fractionation of the alcoholic extract; petroleum ether fraction was the most active fraction that decreased different hyperlipidemia biochemical parameters. After chromatographic analysis; oleamide, ethylpalmitate, ß-amyrin, stigmasterol and ß-sitosterol were identified. The GLC analysis of unsaponifiable matter revealed the presence of; lignoceric acid as a major constituent in the most bioactive fraction. In conclusion, petroleum ether fraction possessed a hypolipidemic effect in STZ-induced diabetic rats, which may be attributed to its phytosterols, fatty acid and amide compounds. The finding of the present investigation strongly demonstrates the potential of non-polar fraction of P. oleracea L. seed in combating hyperlipidemia in diabetic condition. So the petroleum ether fractions and its constituents can be used as hypolipdemic supplement in the developing countries towards the development of new therapeutic agents.

High-performance thin-layer chromatography HPTLC-direct bioautography as a method of choice for alpha-amylase and antioxidant activity evaluation in marine algae.

High-Performance Thin-layer chromatography (HPTLC) combined with DPPH free radical method and α-amylase bioassay was used to compare antioxidant and antidiabetic activities in ethanol and ethyl acetate extracts from 10 marine macroalgae species (3 Chlorophyta, 4 Phaeophyta and 3 Rhodophyta) from Blue Lagoon beach (Malaysia). Samples were also evaluated for their phenolic and stigmasterol content. On average, higher antioxidant activity was observed in the ethyl acetate extracts (55.1mg/100g gallic acid equivalents (GAE) compared to 35.0mg/100g GAE) while, as expected, phenolic content was higher in ethanol extracts (330.5mg/100g GAE compared to 289.5mg/100g GAE). Amounts of fucoxanthin, stigmasterol and α-amylase inhibitory activities were higher in ethyl acetate extracts. Higher enzyme inhibition is therefore related to higher concentrations of triterpenes and phytosterols (Note: these compounds are more soluble in ethyl acetate). Ethyl acetate extracts from Caulerpa racemosa and Padina minor, had the highest α-amylase inhibitory activity, and also showed moderately high antioxidant activities, stigmasterol content and polyphenolic content. Caulerpa racemose, being green algae, does not contain fucoxanthin, while Padina minor, being brown algae, contains high amounts of fucoxanthin. Therefore, it is very unlikely that fucoxanthin contributes to α-amylase inhibitory activity as previously reported.

Metal ions accelerated phytosterol thermal degradation on Ring A & Ring B of steroid nucleus in oils.

This study aimed to investigate the effect of metal ions on the degradation of phytosterols in oils. The oil was heated at 180°C for 1h with/without addition of Fe3+, Fe2+, Cu2+, Mn2+, Zn2+, Na+, Al3+ and Mg2+. Variations of ß-sitosterol, stigmasterol, campesterol, brassicasterol and their degradation products were confirmed by the GC-MS analysis. In general, the increase of the metal ion concentration resulted in more phytosterol degradation, and the ability of metal ions following decreasing order: Fe3+>Fe2+>Mn2+≥Cu2+≥Zn2+>Na+≥Mg2+>Al3+. Metal ions significantly induced phytosterol autoxidation on C5, C6 and C7 on Ring B of steroid nucleus at even a low concentration, and induced dehydration on the C3 hydroxyl to form dienes and trienes at high concentration. The metal ions in oils are accounted for increasing phytosterol degradation, which decreases food nutritional quality and gives rise to the formation of undesirable compounds.

Buddleja thyrsoides Lam. crude extract presents antinociceptive effect on an arthritic pain model in mice.

Arthritis is a chronic inflammatory disease which reduces the life quality of affected individuals. Therapeutic tools used for treating inflammatory pain are associated with several undesirable effects. Buddleja thyrsoides Lam., known as 'Barbasco' or 'Cambara', is mostly used in several disorders and possesses antirheumatic, anti-inflammatory, and analgesic properties. Here, we investigated the antinociceptive and anti-inflammatory effects of the B. thyrsoides crude extract applied orally and topically in acute pain models and an arthritic pain model induced by complete Freund's adjuvant (CFA) paw injection in male mice (25-30 g). The high-performance liquid chromatography (HPLC) of the B. thyrsoides extract crude revealed the presence of the lupeol, stigmasterol, and ß-sitosterol. The stability study of the B. thyrsoides gel did not show relevant changes at low temperatures. The oral treatment with the B. thrysoides extract prevented the capsaicin-induced spontaneous nociception and the acetic acid-induced abdominal writhing, but did not alter the thermal threshold in the tail immersion test. The B. thyrsoides antinociceptive effect was not reversed by naloxone in the capsaicin test. The B. thyrsoides oral or topical treatment reversed the CFA-induced mechanical allodynia and thermal hyperalgesia with maximum inhibition (Imax) of 69 ± 6 and 68 ± 5% as well as 78 ± 15 and 87 ± 12%, respectively. Moreover, the topical but not oral treatment inhibited the CFA-induced cell infiltration, but did not reduce the paw edema significantly. The oral treatment with B. thyrsoides did not cause adverse effects. These findings suggest that the oral or topical treatment with B. thyrsoides presents antinociceptive actions in an arthritic pain model without causing adverse effects.

Oil Characteristics of Four Palestinian Olive Varieties.

Olive oil represents an important source of income for Palestinian farmers in local, national and international markets. Sometimes, olive oil produced in local climatic conditions, does not achieve the International Olive Council (IOC) trade standards so that international markets are precluded. The oil chemical composition and sensory profile of four Palestinian olive varieties (Nabali Baladi, Nabali Mohassan, Souri and K18) were characterized in 2010 throughout an in situ evaluation. Most of the physicchemical characteristics and the fatty acid composition of the varieties met the International Olive Council trade standards (IOC-TS) for extra virgin olive oils. Values of K270 for Nabali Baladi and linolenic acid for Souri slightly exceeded the limit. Eicosanoic acid exceeded the IOC-TS limits in the oils of all considered varieties. Among the sterols, the Δ-7-stigmastenol resulted too high for Nabali Baladi and Souri. Sensory profile for the tested varieties showed a reminiscence of tomato or artichoke and light to medium bitter and pungent sensations. Results represent an important baseline reference for further studies about oil composition and quality of the main Palestinian olive germplasm and provide indication of potential critical points to be controlled in order to ensure the full achievement of IOC-TS and access international markets.