Plant Sterol-Poor Diet Is Associated with Pro-Inflammatory Lipid Mediators in the Murine Brain.

Plant sterols (PSs) cannot be synthesized in mammals and are exclusively diet-derived. PSs cross the blood-brain barrier and may have anti-neuroinflammatory effects. Obesity is linked to lower intestinal uptake and blood levels of PSs, but its effects in terms of neuroinflammation-if any-remain unknown. We investigated the effect of high-fat diet-induced obesity on PSs in the brain and the effects of the PSs campesterol and ß-sitosterol on in vitro microglia activation. Sterols (cholesterol, precursors, PSs) and polyunsaturated fatty acid-derived lipid mediators were measured in the food, blood, liver and brain of C57BL/6J mice. Under a PSs-poor high-fat diet, PSs levels decreased in the blood, liver and brain (>50%). This effect was reversible after 2 weeks upon changing back to a chow diet. Inflammatory thromboxane B2 and prostaglandin D2 were inversely correlated to campesterol and ß-sitosterol levels in all brain regions. PSs content was determined post mortem in human cortex samples as well. In vitro, PSs accumulate in lipid rafts isolated from SIM-A9 microglia cell membranes. In summary, PSs levels in the blood, liver and brain were associated directly with PSs food content and inversely with BMI. PSs dampen pro-inflammatory lipid mediators in the brain. The identification of PSs in the human cortex in comparable concentration ranges implies the relevance of our findings for humans.

Imidazole-modified C6 -chitosan derivatives used to extract ß-sitosterol from edible oil samples with a microwave-assisted solid phase extraction method.

ß-Sitosterol is a major bioactive constituent in plants with potent anticancer effects against many human cancer cells, but its bioavailability and therapeutic efficacy are limited by its poor solubility in water. In this study, C6 -imidazole chitosan, C6 -1-methylimidazole chitosan, C6 -1-ethylimidazole chitosan, C6 -1-vinylimidazole chitosan, C6 -1-allylimidazole chitosan, and C6 -1-butylimidazole chitosan were prepared to extract ß-sitosterol from edible oil samples via ultrasonic-assisted solid liquid extraction. The structures and properties of the newly synthesized products were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and elemental analysis. The extraction abilities of the derivatives were tested in the experiment with high-performance liquid chromatography (limit of detection 0.21 µg/g and limit of quantification 0.67 µg/g), and the % relative standard deviation (<3.25%) and recovery values of the prepared chitosan derivatives toward ß-sitosterol (average: 100.20%) were acceptable. The spiked interday and intraday recoveries of ß-sitosterol were 102.60 ± 2.78 and 103.90 ± 3.04%, respectively. The actual amounts of ß-sitosterol extracted from three real samples using C6 -imidazole chitosan according to the solid phase extraction method were 3302.40, 901.70, and 2045.60 mg/kg for corn oil, olive oil, and pea oil, respectively.

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.

Phenolic acids and phytosterols in rice grains and wheat flours consumed in five regions of China.

Phenolic acids and phytosterols, the main functional compounds in cereals, could promote wellbeing and reduce the risks of diet-related diseases. This study aimed to demonstrate phenolic acid and phytosterol profiles in rice grains and wheat flours, and estimate their intakes in five geographical regions and among different age groups. Phenolic acids and phytosterols mainly existed in bound form, and the whole rice grain had high amount of 161.39 to 368.74 µg/g and 37.50 to 93.31 mg/ 100 g, respectively. In total, nine phenolic acids and six phytosterols were detected with ferulic and p-coumaric acid, and ß-sitosterol the most abundant. The dietary intakes of phenolic acids and phytosterols were calculated combined with the dietary foods intake data of Chinese people. The intakes of total phenolic acids and phytosterols from rice grains and wheat flours varied across different regions with Beijing the highest among the five regions. At the age of 2 to 70 years, the average intakes of phenolic acids and phytosterols from rice and wheat flours were 7.74 to 17.52 and 58.02 to 135.61 mg/sp/day, respectively. If 3-ounce of polished rice was replaced by black rice grain, the predicted intakes of total phenolic acids and phytosterols from rice grains and wheat flours would increase by at least 196% and 68%, respectively, especially for free phenolic acids and phytosterols. PRACTICAL APPLICATION: This study would help the consumers know how much phenolic acids and phytosterols they would get from 3 ounces of black rice in a reasonable intake of staple food but shift away other kinds of foods. It could also provide inspirations for food industries to explore the functional cereal foods that are rich in phenolic acids and phytosterols for different regions and different age groups.

Supercritical Fluid Extract of Putranjiva roxburghii Wall. Seeds Mitigates Fertility Impairment in a Zebrafish Model.

Putrajeevak (Putranjiva roxburghii Wall.; synonym Drypetes roxburghii (Wall.) Hurus) seeds have been used since ancient times in the treatment of infertility in the Ayurvedic system of medicine in India. In this study, the oil component of Putrajeevak seeds (PJSO) was extracted using the supercritical fluid extraction (SCFE) method using liquid CO2 and the constituents were analyzed using gas chromatography-flame ionized detectorand high-performance thin-layer chromatography. PJSO contained trace amounts of ß-sitosterol with oleic and linoleic acids as the major fatty acid constituents. Male and female zebrafish were mutagenized with N-ethyl-N-nitrosourea (ENU) and fish that produced less than 20 viable embryos were selected for the study. SCFE oil extracts from the P. roxburghii seeds were used in this study to reverse fertility impairment. The mutant fish were fed with PJSO for a period of 14 days and the rates of fertility, conception, and fecundity were determined with wild-type healthy fish as a breeding partner. Treatment with PJSO increased the ovarian follicle count as well as the number of mature eggs, while reducing the number of ovarian cysts. Sperm count as well as sperm motility were greatly enhanced in the ENU-mutagenized male zebrafish when treated with PJSO. The results obtained in this study demonstrate the effectiveness of P. roxburghii seed oil in reversing impaired fertility in both male and female zebrafish models.

Improvement of physicochemical properties of reduced-cholesterol butter by the addition of ß-sitosteryl oleate.

Beta-cyclodextrin (ß-CD) has been shown to successfully lower the cholesterol content of dairy products, such as butter, but the process tends to negatively impact the overall quality and consistency. In this study, ß-sitosterol, which is similar in structure as cholesterol, was reacted with oleic acid to form ß-sitosteryl oleate (BSO), and this was used to improve the consistency of reduced-cholesterol butter. The reaction was catalyzed by sodium bisulfate (2%, w/w) at 140 °C, and the highest degree of esterification (94.3%) was obtained after 9 hr of reaction using a ß-sitosterol-oleic acid molar ratio of 1:5. Ultra-pasteurized cream was then treated with 15% (w/v) ß-CD at 40 °C with stirring (100 rpm), for 30 min. Results indicated a 95.4% reduction in cholesterol content. Finally, the reduced-cholesterol cream was constituted to contain 3, 5, and 10% (w/w) BSO, after which fat was extracted from the three formulations and their melting profiles compared to that of milk fat. The cream containing 3% BSO showed a profile similar to milk fat and was, therefore, used to formulate BSO-incorporated reduced-cholesterol butter (BSOB). Instrumental analyses showed that BSOB was comparable to the control butter with respect to physical properties, such as hardness/firmness and adhesiveness. PRACTICAL APPLICATION: A modified plant sterol, beta-sitosteryl oleate, was incorporated into a reduced-cholesterol butter to improve its physicochemical properties. The reduced-cholesterol butter was comparable to regular butter with respect to its consistency and melting properties and could be made into sticks. In addition to the reduced-cholesterol butter, this product could provide the foundation for new products blending butter and oils to create other low-cholesterol, reduced saturated-fat products, possibly in stick form.

Quality Properties, Fatty Acid and Sterol Compositions of East Mediterranean Region Olive Oils.

In this study, important physicochemical properties, fatty acid and sterol compositions of olive oils from the olives which harvested from Mersin (Buyuk Topak Ulak, Gemlik, Sari Ulak), Adana (Gemlik), Osmaniye (Gemlik) and Hatay (Gemlik, Kargaburun, Hasebi, Halhali) in the Eastern Mediterranean region of Turkey have been investigated. Ripening index and oil yield analysis of the olives and free fatty acids, peroxide value, UV absorbance (K232, K270), fatty acid composition, sterol composition, erythrodiol+uvaol content, and total sterol analysis of the olive oil samples were carried out. The levels of free acidity in the olive oil samples ranged from 0.39% (Hatay Gemlik: HG3) to 2.23% (Mersin Gemlik: MG). Peroxide value ranged from 8.87 to 18.87 meq O2/kg. As K232 values in the oils fluctuated between 1.4370 and 2.3970, K270 values varied between 0.1270 and 0.1990. The results showed that all ΔK values were lower than the maximum legal limit of 0.01. The main fatty acid in all oil samples was oleic acid, ranging from 58.72% (Hatay Hasebi: HHs) to 74.54% (Hatay Gemlik: HG2). Palmitic acid values were within the percentage of 12.83% (Hatay Kargaburun: HK) to 18.50% (HHs). Total sterol content varied from 720.41 mg/kg (Hatay Kargaburun: HK) to 4519.17 mg/kg (Buyuk Topak Ulak: BTU). The ß-sitosterol percentage of olive oils ranged from 76.12% (Adana Gemlik: AG) to 94.23% (Buyuk Topak Ulak: BTU). The results of this study indicated that variety significantly affect the quality indices, fatty acid and sterol compositions of olive oils significantly varied among varieties.

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.

Network Pharmacology Approach to Uncover the Mechanism Governing the Effect of Simiao Powder on Knee Osteoarthritis.

OBJECTIVE: To explore the molecular mechanism of Simiao powder in the treatment of knee osteoarthritis. METHODS: Based on oral bioavailability and drug-likeness, the main active components of Simiao powder were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). GeneCard, OMIM, DisGeNET, DrugBank, PharmGkb, and the Therapeutic Target Database were used to establish target databases for knee osteoarthritis. Cytoscape software was used to construct a visual interactive network diagram of "active ingredient - action target - disease." The STRING database was used to construct a protein interaction network and analyze related protein interaction relationships. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) biological process enrichment analysis were performed on the core targets. Additionally, Discovery Studio software was used for molecular docking verification of active pharmaceutical ingredients and disease targets. RESULTS: Thirty-seven active components of Simiao powder were screened, including 106 common targets. The results of network analysis showed that the targets were mainly involved in regulating biological processes such as cell metabolism and apoptosis. Simiao powder components were predicted to exert their therapeutic effect on the AGE-RAGE signaling pathway in diabetic complications, IL-17 signaling pathway, TNF signaling pathway, Toll-like receptor signaling pathway, and HIF-1 signaling pathway. The molecular docking results showed that the active components of Simiao powder had a good match with the targets of IL1B, MMP9, CXCL8, MAPK8, JUN, IL6, MAPK1, EGF, VEGFA, AKT1, and PTGS2. CONCLUSION: Simiao powder has multisystem, multicomponent, and multitarget characteristics in treating knee osteoarthritis. Its possible mechanism of action includes inhibiting the inflammatory response, regulating immune function, and resisting oxidative stress to control the occurrence and development of the disease. Quercetin, wogonin, kaempferol, beta-sitosterol, and other active ingredients may be the material basis for the treatment of knee osteoarthritis.

LC-MS/MS and GC-MS profiling as well as the antimicrobial effect of leaves of selected Yucca species introduced to Egypt.

Few studies thoroughly investigated different Yucca species introduced to Egypt. As a part of our ongoing investigation of the Yucca species; Yucca aloifolia and its variety Yucca aloifolia variegata, Yucca filamentosa, and Yucca elephantipes (Asparagaceae) were extensively subjected to phytochemical and antimicrobial investigation. Yucca species cultivated in Egypt showed no antimicrobial effect. GC/MS of the lipoid contents of Y. aloifolia variegata was carried out. Twenty-six fatty acids were identified. Saturated fatty acids established almost twice the unsaturated ones and constituted 64.64% of which palmitic acid and palmitoleic acid signifying 58.28% and 30.98%, respectively. Hydrocarbons were 21 constituting 39.64% of the unsaponifiable fraction. Only three sterols 42.36% were detected, major was γ-sitosterol. LC-MS/MS comparison of the 4 plant extracts imply that Y.aloifolia variegata L extract was the richest, which was apparent through its superior biological activity. LC-MS/MS analysis of the total alcoholic extract (Alc) of the leaves of Y.aloifolia variegata L. was performed using MS-techniques at different voltages; equal to 35 and 135 eV. Negative and positive-ion modes analyses at low fragmentation energy allowed the tentative identification of 41 and 34 compounds, respectively. The LC-ESI-MS/MS analysis in the positive mode proved to be better in the identification of saponins.

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).

Chemical and Physical Characterization of the Hackberry (Celtis australis) Seed Oil: Analysis of Tocopherols, Sterols, ECN and Fatty Acid Methyl Esters.

For the very first time, the nutritional and physicochemical properties of the oil extracted from hackberry Celtis australis fruit were investigated with the aim of possible applications of such wild fruit oil. The physicochemical properties such as peroxide value, acidity, saponification, iodine value and total fat content of the extracted oil were examined extensively. The obtained results showed that peroxide value, acidity, saponification, iodine value and total fat content of the extracted oil were found to be 4.9 meq O2/kg fat, 0.9 mg KOH/g fat, 193.6 mg KOH/g fat, 141.52 mg I2/g fat and ~5%, respectively. The predominant fatty acid found in this wild fruit is linoleic acid which was calculated to be 73.38%±1.24. In addition, gamma-tocopherol (87%) and ß-sitosterol (81.2%±1.08) were the major tocopherol and sterol compositions found in Celtis australis seed oil. Moreover, equivalent carbon number (ECN) analysis has indicated that the three linoleic acids are the main composition of the triacylglycerols extracted from Celtis australis. Also, the high value of omega 6 and ß-sitosterol make this oil applicable in cosmetics and pharmaceutical applications.

Application of a 2D Molybdenum Telluride in SERS Detection of Biorelevant Molecules.

Two-dimensional (2D) transition-metal dichalcogenides have become promising candidates for surface-enhanced Raman spectroscopy (SERS), but currently very few examples of detection of relevant molecules are available. Herein, we show the detection of the lipophilic disease marker ß-sitosterol on few-layered MoTe2 films. The chemical vapor deposition (CVD)-grown films are capable of nanomolar detection, exceeding the performance of alternative noble-metal surfaces. We confirm that the enhancement occurs through the chemical enhancement (CE) mechanism via formation of a surface-analyte complex, which leads to an enhancement factor of ≈104, as confirmed by Fourier transform infrared (FTIR), UV-vis, and cyclic voltammetry (CV) analyses and density functional theory (DFT) calculations. Low values of signal deviation over a seven-layered MoTe2 film confirms the homogeneity and reproducibility of the results in comparison to noble-metal substrate analogues. Furthermore, ß-sitosterol detection within cell culture media, a minimal loss of signal over 50 days, and the opportunity for sensor regeneration suggest that MoTe2 can become a promising new SERS platform for biosensing.

High prevalence of increased sitosterol levels in hypercholesterolemic children suggest underestimation of sitosterolemia incidence.

BACKGROUND: Sitosterolemia is an inherited lipid disorder which presents with elevated serum sitosterol and can result in an increased risk of premature cardiovascular disease. However, sitosterol cannot be accurately measured by routine diagnostic assays, meaning that sitosterolemia diagnosis can often be difficult, especially with many clinical features overlapping with familial hypercholesterolemia. With such complications resulting in increasing reports of misdiagnosis, the prevalence of sitosterolemia is predicted to be much higher than previously reported. METHODS: Gas chromatography-mass spectrometry was utilized to measure sitosterol levels of normocholesterolemic and hypercholesterolemic children. Subsequently, an epidemiologically determined cutoff level of sitosterol was calculated and applied to estimate the prevalence of children with increased sitosterol and identify potential sitosterolemia patients. Massively parallel sequencing was used to confirm the diagnosis in suspected patients. RESULTS: Samples from 109 normocholesterolemic and 220 hypercholesterolemic were tested for phytosterols. Sitosterol and campesterol levels were significantly increased in hypercholesterolemic children (mean 22.0±45.9 µmol/L for sitosterol and 26.0±32.8 µmol/L for campesterol) compared to normocholesterolemic children (mean 12.1±4.9 µmol/L for sistosterol and 14.8±6.7 µmol/L for campesterol). Via application of a cutoff of 35.9 µmol/L, the prevalence rates for increased and overtly increased sitosterol in hypercholesterolemic children were 6.4% and 1.4% respectively. Furthermore, 3 suspected sitosterolemia patients were identified, with 2 patients receiving molecular confirmation for sitosterolemia diagnosis. CONCLUSIONS: Our findings reaffirm that the prevalence of sitosterolemia is probably much higher than previously reported, which also indicates the significant risk of misdiagnosis of sitosterolemia with familial hypercholesterolemia. Special lipid testing including sitosterol, especially in children with uncontrolled hypercholesterolemia, is recommended in children in order to identify potential sitosterolemia patients that would otherwise be neglected.

ß-sitosterol ameliorates influenza A virus-induced proinflammatory response and acute lung injury in mice by disrupting the cross-talk between RIG-I and IFN/STAT signaling.

ß-Sitosterol (24-ethyl-5-cholestene-3-ol) is a common phytosterol Chinese medical plants that has been shown to possess antioxidant and anti-inflammatory activity. In this study we investigated the effects of ß-sitosterol on influenza virus-induced inflammation and acute lung injury and the molecular mechanisms. We demonstrate that ß-sitosterol (150-450 µg/mL) dose-dependently suppresses inflammatory response through NF-κB and p38 mitogen-activated protein kinase (MAPK) signaling in influenza A virus (IAV)-infected cells, which was accompanied by decreased induction of interferons (IFNs) (including Type I and III IFN). Furthermore, we revealed that the anti-inflammatory effect of ß-sitosterol resulted from its inhibitory effect on retinoic acid-inducible gene I (RIG-I) signaling, led to decreased STAT1 signaling, thus affecting the transcriptional activity of ISGF3 (interferon-stimulated gene factor 3) complexes and resulting in abrogation of the IAV-induced proinflammatory amplification effect in IFN-sensitized cells. Moreover, ß-sitosterol treatment attenuated RIG-I-mediated apoptotic injury of alveolar epithelial cells (AEC) via downregulation of pro-apoptotic factors. In a mouse model of influenza, pre-administration of ß-sitosterol (50, 200 mg·kg-1·d-1, i.g., for 2 days) dose-dependently ameliorated IAV-mediated recruitment of pathogenic cytotoxic T cells and immune dysregulation. In addition, pre-administration of ß-sitosterol protected mice from lethal IAV infection. Our data suggest that ß-sitosterol blocks the immune response mediated by RIG-I signaling and deleterious IFN production, providing a potential benefit for the treatment of influenza.

Fast Quantification Without Conventional Chromatography, The Growing Power of Mass Spectrometry.

Mass spectrometry (MS) in hyphenated techniques is widely accepted as the gold standard quantitative tool in life sciences. However, MS possesses intrinsic analytical capabilities that allow it to be a stand-alone quantitative technique, particularly with current technological advancements. MS has a great potential for simplifying quantitative analysis without the need for tedious chromatographic separation. Its selectivity relies on multistage MS analysis (MSn), including tandem mass spectrometry (MS/MS), as well as the ever-growing advancements of high-resolution MS instruments. This perspective describes various analytical platforms that utilize MS as a stand-alone quantitative technique, namely, flow injection analysis (FIA), matrix assisted laser desorption ionization (MALDI), including MALDI-MS imaging and ion mobility, particularly high-field asymmetric waveform ion mobility spectrometry (FAIMS). When MS alone is not capable of providing reliable quantitative data, instead of conventional liquid chromatography (LC)-MS, the use of a guard column (i.e., fast chromatography) may be sufficient for quantification. Although the omission of chromatographic separation simplifies the analytical process, extra procedures may be needed during sample preparation and clean-up to address the issue of matrix effects. The discussion of this manuscript focuses on key parameters underlying the uniqueness of each technique for its application in quantitative analysis without the need for a chromatographic separation. In addition, the potential for each analytical strategy and its challenges are discussed as well as improvements needed to render them as mainstream quantitative analytical tools. Overcoming the hurdles for fully validating a quantitative method will allow MS alone to eventually become an indispensable quantitative tool for clinical and toxicological studies.

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.

Phytochemical studies of Jurinea macrocephala roots from Western Himalaya.

Phytochemical investigation of the Jurinea macrocephala roots afforded six compounds namely ß-sitosterol (1), lupenone (2), physcion (3), ptiloepoxide (4), 20, 21α-epoxytaraxastan-3ß-ol (5) and chlorogenic acid (6). All the compounds were isolated for the first time in roots. The structures of the compounds were established by analysis of their spectroscopic (1H and 13C NMR) and spectrometric (MS) data, as well as by comparison of these with those reported in the literature. Metabolic profiling of chloroform and ethyl acetate fractions were also accomplished using NMR. In NMR analysis, ERETIC (electronic reference to access in-vivo concentration) 2 method was used for the quantification of identified metabolites. High quantity of chlorogenic acid (6, 130 mg/g) lupenone (2, 33.4 mg/g) and amyrins (α, ß) (170.6 mg/g) were detected in ethyl acetate and chloroform fractions. Further studies on the biological evaluation of phenolic-rich and chloroform fractions could be beneficial to explore its pharmaceutical potential.

Chemoprotective and antiobesity effects of tocols from seed oil of Maqui-berry: Their antioxidative and digestive enzyme inhibition potential.

Maqui-berry (Aristotelia chilensis) is the emerging Chilean superfruit with high nutraceutical value. Until now, the research on this commodity was focused on the formulations enriched with polyphenols from the pulp. Herein, contents of tocols were compared in the seed oil of Maqui-berry obtained through three different extraction methods followed by determining their antioxidative and enzyme inhibitions in-vitro. Firstly, oilseed was extracted with n-hexane (Soxhlet method), chloroform/methanol/water (Bligh and Dyer method) and pressing (industrial). These samples were used to access their effects against DPPH, HORAC, ORAC, FRAP, Lipid-peroxidation (TBARS), α-amylase, α-glucosidase, and pancreatic lipase. All the isomers of tocopherol and tocotrienol were identified, and ß-sitosterol was the only sterol found in higher amounts than other vegetable oils. The Bligh and Dyer method could lead to the highest antioxidative capacity compared to Soxhlet and press methods likely because the latter have a higher amount of tocopherols. Further, seed oil from Maqui berry and their tocols (α, ß, γ, δ-tocopherols, tocotrienols, and ß-sitosterol) warrant clinical investigation for their antioxidative and antiobesity potential. Taken together, these findings provide relevant and suitable conditions for the industrial processing of Maqui-berry.