Antiproliferative Acylated Glycerols from New Zealand Propolis.

Previous work has shown that a number of phenolic components of NZ propolis possess antiproliferative activity against certain human gastrointestinal cancer cell lines. Here we report on a series of acylglycerols isolated from the nonpolar fraction of propolis resin, which represent further bioactive constituents unrelated to the more usual phenolic compounds generally found in propolis. NZ propolis is sourced from poplar trees, and the acylglycerols have been shown to be present in the leaves and buds of some common poplars. The compounds are a series of monoglycerides containing 3,8-dihydroxy fatty acids, many of which are further acylated with acetic acid residues. The dihydroxy fatty acids are C18 to C24, with the most abundant being C20 and C22. These acylglycerols were found to have strong antiproliferative activity against three human gastrointestinal cell lines, particularly gastric cancer cell line NCI-N87, where one example shows an IC50 of less than 50 µM.

Potential of Fatty Oils from Traditional Chinese Medicine in Cancer Therapy: A Review for Phytochemical, Pharmacological and Clinical Studies.

Cancer management is a worldwide challenge. In addition to effective cancer therapies like chemotherapy, radiotherapy and surgery, treatment based on traditional Chinese medicine (TCM) and combined TCM with western medicine has gradually gained attention in Oriental countries. One potential TCM approach using extracted fatty oils, containing fatty acids which are important active ingredients with a variety of pharmacological activities, makes significant contributions to cancer treatment. The strategies of treating cancer with the fatty oils of TCM were classified into "Fuzheng", which usually associates with improving immunity, represented by coix seed oil. The other classification is "Quxie", which relates to inducing apoptosis of cancer cells, and is represented by Brucea javanica oil. Compared with other active substances, the literature about anticancer fatty oils is relatively limited, and most of them focus on the composition and other biological activities without a systematic review. Therefore, based on the theories of "Fuzheng" and "Quxie" in TCM, in this paper, the anticancer effects of fatty oils have been reviewed. The chemical composition, anticancer mechanism, listed drugs, studying dosage form and clinical application of fatty oils have also been discussed. In summary, since there are different types and abundance of fatty oils among botanicals, anticancer effects of fatty oils can be achieved through two TCM theory-based strategies. We hoped that this review paper can reveal the anticancer potential of fatty oils and provide a reference for future related studies.

Cycloartane-type triterpene glycosides anopanins A-C with monoacyldigalactosylglycerols from Anodendron paniculatum.

Three cycloartane-type triterpene glycosides (anopanins A-C), together with three known monoacyldigalactosylglycerols gingerglycolipids A-C and (2S)-1-O-palmitoyl-3-O-[α-d-galactopyranosyl-(1→6)-ß-d-galactopyranosyl]-sn-glycerol, were isolated from the aerial parts of Anodendron paniculatum collected in Viet Nam. The chemical structures of the present compounds were elucidated by means of 1D and 2D NMR and HRESIMS spectroscopy, and by comparing to the reported data in the literature. These compounds did not show significant growth inhibitory activities (IC50 > 100 µg/mL) against the tested cancer cell lines LU-1 (lung adenocarcinoma), KB (epidermoid carcinoma), Hep-G2 (hepatoma cancer), MKN-7 (stomach cancer), and SW-480 (colon adenocarcinoma).

E-cigarette liquids: Constancy of content across batches and accuracy of labeling.

AIMS: To assess whether bottles of refill liquids for e-cigarettes were filled true to label, whether their content was constant across two production batches, and whether they contained impurities. METHODS: In 2013, we purchased on the Internet 18 models from 11 brands of e-liquids. We purchased a second sample of the same models 4months later. We analyzed their content in nicotine, anabasine, propylene glycol, glycerol, ethylene glycol and diethylene glycol, and tested their pH. RESULTS: The median difference between the nicotine value on the labels and the nicotine content in the bottles was 0.3mg/mL (range -5.4 to +3.5mg/mL, i.e. -8% to +30%). For 82% of the samples, the actual nicotine content was within 10% of the value on the labels. All models contained glycerol (median 407mg/mL), and all but three models contained propylene glycol (median 650mg/mL). For all samples, levels of anabasine, ethylene glycol and diethylene glycol were below our limits of detection. The pH of all the e-liquids was alkaline (median pH=9.1; range 8.1 to 9.9). The measured content of two batches of the same model varied by a median of 0% across batches for propylene glycol, 1% for glycerol, 0% for pH, and 0.5% for nicotine (range -15% to +21%; 5th and 95th percentiles: -15% and +10%). CONCLUSIONS: The nicotine content of these e-liquids matched the labels on the bottles, and was relatively constant across production batches. The content of propylene glycol and glycerol was also stable across batches, as was the pH.

New mono-ether of glycerol and triterpenes with DPPH radical scavenging activity from Cameroonian propolis.

The extracts of some propolis samples were analysed by GC-MS and then purified by column chromatography. The latter led to the isolation of a new mono-ether of glycerol, 1'-O-eicosanyl glycerol and a new triterpene, methyl-3ß,27-dihydroxycycloart-24-en-26-oate together with known triterpenoids namely betulin, 3ß-hydroxylanostan-9,24-dien-21-oic acid, mangiferonic acid, a mixture of ambolic acid and ß-sitosterol, 3ß-hydroxycycloartan-12,24(25)-diene and 27-hydroxymangiferonic acid. The DPPH radical scavenging potential of some extracts and compounds were measured. The radical scavenging activity varied from Hexane extract of Foumban propolis (IC50 = 5.6 mg/mL) to Methanol extract of Foumban propolis (IC50 = 1.07 mg/mL) for the extracts and from 3ß-hydroxylanostan-9,24-dien-21-oic acid (IC50 = 1.22 mg/mL) to 1'-O-eicosanyl glycerol (IC50 = 0.93 mg/mL) for the compounds. Activities of samples were moderate as they remained closer to those of the standard antioxidants Gallic acid (IC50 = 0.30 mg/mL) and vitamin C (IC50 = 0.80 mg/mL), especially 1'-O-eicosanyl glycerol, the most active compound.

Alkylglycerols reduce serum complement and plasma vascular endothelial growth factor in obese individuals.

Alkylglycerols (AKGs), isolated or present in shark liver oil have anti-inflammatory properties. Complement 3 (C3) and 4 (C4) participate in lipid metabolism and in obesity, contributing to the metabolic syndrome and to the low-grade inflammation associated with obesity. In a randomized, controlled, crossover study, 26 non-diabetes obese individuals were assigned two preparations with low (LAC, 10 mg AKGs) and high (HAC, 20 mg AKGs) AKG content. Intervention periods were of 3 weeks preceded by 2-week washout periods in which shark liver oil was avoided. Cholesterol, C3, C4, and vascular endothelial growth factor (VEGF) decreased in a linear trend (P < 0.01) from baseline (control) to LAC and HAC. Values after HAC were significantly lower (P < 0.05) versus both baseline and after LAC. No adverse effects were observed or reported. Data from this pilot study open a promising field for the study of the beneficial effects of AKGs on cardiovascular risk factors in obese individuals.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the direct detection of 2-monochloropropanediol (2-MCPD) esters in edible oils.

A new analytical method has been developed and validated for the detection and quantification of 2-monochloropropanediol (2-MCPD) esters in edible oils. The target compounds are potentially carcinogenic contaminants formed during the processing of edible oils. As the 2-MCPD esters that occur most frequently in refined edible oils were not commercially available, standards were synthesized with identity and purity (95+%) confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and (1)H NMR. Target analytes are separated from edible oil matrices using a two-step solid-phase extraction (SPE) procedure. The extracts are then analyzed using LC-MS/MS with electrospray ionization (ESI). The method has been validated for 11 2-MCPD diesters and 3 2-MCPD monoesters in soybean oil, olive oil, and palm oil using an external calibration curve. The ranges of average recoveries and relative standard deviations (RSD) across the three oil matrices at three spiking concentrations are 79-106% (3-13% RSD) for the 2-MCPD diesters and 72-108% (4-17% RSD) for the 2-MCPD monoesters, with limits of quantitation at or below 30 ng/g for the diesters and 90 ng/g for the monoesters.

Adsorption of glycerol, monoglycerides and diglycerides present in biodiesel produced from soybean oil.

The most common methods currently used for the removal of waste glycerol, monoglycerides and diglycerides remaining after phase separation during biodiesel production involve wet processes. These procedures are not environmentally viable because they require large volumes of water and thus generate significant quantities of effluent. In this study, adsorption was employed to replace this purification step. Some commercial activated carbons were tested along with adsorbents chemically modified with HNO3. A kinetics study was conducted at 30 degrees C and adsorption isotherms were obtained at 20 degrees C, 30 degrees C and 40 degrees C. The results indicated that the adsorption of glycerol increased with the use of chemically-modified activated carbon, showing that pH has a strong influence on glycerol adsorption. The pseudo-first-order kinetic model provided the best fit with the experimental data for the monoglycerides while the pseudo-second-order model showed a better fit for the glycerol and diglycerides. The Freundlich model had the best fit with experimental data on the adsorption equilibrium for all temperatures. The thermodynamic study indicated that the adsorption process is endothermic and thus adsorption is favoured by increasing the temperature. The adsorption process using chemically-modified activated carbon was therefore very effective for the removal of waste glycerol resulting from biodiesel production, which is of considerable significance given the legal limits imposed.

Simultaneous determination of 3-monochloropropane-1,2-diol and acrylamide in food by gas chromatography-triple quadrupole mass spectrometry with coupled column separation.

Both 3-monochloropropane-1,2-diol (3-MCPD) and acrylamide are contaminants found in heat-processed foods and their related products. A quantitative method was developed for the simultaneous determination of both contaminants in food by gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS). The analytes were purified and extracted by the matrix solid-phase dispersion extraction (MSPDE) technique with Extrelut NT. A coupled column (a 3 m Innowax combined with a 30 m DB-5 ms) was developed to separate both compounds efficiently without derivatization. Triple quadrupole mass spectrometry in multiple reaction monitoring mode (MRM) was applied to suppress matrix interference and obtain good sensitivity in the determination of both analytes. The limit of detection (LOD) in the sample matrix was 5 µg kg(-1) for 3-MCPD or acrylamide. The average recoveries for 3-MCPD and acrylamide in different food matrices were 90.5-107% and 81.9-95.7%, respectively, with the intraday relative standard deviations (RSDs) of 5.6-13.5% and 5.3-13.4%, respectively. The interday RSDs were 6.1-12.6% for 3-MCPD and were 5.0-12.8% for acrylamide. Both contaminants were found in samples of bread, fried chips, fried instant noodles, soy sauce, and instant noodle flavoring. Neither 3-MCPD nor acrylamide was detected in the samples of dairy products (solid or liquid samples) and non-fried instant noodles.

Effects of purified glycerol from biodiesel on miRNAs compared to the expression profile of selected mRNAs in Balb/c mice.

BACKGROUND: We investigated the effect of corn-derived biodiesel glycerol on microRNAs (miRNAs) and mRNAs, which play a central role in regulating cell survival, apoptosis and carcinogenesis. MATERIALS AND METHODS: Inbred Balb/c mice were treated with purified glycerol from biodiesel for 24 hours. After administration, we determined the expressions of miR-21, miR-27a, miR-34a, miR-93, miR-143, miR-146a, miR-148a, miR-155, miR-196a, miR-203, miR-205, miR-221 and nuclear factor kappa-light-chain enhancer of activated B-cells-1 (Nfκb1), mitogen-activated protein kinase-8 (Mapk8) and v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (K-ras) genes in the liver of mice. RESULTS: We found a parallel altered expression of miRNAs and mRNAs in animals consuming biodiesel glycerol that compared to control mice; these alterations reached significant levels only in few cases. CONCLUSION: Biodiesel glycerol presents no higher risk for carcinogenicity or toxicity.

Isolation and characterization of five glycerol esters from Wuhan propolis and their potential anti-inflammatory properties.

Five new glycerol esters including 2-acetyl-1-coumaroyl-3-cinnamoylglycerol (13), (+)-2-acetyl-1-feruloyl-3-cinnamoylglycerol (14), (-)-2-acetyl-1-feruloyl-3-cinnamoylglycerol (15), 2-acetyl-1,3-dicinnamoylglycerol (16), and (-)-2-acetyl-1-(E)-feruloyl-3-(3″(ζ),16″)-dihydroxy-palmitoylglycerol (17) were isolated from methanolic extract of Wuhan propolis. The chemical structures of the five new compounds were confirmed by (1)H and (13)C NMR and HR-MS spectra. Twelve minor constituents in the Wuhan propolis extract were tentatively identified by UPLC-Q-TOF-MS, according to their characteristic UV spectrum, retention times, and accurate MS data. The anti-inflammatory activities of the five new compounds were studied in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophage cells. All five compounds exhibited remarkable inhibitory effects on interleukin (IL)-1ß, IL-6, and cyclooxygenase (COX)-2 mRNA expressions at 10 and 100 µM, suggesting that Wuhan propolis may serve as a potential anti-inflammatory functional food ingredient or nutraceutical.

Optimization of ultrasonic-assisted extraction of 3-monochloropropane-1,2-diol (MCPD) and analysis of its esters from edible oils by gas chromatography-mass spectrometry.

In this paper, ultrasonic-assisted extraction of 3-chloropropane-1,2-diol and its esters from edible oils was studied with isotope dilution GC-MS. Effects of several experimental parameters, such as types and concentrations of extracting solvent, ratios of liquid to material, extraction temperature, time of ultrasonic treatment on the extraction efficiency of 3-chloropropane-1,2-diol and its esters from edible oils and sample preparation for calibration were compared and optimized. The optimal extraction conditions were suggested as 66 mg oil sample in mixture of 0.5 mL MTBE/ethyl acetate (20% v/v) and 0.5 mL of sulfuric acid/n-propanol (0.3% v/v), being extracted for 30 min at 45°C under ultrasonic irradiation. Good linearity was gained in the range of 0.020-5.000 µg/g with the limit of detection (LOD) of 0.006 µg/g (S/N = 3) and the limit of quantification (LOQ) of 0.020 µg/g (S/N = 10). The recoveries at five spiked concentrations were ranged from 91.9 to 109.3% with RSD less than 9.4%. The method was successfully applied to the determination of 3-chloropropane-1,2-diol and its esters amounts in rapeseed, sesame, peanut, camellia, and soybean oils.

Evaluation of the composition of culture medium for yeast biomass production using raw glycerol from biodiesel synthesis

The work herewith investigated the production of yeast biomass as a source of protein, using Yarrowia lipolytica NRRL YB-423 and raw glycerol from biodiesel synthesis as the main carbon source. A significant influence of glycerol concentration, initial pH and yeast extract concentration on biomass and protein content was observed according to the 2v5-1 fractional design. These factors were further evaluated using a central composite design and response surface methodology, and an empirical model for protein content was established and validated. The biomass of Yarrowia lipolytica NRRL YB-423 reached 19.5 ± 1.0 g/L in shaken flasks cultivation, with a protein content of 20.1 ± 0.6% (w/w).

Experimental assessment of toxic phytochemicals in Jatropha curcas: oil, cake, bio-diesel and glycerol.

BACKGROUND: Jatropha curcas seed is a rich source of oil; however, it can not be utilised for nutritional purposes due to presence of toxic and anti-nutritive compounds. The main objective of the present study was to quantify the toxic phytochemicals present in Indian J. curcas (oil, cake, bio-diesel and glycerol). RESULTS: The amount of phorbol esters is greater in solvent extracted oil (2.8 g kg⁻¹) than in expeller oil (2.1 g kg⁻¹). Liquid chromatography-mass spectroscopy analysis of the purified compound from an active extract of oil confirmed the presence of phorbol esters. Similarly, the phorbol esters content is greater in solvent extracted cake (1.1 g kg⁻¹) than in cake after being expelled (0.8 g kg⁻¹). The phytate and trypsin inhibitory activity of the cake was found to be 98 g kg⁻¹ and 8347 TIU g⁻¹ of cake, respectively. Identification of curcin was achieved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the concentration of curcin was 0.95 g L⁻¹ of crude concentrate obtained from cake. CONCLUSION: Higher amounts of phorbol esters are present in oil than cake but bio-diesel and glycerol are free of phorbol esters. The other anti-nutritional components such as trypsin inhibitors, phytates and curcin are present in cake, so the cake should be detoxified before being used for animal feed.

Optimization of biomass production of a mutant of Yarrowia lipolytica with an increased lipase activity using raw glycerol

The yeast Yarrowia lipolytica accumulates oils and is able to produce extracellular lipases when growing in different carbon sources including glycerol, the principal by-product of the biodiesel industry. In this study, biomass production of a novel mutant strain of Y. lipolytica was statistically optimized by Response Surface Methodology in media containing biodiesel-derived glycerol as main carbon source. This strain exhibited distinctive morphological and fatty acid profile characteristics, and showed an increased extracellular lipase activity. An organic source of nitrogen and the addition of 1.0 g/l olive oil were necessary for significant lipase production. Plackett-Burman and Central Composite Statistical Designs were employed for screening and optimization of fermentation in shaken flasks cultures, and the maximum values obtained were 16.1 g/l for biomass and 12.2 Units/ml for lipase, respectively. Optimized batch bioprocess was thereafter scaled in aerated bioreactors and the values reached for lipase specific activity after 95 % of the glycerol had been consumed, were three-fold higher than those obtained in shaken flasks cultures. A sustainable bioprocess to obtain biomass and extracellular lipase activity was attained by maximizing the use of the by-products of biodiesel industry.

Optimización de la producción de biomasa usando glicerol crudo, de una cepa mutante de Yarrowia lipolytica con actividad incrementada de lipasa. La levadura Yarrowia lipolytica acumula aceites y produce una lipasa extracelular al crecer en diferentes fuentes de carbono, entre ellas el glicerol, principal subproducto de la creciente industria del biodiésel. En el presente trabajo, se optimizó mediante la metodología de superficies de respuesta la producción de biomasa de una nueva cepa mutante de Y. lipolytica, empleando medios con glicerol derivado de la industria del biodiésel como principal fuente de carbono. Esta cepa presentó características morfológicas y perfil de ácidos grasos distintivos, y una mayor actividad de lipasa extracelular. Para obtener una producción significativa de lipasa extracelular, fue necesario el agregado de una fuente orgánica de nitrógeno y de 1 g/l de aceite de oliva. Se utilizaron los diseños estadísticos de Plackett-Burman y central compuesto para la selección y la optimización de las fermentaciones en frascos agitados; los máximos valores de biomasa y de lipasa obtenidos fueron de 16,1 g/l y 12,2 unidades/ml, respectivamente. Luego, el bioproceso en lote optimizado se escaló a biorreactores aireados, y los valores de actividad específica de lipasa alcanzados después de haberse consumido el 95 % del glicerol fueron tres veces más altos que los obtenidos en los cultivos en frascos agitados. En suma, se desarrolló un bioproceso sostenible para la obtención de biomasa y de una actividad de lipasa extracelular, que a la vez maximiza el uso de subproductos de la industria del biodiésel.

Combined solid-phase extraction and gas chromatography-mass spectrometry used for determination of chloropropanols in water.

A sensitive and rapid derivatization method for the simultaneous determination of 1,3-dichloro-2-propanol (1,3-DCP) and 3-chloropropane-1,2-diol (3-MCPD) in water samples has been developed. The aim was to research the optimal conditions of the derivatization process for two selected reagents. A central composite design was used to determine the influence of derivatization time, derivatization temperature and reagent volume. A global desirability function was applied for multi-response optimization. The analysis was performed by gas chromatography-mass spectrometry. During the optimization of the extraction procedure, four different types of solid-phase extraction (SPE) columns were tested. It was demonstrated that the Oasis HLB cartridge produced the best recoveries of the target analytes. The pH value and the salinity were investigated using a Doehlert design. The best results for the SPE of both analytes were obtained with 1.5 g of NaCl and pH 6. The proposed method provides high sensitivity, good linearity (R(2)≥0.999) and repeatability (relative standard deviations % between 2.9 and 3.4%). Limits of detection and quantification were in the range of 1.4-11.2 ng/mL and 4.8-34.5 ng/mL, respectively. Recoveries obtained for water samples were ca. 100% for 1,3-DCP and 3-MCPD. The method has been successfully applied to the analysis of different samples including commercially bottled water, an influent and effluent sewage.

[Two new phenolic acids from Drynariae rhizoma].

To study the chemical constituents of Drynariae Rhizoma, nine phenolic acids were isolated from a 70% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, 2D NMR, and HR-ESI-MS, these compounds were identified as 4, 4'-dihydroxy-3, 3'-imino-di-benzoic acid (1), protocatechuic acid (2), gallic acid (3), p-hydroxybenzoic acid (4), (E)-caffeic acid (5), ethyl trans-3, 4-dihydroxycinnamate (6), caffeic acid 4-O-beta-D-glucopyranoside (7), p-coumaric acid 4-O-beta-D-glucopyranoside (8), and 23(S)-12-O-caffeoyl-12-hydroxyllauric acid glycerol ester (9), separately. Among them, 1 and 9 are new compounds, and 3, 4, and 6 were isolated from Drynaria species for the first time.

New chemical constituents from the rhizomes of Sparganium stoloniferum.

A new sucrose ester named beta-D-(1-O-acetyl-3-O-cis-feruloyl)fructofuranosyl alpha-D-2',3',6'-O-triacetylglucopyranoside 1 and a novel phenylpropanoid glycerol named 1-O-cis-feruloyl-3-O-trans-p-coumaroylglycerol 2, have been isolated from Sparganium stoloniferum. Their structures were elucidated by spectroscopic methods.

1,3-Diferuloyl-sn-glycerol from the biocatalytic transesterification of ethyl 4-hydroxy-3-methoxy cinnamic acid (ethyl ferulate) and soybean oil.

1,3-Diferuloyl-sn-glycerol is found ubiquitously throughout the plant kingdom, possessing ultraviolet adsorbing and antioxidant properties. Diferuloyl glycerol was synthesized and isolated as a byproduct in up to 5% yield from a pilot plant scale packed-bed, biocatalytic transesterification of ethyl ferulate with soybean oil or mono- and diacylglycerols from soybean oil. The yield of the diferuloyl glycerol byproduct was directly proportional to the overall water concentration of the bioreactor. The isolated diferuloyl glycerol exhibited good ultraviolet adsorbing properties, 280-360 nm with a lambda(max) 322 nm, and compared well to the efficacy of commercial sunscreen active ingredients. The antioxidant capacity of diferuloyl glycerol (0.25-2.5 mM) was determined by its ability to scavenge 2,2-diphenyl-1-picrylhydrazyl radicals and was comparable to that of ferulic acid. At current pilot plant scale production capacity, 120 kg diferuloyl glycerol byproduct could be isolated per year.

Supercritical carbon dioxide fractionation of nonesterified alkoxyglycerols obtained from shark liver oil.

Ethanolysis of shark liver oil was carried out to generate a product enriched in nonesterified alkoxyglycerols and fatty acid ethyl esters. For the present study, the original oil contained very low amounts of squalene, and thus, unsaponifiable matter was mainly constituted by nonesterified alkoxyglycerols (NEAKG). A small percentage of monoesterified alkoxyglycerols (MEAKG) was also detected. Supercritical fluid extraction was employed to fractionate the mixture, achieving a complete elimination of esters and concentrating the alkoxyglycerol compounds in the raffinate product. Extractions were carried out in a countercurrent packed column, using extraction pressures in the range of 140-180 bar, temperatures from 45 to 65 degrees C, and a solvent-to-feed ratio of 15. NEAKG + MEAKG purity obtained in the raffinate at the best extraction conditions was around 78% w/w, and satisfactory yield (>60%) was also achieved. Therefore, the raffinate product can be re-esterified to design highly valuable ether lipid compounds.