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Vitamin E (VitE) production from crude palm oil (CPO) has been extensively studied and industrially conducted. VitE in CPO is in the range of 600 to 1,000 ppm, and is usually produced from one of the main by-products of edible palm oil production, namely palm fatty acid distillate (PFAD). PFAD contains 4,000 to 5,500 ppm of VitE, and is produced from deodorization process of palm oil purification. This paper presents an innovative process of VitE concentrate production from CPO. A scrubber was designed and installed between the deodorizer and conventional PFAD scrubber. The main objective of this new scrubber was a recovery of glycerides from PFAD. This new scrubber is operated at 150 to 160℃. The scrubbed oil is named as Scrubbed Palm Fatty Acid Distillate (S-PFAD). This simple and efficient modified process can retrieve glycerides as S-PFAD at 0.3% recovery and it enhances VitE concentration in S-PFAD to the range of 28,000 to 32,000 ppm, which is the highest concentration of VitE that has ever been produced in the palm oil production. Fatty acids and glycerides in S-PFAD were esterified and transesterified to methyl esters. The methyl esters were evaporated from S-PFAD, and S-PFAD residue oil contained 24.7% VitE.

To evaluate the endocannabinoid system in an animal model of Parkinson's disease, in-tube solid-phase microextraction (in-tube SPME) was directly coupled to a tandem mass spectrometry (MS/MS) system for determination of the endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG) in rat brain samples. In-tube SPME-which consisted of a microtube of restricted access material (RAM) with a hydrophilic diol external surface and a hydrophobic octyl inner surface-efficiently excluded (up to 95%) macromolecules from the biological samples and selectively pre-concentrated the analytes. In-tube SPME parameters, such as sample volume, mobile phases, flow rate, and pre-concentration time, were evaluated to improve the extraction efficiency and throughput performance. The selectivity of the in-tube SPME and MS/MS (MRM mode) techniques allowed them to be directly coupled online, which dismissed the need for the chromatographic separation step. The in-tube SPME-MS/MS method was validated and shown to be linear from 6.0 to 30.0 ng mL -1 for AEA and from 10.0 to 100.0 ng mL -1 for 2-AG; the intra- and inter-assay accuracy and precision were lower than 15%. Parallelism between the calibration curves constructed in the matrix and aqueous solution confirmed that there was no matrix effect. The method allowed endogenous concentrations of AEA and 2-AG to be determined in rat brain striatum from unilaterally 6-hydroxydopamine-lesioned animals. The concentrations of these endocannabinoids in striatum ipsilateral and contralateral to the lesion differed significantly (p<0.001)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

A simple and reliable method was developed and validated to determine the endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG) in rat brain samples by micro salting-out assisted liquid-liquid extraction combined with ultra-high performance liquid chromatography tandem mass spectrometry (SALLLE/UHPLC-MS/MS). The SALLE parameters (brain homogenate volume, salting-out agent, salt concentration, salt solution volume, organic solvent, organic solvent volume, and centrifugation temperature) were optimized to improve sensitivity and selectivity of the method. The SALLE/UHPLC-MS/MS method presented linear ranges from 2.00 to 20.00 ng mL -1 for AEA and from 0.300 to 10.00 μg mL -1 for 2-AG, no significant matrix effect, and inter- and intra-assay precision and accuracy with CV and RSE values lower than 15%, respectively. This innovative method was successfully applied to determine AEA and 2-AG in brain hemispheres from a 6-OHDA animal model of Parkinson's disease (PD)., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In this paper, we report a method for the separation of hydroxy fatty acid and non-hydroxy fatty acid containing neutral lipid classes via normal phase HPLC with UV detection on a PVA-Sil column. The hexane/isopropanol/methanol/water based method separates all the neutral lipids in 21 min, and subsequently flushes through the polar lipids by 27 min such that prefractionation of neutral and polar lipids are not required, and the column is re-equilibrated for the next run in 15 min, for a total run time of 45 min per sample. The separation was demonstrated at both 1.0 mL/min and 1.5 mL/min for added applicability for fraction collection or inline analysis. Separation of various hydroxy fatty acid containing lipids was demonstrated from three different plant species Ricinus communis, Physaria fendleri, and engineered Arabidopsis thaliana. Additionally, we have combined this method with an in-line liquid scintillation counter for the separation and quantification of 14 C labeled lipids obtained from in vivo metabolic flux experiments conducted in the developing seeds of Arabidopsis thaliana., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Massa Medicata Fermentata (MMF), known as Shenqu, is an important traditional Chinese medicine widely used to treat indigestion, vomiting, and diarrhea. In this study, a new benzochroman, 3( S )-3,4-dihydro-5,10-di- β -d-glucopyranoside-2,2-dimethyl-2 H -naphtho(2,3- b )pyran-3-ol ( 1 ), and five known galactosyl acylglycerols ( 2 ⁻ 6 ) were isolated from a methanol extract from MMF. In addition, their chemical structures were determined by chemical and spectroscopic methods, which were compared with the previously reported data. Furthermore, the effects of isolated compounds on lipopolysaccharide (LPS)-stimulated bone marrow-derived dendritic cells were investigated. Compounds 1 ⁻ 3 exhibited significant inhibitory effects on the LPS-induced production of IL-6 and IL-12 p40, with IC 50 values ranging from 1.6 to 10.2 μM. Compounds 2 and 3 also exhibited strong inhibitory effects on the LPS-stimulated production of TNF-α with IC 50 values of 12.0 and 11.2 μM, respectively. The results might provide a scientific basis for the development of the active components in MMF, as well as for novel anti-inflammatory agents.

A new glycosyl glyceride (5) along with twelve known ones (1-4 and 6-13) including two sulfoquinovosyl glycerides (1 and 2) were isolated from the aerial parts of Malva verticillata. Based on several spectroscopic methods, compound 5 was identified to be (2S)-1-O-β-d-galactopyranosyl-3-O-isostearoyl glyceride, and named malvaglycolipid A. Compounds 1 and 2 contained a unique sugar, (6-deoxy-6-sulfo)-α-d-glucopyranose, which very rarely occurs in natural sources. This is the first report for the isolation of compounds 1 and 2 from natural sources and the structure determination using NMR experiment. It was also of note that no glycosyl glyceride has previously been isolated from the family of Malvaeae. Most glycosyl glycerides showed cytotoxicity to HepG2, AGS, HCT-15, and A549 human cancer cells. Especially, compounds 1, 2, and 11 exhibited significant cytotoxicity to AGS cells, with IC 50 values of 33.7 ± 0.64 μM, 11.1 ± 0.07 μM, and 10.6 ± 0.10 μM, respectively. The n-BuOH fraction and compounds 1, 2, and 11 increased the number of apoptotic cells in the Tali assay and had a significant effect on the levels of proteins related to apoptosis including PARP, caspase-3, Bcl-2, Bax, and β-actin., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The tree Aquilaria malaccensis is a valuable source of agarwood, which is used in herbal medicinal preparations. Phytochemical research on A. malaccensis seeds has led to the isolation of four new phorbol esters (1-4), two known phorbol esters (5, isolated from Nature for the first time, and 6), and two known glycerides (7 and 8). The structures of these isolates were elucidated by means of spectroscopic data interpretation. The inflammation-modulatory activities of the isolates on elastase release and superoxide anion generation in human neutrophils were evaluated. Interestingly, phorbol esters 1, 5, and 6 showed potent inhibitory activity on elastase release in human neutrophils, with IC 50 values of 2.7, 0.8, and 2.1 μM, respectively. All isolated phorbol esters exerted enhancing activity on superoxide anion generation. The results indicated that phorbol esters may play a bilateral modulatory role in the processes of inflammation. In addition, the compounds were evaluated for their cytotoxic properties against HepG2 (hepatoma), MDA-MB-231 (breast), and A549 (lung) cancer cells, but all compounds were inactive for all cell lines used (IC 50 > 10 μM).

The current study aimed to evaluate the antioxidant efficiency of different extracts of corn silk. In addition, the impact of corn silk extract on oxidative stability of neem biodiesel during storage was studied. The highest phenolics, DPPH radical scavenging and reducing power activities were recorded for methanol-water extract. The longest oxidation stability (10 h) was observed for biodiesel samples blended with 1000 ppm of corn silk extract (CSE). At the end of storage period the induction time of biodiesel samples mixed with 1000 ppm of CSE or butylated hydroxytoluene (BHT) were about 6.72 and 5.63 times as high as in biodiesel samples without antioxidants. Biodiesel samples blended with 1000 ppm of CSE had the lowest acidity at the end of storage period. Peroxide value of biodiesel samples containing 1000 ppm of CSE was about 4.28 times as low as in control sample without antioxidants.

Microbes belonging to the genus Verrucosispora possess significant chemical diversity and biological properties. They have attracted the interests of many researchers and are becoming promising resources in the marine natural product research field. A bioassay-guided isolation from the crude extract of Verrucosispora sp. strain MS100047, isolated from sediments collected from the South China Sea, has led to the identification of a new salicylic derivative, glycerol 1-hydroxy-2,5-dimethyl benzoate (1), along with three known compounds, brevianamide F (2), abyssomicin B (3), and proximicin B (4). Compound 1 showed selective activity against methicillin-resistant Staphylococcus aureus (MRSA) with a minimum inhibitory concentration (MIC) value of 12.5 μg/mL. Brevianamide F (2), which was isolated from actinomycete for the first time, showed a good anti-BCG activity with a MIC value of 12.5 μg/mL that has not been reported previously in literatures. Proximicin B (4) showed significant anti-MRSA (MIC = 3.125 μg/mL), anti-BCG (MIC = 6.25 μg/mL), and anti-tuberculosis (TB) (MIC = 25 μg/mL) activities. This is the first report on the anti-tubercular activities of proximicins. In addition, Verrucosispora sp. strain MS100047 was found to harbor 18 putative secondary metabolite gene clusters based on genomic sequence analysis. These include the biosynthetic loci encoding polyketide synthase (PKS) and non-ribosomal peptide synthetase (NRPS) consistent with abyssomicins and proximicins, respectively. The biosynthetic pathways of these isolated compounds have been proposed. These results indicate that MS100047 possesses a great potential as a source of active secondary metabolites.

As a part of our continuing work to find out bioactive lead molecules from marine invertebrates, the CHCl₃ fraction of the organic extract of the Red Sea sponge Theonella mirabilis showed cytotoxic activity in our primary screen. Bioassay-guided purification of the active fractions of the sponge's extract resulted in the isolation of two new glycerides, mirabolides A and B (1 and 2), together with the reported 4-methylene sterols, conicasterol (3) and swinhosterol B (4). The structures of the compounds were assigned by interpretation of their 1D (¹H, (13)C), 2D (COSY, HSQC, HMBC, ROESY) NMR spectral data and high-resolution mass determinations. Compounds 1-4 displayed marked cytotoxic activity against human breast adenocarcinoma cell line (MCF-7) with IC50 values of 16.4, 5.18, 6.23 and 3.0 μg/mL, respectively, compared to 5.4 μg/mL observed by doxorubicin as reference drug.

The glyceride in oil food simulant usually causes serious interferences to target analytes and leads to failure of the normal function of the RP-HPLC column. In this work, a convenient HPLC-UV method for the determination of the total specific migration of nine ultraviolet (UV) absorbers in food simulants was developed based on 1,1,3,3-tetramethylguanidine (TMG) and organic phase anion exchange (OPAE) SPE to efficiently remove glyceride in olive oil simulant. In contrast to the normal ion exchange carried out in an aqueous solution or aqueous phase environment, the OPAE SPE was performed in the organic phase environments, and the time-consuming and challenging extraction of the nine UV absorbers from vegetable oil with aqueous solution could be readily omitted. The method was proved to have good linearity (r≥0.99992), precision (intra-day RSD≤3.3%), and accuracy(91.0%≤recoveries≤107%); furthermore, the lower limit of quantifications (0.05-0.2mg/kg) in five types of food simulants(10% ethanol, 3% acetic acid, 20% ethanol, 50% ethanol and olive oil) was observed. The method was found to be well suited for quantitative determination of the total specific migration of the nine UV absorbers both in aqueous and vegetable oil simulant according to Commission Regulation (EU) No. 10/2011. Migration levels of the nine UV absorbers were determined in 31 plastic samples, and UV-24, UV-531, HHBP and UV-326 were frequently detected, especially in olive oil simulant for UV-326 in PE samples. In addition, the OPAE SPE procedure was also been applied to efficiently enrich or purify seven antioxidants in olive oil simulant. Results indicate that this procedure will have more extensive applications in the enriching or purification of the extremely weak acidic compounds with phenol hydroxyl group that are relatively stable in TMG n-hexane solution and that can be barely extracted from vegetable oil., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In this study, the effect of Mougeotia nummuloides and Spirulina major on Vero cells (African green monkey kidney), C6 cells (rat brain tumor cells) and HeLa cells (human uterus carcinoma) was investigated in vitro. The antiproliferative effect of the methanol extract of M. nummuloides and S. major compared with 5-fluorourasil (5-FU) and cisplatin was tested at various concentrations using the BrdU Cell Proliferation ELISA. Both M. nummuloides and S. major extracts significantly inhibited the proliferation of Vero, HeLa and C6 cancer cell lines with IC50 and IC75 values. The M. nummuloides extract exhibited higher activity than 5-FU and cisplatin on Vero and C6 cells at high concentrations. The S. major extract revealed better antifproliferative activity than standards against Vero cells at 500 μg/mL. The compounds of methanol extracts were determined by GC-MS after the silylation process. Trehalose, monostearin and 1-monopalmitin were detected as major products in the M. nummuloides extract where as in the S. major extract; monostearin, 1-monopalmitin and hexyl alcohol were the main constituents.

LC-MS/MS is a powerful analytical technique that provides unequivocal identification and reliable quantification of the analytes, using Selected Reaction Monitoring or Multi Reaction Monitoring acquisition mode.2-Arachidonoylglycerol (2-AG) is the most abundant endocannabinoid, which plays a major role in a wide variety of physiological and pathological processes. Analysis of 2-AG by means of LC-MS/MS allows the detection of very low concentrations in biological samples. Here, we describe how to determine 2-AG levels in tiny samples of tissues and plasma through LC-MS/MS, by using very quick and easy to perform extraction procedures, with reduced solvent consumption.

Porcine liver carboxylesterase (PLE) belongs to carboxylesterase family (EC 3.1.1.1) as a serine-type esterase. The PLE-catalyzed esterification of capric acid with glycerol in reverse micelles was investigated on the catalytic performance and enzyme kinetics. The most suitable structure of reverse micelles was comprised of isooctane (reaction medium) and bis(2-ethylhexyl) sodium sulfosuccinate (AOT, anionic surfactant) with 0.1 of R-value ([water]/[surfactant]) and 3.0 of G/F-value ([glycerol]/[fatty acid]) for the PLE-catalyzed esterification. In the aspect of regio-selectivity, the PLE mainly produced 1-monocaprin without any other products (di- and/or tricaprins of subsequent reactions). Furthermore, the degree of esterification at equilibrium state (after 4 h from the initiation) was 62.7% under the optimum conditions at pH 7.0 and 60 °C. Based on Hanes-Woolf plot, the apparent Km and Vmax values were calculated to be 16.44 mM and 38.91 μM/min/mg protein, respectively., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Truffles are the fruiting body of fungi, members of the Ascomycota phylum endowed with major gastronomic and commercial value. The development and maturation of their reproductive structure are dependent on melanin synthesis. Since anandamide, a prominent member of the endocannabinoid system (ECS), is responsible for melanin synthesis in normal human epidermal melanocytes, we thought that ECS might be present also in truffles. Here, we show the expression, at the transcriptional and translational levels, of most ECS components in the black truffle Tuber melanosporum Vittad. at maturation stage VI. Indeed, by means of molecular biology and immunochemical techniques, we found that truffles contain the major metabolic enzymes of the ECS, while they do not express the most relevant endocannabinoid-binding receptors. In addition, we measured anandamide content in truffles, at different maturation stages (from III to VI), through liquid chromatography-mass spectrometric analysis, whereas the other relevant endocannabinoid 2-arachidonoylglycerol was below the detection limit. Overall, our unprecedented results suggest that anandamide and ECS metabolic enzymes have evolved earlier than endocannabinoid-binding receptors, and that anandamide might be an ancient attractant to truffle eaters, that are well-equipped with endocannabinoid-binding receptors., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Mosquitoes (Diptera: Culicidae) represent an important threat to millions of people worldwide, since they act as vectors for important pathogens, such as malaria, yellow fever, dengue and West Nile. Control programmes mainly rely on chemical treatments against larvae, indoor residual spraying and insecticide-treated bed nets. In recent years, huge efforts have been carried out to propose new eco-friendly alternatives, with a special focus on the evaluation of plant-borne mosquitocidal compounds. Major examples are neem-based products (Azadirachta indica A. Juss, Meliaceae) that have been proven as really effective against a huge range of pests of medical and veterinary importance, including mosquitoes. Recent research highlighted that neem cake, a cheap by-product from neem oil extraction, is an important source of mosquitocidal metabolites. In this review, we examined (i) the latest achievements about neem cake metabolomics with special reference to nor-terpenoid and related content; (ii) the neem cake ovicidal, larvicidal and pupicidal toxicity against Aedes, Anopheles and Culex mosquito vectors; (iii) its non-target effects against vertebrates; and (iv) its oviposition deterrence effects on mosquito females. Overall, neem cake can be proposed as an eco-friendly and low-cost source of chemicals to build newer and safer control tools against mosquito vectors.

Coprinus comatus, a novel cultivated edible mushroom, has a various of pharmacological effects due to its many active components. In this study, agaricoglycerides, a new class of fungal secondary metabolites that have strong activity against neurolysin, were isolated from C. comatus mycelia. Simultaneously, a 3-level Box-Behnken factorial design was used, combined with response surface methodology, to optimize the precursor composition of agaricoglycerides for the production of agaricoglyceride A. The model estimated that a maximal yield of agaricoglyceride A (20.105 mg/L) could be obtained when the concentrations of 4-hydroxybenzoic acid, glycerol, and methanol (MeOH) were set at 75 mg/L, 0.75 mL/L, and 0.75 mL/L, respectively. The verified experiments showed that the model was significantly consistent with the model prediction. These results showed that appropriately adding the precursors could increase the production of agaricoglyceride A.

A monoacylglycerol (1) and a 1:1 mixture of two monogalactosyl diacylglycerols (MGDGs) (2 and 3) were isolated from the brown seaweed Fucus spiralis Linnaeus. The structures were elucidated by spectroscopic means (NMR and MS) and by comparison with the literature. Compound 1 was composed of a glycerol moiety linked to oleic acid (C18:1 Ω9). Compounds 2 and 3 contained a glycerol moiety linked to a galactose unit and eicosapentaenoic acid (C20:5 Ω3) combined with octadecatetraenoic acid (C18:4 Ω3) or linolenic acid (C18:3 Ω3), respectively. The isolated compounds were tested for their cytotoxic and anti-inflammatory activity in RAW 264.7 macrophage cells. All of them inhibited NO production at non-cytotoxic concentrations. The fraction consisting of compounds 2 and 3, in a ratio of 1:1, was slightly more effective than compound 1 (IC₅₀ of 60.06 and 65.70 µg/mL, respectively). To our knowledge, this is the first report of these compounds from F. spiralis and on their anti-inflammatory capacity.

Two new glycerides, didemnacerides A (1) and B (2), together with three known sterols, 24-ethyl-25-hydroxycholesterol (3), cholest-6-en-3,5,8-triol (4) and cholestane-3β,5α,6β-26-tetrol (5), were isolated from the Red Sea ascidian Didemnum sp. Their structures were elucidated by using extensive 1D ((1)H, (13)C) and 2D ((1)H-(1)H COSY, HSQC and HMBC) NMR studies and mass spectroscopic data (GC-MS and HR-MS) as well as alkaline hydrolysis followed by GC-MS and NMR spectral analyses of the fatty acid methyl esters. This is the first report of compounds 3-5 from the Red Sea ascidian Didemnum species.

To determine the no-observed-adverse-effect level (NOAEL) of exposure and target organs of neem oil for establishing safety criteria for human exposure, the subchronic toxicity study with neem oil in mice was evaluated. The mice (10 per sex for each dose) was orally administered with neem oil with the doses of 0 (to serve as a control), 177, 533 and 1600 mg/kg/day for 90 days. After the treatment period, observation of reversibility or persistence of any toxic effects, mice were continuously fed without treatment for the following 30 days. During the two test periods, the serum biochemistry, organ weight and histopathology were examined. The results showed that the serum biochemistry and organ coefficient in experimental groups had no statistical difference compared with those of the control group. At the 90th day, the histopathological examinations showed that the 1600 mg/kg/day dose of neem oil had varying degrees of damage on each organ except heart, uterus and ovarian. After 30-day recovery, the degree of lesions to the tissues was lessened or even restored. The NOAEL of neem oil was 177 mg/kg/day for mice and the target organs of neem oil were determined to be testicle, liver and kidneys.

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.

During a large-scale isolation campaign for the heterodimeric phloroglucinyl pyrone arzanol (1a) from Helichrysum italicum subsp. microphyllum, several new phenolics as well as an unusual class of lipids named santinols (5a-c, 6-8) have been characterized. Santinols are angeloylated glycerides characterized by the presence of branched acyl- or keto-acyl chains and represent a hitherto unreported class of plant lipids. The antibacterial activity of arzanol and of a selection of Helichrysum phenolics that includes coumarates, benzofurans, pyrones, and heterodimeric phloroglucinols was evaluated, showing that only the heterodimers showed potent antibacterial action against multidrug-resistant Staphylococcus aureus isolates. These observations validate the topical use of Helichrysum extracts to prevent wound infections, a practice firmly established in the traditional medicine of the Mediterranean area.

Endocannabinoids (ECs) are endogenous compounds that interact with type-1 and type-2 cannabinoid receptors (CB(1) and CB(2)), as well as non-cannabinoid receptors. The multitude of roles attributed to ECs makes them an emerging target of pharmacotherapy for a number of disparate diseases. Here a high-throughput bioanalytical method based on micro SPE (μ-SPE) followed by LC-MS/MS analysis for the simultaneous determination of the two major endocannabinoids 2-arachidonoylglycerol (2-AG) and N-arachidonoylethanolamine (anandamide, AEA) in human plasma is presented. The chromatographic conditions obtained with the fused-core column allowed a good separation in 10 min also of the AG isomers. A very simple and reliable extraction has been optimised by means of C18-modified tips: it requires only 100 μL of plasma and allows the use of minimal volumes of organic solvent. The present method allows a rapid and effective clean-up, which also minimises the isomerisation of 2-AG. The whole procedure has been validated following the FDA guidelines for bioanalytical methods validation: the satisfactory recovery values, the negligible matrix effect and the good values of accuracy and reproducibility make it a simple and high-throughput analytical tool for clinical and biochemical studies on endocannabinoid signaling in humans.

Objective: To investigate the chemical constituents in the roots and rhizomes of Sinopodophyllum emodi., Methods: The compounds were isolated by many different chromatographic methods such as silica gel, Sephadex LH-20, and ODS column. Their structures were identified by their physicochemical properties and spectrascopic data., Results: Nine compounds were isolated and identified as isopicrodeoxypodophyllotoxin(I), 3beta-hydroxy-7alpha-methoxy-24beta-ethyl-cholest-5-ene(II), 5alpha, 8alpha-epidioxy-(22E,24R)-erg-osta-6,22-dien-3beta-ol(III), 7beta-hydroxysitosterol (IV), beta-sitosterol (V), daucosterol (VI), alpha-glyceryl palmitate (VII), alpha-D-glucose (VIII), 5-hydromethyl furaldehyde (IX)., Conclusion: Compounds I - IV, VII - IX are obtained from this genus for the first time.

We show that the characterization of mono-, di- and triglycerols can be readily accomplished by high performance liquid chromatography (HPLC) with silver(I)-mercaptopropyl modified silica gel, or silver thiolate chromatographic material (AgTCM), which can be used with evaporative light scattering detection (ELSD) or atmospheric pressure chemical ionization mass spectroscopy (APCI-MS). Separation of triglycerols varying by degrees of unsaturation and cis/trans configuration in common oil samples can be achieved using a simple linear gradient of hexane and acetone. In addition to double bonds, AgTCM also displays major selectivity for compounds with different levels of polarity, allowing for efficient separation between mono-, di- and triglycerols. In comparison to conventional reversed phase columns, AgTCM produces simple chromatograms for rapid assessment of degrees of unsaturation and the amount of trans fats in triglycerides, which are central issues to food quality determination. In comparison to previous silver-ion based HPLC separations, AgTCM-HPLC based column offers greatly enhanced stability, inertness, durability, and reproducibility allowing routine coupling of the HPLC with a mass spectrometer for detection., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Algal biofuels are a growing interest worldwide due to their potential in terms of sustainable greenhouse gas displacement and energy production. This article describes a comparative survey of biodiesel production and conversion yields of biodiesel via alkaline transesterification of acylglycerols extracted from the microalgae Thalassiosira pseudonana and Phaeodactylum tricornutum, grown under silicate or nitrate limitation, and that of model vegetable oils: soybean, and rapeseed oil. Acylglycerols were extracted with n-hexane and the total yield per biomass was determined by gravimetric assay. Under our conditions, the total acylglycerol yield from the microalgae studied was 13-18% of total dry weight. The biodiesel samples were analyzed using gas chromatography-flame ionization detector to determine quantitative information of residual glycerol, mono-, di-, and tri-acylglycerol concentrations in the biodiesel. All of the algal-based biodiesel demonstrated less mono-, di-, and tri-acylglycerol concentrations than the vegetable-based biodiesel under identical transesterification conditions. The fatty acid compositions of all the feedstock oils and their resultant biodiesel were also analyzed and reported. Based on the fatty acid methyl ester compositions of our samples we qualitatively assessed the suitability of the algal-derived biodiesel in terms of cetane number (CN), cold-flow properties, and oxidative stability., (Copyright © Published 2011. This article is a U.S. Government work and is in the public domain in the USA.)

A 70% ethanol extract of the roots of Livistona chinensis has been investigated, led to the isolation of 13 compounds, including a new ceramide, (2S,3S,4R,9Z)-2-[(2R)-2-hydroxytricosanoylamino]-9-octadecene-1,3,4-triol (2), a new glycosyl ceramide, 1-O-β-D-glucopyranosyl-(2S,3S,4R,9Z)-2-[(2R)-2-hydroxydocosanoylamino]-9-octadecene-1,3,4-triol (3), three new monoacylglycerols, 1-(34-hydroxytetratriacontanoyl)-sn-glycerol (9), 1-[nonadeca-(9Z,12Z)-dienoyl]-sn-glycerol (10), and 1-[12-hydroxypentatriaconta-(13E,15Z)-dienoyl]-sn-glycerol (11), a new diacylglycerol, 1-(heptadeca-6Z,9Z-dienoyl)-3-(octadeca-6Z,9Z,12Z-trienoyl)-sn-glycerol (12), as well as a new diacylglycerol aminoglycoside, 1-octadecanoyl-2-nonadecanoyl-3-O-(6-amino-6-deoxy)-β-D-glucopyranosyl-sn-glycerol (13). The structures of new compounds were elucidated, based on spectroscopic, zymologic and chemical methods. Among the compounds tested, compounds 3, 4 and 13 showed significantly antiproliferative effects against the human tumor cell lines (K562, HL-60, HepG2, and CNE-1) with the IC₅₀ of 10-65 μM. To our knowledge, this is first report of the occurrence of ceramides and acylglycerols in the genus Livistona., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Analysis of the endocannabinoid (EC) system's key molecules 2-arachidonoyl glycerol (2AG) and arachidonoyl ethanolamide (anandamide, AEA) is challenging due to several peculiarities. 2AG isomerizes spontaneously to its biologically inactive analogue 1-arachidonoyl glycerol (1AG) by acyl migration and it is only chromatographically distinguishable from 1AG. Matrix-effects caused primarily by co-extracted phospholipids may further compromise analysis. In addition, 2AG and 1AG are unstable under certain conditions like solvent evaporation or reconstitution of dried extracts. We examined effects of different organic solvents and their mixtures, such as toluene, ethyl acetate, and chloroform-methanol, on 2AG/1AG isomerisation, 2AG/1AG stability, and matrix-effects in the UPLC-MS/MS analysis of 2AG and AEA in human plasma. Toluene prevented, both, 2AG isomerisation to 1AG and degradation of 2AG/1AG during evaporation. Toluene extracts contain only 2% of matrix-effect-causing plasma phospholipids compared to extracts from the traditionally used solvent mixture chloroform-methanol. Toluene and all other tested organic solvents provide comparable 2AG and AEA extraction yields (60-80%). Based on these favourable toluene properties, we developed and validated a UPLC-MS/MS method with positive electrospray ionization (ESI+) that allows for simultaneous accurate and precise measurement of 2AG and AEA in human plasma. The UPLC-MS/MS method was cross-validated with a previously described fully-validated GC-MS/MS method for AEA in human plasma. A close correlation (r(2)=0.821) was observed between the results obtained from UPLC-MS/MS (y) and GC-MS/MS (x) methods (y=0.01+0.85x). The UPLC-MS/MS method is suitable for routine measurement of 2AG and AEA in human plasma samples (1 mL) in clinical settings as shown by quality control plasma samples processed over a period of 100 days. The UPLC-MS/MS method was further extended to human urine. In urine, AEA was not detectable and 2AG was detected in only 3 out of 19 samples from healthy subjects at 160, 180 and 212 pM corresponding to 12.3, 14.5 and 9.9 pmol/mmol creatinine, respectively., (Copyright © 2011 Elsevier B.V. All rights reserved.)

In this study, activated bleaching earth (ABE) was used to eliminate glycidyl esters from both triacyl- and diacylglycerol oils. To investigate the mechanism, glycerol dioleate containing glycidyl palmitate (GP) was treated with ABE and the fate of the GP was monitored by analyzing the feed, treated, and ABE-absorbed oils using a gas-liquid chromatograph equipped with a flame-ionized detector. GP was completely removed from both the treated and absorbed oils. This indicates that this treatment is useful for GE removal from diacylglycerol oil, although it was not achieved by absorption of GE on ABE but rather by modification of GP. The results of composition analysis demonstrate that GP is transformed to glycerol monopalmitate, glycerol palmitate oleate, and glycerol dipalmitate at a recovery rate of 99.1 ± 1.3 %. An increase in glycerol monooleate and trace amounts of free glycerol and fatty acids were also observed after treatment. The transformation is proposed to involve a ring-opening reaction of GP with water contained in the ABE and in the bulk oil followed by an interesterification reaction among the resultant monopalmitate and the glycerol dioleate of the bulk oil. All the generated compounds were simple acylglycerols and glycerol. Therefore, ABE treatment could be useful for GE removal during the manufacture of edible oils.

The eggs (nits) of head and body lice (Pediculus humanus capitis, Pediculus humanus corporis) were incubated for 5, 10, 15, 20, 30 or 45 min into a neem seed extract contained in a fine shampoo formulation (e.g. Wash Away® Louse), which is known for its significant killing effects of larvae and adults of head lice. The aim of the study was to test whether the developmental stages inside the eggs are also killed after the incubation into the shampoo. It was found that an incubation time of only 5 min was sufficient to prohibit any hatching of larvae, whilst 93 ± 4% of the larvae in the untreated controls of body lice hatched respectively about 76% of the controls in the case of head lice. Apparently, the neem-based shampoo blocked the aeropyles of the eggs, thus preventing the embryos of both races of lice from accessing oxygen and from releasing carbon dioxide. Thus, this product offers a complete cure from head lice upon a single treatment, if the lice (motile stages, eggs) are fully covered for about 10 min.

Correlations between the dimensions of a 2-D separation create trend lines that depend on structural or chemical characteristics of the compound class and thus facilitate classification of unknowns. This broadly applies to conventional ion mobility spectrometry (IMS)/mass spectrometry (MS), where the major biomolecular classes (e.g., lipids, peptides, nucleotides) occupy different trend line domains. However, strong correlation between the IMS and MS separations for ions of same charge has impeded finer distinctions. Differential IMS (or FAIMS) is generally less correlated to MS and thus could separate those domains better. We report the first observation of chemical class separation by trend lines using FAIMS, here for lipids. For lipids, FAIMS is indeed more independent of MS than conventional IMS, and subclasses (such as phospho-, glycero-, or sphingolipids) form distinct, often non-overlapping domains. Even finer categories with different functional groups or degrees of unsaturation are often separated. As expected, resolution improves in He-rich gases: at 70% He, glycerolipid isomers with different fatty acid positions can be resolved. These results open the door for application of FAIMS to lipids, particularly in shotgun lipidomics and targeted analyses of bioactive lipids.

In this work we report the purification of a crude acetin mixture into mono-, di- and triacetin by countercurrent chromatography. The process was initially tested on a small, semi-preparative scale (0.5 g) to determine its efficiency. The process was then scaled up to accommodate 2.5 g of crude reaction products containing a mixture of the acetins. The solvent system ethyl acetate/n-butanol/water 1:0.2:1 was used in all separation procedures. Mono-, di- and triacetins were separated similarly in the semi-preparative and preparative runs., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This paper presents a research interest concentrating on aims to establish a feasible industrial process for enzymatic production of highly pure glycerol monooleate (GMO). The synthesis of high oleic glycerol monooleate by enzymatic glycerolysis of high oleic sunflower oil, using Novozyme 435 as the biocatalyst, in a binary solvent mixture of tert-butanol and tert-pentanol (80/20, v/v), at a lab scale has been studied. A yield of 75.31% monoacylglycerol has been achieved at the first stage. A yield of 93.3% GMO was finally reached after further purification at the second stage. To evaluate the possibility of the process for industrialization, production of GMO was performed at a pilot-plant scale under the correspondingly adjusted conditions. A yield of 68.17% and 93.4% of GMO was obtained, respectively, at the end of the three stages., (Copyright © 2010 Elsevier Inc. All rights reserved.)

This paper reports on a method for development and validation for simultaneous characterization and determination of oxygenated polar impurities--free fatty carboxylic acids (FFAs), partial glycerides (monoacylglycerides, MGs), residual glycerol and free sterols--in various biodiesels based on the combination of solid-phase extraction (SPE), silylation and GC/MS technologies. The effects of various SPE and silylation conditions on the method recoveries were evaluated. Using this integrated SPE-GC/MS method, 38 target polar compounds (13 FFAs, 17 glycerides and 8 sterols) in 9 biodiesels derived from 4 different feedstocks were successfully separated and quantified. It was found that the carbon chain length of FFAs was ranged from C(6) to C(24), with C(16) and C(18) being the most abundant in all biodiesels. The total FFAs concentration was consistent with the acid values (AVs) measured by standard method ASTM D974-04. MG congeners with carbon number of 18 (mono-C18) were most abundant in the biodiesel samples, followed by mono-C(16) and free glycerol. β-Sitosterol and campesterol were found to be the prevailing phytosterols in all pure vegetable oil-based biodiesels, while brassicasterol and stigmasterol was only significant in the biodiesel from canola oil and soybean oil, respectively, and abundant cholesterol was only detected in animal fat-based biodiesels., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Chloroplasts of plants contain an intricate membrane system, the thylakoids, which harbor the complexes of the photosynthetic machinery. Chloroplasts are confined by two membranes, the inner and outer envelope. The major glycerolipids of chloroplasts are the glycolipids monogalactosyl diacylglycerol (MGD), digalactosyl diacylglycerol (DGD), and sulfoquinovosyl diacylglycerol (SQD). Furthermore, two phospholipids, phosphatidyl glycerol (PG) and phosphatidyl choline (PC), are found in chloroplast membranes. The photosystems and light-harvesting complexes in the thylakoids are rich in photosynthetic pigments (chlorophyll, carotenoids, and xanthophylls) and contain a unique set of prenylquinol lipids (tocochromanol/vitamin E, plastoquinol, and phylloquinol/vitamin K1). In this chapter, methods for the isolation and quantification of chloroplast and leaf glycerolipids and prenylquinol lipids are presented. Glycerolipids are separated by thin-layer chromatography prior to conversion of the fatty acids into methyl esters. Fatty acid methyl esters are subsequently quantified by gas chromatography. Prenylquinol lipids are separated by HPLC and quantified by UV absorption (plastoquinol) or fluorescence (tocochromanol, phylloquinol).

Matrix effects caused by compounds endogenous to the biological sample are a primary challenge in quantitative LC/MS/MS bioanalysis. Many approaches have been developed to minimize matrix effects such as optimization of sample extraction procedures and use of isotopically labeled internal standards. Unexpected matrix components may still remain undetected, however, because of the selective mass transitions monitored during MS/MS analysis. Glycerophosphocholines are the major phospholipids in plasma that have been widely shown to cause significant matrix effects on electrospray ionization efficiencies for target analytes. The purpose of this work was to investigate potential matrix effects resulting from different endogenous lipid classes, including phospholipids, acylglycerols and cholesterols, in order to establish a library for the relative presence of these components in biological sample extracts obtained by commonly used sample preparation techniques. Thirteen compounds were selected which were representatives of eight phospholipids classes, mono, di, triacylglycerols, cholesterol and cholesterol esters. Post-column infusion experiments were carried out to compare relative ion suppression effects of these compounds. Chlorpheniramine and loratadine were selected as model test analytes. A Concentration Normalized Suppression Factor (%CNSF) was defined to allow comparison of ion suppression effects resulting from different endogenous lipids according to their typical concentrations in human plasma and erythrocytes. A simple LC/MS/MS method was developed to monitor these endogenous components in sample extracts and their extraction recoveries from a plasma pool were compared using protein precipitation, liquid-liquid extraction, supported-liquid extraction, solid phase extraction and Hybrid SPE-precipitation methods. Endogenous lipid components other than GPChos, such as cholesterols and triacylglycerols, may result in significant matrix effects and should be monitored during method development. No single extraction procedure was efficient in removing all of the various lipid components. Use of the results presented here, along with a consideration of analyte chemical structure, the type of matrix and the type of sample preparation procedure, may help a bioanalytical scientist to better anticipate and minimize matrix effects in developing LC/MS/MS-based methods., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Phytochemical investigation of Hylodendron gabunensis resulted in the isolation of two new glyceride derivatives, hyloglyceride (1) and hylodiglyceride (2). The structures of the two new compounds were determined by comprehensive analysis of their 1D and 2D NMR spectroscopic, and HREIMS data. One known compound was also isolated and identified as beta-sitosterol.

The nudibranch Austrodoris kerguelenensis is distributed widely around the Antarctic coast and continental shelves. Earlier collections from McMurdo Sound and the Weddell Sea shelf have afforded a suite of diterpene glyceride esters, a compound class implicated as a chemical defense in nudibranchs. The present chemical investigation of A. kerguelenensis collected near Palmer Station on the Western Antarctic Peninsula has revealed additional examples, palmadorins A-C (1-3), as the first three members of a new series of clerodane diterpenes. In this paper we describe their isolation, structure elucidation, and stereochemical analysis using a combination of one- and two-dimensional NMR spectroscopy and wet chemical methods.

Background: Mosquitoes transmit serious human diseases, causing millions of deaths every year. Use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of botanical origin have been reported as useful for control of mosquitoes. Azadirachta indica (Meliaceae) and its derived products have shown a variety of insecticidal properties. The present paper discusses the larvicidal activity of neem-based biopesticide for the control of mosquitoes., Methods: Larvicidal efficacy of an emulsified concentrate of neem oil formulation (neem oil with polyoxyethylene ether, sorbitan dioleate and epichlorohydrin) developed by BMR & Company, Pune, India, was evaluated against late 3rd and early 4th instar larvae of different genera of mosquitoes. The larvae were exposed to different concentrations (0.5-5.0 ppm) of the formulation along with untreated control. Larvicidal activity of the formulation was also evaluated in field against Anopheles, Culex, and Aedes mosquitoes. The formulation was diluted with equal volumes of water and applied @ 140 mg a.i./m(2) to different mosquito breeding sites with the help of pre calibrated knapsack sprayer. Larval density was determined at pre and post application of the formulation using a standard dipper., Results: Median lethal concentration (LC(50)) of the formulation against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti was found to be 1.6, 1.8 and 1.7 ppm respectively. LC(50) values of the formulation stored at 26 degrees C, 40 degrees C and 45 degrees C for 48 hours against Ae. aegypti were 1.7, 1.7, 1.8 ppm while LC(90) values were 3.7, 3.7 and 3.8 ppm respectively. Further no significant difference in LC(50) and LC(90) values of the formulation was observed against Ae. aegypti during 18 months storage period at room temperature. An application of the formulation at the rate of 140 mg a.i./m(2) in different breeding sites under natural field conditions provided 98.1% reduction of Anopheles larvae on day 1; thereafter 100% reduction was recorded up to week 1 and more than 80% reduction up to week 3, while percent reduction against Culex larvae was 95.5% on day 1, and thereafter 80% reduction was achieved up to week 3. The formulation also showed 95.1% and, 99.7% reduction of Aedes larvae on day 1 and day 2 respectively; thereafter 100% larval control was observed up to day 7., Conclusion: The neem oil formulation was found effective in controlling mosquito larvae in different breeding sites under natural field conditions. As neem trees are widely distributed in India, their formulations may prove to be an effective and eco-friendly larvicide, which could be used as an alternative for malaria control.

Infestations with the poultry red mite Dermanyssus gallinae represent a major ectoparasite problem in poultry and affects egg and meat production worldwide. The effects of the neem seed product Mite-Stop against the red poultry mite were investigated. Five primitive poultry farms in two small villages in the Nile Delta and Giza district were selected for the study. The neem extract was diluted 1:40 and 1:50 with tap water just prior to use. Application of the two dilutions of the provided product was performed to soil, cracks and crevices of the examined area as well as to mite-infested birds on day 0 and day 7. Two hours after treatment soil dust was collected from sprayed regions of the stable and from unsprayed control regions of the same stable. The treated chickens were also checked for mites 2 h after each treatment. The examination of the chickens 2 h after spraying showed that they were free of mites. The examination of treated soil with the Tullgren funnel apparatus 2 h after the first spraying on day 0 already showed a considerable reduction of living mites compared to controls. Seven days after the first treatment of the soil the number of living mites was reduced for 80% in the treated soil and decreased even more after the second spraying, since those larvae that had hatched from eggs in the meantime were killed. The 1:40 dilution of the neem seed extract with tap water was superior to the 1:50 dilution. These results clearly show a very high killing rate of the extract, if the mites come in direct contact with the compound. However, in order to obtain extinction also of hidden and freshly hatched stages repeated spraying should be done three times within 8-10 days.

The endocannabinoids anandamide and 2-arachidonoylglycerol, as well as several anandamide-related N-acylethanolamines, belong to a family of lipid transmitter that regulate fundamental physiological processes, including neurotransmission and neuroinflammation. Their precise quantification in biological matrices can be achieved by gas chromatography-mass spectrometry (GC-MS), but this method typically requires multiple time-consuming purification steps such as solid-phase extraction followed by HPLC. Here we report a novel solid-phase extraction procedure allowing for single-step, and thus higher throughput, purification of endocannabinoids and N-acylethanolamines before GC-MS quantification. We determined the minimal amount of mouse brain tissue required to reliably detect endocannabinoids and N-acylethanolamines when using this approach and provide direct evidence for quantification accuracy by using radioactive and deuterated standards spiked into mouse brain samples. Using this method, we found that mouse brain contains much higher levels of anandamide (>1 nmol/g tissue) than previously reported, whereas levels of 2-arachidonoylglycerol and other N-acylethanolamines are well within the range of previous reports. In addition, we show that mouse brain amounts of endocannabinoids and N-acylethanolamines differ depending on animal gender as well as on whether the tissue was fixed or not. Our study shows that endocannabinoid and N-acylethanolamine levels quantified in mouse brain by GC-MS depend closely on tissue amount and preparation as well as on animal gender and that, depending on such parameters, anandamide levels could be underestimated.

An acylglycerol (AG) containing four acyl chains, (12-ricinoleoylricinoleoyl)diricinoleoylglycerol (RRRR), was positively identified for the first time in a natural source in castor oil using electrospray ionization tandem mass spectrometry (ESI-MS/MS). HPLC-purified RRRR from castor oil was subjected to ion trap and high-resolution ESI-MS/MS. The precursor and fragment ions of [RRRR + Na]+ showed the expected masses, and the sodiated fragment ions of both diacylglycerols and fatty acids were detected. Because fragment ions of fatty acids from [AG + NH4]+ adducts cannot be detected by ESI-MS/MS, [AG + Na]+ adducts are more informative. Radiolabeled triricinolein (RRR) was incorporated into RRRR in castor microsomes, indicating that RRRR is biosynthesized in castor bean. This newly identified and biosynthesized RRRR represents a new AG subclass of tetra-acylglycerols (or acylacyldiacylglycerol).

(R)-Glycerol-monolinoleate 4 and (R)-glycerol-monostearate 5 were isolated from the ethyl acetate extracts of Saururus chinensis roots and (R)- or (S)-fatty acid glycerols 4 and 5 were synthesized for confirming their structures and evaluating their inhibitory activities against Lp-PLA(2). The (R)-4 and (S)-4 exhibited Lp-PLA(2) inhibitory activities with IC(50) values of 45.0 and 52.0 microM, respectively.

Aim: To study the chemical constituents of the rhizome of Matteuccia struthiopteris (L.) The constituents were separated and purified by column chromatography with normal Todaro., Methods: phase silica gel and Sephadex LH-20. Their structures were identified on the basis of physical and spectral data (MS, NMR, HMBC and HMQC)., Results: Four compounds were isolated and identified as 1-O-beta-D-gl ucopyranosyl-(2S,3R,4E, 8Z) -2-N-(2'-hydroxydocosanoyl) eicosasphinga-4,8-dienine (1), 1-O-beta-D-galactosyl-(6-->1)-alpha-D-galactosyl-2,3-O-dihexadecanoyl-glycerol (2), succinic acid (3), D-mannitol (4)., Conclusion: Compounds 1 and 2 are new compounds. Compounds 3 and 4 were isolated from this plant for the first time.

Four compounds have been isolated from the algae Hydroclathrus clathratus collected from the South China Sea. Their structures were established as 1-glyceryl hexadecanoate (1), eicosanamide, N-[2'-hydroxy-1'-(hydroxymethyl) pentadecyl] (2), uracil (3), mannitol (4) by MS, 1HNMR and 13CNMR. Compound 1, 2 and 3 were isolated from this algae for the first time.

The chemical composition of the lipophylic fraction from the halophytic plant Euphorbia peplis L. was investigated. Compared to other terrestrial higher plants an increase of triacylglycerols and especially of glycolipids was observed. The main phospholipid was phosphatidyl choline, followed by almost equal concentrations of phosphatidyl ethanolamine and phosphatidyl glycerol. A relatively high concentration of phosphatidic acids (6.5% of the total phospholipids) was found. The main sterol appeared to be sitosterol and significant amounts of tetracyclic triterpene alcohols were found. The composition of the volatile compounds is relatively simple and only one chlorinated compound, identified as 2,2-diethoxy-1-chloroethane, was found. There was a strong toxicity of the total lipophylic extract towards Artemia salina.

The chemical composition of the oil extracted from the seeds of Paullinia cupana var. sorbilis (Mart.) Ducke (syn. P. sorbilis) was investigated. Cyanolipids constituted 3% of the total oil from guaraná seeds, whereas acylglycerols accounted for 28%. 1H and 13C NMR analyses indicated that type I cyanolipids (1-cyano-2-hydroxymethylprop-2-ene-1-ol diesters) are present in the oil from P. cupana. GC and GC-MS analysis showed that cis-11-octadecenoic (cis-vaccenic acid) and cis-11-eicosenoic acids were the main FA (30.4 and 38.7%) esterified to the nitrile group. Paullinic acid (7.0%) was also an abundant component. Oleic acid (37.4%) was the dominant fatty acyl chain in the acylglycerols.

Two new lysophosphatidylcholines (1, 2) and four new monoglycerides (5-8) were isolated from the marine sponge Stelletta sp. by bioactivity-guided fractionation. The planar structures of the new compounds were established on the basis of NMR and MS analyses. The stereochemistry was defined by comparison of the optical rotation. The compounds were evaluated for cytotoxicity against a small panel of five human tumor cell lines.

A factor stimulating a mitogenic activity of peritoneal macrophages is purified from bovine udder. It is identified as a triglyceride, 1-palmitoyl-2-linoleoyl-3-acetyl-RAC -glycerol (RAC -MADG). In this study, its enantiomers, R-(+)-and S-(-)-1-palmitoyl-2-linoleoyl-3-acetylglycerol (R-(+)-MADG, S-(-)-MADG) are synthesized. Among them, R-(+)-MADG enantiomer turns out to increase a mitogenic activity in mouse peritoneal macrophages. Also, (S)-(-)-MADG shows a low mitogenic activity. Treatment of a macrophage with R-(+)-MADG increases reactive oxygen species(ROS). Furthermore, treatment of macrophages with antioxidant, N-acetyl-L-cysteine (NAC), suppresses the R-(+)-MADG-dependent macrophage proliferation. Results show that the generation of ROS induces in R-(+)-MADG-dependent cell signaling. Treatment of a macrophage with R-(+)-MADG increases the activity of protein kinase C (PKC). Treatment of macrophages with calphostin C inhibits R-(+)-MADG-induced macrophage proliferation. Results suggest that R-(+)-MADG enhances the activity of protein kinase C (PKC) and stimulates the macrophage growth. In conclusions, R-(+)-MADG accelerates the production of ROS and increases the activity of PKC to eventually stimulate macrophage cell growth. The existence of RAC -MADG in bovine udder and milk provides passive protection for the neonate and immunostimulatory capabilities., (Copyright 2003 S. Karger AG, Basel)

Phospholipase A(2) (PLA(2)) was purified to homogeneity from the supernatant fraction of rat testis homogenate. The purified 63-kDa enzyme did not require Ca(2+) ions for activity and exhibited both phosphatidic acid-preferring PLA(2) and monoacylglycerol lipase activities with a modest specificity toward unsaturated acyl chains. Anionic detergents enhanced these activities. Serine-modifying irreversible inhibitors, (p-amidinophenyl) methanesulfonyl fluoride and methylarachidonyl fluorophosphonate, inhibited both activities to a similar extent, indicating a single active site is involved in PLA(2) and lipase activities. The sequence of NH(2)-terminal 12 amino acids of purified enzyme was identical to that of a carboxylesterase from rat liver. The optimal pH for PLA(2) activity (around 5.5) differed from that for lipase activity (around 8.0). At pH 5.5 the enzyme also hydrolyzed bis(monoacylglycerol) phosphate, or lysobisphosphatidic acid (LBPA), that has been hitherto known as a secretory PLA(2)-resistant phospholipid and a late endosome marker. LBPA-enriched fractions were prepared from liver lysosome fractions of chloroquine-treated rats, treated with excess of pancreatic PLA(2), and then used for assaying LBPA-hydrolyzing activity. LBPA and the reaction products were identified by microbore normal phase high performance liquid chromatography/electrospray ionization ion-trap mass spectrometry. These enzymatic properties suggest that the enzyme can metabolize phosphatidic and lysobisphosphatidic acids in cellular acidic compartments.