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Gel Chromatography (gel + chromatography)
Kinds of Gel Chromatography Selected AbstractsExtraction, purification and characterization of wax from flax (Linum usitatissimum) strawEUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY, Issue 7 2009Yasantha Athukorala Abstract The chemical composition and selected physical parameters of wax extracted from flax straw with supercritical CO2 (SC-CO2) and hexane have been determined. From the GC/MS results, clear variations in composition and component distributions were observed between SC-CO2 - and hexane-extracted samples. The major components of the SC-CO2 and hexane extracts from three flax cultivars were: fatty acids (36,49%), fatty alcohols (20,26%), aldehydes (10,14%), wax esters (5,12%), sterols (7,9%) and alkanes (4,5%). Purification of SC-CO2 -extracted wax with silica gel chromatography yielded 0.4,0.5% (dry matter) and was composed primarily of wax esters (C44, C46 and C48) and alkanes (C27, C29 and C31). UV-Vis scans of the purified wax samples exhibited two main peaks indicating the presence of conjugated dienes and carotenoids or related compounds. Fourier transform infrared results showed prominent peaks at 2918 (-C-H), 2849 (-C-H), 1745 (-C=O), 1462 (-C-H), 1169 (-C-O) and 719,cm,1 (-(CH2)n -), with NorLin wax showing a slightly deviating pattern compared to the other samples. Thermal analysis by differential scanning calorimetry revealed a mean melting point of 55,56,°C and oxidation temperatures of 146,153,°C for purified wax from flax straw processed using different procedures. [source] Synthesis of N -Bz-Protected D -Daunosamine and D -Ristosamine by Silica Gel Promoted Intramolecular Conjugate Addition of Trichloroacetimidates obtained from Osmundalactone and Its EpimerEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 11 2010Yoshitaka Matsushima Abstract Trichloroacetimidates prepared from osmundalactone and its epimer unexpectedly undergo intramolecular conjugate addition during silica gel chromatography to produce oxazolines in excellent yields. The novel, simple synthesis of N -Bz-protected D -daunosamine and D -ristosamine from these oxazolines is described in this paper. [source] Structures of two cell wall-associated polysaccharides of a Streptococcus mitis biovar 1 strainFEBS JOURNAL, Issue 24 2000A unique teichoic acid-like polysaccharide, the group O antigen which is a C-polysaccharide in common with pneumococci The cell wall of Streptococcus mitis biovar 1 strain SK137 contains the C-polysaccharide known as the common antigen of a closely related species Streptococcus pneumoniae, and a teichoic acid-like polysaccharide with a unique structure. The two polysaccharides are different entities and could be partially separated by gel chromatography. The structures of the two polysaccharides were determined by chemical methods and by NMR spectroscopy. The teichoic acid-like polymer has a heptasaccharide phosphate repeating unit with the following structure: The structure neither contains ribitol nor glycerol phosphate as classical teichoic acids do, thus we have used the expression teichoic acid-like for this polysaccharide. The following structure of the C-polysaccharide repeating unit was established: where AAT is 2-acetamido-4-amino-2,4,6-trideoxy- d -galactose. It has a carbohydrate backbone identical to that of one of the two structures of C-polysaccharide previously identified in S. pneumoniae. C-polysaccharide of S. mitis is characterized by the presence, in each repeating unit, of two residues of phosphocholine and both galactosamine residues in the N-acetylated form. Immunochemical analysis showed that C-polysaccharide constitutes the Lancefield group O antigen. Studies using mAbs directed against the backbone and against the phosphocholine moiety of the C-polysaccharide revealed several different patterns of these epitopes among 95 S. mitis and Streptococcus oralis strains tested and the exclusive presence of the group O antigen in the majority of S. mitis biovar 1 strains. [source] The identity of the O-specific polysaccharide structure of Citrobacter strains from serogroups O2, O20 and O25 and immunochemical characterisation of C. youngae PCM 1507 (O2a,1b) and related strainsFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 1-2 2003gorzata Miesza Abstract Serological studies using SDS,PAGE and immunoblotting revealed that from five strains that are ascribed to Citrobacter serogroup O2, four strains, PCM 1494, PCM 1495, PCM 1496 and PCM 1507, are reactive with specific anti- Citrobacter O2 serum. In contrast, strain PCM 1573 did not react with anti- Citrobacter O2 serum and, hence, does not belong to serogroup O2. The LPS of Citrobacter youngae O2a,1b (strain PCM 1507) was degraded under mild acidic conditions and the O-specific polysaccharide (OPS) released was isolated by gel chromatography. Sugar and methylation analyses along with 1H- and 13C-NMR spectroscopy, including two-dimensional 1H,1H COSY, TOCSY, NOESY and 1H,13C HSQC experiments, showed that the repeating unit of the OPS has the following structure: NMR spectroscopic studies demonstrated that Citrobacter werkmanii O20 and C. youngae O25 have the same OPS structure as C. youngae O2. Sugar and methylation analyses of the core oligosaccharide fractions demonstrated structural differences in the lipopolysaccharide core regions of these strains, which may substantiate their classification in different serogroups. [source] Enzyme-Catalyzed Synthesis of a Hybrid N-Linked Oligosaccharide using N-Acetylglucosaminyltransferase IADVANCED SYNTHESIS & CATALYSIS (PREVIOUSLY: JOURNAL FUER PRAKTISCHE CHEMIE), Issue 11-12 2008Rui Chen Abstract The soluble catalytic domain of human N-acetylglucosaminyltransferase I was purified from Escherichia coli and utilized in the enzyme-catalyzed conversion of high mannose N-linked oligosaccharide 1 into the rare hybrid oligosaccharide 2. Analysis of the reaction showed that the conversion of high mannose 1 into hybrid oligosaccharide 2 proceeded to 100% completion as assessed by MALDI-TOF-MS. Purification of the large polar oligosaccharide by gel filtration and silica gel chromatography afforded a 42% isolated yield of oligosaccharide 2. This enzyme-catalyzed reaction can be utilized to produce rare hybrid oligosaccharides for biochemical and structural studies. [source] Analyses of Glycolipids in Clove, Red Pepper, and Nutmeg by High-Performance Liquid ChromatographyJOURNAL OF FOOD SCIENCE, Issue 6 2000H. Suzuki ABSTRACT: To determine the existence of glycolipids (neutral glycosphingolipid and glycoglycerolipid) in clove, red pepper, and nutmeg, we performed silica gel chromatography and high-performance liquid chromatography (HPLC) using an Aquasil-SS column and a C8 -reversed-phase silica gel column. HPLC (Aquasil-SS column) with a UV absorption detector was used to analyze neutral glycosphingolipid. These chromatograms showed two typical peaks in clove lipids. UV-HPLC (C8 -reversed phase silica gel column) was also used to analyze glycoglycerolipid. The chromatograms indicated a small peak in clove lipids. Moreover, we observed the same two peaks in the glycolipid fraction of clove lipid when we used HPLC (Aquasil-SS column) with a differential refractometer detector. These results suggest that clove may contain new and plural neutral glycosphingolipids. [source] Compounds from Ageratum conyzoides: isolation, structural elucidation and insecticidal activityPEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 6 2007Márcio D Moreira Abstract This work aimed at identifying plant compounds with insecticidal activity against Diaphania hyalinata (L.) (Lepidoptera: Pyralidae), Musca domestica (L.) (Diptera: Muscidae), Periplaneta americana (L.) (Blattodea: Blattidae) and Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae). The plant species used were: basil (Ocimum selloi Benth.), rue (Ruta graveolens L.), lion's ear (Leonotis nepetaefolia L.), Jimson weed (Datura stramonium L.), ,baleeira' herb (Cordia verbenaceae L.), mint (Mentha piperita L.), wild balsam apple (Mormodica charantia L.) and billy goat weed (Ageratum conyzoides L.). Firstly, the insecticidal activities of hexane and ethanol plant extracts were evaluated against adults of R. dominica. Among them, only the hexane extract of A. conyzoides showed insecticidal activity. The hexane extract of this plant species was therefore fractionated by silica gel column chromatography to isolate and purify its bioactive chemical constituents. Three compounds were identified using IR spectra, 1H NMR, 13C NMR, HMBC and NOE after gel chromatography: 5,6,7,8,3,, 4,, 5,-heptamethoxyflavone, 5,6,7,8,3,-pentamethoxy-4,, 5,-methylenedioxyflavone and coumarin. The complete assignment of 13C NMR to 5,6,7,8,3,-pentamethoxy-4,, 5,-methylenedioxyflavone was successfully made for the first time. 5,6,7,8,3,-Pentamethoxy-4,, 5,-methylenedioxyflavone did not show any insecticidal activity against the four insect species tested. 5,6,7,8,3,, 4,, 5,-Heptamethoxyflavone showed low activity against D. hyalinata and R. dominica and was not toxic to M. domestica or P. americana. In contrast, coumarin showed insecticidal activity against all four insect pest species tested, with the following order of susceptibility: R. dominica < P. americana < D. hyalinata < M. domestica after 24 h exposure. Copyright © 2007 Society of Chemical Industry [source] Role of the N-terminal Region in the Function of the Photosynthetic Bacterium Transcription Regulator PpsR,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2008Yoichi Yamazaki PpsR is a transcription repressor for the gene cluster encoding photosystem genes in Rhodobacter sphaeroides. Repression activity is accomplished by DNA binding on the promoter regions of the photosystem gene clusters, and depends on both the redox potential and the presence of antirepressor protein AppA. To understand DNA repression regulation by PpsR, we investigated the function of PpsR domains in self-association for DNA binding. We constructed domain-deletion mutants and verified DNA-binding activity and dimer formation. Gel shift assay for measuring the DNA-binding activity of three sequential N-terminal deletion mutants revealed that N-terminal deletions (of minimum 121 residues) caused loss of binding activity. Size-exclusion gel chromatography revealed that deletion mutant which lacks the N-terminal 121-amino acid deletion mutant to exist as a dimer, although it was less stable than the intact PpsR. The mutants lacking the adjacent regions, Q-linker region and the first Per-Ant-Sim domain, did not form dimers, suggesting the involvement of the N-terminal region in dimer formation. This region is thus considered to be a functional domain in self-association, although not yet identified as a structural domain. Circular dichroism spectrum of the N-terminal region fragment exhibited a ,/, structure. We conclude that this region is a structural and functional domain, contributing to PpsR repression through dimer stabilization. [source] Nrf2-mediated induction of detoxifying enzymes by alantolactone present in Inula heleniumPHYTOTHERAPY RESEARCH, Issue 11 2008Ji Yeon Seo Abstract Our previous study showed that a methanol extract of Inula helenium had the potential to induce detoxifying enzymes such as quinone reductase (QR) and glutathione S -transferase (GST) activity. In this study the methanol extract was further fractionated using silica gel chromatography and vacuum liquid chromatography, to yield pure compounds alantolactone and isoalantolactone as QR inducers. Alantolactone caused a dose-dependent induction of antioxidant enzymes including QR, GST, , -glutamylcysteine synthase, glutathione reductase, and heme oxygenase 1 in hepa1c1c7 mouse hepatoma cells. The compound increased the luciferase activity of HepG2-C8 cells, transfectants carrying antioxidant response element (ARE)-luciferase gene, in a dose-dependent manner, suggesting ARE-mediated transcriptional activation of antioxidant enzymes. Alantolactone also stimulated the nuclear accumulation of Nrf2 that was inhibited by phosphatidylinositol 3-kinase (PI3K) inhibitors. In conclusion, alantolactone appears to induce detoxifying enzymes via activation of PI3K and JNK signaling pathways, leading to translocation of Nrf2, and subsequent interaction between Nrf2 and ARE in the encoding genes. Copyright © 2008 John Wiley & Sons, Ltd. [source] Proteomic analysis of high-density lipoproteinPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 2 2006Farhad Rezaee Dr. Abstract Plasma lipoproteins, such as high-density lipoprotein (HDL), can serve as carriers for a wide range of proteins that are involved in processes such as lipid metabolism, thrombosis, inflammation and atherosclerosis. The identification of HDL-associated proteins is essential with regards to understanding these processes at the molecular level. In this study, a combination of proteomic approaches including 1-DE and 2-DE MALDI-TOF, isotope-coded affinity tag and Western blot analysis were employed to identify proteins associated with human HDL. To minimize potential losses of HDL-associated proteins during isolation, a one-step ultracentrifugation technique was applied and the quality of purified HDL was confirmed by nephelometry, high-performance gel chromatography, and Western blot analysis. MS analysis revealed the presence of 56 HDL-associated proteins including all known apolipoproteins and lipid transport proteins. Furthermore, proteins involved in hemostasis and thrombosis, the immune and complement system were found. In addition, growth factors, receptors, hormone-associated proteins and many other proteins were found to be associated with HDL. Our approach thus resulted in the identification of a large number of proteins associated with HDL. The combination of proteomic technologies proved to be a powerful and comprehensive tool for the identification of proteins on HDL. [source] | ||||||||||||||||