Complementary analysis of lipids in whole bacteria cells by thermally assisted hydrolysis and methylation-GC and MALDI-MS combined with on-probe sample pretreatment |
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| Institution: | 1. Division of Energy Science, EcoTopia Science Institute, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan;2. Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan;3. Department of Materials Science and Engineering, Graduate School of Engineering, Nagoya Institute of Technology, Nagoya 466-8555, Japan;1. Department of Chemistry, Michigan State University, East Lansing, MI, USA;2. Thermo Fisher Scientific, San Jose, CA, USA;3. School of Chemistry, Department of Biochemistry and Molecular Biology, Bio21 Institute of Molecular Science and Biotechnology, The University of Melbourne, Parkville, VIC, Australia;1. Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, Denmark;2. School of Pharmacy, University of Oslo, P.O. Box 1068, Blindern, 0316 Oslo, Norway;1. School of Physics and Engineering, Qufu Normal University, 273100 Qufu, Shandong, PR China;2. School of Physics and Optoelectronics Engineering, Ludong University, 264025 Yantai, Shandong, PR China;3. School of Energy Engineering, Xinjiang Institute of Engineering, Urumqi 830091, PR China;1. Department of Ophthalmology, Gifu University Graduate School of Medicine, Gifu, Japan;2. Department of Ophthalmology, Toho University Ohashi Medical Center, Tokyo, Japan;3. Laboratory for Clinical Investigation, Gifu University Hospital, Gifu, Japan;4. Department of Microbiology, Tokyo Medical University Graduate School of Medicine, Tokyo, Japan;1. Okazaki Institute for Integrative Bioscience, Institute for Molecular Science, National Institutes of Natural Sciences, Aichi 444-8787, Japan;2. Department of Functional Molecular Science, School of Physical Sciences, The Graduate University for Advanced Studies (SOKENDAI), Kanagawa 240-0193, Japan;3. Department of Physics, Faculty of Science and Engineering, Chuo University, Tokyo 112-8551, Japan;4. Department of Applied Chemistry, The University of Tokyo, Tokyo 113-8656, Japan;5. Department of Chemistry, Graduate School of Science, Chiba University, Chiba 263-8522, Japan;6. JST, PRESTO, Chiba 263-8522, Japan;1. School of Chemistry and Chemical Engineering, Southwest University, No. 2 Tiansheng Road, BeiBei District, Chongqing 400715, China;2. Department of Radiology, University of Minnesota, 420 Delaware Street SE, Minneapolis, MN 55455, USA;3. School of Chemistry, Chemical Engineering and Life Sciences, Wuhan University of Technology, Wuhan 430070, China;4. Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200062, China |
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| Abstract: | The molecular structure of lipids in whole bacteria cells was characterized in detail by using two different and complementarily direct analyses; thermally assisted hydrolysis and methylation-gas chromatography (THM-GC) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) combined with on-probe sample pretreatment. First, THM-GC in the presence of tetramethylammonium hydroxide (TMAH) was applied to compositional analysis of the fatty acid components of lipids in whole bacterial cells. On the resulting chromatograms, a series of fatty acid components in bacterial lipids were clearly observed as resolved peaks of their methyl esters. The fatty acid compositions determined on the basis of the peak intensities were in good agreement with those obtained by the conventional technique involving solvent extraction of the lipids in the samples. Furthermore, MALDI-MS combined with the on-probe sample treatment, using 2,5-dihydroxybenzoic acid (DHB) and sodium iodide (NaI) as matrix and cationization reagents, respectively, was used to detect intact phospholipids directly from whole bacterial cells. The MALDI spectra thus obtained showed an array of ions generated from bacterial phospholipids, such as phosphatidylethanolamines (PEs) and phosphatidylglycerols (PGs). Finally, the bacterial lipids were comprehensively characterized in terms of the acyl groups and the molecular structures by taking both of the results obtained by THM-GC and MALDI-MS into consideration. |
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