{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,11]],"date-time":"2026-11-11T08:55:37Z","timestamp":1794387337924,"version":"4.9.0"},"reference-count":40,"publisher":"American Chemical Society (ACS)","issue":"19","license":[{"start":{"date-parts":[[2020,8,14]],"date-time":"2020-08-14T00:00:00Z","timestamp":1597363200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2020,8,14]],"date-time":"2020-08-14T00:00:00Z","timestamp":1597363200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2020,8,14]],"date-time":"2020-08-14T00:00:00Z","timestamp":1597363200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/100006151","name":"Basic Energy Sciences","doi-asserted-by":"publisher","award":["DE-SC0015662"],"award-info":[{"award-number":["DE-SC0015662"]}],"id":[{"id":"10.13039\/100006151","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,10,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Plant cell wall polysaccharide analysis encompasses the utilization of a variety of analytical tools, including gas and liquid chromatography, mass spectrometry (MS), and nuclear magnetic resonance (NMR) spectroscopy. These methods provide complementary data, which enable confident structural proposals of the many complex polysaccharide structures that exist in the complex matrices of plant cell walls. However, cell walls contain fractions of varying solubilities, and a few techniques are available that can analyze all fractions simultaneously. We have discovered that permethylation affords the complete dissolution of both soluble and insoluble polysaccharide fractions of plant cell walls in organic solvents such as chloroform or acetonitrile, which can then be analyzed by a number of analytical techniques including MS and NMR. In this work, NMR structure analysis of 10 permethylated polysaccharide standards was undertaken to generate chemical shift data providing insights into spectral changes that result from permethylation of polysaccharide residues. This information is of especial relevance to the structure analysis of insoluble polysaccharide materials that otherwise are not easily investigated by solution-state NMR methodologies. The preassigned NMR chemical shift data is shown to be vital for NMR structure analysis of minor polysaccharide components of plant cell walls that are particularly difficult to assign by NMR correlation data alone. With the assigned chemical shift data, we analyzed the permethylated samples of destarched, alcohol-insoluble residues of switchgrass and poplar by two-dimensional NMR spectral profiling. Thus, we identified, in addition to the major polysaccharide components, two minor polysaccharides, namely, &amp;lt;5% 3-linked arabinoxylan (switchgrass) and &amp;lt;2% glucomannan (poplar). In particular, the position of the arabinose residue in the arabinoxylan of the switchgrass sample was confidently assigned based on chemical shift values, which are highly sensitive to local chemical environments. Furthermore, the high resolution afforded by the 1H NMR spectra of the permethylated switchgrass and poplar samples allowed facile relative quantitative analysis of their polysaccharide composition, utilizing only a few milligrams of the cell wall material. The concepts herein developed will thus facilitate NMR structure analysis of insoluble plant cell wall polysaccharides, more so of minor cell wall components that are especially challenging to analyze with current methods.<\/jats:p>","DOI":"10.1021\/acs.analchem.0c02379","type":"journal-article","created":{"date-parts":[[2020,8,14]],"date-time":"2020-08-14T14:45:48Z","timestamp":1597416348000},"page":"13221-13228","source":"Crossref","is-referenced-by-count":28,"title":["Evaluating the Utility of Permethylated Polysaccharide\nSolution NMR Data for Characterization of Insoluble Plant Cell Wall\nPolysaccharides"],"prefix":"10.1021","volume":"92","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2412-6970","authenticated-orcid":true,"given":"Ikenna\nE.","family":"Ndukwe","sequence":"first","affiliation":[{"name":"University of Georgia , , 315 Riverbend Road , , ,","place":["Athens, Georgia, United States, 30602"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian","family":"Black","sequence":"additional","affiliation":[{"name":"University of Georgia , , 315 Riverbend Road , , ,","place":["Athens, Georgia, United States, 30602"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1118-8953","authenticated-orcid":true,"given":"Christian","family":"Heiss","sequence":"additional","affiliation":[{"name":"University of Georgia , , 315 Riverbend Road , , ,","place":["Athens, Georgia, United States, 30602"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parastoo","family":"Azadi","sequence":"additional","affiliation":[{"name":"University of Georgia , , 315 Riverbend Road , , ,","place":["Athens, Georgia, United States, 30602"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,8,14]]},"reference":[{"key":"2026081803341381100_cit1","doi-asserted-by":"publisher","first-page":"831","DOI":"10.1641\/0006-3568(2004)054[0831:TCWTBT]2.0.CO;2","article-title":"The Cell\nWalls That Bind the Tree of Life","volume":"54","author":"Niklas","year":"2004","journal-title":"Bioscience"},{"key":"2026081803341381100_cit2","doi-asserted-by":"publisher","first-page":"567","DOI":"10.1146\/annurev-arplant-042110-103809","article-title":"Evolution and Diversity\nof Plant Cell Walls: From Algae to Flowering Plants","volume":"62","author":"Popper","year":"2011","journal-title":"Annu. Rev. Plant Biol."},{"key":"2026081803341381100_cit3","doi-asserted-by":"publisher","first-page":"1879","DOI":"10.1016\/j.carres.2009.05.021","article-title":"The Structure, Function,\nand Biosynthesis of Plant\nCell Wall Pectic Polysaccharides","volume":"344","author":"Caffall","year":"2009","journal-title":"Carbohydr.\nRes."},{"key":"2026081803341381100_cit4","doi-asserted-by":"publisher","first-page":"1854","DOI":"10.3389\/fpls.2016.01854","article-title":"Emerging\nTechnologies for the Production\nof Renewable Liquid Transport Fuels from Biomass Sources Enriched\nin Plant Cell Walls","volume":"7","author":"Tan","year":"2016","journal-title":"Front. Plant Sci."},{"key":"2026081803341381100_cit5","doi-asserted-by":"publisher","first-page":"347","DOI":"10.1038\/s41467-018-08252-0","article-title":"Lignin-Polysaccharide Interactions\nin Plant Secondary Cell Walls Revealed by Solid-State NMR","volume":"10","author":"Kang","year":"2019","journal-title":"Nat. Commun."},{"key":"2026081803341381100_cit6","doi-asserted-by":"publisher","first-page":"101660","DOI":"10.1016\/j.ssnmr.2020.101660","article-title":"Solid-State\nNMR of Plant and Fungal Cell Walls: A Critical Review","volume":"107","author":"Zhao","year":"2020","journal-title":"Solid State Nucl. Magn. Reson."},{"key":"2026081803341381100_cit7","doi-asserted-by":"publisher","first-page":"1226","DOI":"10.1016\/j.jchromb.2010.11.013","article-title":"Derivatization\nof Carbohydrates\nfor GC and GC\u2013MS Analyses","volume":"879","author":"Ruiz-Matute","year":"2011","journal-title":"J. Chromatogr.,\nB"},{"key":"2026081803341381100_cit8","doi-asserted-by":"publisher","first-page":"987","DOI":"10.1016\/S1044-0305(97)00124-4","article-title":"Tandem Mass Spectrometry\nof Alkali Cationized Polysaccharides\nin a Quadrupole Ion Trap","volume":"8","author":"Asam","year":"1997","journal-title":"J. Am. Soc. Mass Spectrom."},{"key":"2026081803341381100_cit9","doi-asserted-by":"publisher","first-page":"937","DOI":"10.1002\/rcm.1290091014","article-title":"Matrix-Assisted\nLaser Desorption\/Ionization Mass Spectrometry of Polysaccharides","volume":"9","author":"Garrozzo","year":"1995","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"2026081803341381100_cit10","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3389\/fpls.2012.00045","article-title":"Mass Spectrometry\nfor Characterizing Plant Cell Wall Polysaccharides","volume":"3","author":"Bauer","year":"2012","journal-title":"Front. Plant Sci."},{"key":"2026081803341381100_cit11","doi-asserted-by":"publisher","first-page":"4589","DOI":"10.1021\/cr990302n","article-title":"Carbohydrate Structural\nDetermination by NMR Spectroscopy: Modern Methods and Limitations\n\u2020","volume":"100","author":"Duus","year":"2000","journal-title":"Chem. 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Protoc."},{"key":"2026081803341381100_cit15","doi-asserted-by":"publisher","first-page":"1532","DOI":"10.1002\/elps.201500561","article-title":"LC-MS\/MS Analysis of\nPermethylated\nFree Oligosaccharides and N-Glycans Derived from Human, Bovine, and\nGoat Milk Samples","volume":"37","author":"Dong","year":"2016","journal-title":"Electrophoresis"},{"key":"2026081803341381100_cit16","first-page":"83","article-title":"Analysis of Permethylated\nGlycan by Liquid Chromatography (LC) and Mass Spectrometry (MS)","volume-title":"Methods in Molecular Biology","author":"Zhou","year":"2017"},{"key":"2026081803341381100_cit17","doi-asserted-by":"publisher","first-page":"669","DOI":"10.1021\/cr60286a004","article-title":"Nuclear Magnetic\nResonance in Carbohydrate Chemistry","volume":"73","author":"Kotowycz","year":"1973","journal-title":"Chem. 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Binding Specificity and Calorimetric Analysis \u2020","volume":"35","author":"Tomme","year":"1996","journal-title":"Biochemistry"},{"key":"2026081803341381100_cit20","doi-asserted-by":"publisher","first-page":"627","DOI":"10.1002\/(SICI)1521-3935(20000301)201:6&lt;627::AID-MACP627&gt;3.0.CO;2-Y","article-title":"Effective Preparation\nof Cellulose Derivatives in a New Simple Cellulose\nSolvent","volume":"201","author":"Heinze","year":"2000","journal-title":"Macromol. Chem. 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Chem."},{"key":"2026081803341381100_cit23","doi-asserted-by":"publisher","first-page":"7549","DOI":"10.1039\/C3RA46338A","article-title":"A Gel-State 2D-NMR\nMethod for Plant Cell Wall Profiling and Analysis:\nA Model Study with the Amorphous Cellulose and Xylan from Ball-Milled\nCotton Linters","volume":"4","author":"Kim","year":"2014","journal-title":"RSC Adv."},{"key":"2026081803341381100_cit24","doi-asserted-by":"publisher","first-page":"576","DOI":"10.1039\/B916070A","article-title":"Solution-State 2D NMR of Ball-Milled Plant Cell Wall Gels in DMSO-D6\/Pyridine-D5","volume":"8","author":"Kim","year":"2010","journal-title":"Org. Biomol. Chem."},{"key":"2026081803341381100_cit25","doi-asserted-by":"publisher","first-page":"1579","DOI":"10.1038\/nprot.2012.064","article-title":"Whole Plant Cell Wall Characterization\nUsing Solution-State 2D NMR","volume":"7","author":"Mansfield","year":"2012","journal-title":"Nat. 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Spectroscopy of\nPermethylated Gluco-, Galacto-, and Manno-Pyranoses and Their 6-Deoxy\nAnalogues","volume":"39","author":"Haverkamp","year":"1975","journal-title":"Carbohydr. Res."},{"key":"2026081803341381100_cit29","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1016\/S0008-6215(00)82810-4","article-title":"13C-\nand 1H-N.M.R. Spectroscopy of\nPermethylated \u03b1- and \u03b2-D-Galactopyranoses","volume":"33","author":"Haverkamp","year":"1974","journal-title":"Carbohydr. Res."},{"key":"2026081803341381100_cit30","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/S0008-6215(00)87068-8","article-title":"13C-\nand 1H-Nuclear Magnetic Resonance\nSpectroscopy of Permethylated Disaccharides","volume":"37","author":"Haverkamp","year":"1974","journal-title":"Carbohydr. Res."},{"key":"2026081803341381100_cit31","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1016\/0008-6215(84)85242-8","article-title":"A Simple and Rapid\nMethod for the Permethylation of Carbohydrates","volume":"131","author":"Ciucanu","year":"1984","journal-title":"Carbohydr. Res."},{"key":"2026081803341381100_cit32","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1016\/0003-2697(92)90048-C","article-title":"A Comprehensive\nProcedure for Preparation of Partially\nMethylated Alditol Acetates from Glycoprotein Carbohydrates","volume":"203","author":"Anumula","year":"1992","journal-title":"Anal. Biochem."},{"key":"2026081803341381100_cit33","doi-asserted-by":"publisher","first-page":"1514","DOI":"10.1021\/ac1032534","article-title":"Structural Analysis\nof Glycans by NMR Chemical Shift\nPrediction","volume":"83","author":"Lundborg","year":"2011","journal-title":"Anal. Chem."},{"key":"2026081803341381100_cit34","doi-asserted-by":"publisher","first-page":"344","DOI":"10.1002\/mrc.1270110706","article-title":"Spectres RMN Des Polysaccharides\net de Leurs De\u0301rive\u0301s: Influence Des Substituants Sur\nLe De\u0301placement Chimique13C","volume":"11","author":"Gagnaire","year":"1978","journal-title":"Org. Magn.\nReson."},{"key":"2026081803341381100_cit35","doi-asserted-by":"publisher","first-page":"216","DOI":"10.1002\/mrc.1270120410","article-title":"13C-NMR-Untersuchungen Zur Strukturaufkla\u0308rung\nvon Polysacchariden Und Deren Methylderivaten","volume":"12","author":"Friebolin","year":"1979","journal-title":"Org. Magn. Reson."},{"key":"2026081803341381100_cit36","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1016\/0008-6215(86)85069-8","article-title":"Analysis of\nthe 13C-n.m.r. Spectra of Hydrolyzed and Methanolyze O-Methylcelluloses:\nMonomer Compositions and Models for Their Description","volume":"157","author":"Reuben","year":"1986","journal-title":"Carbohydr. Res."},{"key":"2026081803341381100_cit37","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1016\/j.carres.2018.02.002","article-title":"Partially Methylated Galactans Containing\nDifferent Proportions of 3-O-Methyl-Galactose from Pleurotus Citrinopileatus","volume":"458-459","author":"Brito","year":"2018","journal-title":"Carbohydr. Res."},{"key":"2026081803341381100_cit38","doi-asserted-by":"publisher","first-page":"898","DOI":"10.1039\/c3ee23801f","article-title":"Investigating\nPlant Cell Wall Components That Affect\nBiomass Recalcitrance in Poplar and Switchgrass","volume":"6","author":"Demartini","year":"2013","journal-title":"Energy Environ. Sci."},{"key":"2026081803341381100_cit39","doi-asserted-by":"publisher","first-page":"249","DOI":"10.1038\/nbt.4067","article-title":"Sugar\nRelease and Growth of Biofuel Crops Are Improved by Downregulation\nof Pectin Biosynthesis","volume":"36","author":"Biswal","year":"2018","journal-title":"Nat. Biotechnol."},{"key":"2026081803341381100_cit40","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1007\/s00425-012-1588-z","article-title":"Distribution of Glucomannans and Xylans in Poplar Xylem\nand Their Changes under Tension Stress","volume":"236","author":"Kim","year":"2012","journal-title":"Planta"}],"container-title":["Analytical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.analchem.0c02379","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/92\/19\/13221\/8594482\/ac0c02379.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/92\/19\/13221\/8594482\/ac0c02379.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T08:57:07Z","timestamp":1787043427000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/ancham\/article\/92\/19\/13221\/828355\/Evaluating-the-Utility-of-Permethylated"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,14]]},"references-count":40,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2020,8,14]]},"published-print":{"date-parts":[[2020,10,6]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.analchem.0c02379","relation":{"has-part":[{"id-type":"doi","id":"https:\/\/doi.org\/10.1021\/acs.analchem.0c02379.s001","asserted-by":"subject"}]},"ISSN":["0003-2700","1520-6882"],"issn-type":[{"value":"0003-2700","type":"print"},{"value":"1520-6882","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,14]]}}}