DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Determination of hydroxyl groups in biorefinery resources via quantitative 31P NMR spectroscopy [Determination of hydroxyl groups in biorefinery resources via quantitative 31P Nuclear Magnetic Resonance Spectroscopy]

Journal Article · · Nature Protocols
ORCiD logo [1]; ORCiD logo [2];  [3];  [1]; ORCiD logo [4]; ORCiD logo [5]; ORCiD logo [6]
  1. The Univ. of Tennessee, Knoxville, TN (United States)
  2. Ca’ Foscari Univ. of Venice, Venice (Italy)
  3. Qingdao Univ., Qingdao (China)
  4. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  5. The Univ. of Tennessee, Knoxville, TN (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); The Univ. of Tennessee Inst. of Agriculture, Knoxville, TN (United States)
  6. North Carolina State Univ., Raleigh, NC (United States)

The analysis of chemical structural characteristics of biorefinery product streams (such as lignin and tannin) has advanced substantially over the past decade, with traditional wet-chemical techniques being replaced or supplemented by NMR methodologies. Quantitative 31P NMR spectroscopy is a promising technique for the analysis of hydroxyl groups because of its unique characterization capability and broad potential applicability across the biorefinery research community. This protocol describes procedures for (i) the preparation/solubilization of lignin and tannin, (ii) the phosphitylation of their hydroxyl groups, (iii) NMR acquisition details, and (iv) the ensuing data analyses and means to precisely calculate the content of the different types of hydroxyl groups. Compared with traditional wet-chemical techniques, the technique of quantitative 31P NMR spectroscopy offers unique advantages in measuring hydroxyl groups in a single spectrum with high signal resolution. Furthermore, the method provides complete quantitative information about the hydroxyl groups with small amounts of sample (~30 mg) within a relatively short experimental time (~30–120 min).

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC05-00OR22725
OSTI ID:
1558488
Journal Information:
Nature Protocols, Vol. 14, Issue 9; ISSN 1754-2189
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 230 works
Citation information provided by
Web of Science

References (170)

Synthesis and characterization of biobased poly(butylene succinate- ran -butylene adipate). Analysis of the composition-dependent physicochemical properties journal February 2017
A polymer of caffeyl alcohol in plant seeds journal January 2012
1 H– 13 C HSQC NMR Spectroscopy for Estimating Procyanidin/Prodelphinidin and cis / trans -Flavan-3-ol Ratios of Condensed Tannin Samples: Correlation with Thiolysis journal February 2015
Determination of Cellulose Reactivity by Using Phosphitylation and Quantitative 31 P NMR Spectroscopy journal November 2008
Artificial miRNA inhibition of phosphoenolpyruvate carboxylase increases fatty acid production in a green microalga Chlamydomonas reinhardtii journal April 2017
31P NMR spectroscopy in the quality control and authentication of extra-virgin olive oil: A review of recent progress journal January 2007
Characterization and analysis of the molecular weight of lignin for biorefining studies journal June 2014
Thermo-responsive and fluorescent cellulose nanocrystals grafted with polymer brushes journal January 2015
Correlations of the Antioxidant Properties of Softwood Kraft Lignin Fractions with the Thermal Stability of Its Blends with Polyethylene journal January 2015
Revealing the structure and distribution changes of Eucalyptus lignin during the hydrothermal and alkaline pretreatments journal April 2017
A thioacidolysis method tailored for higher-throughput quantitative analysis of lignin monomers journal September 2016
Small-scale and automatable high-throughput compositional analysis of biomass journal October 2010
Determination of the Diglyceride Content in Greek Virgin Olive Oils and Some Commercial Olive Oils by Employing 31 P NMR Spectroscopy journal April 2002
Characterization of Whole Biomasses in Pyridine Based Ionic Liquid at Low Temperature by 31P NMR: An Approach to Quantitatively Measure Hydroxyl Groups in Biomass As Their Original Structures journal March 2018
Chemical Groups and Structural Characterization of Lignin via Thiol-Mediated Demethylation journal December 2013
Determination of Carbonyl Groups of Six Round Robin Lignins by Modified Oximation and FTIR Spectroscopy journal January 1998
Diversity of structure and function in oligomeric flavanoids journal January 1992
Products and Functional Group Distributions in Pyrolysis Oil of Chromated Copper Arsenate (CCA)-Treated Wood, as Elucidated by Gas Chromatography and a Novel 31 P NMR-Based Method journal August 2007
Proanthocyanidin Biosynthesis — Still More Questions than Answers? journal January 2006
A Tale of Two Monasteries. Westminster and Saint-Denis in the Thirteenth Century journal January 2009
Loss of function of cinnamyl alcohol dehydrogenase 1 leads to unconventional lignin and a temperature-sensitive growth defect in Medicago truncatula journal July 2013
Catalytic Oxidation of Lignins into the Aromatic Aldehydes: General Process Trends and Development Prospects journal November 2017
Application of 31P NMR spectroscopy and chemical derivatization for metabolite profiling of lipophilic compounds in human serum journal July 2009
Arylpropane-1,3-diols in Lignins from Normal and CAD-Deficient Pines journal July 1999
Unveiling the structure and ultrastructure of lignin carbohydrate complexes in softwoods journal November 2013
Cellulose Nanocrystal Microcapsules as Tunable Cages for Nano- and Microparticles journal October 2015
The strong association of condensed phenolic moieties in isolated lignins with their inhibition of enzymatic hydrolysis journal January 2016
Determination of Water Content in Olive Oil by 31 P NMR Spectroscopy journal March 2008
The Path Forward for Biofuels and Biomaterials journal January 2006
Structural Characterization of Lignin during Pinus taeda Wood Treatment with Ceriporiopsis subvermispora journal July 2004
Rapid visual estimation and spectrophotometric determination of tannin content of sorghum grain journal November 1977
Imidazolium-Based Ionic Liquids as Efficient Reagents for the C−O Bond Cleavage of Lignin journal December 2017
Fast dissolution pretreatment of the corn stover in gamma-valerolactone promoted by ionic liquids: Selective delignification and enhanced enzymatic saccharification journal December 2018
Determination of Hydroxyl Groups in Lignins Evaluation of 1 H-, 13 C-, 31 P-NMR, FTIR and Wet Chemical Methods journal January 1994
Characteristics of Lignin Fractions from Dilute Acid Pretreated Switchgrass and Their Effect on Cellobiohydrolase from Trichoderma longibrachiatum journal February 2018
A new method for the analysis of phenolic groups in lignins by ʹH NMR spectrometry journal August 1994
Recent innovations in analytical methods for the qualitative and quantitative assessment of lignin journal September 2015
An Unusual Lignin from Kenaf journal January 1996
Hydroxystilbenes Are Monomers in Palm Fruit Endocarp Lignins journal June 2017
Quantitative hsqc Analyses of Lignin: a Practical Comparison journal March 2013
Fast, Easy, and Economical Quantification of Lignin Phenolic Hydroxyl Groups: Comparison with Classical Techniques journal April 2018
Determination of Arylglycerol-β-aryl Ethers and Other Linkages in Lignins Using DFRC/ 31 P NMR journal February 2001
Tannin Structural Elucidation and Quantitative 31 P NMR Analysis. 1. Model Compounds journal September 2013
Quality assessment and authentication of virgin olive oil by NMR spectroscopy: A critical review journal February 2013
Quantitative Phosphorus-31 NMR Analysis of Six Soluble Lignins journal February 1994
Lignin Valorization: Improving Lignin Processing in the Biorefinery journal May 2014
2-Chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane, a Reagent for the Accurate Determination of the Uncondensed and Condensed Phenolic Moieties in Lignins journal June 1995
Quantitative Phosphorus-31 NMR Analysis of Lignins, a New Tool for the Lignin Chemist journal February 1994
A comparative C13 NMR study of polyflavonoid tannin extracts for phenolic polycondensates journal December 1993
Lignin Structural Variation in Hardwood Species journal May 2012
New insights into the structure and composition of technical lignins: a comparative characterisation study journal January 2016
Thermal properties of lignin in copolymers, blends, and composites: a review journal January 2015
Characterization of fractional cuts of co-solvent enhanced lignocellulosic fractionation lignin isolated by sequential precipitation journal January 2019
Structural Characterization of Lignin in Wild-Type versus COMT Down-Regulated Switchgrass journal January 2014
Genetic manipulation of lignin reduces recalcitrance and improves ethanol production from switchgrass journal February 2011
A new method for rapid degree of substitution and purity determination of chloroform-soluble cellulose esters, using 31P NMR journal January 2010
Classification of Edible Oils by Employing 31 P and 1 H NMR Spectroscopy in Combination with Multivariate Statistical Analysis. A Proposal for the Detection of Seed Oil Adulteration in Virgin Olive Oils journal September 2003
Modelling the kinetics of pyrolysis oil hydrothermal upgrading based on the connectivity of oxygen atoms, quantified by 31 P-NMR journal March 2017
Detection and Quantification of Free Glycerol in Virgin Olive Oil by 31P-NMR Spectroscopy journal September 2009
Toward Thermoplastic Lignin Polymers. Part 1. Selective Masking of Phenolic Hydroxyl Groups in Kraft Lignins via Methylation and Oxypropylation Chemistries
  • Sadeghifar, Hasan; Cui, Chengzhong; Argyropoulos, Dimitris S.
  • Industrial & Engineering Chemistry Research, Vol. 51, Issue 51, p. 16713-16720 https://doi.org/10.1021/ie301848j
journal December 2012
Lignin Structural Variation in Hardwood Species journal May 2012
Quantifying Biomass Composition by Gas Chromatography/Mass Spectrometry journal September 2014
Biosynthesis and genetic engineering of proanthocyanidins and (iso)flavonoids journal October 2007
Antioxidant activity and total phenolic content of an isolated Morinda citrifolia L. methanolic extract from Poly-ethersulphone (PES) membrane separator journal January 2015
Milled Wood Lignin: A Linear Oligomer journal November 2011
31 P NMR Spectroscopy in Wood Chemistry. Part VI. Solid State 31 P NMR of Trimethyl Phosphite Derivatives of Chromophores and Carboxylic Acids Present in Mechanical Pulps; a Method for the Quantitative Determination of ortho -Quinones journal January 1994
Lignin genetic engineering for improvement of wood quality: Applications in paper and textile industries, fodder and bioenergy production journal March 2014
Elucidation of Lignin Structure by Quantitative 2D NMR journal June 2011
Universal fractionation of lignin-carbohydrate complexes (LCCs) from lignocellulosic biomass: an example using spruce wood journal March 2013
Detection of Extra Virgin Olive Oil Adulteration with Lampante Olive Oil and Refined Olive Oil Using Nuclear Magnetic Resonance Spectroscopy and Multivariate Statistical Analysis journal April 2005
A new method for the analysis of phenolic groups in lignins by ʹH NMR spectrometry journal August 1994
31 P NMR Spectroscopy in Wood Chemistry. I. Model Compounds journal June 1991
Hydrothermal deoxygenation of pyrolysis oil from Norwegian spruce: Picea abies journal September 2013
Advances in solid-state NMR of cellulose journal June 2014
Determination of phenolic hydroxyl groups in residual lignin using a modified UV-method journal May 1999
31 P NMR Characterization of Tricin and Its Structurally Similar Flavonoids journal April 2017
Application of 31 P NMR Spectroscopy in Food Analysis. 1. Quantitative Determination of the Mono- and Diglyceride Composition of Olive Oils journal March 2000
FTIR-ATR-based prediction and modelling of lignin and energy contents reveals independent intra-specific variation of these traits in bioenergy poplars journal January 2011
Rapid in vivo lipid/carbohydrate quantification of single microalgal cell by Raman spectral imaging to reveal salinity-induced starch-to-lipid shift journal January 2017
Synthesis and characterization of biobased poly(butylene succinate- ran -butylene adipate). Analysis of the composition-dependent physicochemical properties journal February 2017
Tricin, a Flavonoid Monomer in Monocot Lignification journal February 2015
N-Hydroxy Compounds as New Internal Standards for the 31P-NMR Determination of Lignin Hydroxy Functional Groups journal April 2001
Structural characterization of birch lignin isolated from a pressurized hot water extraction and mild alkali pulped biorefinery process journal January 2018
Unveiling the structure and ultrastructure of lignin carbohydrate complexes in softwoods journal November 2013
Application of quantitative 31P NMR in biomass lignin and biofuel precursors characterization journal January 2011
Protein precipitation method for the quantitative determination of tannins journal July 1978
Chemical Transformations of Poplar Lignin during Cosolvent Enhanced Lignocellulosic Fractionation Process journal May 2018
Determination of the Diglyceride Content in Greek Virgin Olive Oils and Some Commercial Olive Oils by Employing 31 P NMR Spectroscopy journal April 2002
Application of 31 P NMR Spectroscopy in Food Analysis. 1. Quantitative Determination of the Mono- and Diglyceride Composition of Olive Oils journal March 2000
Die Verwendung von natürlichen Polyphenolen zur Herstellung von Holzleimen journal July 1980
Proanthocyanidin biosynthesis – still more questions than answers? journal September 2005
31P NMR spectroscopy in the quality control and authentication of extra-virgin olive oil: A review of recent progress journal January 2007
Determination of Hemicellulose, Cellulose and Lignin in Moso Bamboo by Near Infrared Spectroscopy journal November 2015
Recent innovations in analytical methods for the qualitative and quantitative assessment of lignin journal September 2015
Tannin Structural Elucidation and Quantitative 31 P NMR Analysis. 2. Hydrolyzable Tannins and Proanthocyanidins journal September 2013
In-depth investigation on quantitative characterization of pyrolysis oil by 31 P NMR journal January 2016
On the Quantification of Lignin Hydroxyl Groups With 31 P and 13 C NMR Spectroscopy journal April 2015
Structural elucidation of tannins of spent coffee grounds by CP-MAS 13C NMR and MALDI-TOF MS journal July 2015
31P NMR in wood chemistry: A review of recent progress journal March 1995
Synthesis of potentially biobased copolyesters based on adipic acid and butanediols: Kinetic study between 1,4- and 2,3-butanediol and their influence on crystallization and thermal properties journal September 2016
Quality assessment and authentication of virgin olive oil by NMR spectroscopy: A critical review journal February 2013
Tannin Structural Elucidation and Quantitative 31 P NMR Analysis. 1. Model Compounds journal September 2013
Quantitative hsqc Analyses of Lignin: a Practical Comparison journal March 2013
In Situ Determination of Lignin Phenolics and Wood Solubility in Imidazolium Chlorides Using 31 P NMR journal September 2009
A thioacidolysis method tailored for higher-throughput quantitative analysis of lignin monomers journal September 2016
Lignin genetic engineering for improvement of wood quality: Applications in paper and textile industries, fodder and bioenergy production journal March 2014
Solving the interaction of electromagnetic wave with electron by VIM journal January 2015
Revealing the structure of bamboo lignin obtained by formic acid delignification at different pressure levels journal December 2017
Selective precipitation and characterization of lignin–carbohydrate complexes (LCCs) from Eucalyptus journal January 2018
Enzyme adsorption properties on dilute acid pretreated biomass by low vacuum-scanning electron microscopy and structural analysis of lignin journal August 2018
Fractionating Wheat Straw via Phosphoric Acid with Hydrogen Peroxide Pretreatment and Structural Elucidation of the Derived Lignin journal January 2018
Quantifying Biomass Composition by Gas Chromatography/Mass Spectrometry journal September 2014
On the structure of softwood kraft lignin journal January 2017
31 P-NMR analysis of bio-oils obtained from the pyrolysis of biomass journal November 2010
Solution-state 2D NMR of Ball-milled Plant Cell Wall Gels in DMSO-d 6 journal March 2008
The Path Forward for Biofuels and Biomaterials journal January 2006
Detailed Chemical Composition of Condensed Tannins via Quantitative 31 P NMR and HSQC Analyses: Acacia catechu , Schinopsis balansae , and Acacia mearnsii journal August 2016
Quantification of Lignin and Its Structural Features in Plant Biomass Using 13 C Lignin as Internal Standard for Pyrolysis-GC-SIM-MS journal October 2017
Correlations of the Antioxidant Properties of Softwood Kraft Lignin Fractions with the Thermal Stability of Its Blends with Polyethylene journal January 2015
Application of FTIR spectroscopy to the characterization of archeological wood journal January 2016
Lignin Biosynthesis and Structure journal May 2010
Small-scale and automatable high-throughput compositional analysis of biomass text January 2010
Catalytic Oxidation of Lignins into the Aromatic Aldehydes: General Process Trends and Development Prospects journal November 2017
Milled Wood Lignin: A Linear Oligomer journal November 2011
Lignin Valorization: Improving Lignin Processing in the Biorefinery journal May 2014
2-Chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane, a Reagent for the Accurate Determination of the Uncondensed and Condensed Phenolic Moieties in Lignins journal June 1995
A comparative C13 NMR study of polyflavonoid tannin extracts for phenolic polycondensates journal December 1993
Quantitative 31P NMR Spectroscopy of Lignins from Transgenic Poplars journal June 2001
Quantitative 31P NMR analysis of solid wood offers an insight into the acetylation of its components journal November 2014
Lignin Analysis by Permanganate Oxidation. II. Lignins in Acidic Organosolv Pulps journal November 1999
31 P NMR Spectroscopy in Wood Chemistry Part V. Qualitative Analysis of Lignin Functional Groups journal June 1993
Survey of Lignin-Structure Changes and Depolymerization during Ionic Liquid Pretreatment journal September 2017
Determination of Arylglycerol-β-aryl Ethers and Other Linkages in Lignins Using DFRC/ 31 P NMR journal February 2001
Pyrolysis of Wood/Biomass for Bio-oil: A Critical Review journal May 2006
Toward Thermoplastic Lignin Polymers. Part 1. Selective Masking of Phenolic Hydroxyl Groups in Kraft Lignins via Methylation and Oxypropylation Chemistries
  • Sadeghifar, Hasan; Cui, Chengzhong; Argyropoulos, Dimitris S.
  • Industrial & Engineering Chemistry Research, Vol. 51, Issue 51, p. 16713-16720 https://doi.org/10.1021/ie301848j
journal December 2012
Whole plant cell wall characterization using solution-state 2D NMR journal August 2012
P NMR Spectroscopy in Wood Chemistry - Part III. Solid State 31 P NMR of Trimethyl Phosphite Derivatives of Chromophores in Mechanical Pulp journal January 1992
Application of 19 F NMR Spectroscopy for Content Determination of Fluorinated Pharmaceuticals journal January 2017
Universal fractionation of lignin-carbohydrate complexes (LCCs) from lignocellulosic biomass: an example using spruce wood journal March 2013
Biomass accessibility analysis using electron tomography journal December 2015
Characterization of technical lignins by NMR spectroscopy: optimization of functional group analysis by 31P NMR spectroscopy journal August 2015
A novel quantitative 31 P NMR spectroscopic analysis of hydroxyl groups in lignosulfonic acids journal January 2018
Review of NMR Characterization of Pyrolysis Oils journal August 2016
Lignin Biosynthesis and Structure journal May 2010
Coupling P-31 NMR with the Mannich reaction for the quantitative analysis of lignin journal January 1998
FTIR-ATR-based prediction and modelling of lignin and energy contents reveals independent intra-specific variation of these traits in bioenergy poplars journal January 2011
31 P NMR Spectroscopy in Wood Chemistry. Part IV. Lignin Models: Spin Lattice Relaxation Times and Solvent Effects in 31 P NMR journal January 1993
Quantitative 31P NMR analysis of solid wood offers an insight into the acetylation of its components journal November 2014
Evidence for Complex Molecular Architectures for Solvent-Extracted Lignins journal April 2012
Determination of phenolic hydroxyl groups in residual lignin using a modified UV-method journal May 1999
3D Chemical Image using TOF-SIMS Revealing the Biopolymer Component Spatial and Lateral Distributions in Biomass journal October 2012
Solution-state 2D NMR of ball-milled plant cell wall gels in DMSO-d6/pyridine-d5 journal January 2010
The occurrence of tricin and its derivatives in plants journal January 2016
Determination of Water Content in Olive Oil by 31 P NMR Spectroscopy journal March 2008
The Effects on Lignin Structure of Overexpression of Ferulate 5-Hydroxylase in Hybrid Poplar 1 journal April 2009
Facile Quantitative Analysis of Hydroxyl End Groups of Poly(2,6-dimethyl-1,4-phenylene oxide)s by 31P NMR Spectroscopy journal October 1994
Downregulation of pectin biosynthesis gene GAUT4 leads to reduced ferulate and lignin-carbohydrate cross-linking in switchgrass journal January 2019
Chemical Transformations of Buddleja davidii Lignin during Ethanol Organosolv Pretreatment journal April 2010
Elucidation of Lignin Structure by Quantitative 2D NMR journal June 2011
Lignins as macromonomers for polyurethane synthesis: A comparative study on hydroxyl group determination
  • Cateto, Carolina Andreia; Barreiro, Maria Filomena; Rodrigues, Alírio Egídio
  • Journal of Applied Polymer Science, Vol. 109, Issue 5 https://doi.org/10.1002/app.28393
journal January 2008
Bioconversion of hybrid poplar to ethanol and co-products using an organosolv fractionation process: Optimization of process yields journal January 2006
31 P NMR Chemical Shifts of Solvents and Products Impurities in Biomass Pretreatments journal November 2017
31P-N.m.r. spectroscopy in wood chemistry. Phosphite derivatives of carbohydrates journal November 1991
Tannin Structural Elucidation and Quantitative 31 P NMR Analysis. 2. Hydrolyzable Tannins and Proanthocyanidins journal September 2013
Phosphorus-31 NMR spectroscopic analysis of coal pyrolysis condensates and extracts for heteroatom functionalities possessing labile hydrogen journal November 1988
Determination of Carbonyl Groups of Six Round Robin Lignins by Modified Oximation and FTIR Spectroscopy journal January 1998
Chemical characterization and water content determination of bio-oils obtained from various biomass species using 31 P NMR spectroscopy journal March 2012
3D Chemical Image using TOF-SIMS Revealing the Biopolymer Component Spatial and Lateral Distributions in Biomass journal October 2012
Elucidating Structural Characteristics of Biomass using Solution-State 2 D NMR with a Mixture of Deuterated Dimethylsulfoxide and Hexamethylphosphoramide journal April 2016
Chemistry of delignification journal December 1985

Cited By (5)

Advancements and Complexities in the Conversion of Lignocellulose Into Chemicals and Materials journal August 2020
A sustainable wood biorefinery for low–carbon footprint chemicals production journal February 2020
Accelerating Thermal Stabilization by Pyrolytic Lignin for Partially Bio‐Based Carbon Fiber Precursor journal October 2019
Synthesis, Characterization, and Utilization of a Lignin-Based Adsorbent for Effective Removal of Azo Dye from Aqueous Solution journal February 2020
Characterization of Eucalyptus nitens Lignins Obtained by Biorefinery Methods Based on Ionic Liquids journal January 2020

Figures / Tables (11)