Abstract

We compared the amount of lignin as determined by the three most traditional methods for lignin measurement in three tissues (sugarcane bagasse, soybean roots and soybean seed coat) contrasting for lignin amount and composition. Although all methods presented high reproducibility, major inconsistencies among them were found. The amount of lignin determined by thioglycolic acid method was severely lower than that provided by the other methods (up to 95%) in all tissues analyzed. Klason method was quite similar to acetyl bromide in tissues containing higher amounts of lignin, but presented lower recovery of lignin in the less lignified tissue. To investigate the causes of the inconsistencies observed, we determined the monomer composition of all plant materials, but found no correlation. We found that the low recovery of lignin presented by the thioglycolic acid method were due losses of lignin in the residues disposed throughout the procedures. The production of furfurals by acetyl bromide method does not explain the differences observed. The acetyl bromide method is the simplest and fastest among the methods evaluated presenting similar or best recovery of lignin in all the tissues assessed.

Highlights

  • Lignin is a complex heteropolymer that primarily consists of phydroxyphenyl (H), guaiacyl (G), and syringyl (S) units formed by the oxidative coupling of p-coumaryl, coniferyl, and sinapyl alcohols, respectively, which are products of the phenylpropanoid pathway [1]

  • The lignin contents determined by each method were 266, 242, and 28 mg g21 cell wall by the acetyl bromide, Klason, and thioglycolic acid methods, respectively

  • Acetyl bromide method produced the highest recover of lignin, but was quite similar to the value of Klason, whereas the thioglycolic acid method provided the lowest recovery of lignin

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Summary

Introduction

Lignin is a complex heteropolymer that primarily consists of phydroxyphenyl (H), guaiacyl (G), and syringyl (S) units formed by the oxidative coupling of p-coumaryl, coniferyl, and sinapyl alcohols, respectively, which are products of the phenylpropanoid pathway [1]. After complete deposition in the secondary cell wall, lignin provides a coating for cellulose-hemicellulose microfibrils, ensuring stiffness, strength, and impermeability for the lignified tissues. These properties protect the energetic polysaccharides against attack from pathogens and herbivores [2]. Several methods have been described to measure lignin levels in biomass samples [4,5,6,7,8]. There is no universal method to determine lignin. This is a grave drawback because each technique can give different results for a same sample

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