Determination of a suitable molar absorption coefficient (ε) for lignin analysis of fibrous plants using the CASA method

Bozek, Antoine; Mathis, Fabienne; Guillaumat, Laurent; Müssig, Jörg; Boizot, Nathalie; Ismael, Alaa; Ralph, John; Lu, Fachuang; Sibout, Richard; Beaugrand, Johnny

doi:10.1016/j.ijbiomac.2025.142324

Determination of a suitable molar absorption coefficient (ε) for lignin analysis of fibrous plants using the CASA method

Journal Article · Wed Mar 19 04:00:00 EDT 2025 · International Journal of Biological Macromolecules

DOI:https://doi.org/10.1016/j.ijbiomac.2025.142324· OSTI ID:2567048

Bozek, Antoine ^[1]; Mathis, Fabienne ^[2]; Guillaumat, Laurent ^[3]; Müssig, Jörg ^[4]; Boizot, Nathalie ^[5]; Ismael, Alaa ^[6]; Ralph, John ^[7]; Lu, Fachuang ^[7]; Sibout, Richard ^[6]; Beaugrand, Johnny ^[6]

INRAE Nantes (France); HEMP-IT-ADN, Beaufort en Anjou (France)
HEMP-IT-ADN, Beaufort en Anjou (France)
ENSAM Angers (France)
HSB - City University of Applied Sciences, Bremen (Germany)
INRAE, Orleans (France)
INRAE Nantes (France)
University of Wisconsin, Madison, WI (United States)

In this study, the benchmarking and applicability of the CASA method for the analysis of fibrous plants, such as flax, hemp, and jute, is evaluated. Lignin is a phenolic biopolymer present in plant cell walls and is composed of three primary monomeric units, designated G, S, and H. Various factors, such as the genetic variability, influence the relative proportions of these units in plant samples. Recently, the cysteine-assisted sulfuric acid (CASA) method has been introduced as a rapid method for the quantification of lignin in wood samples. The aim of this study is to establish a suitable molar absorption coefficient (ε, L·g^–1·cm^–1) to adapt the CASA method for use with annual plant fibers. This investigation was motivated by the technical advantages of the CASA method, including higher throughput, lower reaction temperatures, and ecological benefits due to the absence of carcinogenic, mutagenic, or reprotoxic (CMR) substances and the need for minimal sample quantities. In this method, lignin solubilization is facilitated by cysteine, which is an amino acid that enhances the reaction kinetics, using a one-hour incubation period. However, as with any spectrophotometric technique, the CASA method depends on a molar absorption coefficient (ε) that varies according to the ratio of the aromatic units within the polymer. To evaluate the suitability of CASA for quantifying lignin in economically significant plant fibers, we investigate the impact of the unit ratios on the accuracy of ε. The results are compared with those of two widely recognized lignin analytical methods: the Klason method, which is a gravimetric reference method, and the acetyl bromide soluble lignin method, which is the most commonly used spectrophotometric approach. The final ε obtained in this study reveals a relative difference in lignin content ranging from –8 % to +9 % based on a comparison between the CASA and Klason methods across different industrial hemp varieties. As a result, using known G:S ratios in annual fibrous plants, ε can be estimated from our results.

View Accepted Manuscript (DOE)

Research Organization:: Great Lakes Bioenergy Research Center (GLBRC), Madison, WI (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018409

OSTI ID:: 2567048

Journal Information:: International Journal of Biological Macromolecules, Journal Name: International Journal of Biological Macromolecules Vol. 308; ISSN 0141-8130

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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