A thioacidolysis method tailored for higher-throughput quantitative analysis of lignin monomers

Biotechnol J. 2016 Oct;11(10):1268-1273. doi: 10.1002/biot.201600266. Epub 2016 Sep 14.

Abstract

Thioacidolysis is a method used to measure the relative content of lignin monomers bound by β-O-4 linkages. Current thioacidolysis methods are low-throughput as they require tedious steps for reaction product concentration prior to analysis using standard GC methods. A quantitative thioacidolysis method that is accessible with general laboratory equipment and uses a non-chlorinated organic solvent and is tailored for higher-throughput analysis is reported. The method utilizes lignin arylglycerol monomer standards for calibration, requires 1-2 mg of biomass per assay and has been quantified using fast-GC techniques including a Low Thermal Mass Modular Accelerated Column Heater (LTM MACH). Cumbersome steps, including standard purification, sample concentrating and drying have been eliminated to help aid in consecutive day-to-day analyses needed to sustain a high sample throughput for large screening experiments without the loss of quantitation accuracy. The method reported in this manuscript has been quantitatively validated against a commonly used thioacidolysis method and across two different research sites with three common biomass varieties to represent hardwoods, softwoods, and grasses.

Keywords: Cell wall structure; Lignin; S/G ratio; Thioacidolysis.

MeSH terms

  • Gas Chromatography-Mass Spectrometry / methods
  • Glycerol / chemistry
  • High-Throughput Screening Assays / methods*
  • Lignin / analysis*
  • Lignin / chemistry
  • Lignin / isolation & purification
  • Poaceae / chemistry
  • Sulfhydryl Compounds / chemistry*
  • Wood / chemistry

Substances

  • Sulfhydryl Compounds
  • Lignin
  • Glycerol