A method for the high-throughput analysis of the relative lignin contents of Cryptomeria japonica samples over a wide concentration range (3-73%), independent of the type of chemical pretreatment, was developed by using Fourier transform infrared spectroscopy. First, the assignments of the infrared absorbance related to lignin were reviewed. Then, various chemical treatments, including alkaline, acid, and hydrothermal processes, and a sodium chlorite oxidation treatment, were performed to prepare samples containing a wide range of different lignin contents. Principal component analysis indicated high variability among the chemical treatments in terms of the corresponding lignin contents as well as the resulting changes in the chemical structure of hemicellulose; this conclusion was supported by the loading vectors. The intensity of the key band of lignin at 1508 cm-1 was calculated using the absorbance at 2900 cm-1 as a reference; a reliable calibration curve with an R2 of 0.968 was obtained independent of the chemical treatment performed. This simple and rapid method for determining the lignin content is expected to be widely applicable for optimizing bioethanol production, as well as monitoring biomass degradation processes.
Keywords: Biomass; Fourier transform infrared spectroscopy; High-throughput analysis; Lignin; Principal component analysis.