Effect of Two-Step Formosolv Fractionation on the Structural Properties and Antioxidant Activity of Lignin

The formosolv fractionation process has been demonstrated to be an effective approach toward lignin recovery as an antioxidant from lignocellulosic biomass. In this study, four lignin fractions, FL-88%, FSL-70%, FIL-70% and FL-EtAc, were isolated from Phragmites australis biomass through two-step formosolv fractionation (88% formic acid delignification followed by 70% aqueous formic acid fractionation). To better understand the structural properties of the lignin obtained from this fractionation process, four isolated lignins were successfully characterized by gel permeation chromatography (GPC), Fourier transform infrared (FT-IR), two-dimensional heteronuclear single quantum coherence nuclear magnetic resonance (2D-HSQC NMR) spectroscopy, thermogravimetric analysis (TGA) and gas chromatograph-mass spectroscopy (GC/MS). It was found that lignin depolymerization via β-O-4 cleavage occurred via a formylation, elimination and hydrolysis mechanism, accompanied by a competitive condensation reaction. Noteworthily, two-step formosolv fractionation can produce specific lignin fractions with different ABTS and DPPH radical scavenging activities. The FL-EtAc fraction with low molecular weight (M_w = 2748 Da) and good homogeneity (PDI = 1.5) showed excellent antioxidant activity, compared with the other three isolated lignin fractions, even equal to that of commercial antioxidant BHT at the same concentration of 2.0 mg·mL⁻¹. These findings are of great help for specific lignin from biomass as a natural antioxidant in the future.