Supplementary Materials

Characterization of Pyrolytic Lignin and Insight into its Formation Mechanisms

using Novel Techniques and DFT Method

Simin Li a, Zhongyang Luo a,*,Wenbo Wang a, Kongyu Lu a, Yi Yang a, Xiaorui Liang a

a State Key Laboratory of Clean Energy Utilization, Zhejiang University, Hangzhou,

310027, PR China

* Corresponding author: zyluo@zju.edu.cn

Journal: Fuel

Methods: 3

Figures: 6

Tables: 1

* Corresponding authorZhongyang Luo. E-mail: zyluo@zju.edu.cn

Sample test methods:

1. HPLC/Qtof-MS

The mobile phases were 0.1% formic acid-water (A) and 0.1% formic acid-

acetonitrile (B) with a flow rate of 0.5mL min−1. The linear gradient programs were as

follows: 0/5, 35/95, 36/5 (min/B%). Injection volume of each sample was 3μL, and

column oven temperature was at 30 . AB TripleTOF 5600plus System (AB SCIEX,℃

Framingham, USA) with an electrospray ionization (ESI) source (Micromass,

Manchester, UK) was applied here for mass spectra. The analysis was performed in ESI

negative ion mode with source voltage set at -4.5kV and source temperature was 550 .℃

The typical MS conditions was as follow: curtain gas (N2) flow rate, 0.2mL min-1;

declustering potential (DP), 100V; collision energy (CE), 10V. As for MS/MS

acquisition mode, the IDA-based auto-MS2 was performed on the 8 most intense ions,

the parameters was almost the same expect that the collision energy (CE) was set at -

40±20V. For a cycle of full scan (1s), the scan range of m/z of precursor ions and

product ions were set at 100-2000Da and 50-1500Da, respectively. The exact mass

calibration was performed automatically before each analysis employing the Automated

Calibration Delivery System. Data processing was carried out using PeakView Software

(version 1.2.0.3) which could automatically obtain Total Ion Chromatogram (TIC) and

Extracted Ion Chromatogram (XIC).

2. GPC tests

All the samples was dissolved in THF (5mg mL-1), and then filtered using a 0.45 μm

filter before test. The temperature of test was 50 , and the volume of 1mL was℃

injected of each sample. THF was used as eluent with a flow rate of 1mL min-1. A UV

detector (254nm) as well as a differential refractometer (RI) was applied as detectors.

Polystyrene samples with various molecular weights were used as standards for

calibration.

3. 2D HSQC NMR

DMSO-d6 with 0.03% (v/v) TMS was used as solvent for all samples. MestReNove

Software (version 6.1.0-6224) was applied for data processing. For R-PLs' test, 50mg

samples and 5mg benzaldehyde as internal standard were dissolved in 0.6mL solvent,

respectively; for M-PL's test, 30mg sample and 3mg benzaldehyde as internal standard

was dissolved in 0.6mL solvent. The HSQC experimental parameters was listed as

follow: 16 ppm sweep width in F2 (1H), 200 ppm sweep width in F1 (13C), 96 t1

increments, a 1.0s relaxation delay, and the evolution time set for a One-Bond 1H-13C

coupling constant of 146Hz, with a total acquisition time of ~8h.

Figure Captions:

Fig. S1 Equipment for pyrolysis of simple model compounds.

Fig. S2 Total Ion Chromatogram (TIC) of RS-PL, CS-PL, WS-PL and M-PL.

Fig. S3 H number vs C number Plot.

Fig. S4 GPC chromatograms and divided peaks (RI detector). Molecular Weight is

relative to polystyrene standard. The process of peaks division is based on original GPC

chromatogram.

Fig. S5 The full 2D HSQC NMR spectra of RS-PL, CS-PL, WS-PL and M-PL

Fig. S6 MS^2 spectra of C12H10O2 (m/z 185.0608, RDB 8) and C14H14O4 (m/z 245.0819,

RDB 8). The spectras from top to bottom are of RS-PL, CS-PL, WS-PL and M-PL.

Fig. S1 Equipment for pyrolysis of simple model compounds.

Fig. S2 Total Ion Chromatogram (TIC) of RS-PL, CS-PL, WS-PL and M-PL.

The three R-PLs were nearly identical, indicating that phenolic oligomers in the

bio-oil derived from different kinds of biomass were almost same in components,

which is of great benefit for the subsequent upgrading process. The TIC of M-PL was

compared with that of R-PLs, and some overlapping peaks could be found. Some

obvious overlapping peaks were marked by dotted lines in the figure, and the retain

time (RT) of which were mostly before 13 min.

Fig. S3 H number vs C number Plot.

Fig. S4 GPC chromatograms and divided peaks (RI detector). Molecular Weight is

relative to polystyrene standard. The process of peaks division is based on original

GPC chromatogram.

Fig. S5 The full 2D HSQC NMR spectra of RS-PL, CS-PL, WS-PL and M-PL

Fig. S6 MS^2 spectra of C12H10O2 (m/z 185.0608, RDB 8) and C14H14O4 (m/z

245.0819, RDB 8). The spectras from top to bottom are of RS-PL, CS-PL, WS-PL

and M-PL.

Table Captions:

Table S1 Summary of GPC analysis of R-PLs & M-PL (RI detector).

Table S1 Summary of GPC analysis of R-PLs & M-PL (RI detector)

Sample

Peak 1 Peak 2 Area Radio

P1/P2Mw Mn d Area Mw Mn d Area

M-PL 400.24 333.33 1.20 4.55 178.53 171.23 1.04 12.88 0.35

RS-PL 965.59 578.03 1.67 21.80 189.48 175.75 1.08 6.19 3.52

CS-PL 961.45 588.24 1.63 20.49 188.52 173.91 1.08 4.97 4.12

WS-PL 954.97 581.40 1.64 20.64 192.46 178.89 1.08 4.64 4.45