1g of extractive-free sample was placed in the conical flask and 12 ml of 72% sulfuric acid was added. 100 mL of distilled water was added to the mixture. After distilled water addition, the mixture was boiled for 2 hours and cooled at room temperature. The insoluble material (lignin) was filtered by using Whatman paper (no. 42). The lignin was washed with distilled water until neutralized with H2SO4 and then oven dried at 105°C for 10 hours at a constant weight and cooled down in a desiccator and weighed [7, 24]. Lignin content was calculated as follows:

where W1 = oven dried sample (1 g); W2 = extracted residues.

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