I' trying to quantify cellulose in rice leaf, using Acetic/nitric acid digestion, followed by 67% sulfuric acid digestion and lastly anthrone analysis.
After acetic/nitric acid digestion, the leaf sample become whitish in color, however, during sulfuric acid digestion the leaf didn't fully hydrolyzed, i could still see tiny fibers in the sulfuric acid solution even after 3 hours, is this normal? I thought the sample should be digested to become a clear solution after 1 hr of 67% sulfuric acid digestion, even the filter paper i used as positive control was fully hydrolyzed into clear solution, but not my leaf sample. What went wrong?
Below is my method:
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Dear Vikki,
I would suggest you to follow the standard biomass compositional analysis procedure put forth by NREL. The method recommends subjecting 0.3 g of your leaf (ground) to concentrated sulphuric acid treatment. Later, you can assess the released sugars (beta-D-glucose and hydrolyzed hemicellulose) using a HPLC.
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