Lab Presentation Preliminary Results Mythreyi Chandoor June 12th ,2009

Lignin coniferyl- sinapyl- (III) alcohols p-coumaryl

Solid State Carbon-13 Nuclear Magnetic Resonance Spectroscopy

Control sample

Control Versus Four Weeks.

Control Versus Eight Weeks

Control versus Twelve Weeks

Overlay of all the Time intevals

FTIR 750 1000 1250 1500 1750 2000 2500 3000 3500 4000 1/cm 0.005 0.01 0.015 0.02 0.025 Abs 3998.44 3408.22 3388.93 3348.42 3331.07 1647.21 1625.99 1512.19 1460.11 1425.40 1265.30 1033.85 798.53 FTIR 1000 1250 1500 1750 2000 2500 3000 3500 1/cm 0.005 0.01 0.015 0.02 0.025 Abs 3358.07 2920.23 1732.08 1510.26 1259.52 1159.22 1039.63 Biodegradation 750 1000 1250 1500 1750 2000 2500 3000 3500 4000 1/cm -0.005 0.005 0.01 0.015 0.02 0.025 Abs 3996.51 3892.35 3869.20 3859.56 3840.27 3738.05 1647.21 1546.91 1512.19 1460.11 1425.40 1267.23 1224.80 1031.92 792.74 FTIR

FTIR wheat Wood Biomass Four Weeks 8 weeks 12 weeks A 3430 0.010194 0.002243 0.002231 0.006792 0.001655 0.2185 A1370 0.000797 0.00192 0.001006 0.010609 0.000639 0.010579 A1165 0.007221 0.002876 0.002087 0.0175 0.009712 0.26779 A1043 0.046237 0.032220 0.026448 0.0475 0.026851 0.0334 A1510 0.0053 0.004679 0.001913 0.009116 0.003959 0.0157 A2890 0.0123 0.002084 0.001621 0.00412 0.000232 0.01547 A1460 0.001995 0.001564 0.000906 0.009162 0.001599 0.012811 A1420 0.00171 0.001682 0.000521 0.010127 0.001323 0.01368 A1720 0.0095 0.001916 0.001055 0.004775 0.00196 0.009149

FTIR wheat Wood Biomass Four Weeks 8 weeks 12 weeks A1600 0.0029 0.00105 0.001279 0.007021 0.001079 0.011057 A844 0.0057 0.000717 0.000068 0.008834 0.000194 0.22955 A2936 0.0121 0.002375 0.001652 0.004980 0.000763 0.015227 A1330 0.00147 0.000447 0.000229 0.010201 0.000448 0.0352 A1269 0.0027 0.004532 0.003650 0.014654 0.002325 0.012736

Functional analysis CONTROL 4 WEEKS 8 WEEKS 12 WEEKS Mean value of OH Groups 16.608 9.0499 10.25 21.25 Mean value of Phenolic OH groups 0.525 1.16 O.16 0.67 Mean value of -OCH3 GROUPS 1.59 2.56 0.796 2.672

Method of Determination. % Total Solids (T final) = ( w2 –w / w1)* 100 W = Dry dish weight ,g W1 = Initial sample weight ,g W2 = Sample weight plus dish weight after drying , g. % of ash content = (w2 – w/ w1) * 100 W= Ignited dish weight ,g W1 = Initial moisture- free sample weight ,g W2 = sample weight plus dish weight after removal from furnace.

Composition Analysis Determination of Total solids Determination of Ash content Determination of Lignin

Determination of Lignin % Acid –Insoluble Lignin = [(w2 – w3) / w1 *( T final / 100% )] * 100 W1 = Initial weight of the sample W2 = Weight of the crucible, acid soluble lignin and acid insoluble lignin after drying in the oven W3 = weight of crucible and acid –insoluble ash after removal from furnace T final = % total solids content of shredded sample on a 105 0 C dry weight basis.

Determination of Lignin %Acid –Soluble lignin = [[(A/ b*a) * df * v/1000ml ] /[ (w * T final) /100] ] * 100 A = Absorbance Df = Dilution factor b = cell path ,1 cm a = absorptivity ,equal to 110 L/g-cm V = filtrate volume W = Initial Biomass sample weight T final = % total solids content of Biomass sample.

Hypothesis Based on the Microscopy results ,The presence of fungal mycelia can be found .Based on the analytical techniques for the analysis of the sugars in the cell wall of the wheat straw and pine wood we can analyze the change in the structure of lignin in the twelve week compounds. when compared to the Control samples. The further analysis of the results of different analytical methods such as FTIR and Solid state NMR ,GC –MS Pyrolysis and HPLC would basically give us information about the different kinds of compounds formed at different intervals of time and thus we can get an rough idea of how and where the bond breakage is occurring in the lignin structure. Though the natural system is a slow process , the Study of the Soil degradation mechanism at 210 c provides an a very important data which would be useful in determination of the deconstruction of lignin in the cell wall as this is occurring at extreme conditions and thus degradation mechanism would be the most convenient way and feasible approach when conditions are optimized .