Print: ISSN 2278-3849 CCSD Per ad NAAS Rating : 3.56 PANN chs in Life Sciences Volume 5 | Number 16 | August, 2016 An International Fortnightly Journal COTTON EC aaa ER) Dheerpura SOCiety for Advancement of Science and Rural Development Re emcee ues ar RecAdvances in Life Sciences 5(16), Print : ISSN 2278-3849, 3930-5933, 2016 Dilute Acid Pretreatment of Paddy Straw Using Orthophosphoric Acid . VIJAYANAND* AND S.KAMARAJ “Department of Bioenergy, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadw email: viayeceanand@gmail com ABSTRACT ‘The vegetative part of Oryza sativa L exscinded during grain harvest is the straw of rice or paddy. ‘Orthophosphoric acid was used in acid hydrolysis of paddy straw to release fermentable sugars which was the hydrolysate. The hydrolysis was carried out for 60, 120 and 180 minutes with acid concentrations of 2.5,5.0, 75 and 10.0 percent. The amount of hydrolysate obtained for each trial was measured and the corresponding solid residue of pretreated biomass was also weighed and recorded. Keywords Paddy straw, Orthophosphoric acid, Hydrolysis, Hydrolysate, Biomass residue Even in the current world scenario, conventional open field burning of paddy straw still persists as the disposal technique used by farmers. Paddy straw is an appealing lignocellulosic biomass material from which biofuel production can be carried out since rice is one of the most staple food consumed globally and is straw can serve as @ perennial resource for biofuel production. Large amounts of rice straw produced are disposed of by buming, which is an environmental hazard, and can cause severe impacts on human health, (Gadde et al. 2009; Nori ef al. 2008). Hence, alternative uses of rice straw have bbeen suggested, and one of them is utilizing the rice straw as a substrate for alternative biofuels (Binod et al. 2010). Bioconversion of rice straw into biobutanol is @ multistep process consisting of pretreatment, enzymatic hydrolysis, and fermentation (Zhang and Cai 2008; Lee et al. 2008), To initiate the production of biobutanol from cellulosic biomass, bioconversion of the cellulosic components into fermentable sugars is necessary (Kumar et al. 2008). The key obstacle for fermentable sugars production is the recalcitrant nature of the raw biomass, and therefore pretreatment is particularly ctucial to alter the cellulosic biomass by physica, thermal, o chemical means to facilitate rapid and efficient enzymatic hydrolysis of carbohydrates to fermentable sugars (Chang and Holtzapple 2000). Despite being considered as a crucial step for the conversion of biomass to liquid fuels, biomass pretreatment is one of the main economic costs in the process, In fact, it hhas been deseribed as the second most expensive unit cost in the conversion of lignocellulose to ethanol and butanol fuels. (Yang and Wyman 2008). The common methods currently used or in development consist of alkali hydrolysis, dilute acid hydrolysis, uncatalysed steam explosion, acid-activated steam explosion, liquid hot water, ammonia fiber explosion, ozonolysis, and CO, explosion (Ruiz et al, 2008; Garlock er al. 2012). MATERIAL AND METHODS Paddy straw was dried and segregated with three 4ifferent total solids of 7.5, 10.0 and 12.5 9%. The biomass ‘was milled and grinded to powder form of <850 um size and was pretreated with orthophosphoric acid having a purity of 85 %, its molecular weight was 92 g and specific gr was 1.82, The hydrolysis was carried out in a autoclave at 100°C for time periods of 60, 120 and 180 minutes, The acid hydrolysed biomass was filtered in a Whatman filter of 100 jum size and the liquid product of hydrolysis, hydrolysate was obtained. The volume of hydrolysate obtained for each trial was recorded and the solid residue of hydrolysed biomass was weighed. The data was recorded for each trial conducted and based on these results inferences was made. RESULTS AND DISCUSSION The acid hydrolysis was carried out using dilute corthophosphoric acid in the concentrations of 2.5, 5.0, and 7.5. The acidified paddy straw hydrolysate slurry was filtered to remove the residual biomass and hydrolysate separately. ‘The quantity of biomass in wet basis and the volume of hydrolysate obtained for each trial are were recorded and the average reading of the each trial, (average of three replicas) is tabulated in the tables 1 to 6 . Similarly the figures 1 to 6 illustrate the amount of solid residues and volume of fermentable hydrolysate sugars obtained from the acid hydrolysis of paddy straw biomass for acetone butanol ethanol (A.B.E.) fermentation, The tables 1, 2 and 3 depict the amount of hydrolysate obtained and the residual paddy straw biomass obtained after filtration for the total solids contents of 7.5, 10 and 12.5% at 100 °C for different acid concentrations at different time, Similarly the figures 4.1 to 46 illustrate the average ‘maximum hydrolysate obtained for the each trial and the pretreated filtate biomass retained after filtration. From the figures 1, 3 and are bar graphs illustrating the weight of wet biomass after each hydrolysis tial for the total solids of 75, 10.0 and 12.5% for orthophosphoric acid concentration of 2.5, $.0, 7.5 and 10.0% for 60, 120 and 180 ‘minutes autoclaving at 100°C. Prom the graphs in figs 4.2, 43 and 45 it can be observed that during the 60, 120 and 180 minutes tials, the weight of the biomass varied for paddy straw having total solid peroent of 7.5, 10.0 and 12.5, however the highest weight of biomass was obtained for the 60 minutes trials. The maximum wet basis weight of biomass was 134, 131 and 129g for2.5 %of orthophosphoric acid likewise the lowest values of hydrolysed wet biomass weight was 120, 121 and 121 g for the acid concentration of 10%.VUAYANAND and KAMARAJ, Dilute Acid Pretreatment of Paddy Straw Using Orthophosphoric Acid 5931 Table 1. Data of solid residue biomass and liquid hydrolysate after acid hydrolysis with paddy straw of 7.5% total solids at 100°C tuPOsconcetraton, o_o ere (wet basis)" Volume of liquid hydrolysate obtained, mL. 0 min 180) 0 min 120 min 180 min 25 ae ira 126 940 930 950 50 127 126 123 930 935 340 15 126 12s 122 932 om a2 100 1s i 120 926 935 936 Table 2. Data of solid residue biomass and liquid hydrolysate after acid hydrolys of 10% total solids at 100°C with paddy straw ‘Weight of filtered solid residue, (wet basis) HPO4 concentration, # ‘Volume of ti id hydrolysate obtained, mL. 60 min 120min—180min 60 min 120 min 25 13 126 126 925 935 935 50 130 125 1s 930 940 os 18 129 14 13 our 935 oor 100 a7 121 a1 930 935 921 Table 3. Dat of 12.5 % total solids at 100°C of solid residue biomass and liquid hydrolysate after acid hydrolysis with paddy straw Weight of itered solid r H:PO, concentration, % ide, g (wet basis) Volume of liquid hydrolysate obtained, mL 60 min 120 min ——‘180min—_— 60 min 10min ‘180 min 25 1B 1B 1s 945 920 940 50 3 130 3 93 925 930 15 BI 129 3 937 92 9x2 100 130 bar RI 931 925 926 ‘The figures 2, 4 and 6 represent the volume of fermentable sugars or the hydrolysate, (the product of the hydrolysis) for trials conducted with total solids of 7.5, 10.0, and 12.5% for orthophosphoric acid concentration of 2.5, 5.0, 75 and 10.0% for 60, 120 and 180 minutes autoclaving at 100°C. The volume of hydrolysate obtained was different for cach trial, the trials conducted for the different acid concentration with the total solids percent of 7.5 with 10% acid concentration yielded the highest volumes of hydrolysate irrespective of the treatment time. The hydrolysate volume of 950 ml of obtained from the acid hydrolysis of 180 minutes with 2.5% acid with raw biomass having total solids of 7.5 %. However the maximum hydrolysate was obtained for the 5 during 120 minutes trial with 10 percent total solids, the volume of hydrolysate obtained was 950 ml, unlike these results the maximum volume of hydrolysate obtained from hydrolysis of paddy straw with 12.5 percent total solids ‘was 945 ml for 2.5 percent acid concentration ‘The lowest values of the hydrolysate obtained were 922 ml during 120 minutes trial for 7.5 % total solids and acid concentrations among all the trials conducted with 7.5 % total solids at 100 °C. Similarly, 917 ml of hydrolysate obtained ffom 60 minutes tral was the lowest value recorded fiom the hydrolysis of paddy straw with 10 percent total ‘acid concentration solids in 10 % acid. The hydrolysate volume of 912 ml was recorded for the hydrolysis with 7.5% acid during 120 ‘minutes tial with total solids of 12.5%, The pretreatment using dilute H,PO, acid was required to obtain improved form of sugars, degradation ot loss of carbohydrates, removing lignin, removing hemicetluloses and inerease the biomass material porosity. From the acid hydrolysis of paddy straw biomass, it was observed that the acid breaks down the covalent bonds, hydrogen bonds and also the vanderwaals force in cellulose, lignin and hemicelluloses. The acid hydrolysis enabled an increase in the porosity and accessibility of the cellulose to enzymes synthesized by the microbes. Further the acid hydrolysis acted in breaking dawn the heterocyclic ether bonds between sugars monomers in polymeric chains of cellulose of hemicelluloses, From the above stated results| fon an average the volume of hydrolysate obtained was 950 ml and the average weight of biomass residue filtered out was 131 g (wet weight). From the results obtained the analysis of sugar concentration in the hydrolysate from tach (ial has fo be carried out for zeroing on the treatment combination attributing highest fermentable sugar concentration, The acid hydrolysis of the paddy straw biomass was cartied out using different concentrations of5932 Advances in Life Sciences (16), 2016 oe fe Fig. Weight of residues obtained during acid hydrolysis of paddy straw with 7.5% total solids 100°C sneohyiekare nt Fig2, Volume of hydrolysate obtained during acid hydrolysis of paddy straw with 7.5% total solids 100°C 13400 13200 13000 dis $isnoo Bizoao 1180 11600 11400 1047S. at 100° Ja220min 120 in, 2380 4 100 ‘Acid Concentration, 9 Fig3. Weight of residues obtained during acid ous 0%, st 200°C | | } | a. 5180 min 0s Acts concentration, % ‘Volum e yarns mi Fig4. Volume of hydrolysate obtained during acid hydrolysis of paddy straw with 10% total solids 100°C hydrolysis of paddy straw with 10% total solids 100°C ‘ifdconennl Fig. Weight of residues obtained during acid hydrolysis of paddy straw with 12.5% total solids at 100°C orthophosphoric acid, total solids, time and temperature ‘The hydrolysis carried out in this study was a dilute acid hydrolysis, the liquid product obtain from the filtration of acidified biomass was hydrolysate. The volume of hydrolysate obtained and the biomass weight were Wale 00 ‘elu hyo,» Fig6. Volume of hydrolysate obtained during acid hydrolysis of paddy straw with 12.5% total solids 100°C measured to quantify the results of each trial. Graphical illustration in which lengths are proportional to quant were derived and discussed. The dilute acid hydrolysis of paddy straw biomass was conducted as a preliminary study to use the hydrolysate as substrate for acetone butanol\VUAYANAND and KAMARAJ, Dilute Acid Pretreatment of Paddy Straw Using Orthophosphoric Acid 5933 ethanol (A.B.E) fermentation, ACKNOWLEDGEMENTS ‘The author would like to acknowledge University Grants Commission (UGC) for the support rendered in pursuing PhD. LITERATURE CITED Binod, P, Sindh, R, Singhania, RR. Vikram, S., Dev, ‘Nagalakshmi S, Kusen, N., Sokumaran,R.., and Pande, 2010. Bioethanol production fom nice straw: An overview, Bioresource Technology 101: 467-4774 ‘Chang V. and Toizapple, M. 2000, Fundamental factors acting biomass enzymatic reactivity, Applied Biochemistry and Biotechnology 8886) : 5-37 ade, Bonne, Menke, and Garvit, 8.2008. Airplluon misions ffm ree Saw open ld burning In, haland and the Philippines, Envvonmental Pollution 187, 1554558 Garlock, RJ, Balan, V, and Dale, B.E. 2012. 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