Bastos, Francisca

Costa, Eduardo

Freixo, Ricardo

Pereira, Carla

Ramos, Oscar

Ribeiro, Alessandra

English

Repositório Institucional da Universidade Católica Portuguesa

Evaluation of different acid hydrolysis in sugarcane bagasse for cellulose-rich extract productionRicardo Freixo, Francisca Bastos, Alessandra Ribeiro, Carla Pereira*, Eduardo Costa, & Oscar Ramos*Universidade Católica Portuguesa, CBQF - Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Rua Diogo Botelho 1327, 4169-005 Porto, Portugal*oramos@porto.ucp.pt; cpereira@porto.ucp.ptIntroduction/Resume ObjectivesMaterial and MethodsConclusionsSugarcane bagasse, the major by-product of the sugarcane industry and a verypromising renewable and low-cost raw material, can be used to produce valuablematerials such as cellulose or cellulose-based materials. This lignocellulosic biomassconstituted mainly by cellulose, hemicellulose and lignin, 42, 28 and 20%, respectively,can be cleaved by enzymatic or acid/alkaline hydrolysis, thus releasing compounds suchas glucose, xylose, furfural or xylo-oligosaccharides (Kim and Day, 2011). One of themajor barriers to lignocellulosic biomass valorization is its recalcitrance to hydrolysiscaused by the hierarchy structure of the plant cell wall (i.e. the strong, rigid and complexmicrofibril structure of the cellulose-lignin-hemicellulose matrix) (Wyman, Cai et al. 2017).Therefore, to address this and facilitate the access to isolated compounds as celulose forfurther production of high-value products, different treatments have been tested to obtainfractions rich in cellulose.The aim of this work was to obtain a fraction rich in cellulose from sugarcane bagassethrough cost-effective process. For this purpose, the impact of different acid hydrolysistreatments were evaluated in the remotion of lignin and hemicellulose in order to producesolid fractions rich in cellulose. Thus, the structural composition of solid fractions rich incellulose obtained by 4 different acid hydrolysis treatments were characterized in terms oflevel of cellulose, hemicellulose, Klason lignin and acid soluble lignin by NREL procedure(Sluiter et al., 2008). Also, the functional groups and crystallinity were assessed byFourier-Transform Infrared spectroscopy (FT-IR) and Powder X-Ray Diffraction (PXRD)analysis, respectively.The sugarcane bagasse was dried and sieved in a blade mill and the fraction greater than 315µm was selected. After this, the biomass was subjected to four different acid treatments, HA1,HA2, HA3 and HA4 according to Guerra-Rodríguez et al. (2012), Laopaiboon, Thani et al.(2010) and Kumar, Dheeran et al. (2015), respectively. The procedures were summarizedbelow:Acid hydrolysis 1 (HA1)Sulfuric acid solution prepared at 2% (w/v) was added to bagasse at a 12:1 ratio (w/w). Then,the solution was heated in a water bath at 100 ºC for 30 min. The solid fraction obtained afterfiltration was washed with water until neutrality and then dried at 55 ºC overnight in an oven.Acid hydrolysis 2 (HA2)Hydrochloric acid solution prepared at 0.5% (v/v) and preheated, was added to bagasse at a15:1 (v/w) ratio. Then, the solution was heated in a water bath at 100 ºC for 4h underagitation. The solid fraction obtained after filtration was washed with water until neutrality, andthen dried at 55 ºC overnight in an oven.Acid hydrolysis 3 (HA3)Sodium hydroxide solution prepared at 5% (w/v) using a 10 M NaOH solution, was added tobagasse at a ratio of 1:20 (v/w). Then, the solution was heated in a water bath at 80 ºC for 60min under agitation. The solid fraction obtained after filtration was washed with water untilneutrality and dried at 50 ºC until constant weight. Afterwards, hydrochloric acid solutionprepared at 0.5% (v/v) and preheated, was added to the solid fraction previously obtained at a15:1 (v/w) ratio. The solid fraction obtained after filtration was washed with water untilneutrality, and then dried at 55 ºC overnight in an oven.Acid hydrolysis 4 (HA4)Sulfuric acid solution prepared at 8% (v/v) was added to bagasse at a ratio of 8:1 (w/w), Then,the solution was heated in autoclave at 100 ºC for 90 min. The solid fraction was washed withwater until neutrality, and then dried at 55 ºC overnight in an oven. Afterwards, a sulfuric acidsolution prepared at 40% (v/v) was added to the solid fraction previously obtained, at a ratio of1:4 ratio (w/w). The solution was heated in a water bath at 80 ºC for 90 min under agitation,while the solid fraction was washed with water until neutrality, and then dried at 55 ºCovernight in an oven.Chemical characterizationThe solid fractions obtained from acid treatments were characterized in terms oflignocellulosic composition by National Renewable Energy Laboratory (NREL) following theanalytical procedures for standard biomass analysis (Sluiter et al., 2008). The structuralcharacterization was assessed by FT-IR and PXRD analysis. The FT-IR spectra wererecorded using the Frontier™ MIR/FIR spectrometer from PerkinElmer in a scanning range of550-4000 cm−1 for 16 scans at a spectral resolution of 4 cm−1, while the PXRD analyses wereperformed on Rigaku MiniFlex 600 diffractometer with Cu kα radiation, with a voltage of 40 kVand a current of 15 mA (3º≤ 2θ ≥ 60º; step of 0.01 and speed rate of 3.0º/min).The data obtained from NREL, photos from the solid fraction, FT-IR and XRDcharacterization for solid fractions rich in cellulose obtained from different acid treatments(HA1-HA4) were presented in Figure 1, 2, 3 respectively and Table 1.The treatment described as acid hydrolysis 3 (HA3), comprising two steps hydrolysis i.e., an alkaline hydrolysis with sodium hydroxide followed by an acid hydrolysis with hydrochloricacid, showed to be the most promising one in lignin removal, producing a solid fraction rich in cellulose (i.e., 59.9%) (Fig. 1) exhibiting the highest crystallinity (i.e., 69.3%) (Tab. 1) andwhitter color (Fig. 2) compared to the other treatments. On other hand, the treatment 4 (HA4) allowed a greater removal of hemicellulose, but the lowest lignin removal proved by thedarker color exhibited by the solid fraction (Fig. 2).ResultsAcknowledgementsThis research was funded by Amyris Bio Products Portugal Unipessoal Lda and Escola Superior de Biotecnologia – Universidade Católica Portuguesa through Alchemyproject- Capturing high value from industrial fermentation bio products (POCI−01−0247−FEDER−027578). We would also like to thank the scientific collaboration under theFundação para a Ciência e a Tecnologia (FCT) project UIDB/50016/2020.ReferencesGuerra-Rodríguez, E., Portilla-Rivera, O., Jarquín-Enríquez, L., Ramírez de León, J., & Vázquez, M. (2012). Acid hydrolysis of wheat straw: A kinetic study. Biomass and Bioenergy, 36, 346-355. doi: 10.1016/j.biombioe.2011.11.005Kumar, S., Dheeran, P., Singh, S. P., Mishra, I. M., & Adhikari, D. K. (2015). Kinetic studies of two-stage sulphuric acid hydrolysis of sugarcane bagasse. Renewable Energy, 83, 850-858. doi: https://doi.org/10.1016/j.renene.2015.05.033Kim, M. & Day, D. F. (2011). Composition of sugar cane, energy cane, and sweet sorghum suitable for ethanol production at Louisiana sugar mills. Journal of Industrial Microbiology and Biotechnology, 38, 803-807.Laopaiboon, P., Thani, A., Leelavatcharamas, V., & Laopaiboon, L. (2010). Acid hydrolysis of sugarcane bagasse for lactic acid production. Bioresource Technology, 101(3), 1036-1043. doi: https://doi.org/10.1016/j.biortech.2009.08.091Sluiter, A., Hames, B., Ruiz, R., Scarlata, C., Sluiter, J., Templeton, D., & Crocker, D. (2008). Determination of Structural Carbohydrates and Lignin in Biomass—NREL/TP-510-42618. Laboratory Analytical Procedure (LAP). Wyman, C. E., Cai, C. M., & Kumar, R. (2017). Bioethanol from Lignocellulosic Biomass. In R. A. Meyers (Ed.), Encyclopedia of Sustainability Science and Technology (pp. 1-27). New York, NY: Springer New York.Treatment Crystallinityindex (%)HA 1 57.0HA 2 59.4HA 3 69.3HA 4 65.9Table 1. Crystallinity index of solid fractions rich in cellulose by the different treatments (HA1-HA4) Figure 3. FT-IR spectra of solid fractions rich in celluloseaffected by the different acid treatments (HA1-HA4)applied in comparison with the data obtained for acommercial cellulose from Sigma-Aldrich (CC).Figure 2. Visual aspect of the solid fractions rich in cellulose affected by the different acid treatments (HA1-HA4) applied.Figure 1. Chemical composition of solid fractions rich in cellulose affected by the different acid treatments(HA1-HA4) applied.

Evaluation of different acid hydrolysis in sugarcane bagasse for cellulose-rich extract production

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Evaluation of different acid hydrolysis in sugarcane bagasse for cellulose-rich extract production

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