Optimisation of two-step dilute acid hydrolysis of spruce for ethanol production

Adam Jomaa
Department of Chemical Engineering, Lund University February 2008
Abstract Ethanol produced from biomass has during recent years become an interesting alternative to petrol in the struggle against raised levels of fossil carbon dioxide in the atmosphere. In Sveg (Hrjedalen, Sweden), an integrated biofuel plant is planned to be constructed and is intended to produce bioethanol, pellets, briquettes, electricity and district heating. This master thesis deals with optimisation of the process conditions in the hydrolysis steps of the ethanol part of the biofuel plant. The hydrolysis method in this work was decided to be two-step dilute acid hydrolysis and the variable process parameters were the temperature, the concentration of the sulphuric acid and the residence time in the hydrolysis vessel. The raw material, spruce chopped into wood chips, was supplied from a sawmill in Sveg. Firstly, different hydrolysis conditions were tested for the first hydrolysis step. The setup that was chosen as the best one was 190C using 0.75% acid and a residence time of 10 minutes. Thereafter, material for experiments on the second hydrolysis step was produced with these conditions. The setup that was chosen as optimal in the second step was 210C using 2% acid for 4 minutes. If the maximum monosaccharide yields from the two hydrolysis step are added together, the yields of glucose and mannose are 36% and 76% respectively (corresponding to 18 g/100 g DM and 11 g/100 g DM). The maximum theoretical ethanol yield that can be obtained is 150 g/kg DM (corresponding to 0.19 L/kg DM). Key words: Dilute acid hydrolysis; ethanol; lignocellulose; spruce; softwood Introduction Since the late 19th century the mean temperature on Earth has increased with 0.8C and the major part of this increase is likely due to anthropogenic emissions of greenhouse gases. Carbon dioxide is the type of greenhouse gas with largest emissions and these originate from the combustion of fossil fuels as coal, oil and natural gas [1]. The transport sector stands for about 25% of the total energy consumption in Sweden and its energy utilisation has increased by 80% since 1970. More than 95% of the Swedish transport sectors sources of energy come from fossil fuels [2]. Thus, a decrease in the use of fossil fuels in the transport sector will have a significant effect on the emissions of fossil carbon dioxide. The European Union (EU) has set up a goal for increased use of renewable biofuels in the transport sector. In 2010, 5.75% of all transportation fuels within EU should be renewable fuels and in 2020 the corresponding goal is 10% [3]. One way to increase the use of renewable fuels is to replace petrol with ethanol produced from biomass. Today, ethanol is mainly produced from agricultural products rich of sugar (e.g. sugar cane and sugar beet) or starch (e.g. corn and wheat). Since the ethanol price is strongly correlated to the raw material costs [4] these feedstocks give a rather expensive product. The increasing demand of biofuels and hence ethanol requires raw materials that are less expensive than agricultural products and that are not used for production of food. Therefore, lignocellulosic biomass (e.g. wood, straw, forestry residues and agricultural residues) is suitable as raw material due to its lower cost and abundant supply [5]. One drawback with ethanol from lignocellulose is that the process is more complex than for sugar and starch and hence the production cost is higher. In addition to this, the ethanol yield is lower for lignocellulose. Therefore, it is crucial to maximise the yield of the raw material and also to utilise the part of the material that does not turn into ethanol. One way to increase the utilisation is to integrate the ethanol production with processes that provide energy or other products that either can be sold or used in the plant itself [6]. In Sveg (Hrjedalen, Sweden) an integrated biofuel plant is planned to be constructed and is intended to produce not only ethanol, but also pellets, briquettes, electricity and district heating.

Adam Jomaa Optimisation of two-step dilute acid hydrolysis of spruce for ethanol production

Furthermore, waste heat and produced carbon dioxide will be used in adjacent greenhouses where cultivation of for example lettuce, tomatoes and spices can be carried out. The raw material for the processes will consist of raw timber products, byproducts from the timber industry and peat, all collected from local areas. The Department of Chemical Engineering at Lund University (Sweden) has been engaged in the design of the ethanol process of the biofuel plant and this master thesis is a part of that work. The aim of this thesis was to optimise the process conditions in the hydrolysis steps for softwood from spruce, one of the raw materials used in the Sveg project. The hydrolysis method in this work was decided to be two-step dilute acid hydrolysis and the variable process parameters were the temperature, the concentration of the acid and the residence time in the hydrolysis vessel. Hydrolysis of lignocellulosic materials Lignocellulose refers to plant biomass that consists mainly of the three components cellulose, hemicellulose and lignin [7]. When lignocellulosic materials are exposed to a severe environment, cellulose and hemicellulose are degraded (hydrolysed) into shorter oligosaccharides and then monosaccharides. At the same time, unwanted reactions occur where these sugars and lignin are converted into degradation products which affect the fermentation step negatively by inhibiting the yeast cells. The most important degradation products that have been studied in this work are hydroxymethylfurfural (HMF) and furfural. HMF is a by-product from hexoses and furfural originates from pentoses. They, in turn, can degrade further to levulinic acid and formic acid [8]. Hemicellulose is hydrolysed at less severe conditions than cellulose. Therefore, with only one hydrolysis step, it can be hard to optimise the amount of produced sugar without getting too much degradation products. One solution to this problem is to perform the hydrolysis in two steps and take out the already solubilised sugars between the two steps. The first step is then performed at milder conditions optimised for hydrolysis of most of the hemicellulose and just a small part of the cellulose. The second step is more severe with the aim to hydrolyse most of the cellulose [6]. This hydrolysis method is called two-step dilute acid hydrolysis, and has been studied in this work. Material and methods Raw material Softwood from spruce (Picea abies), free from bark, was used in the experiments. The wood was provided as wood chips, about 4 to 8 mm thick. The composition of the raw material is given in Table 1.

Table 1. Composition of the raw material (spruce) as percentage of dry material Glucan 43.7 Xylan 6.2 Galactan 2.8 Arabinan 1.6 Mannan 13.0 Lignin 31.9

Impregnation Before the first hydrolysis step the wood chips were impregnated with dilute sulphuric acid (H2SO4). The chips were soaked during one hour at room temperature in buckets containing 0.75 kg of dry material (about 1.0 kg chips) and 10 kg dilute acid (either 0.75 wt-% or 1.5 wt-%). After one hour the excess fluid was removed by means of a strainer. The wet chips were let to drain on the strainer for some minutes. Before the second hydrolysis step the material was soaked during one hour at room temperature in buckets containing 0.75 kg of dry material (about 2.35 kg cake) and 10 kg dilute acid (either 2 wt-% or 4 wt-%). After one hour, the excess fluid was pressed out by hand by means of a strainer covered with a nylon cloth. The material did not drain efficiently by itself and was therefore compressed by hand against the strainer. Hydrolysis The hydrolysis was performed in a 10 litre reactor into which the raw material was fed manually from the top. The hydrolysis began when steam (10-25 bar), supplied by an electric boiler, flowed into the reactor. The hydrolysis process was controlled by a computer and when the set time had elapsed and the pressure was released, the hydrolysed material was collected in a cooled flash-off vessel. The two hydrolysis steps were optimised separately. Firstly, twelve different conditions were tested in the first hydrolysis step (see Table 2). The results were analysed and one condition was chosen as optimal. After that a large amount of raw material was hydrolysed with the chosen parameters and then fourteen different second step hydrolysis conditions were tested (see Table 3).
Table 2. Experimental setup of the first hydrolysis step Batch 1 2 3 4 5 6 7 8 9 10 11 12 Temp. (C) 180 180 180 180 190 190 190 190 200 200 200 200 H2SO4 (%) 0.75 0.75 1.5 1.5 0.75 0.75 1.5 1.5 0.75 0.75 1.5 1.5 Time (min) 5 10 5 10 5 10 2 5 2 5 2 5

Adam Jomaa Optimisation of two-step dilute acid hydrolysis of spruce for ethanol production

Table 3. Experimental setup of the second hydrolysis step Batch 16 17 18 19 20 21 22 23 24 25 26 27 28 29 Temp. (C) 200 200 200 200 210 210 210 210 210 220 220 220 220 220 H2SO4 (%) 2 4 4 4 2 2 2 4 4 2 2 2 4 0 Time (min) 4 1 2 4 1 2 4 1 2 1 2 4 4 4

Concentrations of degradation products in the hydrolysates are shown in Figure 2.

Yield (g/100g DM in raw material) 20,0 18,0 16,0 14,0 12,0 10,0 8,0 6,0 4,0 2,0 0,0
% ;5 m 0C %; 18 ; 1 10 m 0C ,5 % ; 19 1 ,5 ; 5 m 0C % 19 ; 0 ; 1 0 0C ,7 5 m ;0 % ; , 19 7 5 % 5m 0C ; ; 1 10 19 m , 0C 5 % 20 ; 1 ; 2 m 0C ,5 % ; 20 0 ,7 ; 5 m 0C 5% ;0 ; 20 ,7 5 2m 0C % ;1 ; 5 20 m , 0C 5 % ;1 ;2 ,5 m % ;5 m

Mannose Glucose

,7 5 ;0 0C 18 0C

;0

Figure 1. Yields of glucose and mannose in the slurries obtained after the first hydrolysis step experiments.

Pressing and washing The liquid part of the hydrolysed slurry (hydrolysate) was separated from the solid part (cake) by means of a manually operated hydraulic press. A nylon cloth was used as filter and the volume of the press was 3 litres. The pressing operation began with that the slurry was put in the press and the hydrolysate was pressed out and collected. Then the cake was washed with 1.0 litre of hot water (washing liquid) that later was pressed out and collected. All liquids and cakes were stored in a cold-storage room awaiting the analyses. Analyses Concentrations of monomeric sugars were determined by means of HPLC (Shimadzu LC10AD, Kyoto, Japan) with a refractive index detector (Shimadzu RID-10A, Kyoto, Japan). Glucose, xylose, galactose, arabinose and mannose were separated using an Aminex HPX-87P column (Bio-Rad, Hercules, USA) at 85C, with water as eluent at a flow rate of 0.5 ml/min. When analysing the liquids from the second hydrolysis step optimisation, another column was used, Phenomenex Rezex RPM Monosaccharide Pb++ (8%). The temperature for this column was 80C and the flow rate was 0.6 ml/min. Concentrations of degradation products were determined by means of HPLC (Shimadzu LC10AT, Kyoto, Japan) with a refractive index detector (Shimadzu RID-6A, Kyoto, Japan). HMF and furfural were separated using an Aminex HPX87H column (Bio-Rad, Hercules, USA) at 65C, with 5 mM H2SO4 as eluent at a flow rate of 0.5 ml/min. When analysing the hydrolysates from the second hydrolysis step, levulinic acid and formic acid were also separated and quantified. Results Yields of the fermentable monosaccharides glucose and mannose in the slurries obtained after the first hydrolysis step experiments are shown in Figure 1.

Concentration (g/L)

18

3,50 3,00 2,50 2,00 1,50 1,00 0,50 0,00

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Figure 2. Concentrations of degradation products in the hydrolysates obtained after the first hydrolysis step experiments.

The hydrolysis conditions that were chosen as the best for the first hydrolysis step was 190C with 0.75% acid for 10 minutes (batch 6). The yield of fermentable sugars was the highest among batches with low acid concentration and the concentration of degradation products was believed to be acceptable. Yields of monomeric glucose and mannose in the slurries obtained after the second hydrolysis step experiments are shown in Figure 3. Due to problems with the HPLC quantification, small amounts of arabinose are included in the yield for mannose. Concentrations of degradation products in the hydrolysates are shown in Figure 4.
16,0

Yield (g/100 g DM in raw m aterial)

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19

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20

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Figure 3. Yields of glucose and mannose in the slurries obtained after the second hydrolysis step experiments.

20

m i ,1 n m 20 4 0 % , i n C, 2 m 21 4 0 % , i n C 4 m 21 , 2 0 % , i n C 1 m 21 , 2 0 % , i n C 2 m 21 , 2 0 % , i n C 4 m 21 , 4 0 % , i n C, 1 m 22 4 0 % , i n C 2 m 22 , 2 0 % , i n C 1 m 22 , 2 0 % , i n C 2 m 22 , 2 in % 0 C ,4 m 22 , 4 0 % , i n C, 4 0% min ,4 m in 20 0 C, 4%

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Adam Jomaa Optimisation of two-step dilute acid hydrolysis of spruce for ethanol production

Concentration (g/L)

12,0 10,0 8,0 6,0 4,0 2,0 0,0 HMF Furf ural Levulinic acid Formic acid

Figure 4. Concentrations of degradation products in the hydrolysates obtained after the second hydrolysis step experiments.

The hydrolysis conditions that were chosen as the best for the second hydrolysis step was 210C with 2% acid for 4 minutes (batch 22). The yield of fermentable sugars was the second highest among the batches and the concentration of degradation products was believed to be acceptable. The yield of monomeric glucose and mannose from both hydrolysis steps and the sum of them are given in Table 4.
Table 4. Yields of monomeric glucose and mannose. Yield in liquid of the slurry (g/100 g DM) After 1 step Glucose Mannose After 2 step Glucose Mannose Combined two-step Glucose Mannose
nd st

Discussion and conclusions It is difficult to determine which hydrolysis conditions that are optimal after this work. Firstly, only a dozen conditions have been tested in each step and hence leave many temperatures, acid concentrations and residence times untested. Secondly, there is an interplay between the setups of the first and the second step that have not been tested in this work since only one feedstock was used when trying different second hydrolysis step conditions. Thirdly, no fermentation studies have been performed on obtained liquids. So even if one hydrolysis setup results in high yields of sugar, it is not certain that this will correspond to high yields or productivities of ethanol. But the conditions which have been considered as optimal in this work gave rather good yields of monosaccharides compared to the expectations. If the maximum yields from the two hydrolysis steps are summed, the yields of monomeric glucose and mannose will be 36% and 76% respectively. The maximum ethanol yield is 150 g/kg DM (corresponding to 0.19 L/kg DM). Acknowledgements Guido Zacchi, Mats Galbe, Ola Wallberg, Marie Linde and Christian Roslander are gratefully acknowledged for their support and guidance during this thesis work. References

20 0 C 20 , 2 0 % C , 20 , 4 4 m 0 % in C , 20 , 4 1 m 0 % i C ,2 n 21 , 4 0 % min C , 21 , 2 4 m 0 % in C , 21 , 2 1 m 0 % i C ,2 n ,2 21 0 % min C , 21 , 4 4 m 0 % in C , 22 , 4 1 m 0 % in C , 22 , 2 2 m 0 % in C , 22 , 2 1 m 0 % i C ,2 n 22 , 2 0 % min C ,4 22 , 4 0 % min C ,4 ,0 % min ,4 m in

(% of max) 13 58 23 18* 36 76*

6.3 8.4 11 2.6* 18 11*

1.

* Includes small amount of arabinose.

The liquid fed into the fermentation step consists of the hydrolysates and the washing liquids from the two hydrolysis steps. The concentrations of monosaccharides and degradation product in this liquid are shown in Figure 5. The concentrations of HMF, furfural, levulinic and formic acid are 1.0 g/L, 0.73 g/L, 1.7 g/L and 1.0 g/L respectively. The maximum theoretical ethanol yield that can be obtained is calculated by multiplying the combined two-step yield of the fermentable sugars with 0.51. The maximum ethanol yield is 150 g/kg DM (corresponding to 0.19 L/kg DM).
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Figure 5. Concentrations in the liquid consisting of the hydrolysates and washing liquids from the two hydrolysis steps.

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Naturvrdsverket, Klimat i frndring, 2007. S fungerar vxthuseffekten. Available from: http://www.naturvardsverket.se/sv/Klimat-iforandring/Vaxthuseffekten/Sa-fungerarvaxhuseffekten/ [Accessed January 17 2008] Energimyndigheten, 2007. Energilget 2007. (ET2007:49) Energimyndigheten, 2007. Den svenska klimatstrategins utveckling. (ET2007:29) Wingren, A., Galbe, M., Zacchi, G., 2003. Technoeconomic evaluation of producing ethanol from softwood: Comparison of SSF and SHF and identification of bottlenecks. Biotechnology progress, 19, 1109-1117. Kim, S., Dale, B., 2004. Global potential bioethanol production from wasted crops and crop residues. Biomass and Energy, 26 (4), 361-375. Galbe, M., Zacchi, G., 2002. A review of the production of ethanol from softwood. Applied Microbiology and Biotechnology, 59, 618-628. Sjstrm, E., 1993. Wood Chemistry. Fundamentals and Applications. 2nd ed. London: Academic Press. Larsson, S., Palmqvist, E., Hahn-Hgerdahl, B., Tengborg, C., Stenberg, K., Zacchi, G., Nilvebrant, N-O., 1999. The generation of fermentation inhibitors during dilute acid hydrolysis of softwood. Enzyme and Microbial Technology, 24, 151-159.

Concentration (g/L)

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