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A thesis submitted in partial fulfillment of the requirements for the degree of Bachelor of Technology in Chemical Engineering By Nirbhay Gupta Roll no. 10500027 Under guidance of
CERTIFICATE
This is to certify that the thesis entitled HYDROLYSIS OF LIGNOCELLULOSIC MATERIALS FOR ETHANOL PRODUCTION submitted by Mr. Nirbhay Gupta in partial fulfillments for the requirements for the award of Bachelor of Technology Degree in Chemical Engineering at National Institute of Technology, Rourkela is an authentic work carried out by him under my supervision and guidance. To the best of my knowledge the matter embodied in the thesis has not been submitted by any other University/Institute for the award of any Degree or Diploma.
Date:
Dr. A. Kumar
Department of Chemical Engineering National Institute of Technology Rourkela-769008
ACKNOWLEDGEMENT
I would like to make my deepest appreciation and gratitude to Prof. A. Kumar for his invaluable guidance, constructive criticism and encouragement during the course of this project.
I would like to thank Prof. R.K.Singh for being a uniformly excellent advisor. He was always open minded, helpful and provided us with a strong broad idea.
Grateful acknowledgement is made to all the staff and faculty members of Chemical Engineering Department, National Institute of Technology, Rourkela for their consistent encouragement.
In spite of the numerous citations above the author accepts full responsibility for the contents that follow.
DATE:
CONTENTS
Contents List of figures List of Tables Abstract vi v iii v
Chapter 1
INTRODUCTION
Chapter 2
2.1 2.2
LITERATURE REVIEW
Ethanol from Cellulose Ethyl Alcohol: Overview
3
4 7
Chapter 3
3.1
12 13 14 14 17 18
3.2 Physical Pretreatment 3.3 Chemical Pretreatment 3.4 Biological Pretreatment 3.5 Hydrolysis Process
Chapter 4
MATERIALS & EQUIPMENT USED 4.1 Materials Used 4.2 Equipments Required
21 22 22
Chapter 5
PROCEDURE OF EXPERIMENT 5.1 Pretreatment of bagasse 5.2 Hydrolysis 5.3 pH adjustment 5.4 Fermentation
25 26 26 27 27
Chapter 6
RESULTS 6.1 Standard plot for ethanol 6.2 Analysis of ethanol for different days 6.3 Comparison of ethanol concentration with increasing no. of days.
29 30 33
40
Chapter 7
DISCUSSION
43
Chapter 8
REFERENCES
45
LIST OF GRAPHS 6.1 6.2 6.3 6.4 6.5 6.6 6.7 6.8 6.9 6.10 6.11 Standard plot for ethanol Absorbance vs Molarity curve for day 1 Absorbance vs Molarity curve for day 2 Absorbance vs Molarity curve for day 3 Absorbance vs Molarity curve for day 7 Absorbance vs Molarity curve for day 8 Absorbance vs Molarity curve for day 9 Absorbance vs Molarity curve for day 10 Ethanol conc. Vs no. of days curve for 0.2 M Ethanol conc. Vs no. of days curve for 0.3 M Ethanol conc. Vs no. of days curve for 0.5 M 32 33 34 35 36 37 38 39 40 41 42
LIST OF TABLES
Standard data for ethanol concentration Standard for ethanol concentration (revised) Data for molarity vs absorbance for 10 days Ethanol conc. Vs no.of days table for 0.2 M Ethanol conc. Vs no. of days table for 0.3 M Ethanol conc. Vs no. of days table for 0.5 M
30 31 33-39 40 41 42
ABSTRACT
Chapter 1
INTRODUCTION
Energy consumption has increased steadily over the last century as the world population has grown and more countries have become industrialized. Crude oil has been the major resource to meet the increased energy demand. Scientists used several different techniques to estimate the current known crude oil reserves and the reserves as yet undiscovered and concluded that the decline in worldwide crude oil production will begin before 2010. They also predicted that annual global oil production would decline from the current 25 billion barrels to approximately 5 billion barrels in 2050. Because the economy in the US and many other nations depends on oil, the consequences of inadequate oil availability could be severe. Therefore, there is a great interest in exploring alternative energy sources Unlike fossil fuels, ethanol is a renewable energy source produced through fermentation of sugars. Ethanol is widely used as a partial gasoline replacement in the US. Fuel ethanol that is produced from corn has been used in gasohol or oxygenated fuels since the 1980s. These gasoline fuels contain up to 10% ethanol by volume. As a result, the US transportation sector now consumes about 4540 million liters of ethanol annually, about 1% of the total consumption of gasoline. Recently, US automobile manufacturers have announced plans to produce significant numbers of flexible-fueled vehicles that can use an ethanol blend E85 (85% ethanol and 15% gasoline by volume) alone or in combination with gasoline. Using ethanol-blended fuel for automobiles can significantly reduce petroleum use and exhaust greenhouse gas emission. However, the cost of ethanol as an energy source is relatively high compared to fossil fuels. A dramatic increase in ethanol production using the current cornstarch-based technology may not be practical because corn production for ethanol will compete for the limited agricultural land needed for food and feed production. A potential source for low-cost ethanol production is to utilize lignocellulosic materials such as crop residues, grasses, sawdust, wood chips, and solid animal waste.
Chapter 2
LITERATURE REVIEW
10
Cellulose molecules consist of long chains of glucose molecules as do starch molecules, but have a different structural configuration. These structural characteristics plus the encapsulation by lignin makes cellulosic materials more difficult to hydrolyze than starchy materials. Hemicellulose is also comprised of long chains of sugar molecules; but contains, in addition to glucose (a 6-carbon or hexose sugar), contains pentoses (5-carbon sugars). To complicate matters, the exact sugar composition of hemicellulose can vary depending on the type of plant. Since 5-carbon sugars comprise a high percentage of the available sugars, the ability to recover and ferment them into ethanol is important for the efficiency and economics of the process. Recently, special microorganisms have been genetically engineered which can ferment 5-carbon sugars into ethanol with relatively high efficiency 2.1 ETHANOL-FROM-CELLULOSE In times of fuel shortages, fermentation ethanol has been commercially manufactured in the US from cellulosic biomass feedstock using acid hydrolysis techniques. However, it is only recently that cost-effective technologies for producing ethanol-from-cellulose (EFC) in the US have started to emerge. There are three basic types of EFC processesacid hydrolysis, enzymatic hydrolysis, and thermo chemicalwith variations for each. The most common is acid hydrolysis. Virtually any acid can be used; however, sulfuric acid is most commonly used since it is usually the least expensive
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12
13
2.2 ETHYL ALCOHOL: OVERVIEW Ethanol, also called ethyl alcohol, pure alcohol or drinking alcohol, is a volatile, flammable, colorless liquid. It is a psychoactive drug, best known as the type of alcohol found in alcoholic beverages and in modern thermometers. Ethanol is one of the oldest recreational drugs known to man. In common usage, it is often referred to simply as alcohol or spirits. Ethanol has widespread use as a solvent of substances intended for human contact or consumption, including scents, flavorings, colorings, and medicines. In chemistry, it is both an essential solvent and a feedstock for the synthesis of other products. It has a long history as a fuel for heat and light and also as a fuel for internal combustion engines.
Properties Molecular formula Molar mass Appearance Density
C2H6O 46.07 g mol1 colorless clear liquid 0.789 g/cm3 114.3 C, 159 K, 174 F 78.4 C, 352 K, 173 F
Melting point
Fully miscible
Viscosity
Dipole moment
14
2.2.1 PHYSICAL PROPERTIES Ethanol is a volatile, colorless liquid that has a strong characteristic odor. It burns with a smokeless blue flame that is not always visible in normal light. It is also used in finger nail polish remover. The physical properties of ethanol stem primarily from the presence of its hydroxyl group and the shortness of its carbon chain. Ethanols hydroxyl group is able to participate in hydrogen bonding, rendering it more viscous and less volatile than less polar organic compounds of similar molecular weight. Ethanol is a versatile solvent, miscible with water and with many organic solvents
2.2.2 CHEMICAL PROPERTIES Ethanol is classified as a primary alcohol, meaning that the carbon to which its hydroxyl group is attached has at least two hydrogen atoms attached to it.
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2.2.2.2 Halogenation
Ethanol reacts with hydrogen halides to produce ethyl halides such as ethyl chloride and ethyl bromide: CH3CH2OH + HCl CH3CH2Cl + H2O HCl reaction requires a catalyst such as zinc chloride. Hydrogen chloride in the presence of their respective zinc chloride is known as Lucas reagent. CH3CH2OH + HBr CH3CH2Br + H2O HBr requires refluxing with a sulfuric acid catalyst. Ethyl halides can also be produced by reacting ethanol with more specialized halogenating agents, such as thionyl chloride for preparing ethyl chloride, or phosphorus tribromide for preparing ethyl bromide. CH3CH2OH + SOCl2 CH3CH2Cl + SO2 + HCl
16
Ethanol can also form esters with inorganic acids. Diethyl sulfate and triethyl phosphate, prepared by reacting ethanol with sulfuric and phosphoric acid respectively, are both useful ethylating agents in organic synthesis. Ethyl nitrite, prepared from the reaction of ethanol with sodium nitrite and sulfuric acid, was formerly a widely-used diuretic.
2.2.2.5 Dehydration
Strong acid desiccants, such as sulfuric acid, cause ethanol's dehydration to form either diethyl ether or ethylene: 2 CH3CH2OH CH3CH2OCH2CH3 + H2O CH3CH2OH H2C=CH2 + H2O Which product, diethyl ether or ethylene, predominates depends on the precise reaction conditions
2.2.2.6 Oxidation
Ethanol can be oxidized to acetaldehyde, and further oxidized to acetic acid. In the human body, these oxidation reactions are catalyzed by enzymes. In the laboratory, aqueous solutions of strong oxidizing agents, such as chromic acid or potassium permanganate, oxidize ethanol to acetic acid, and it is difficult to stop the reaction at acetaldehyde at high yield. Ethanol can be oxidized to acetaldehyde, without over oxidation to acetic acid, by reacting it with pyridinium chromic chloride.[19] The direct oxidation of ethanol to acetic acid using chromic acid is given below. C2H5OH + 2[O] CH3COOH + H2O The oxidation product of ethanol, acetic acid, is spent as nutrient by the human body as acetyl CoA, where the acetyl group can be spent as energy or used for biosynthesis.
2.2.2.7 Chlorination
When exposed to chlorine, ethanol is both oxidized and its alpha carbon chlorinated to form the compound, chloral. 4Cl2 + C2H5OH CCl3CHO + 5HCl
17
2.2.2.8 Combustion
Combustion of ethanol forms carbon dioxide and water: C2H5OH(g) + 3 O2(g) 2 CO2(g) + 3 H2O(l); (Hr = 1409 kJ/mol[21])
18
Chapter 3
PRODUCTION METHODS
19
The basic five stages of this process are: 1. A pre- treatment phase to make the raw material such as wood or straw amenable to hydrolysis. 2. Hydrolysis, to break down the molecules of cellulose into sugars. 3. Separation of the sugar solution from the residual materials, notably lignin. 4. Yeast fermentation of the sugar solution.
20
Waste materials can be comminuted by a combination of chipping, grinding and milling to reduce cellulose crystallinity. The size of the materials is usually 1030 mm after chipping and 0.22 mm after milling or grinding. Vibratory ball milling has been found to be more effective in breaking down the cellulose crystallinity of spruce and aspen chips and improving the digestibility of the biomass than ordinary ball milling. The power requirement of mechanical comminution of agricultural materials depends on the final particle size and the waste biomass characteristics.
3.2.2. Pyrolysis
Pyrolysis has also been used for pretreatment of lignocellulosic materials. When the materials are treated at temperatures greater than 300 C, cellulose rapidly decomposes to produce gaseous products and residual char. The decomposition is much slower and less volatile products are formed at lower temperatures. Mild acid hydrolysis (1 N H2SO4, 97 C, 2.5 h) of the residues from pyrolysis pretreatment has resulted in 8085% conversion of cellulose to reducing sugars with more than 50% glucose .The process can be enhanced with the presence of oxygen When zinc chloride or sodium carbonate is added as a catalyst, the decomposition of pure cellulose can occur at a lower temperature
removal of the lignin from wheat straw in ozone pretreatment. Enzymatic hydrolysis yield increased from 0% to 57% as the percentage of lignin decreased from 29% to 8% after ozonolysis pretreatment of poplar sawdust. Ozonolysis pretreatment has the following advantages: (1) it effectively removes lignin; (2) it does not produce toxic residues for the downstream processes; and (3) the reactions are carried out at room temperature and pressure. However, a large amount of ozone is required, making the process expensive.
22
of the lignocellulosic materials increases with the removal of the crosslinks. Dilute NaOH treatment of lignocellulosic materials caused swelling, leading to an increase in internal surface area, a decrease in the degree of polymerization, a decrease in crystallinity, separation of structural linkages between lignin and carbohydrates, and disruption of the lignin structure ( Fan et al., 1987). The digestibility of NaOH-treated hardwood increased from 14% to 55% with the decrease of lignin content from 2455% to 20%. However, no effect of dilute NaOH pretreatment was observed for softwoods with lignin content greater than 26%. Dilute NaOH pretreatment was also effective for the hydrolysis of straws with relatively low lignin content of 1018% used the combination of irradiation and 2% NaOH for pretreatment of corn stalk, cassava bark and peanut husk. The glucose yield of corn stalk was 20% in untreated samples compared to 43% after treatment with electron beam irradiation at the dose of 500 kGy and 2% NaOH, but the glucose yields of cassava bark and peanut husk were only 3.5% and 2.5%, respectively. Ammonia was also used for the pretreatment to remove lignin. Iyer et al. (1996) described an ammonia recycled percolation process (temperature, 170 C; ammonia concentration, 2.520%; reaction time, 1 h) for the pretreatment of corn cobs/stover mixture and switchgrass. The efficiency of delignification was 6080% for corn cobs and 6585% for switchgrass.
23
enzymes including polyphenol oxidases, laccases, H2O2 producing enzymes and quinonereducing enzymes can also degrade lignin. The advantages of biological pretreatment include low energy requirement and mild environmental conditions. However, the rate of hydrolysis in most biological pretreatment processes is very low
studied, and the effectiveness of the hydrolysis was evaluated in terms of hemicellulose solubilization.
Yeast are able to metabolize some foods, but not others. In order for an organism to make use of a potential source of food, it must be capable of transporting the food into its cells. It must also have the proper enzymes capable of breaking the foods chemical bonds in a useful way. Sugars are vital to all living organisms. Yeast are capable of using some, but not all sugars as a food source. Yeast can metabolize sugar in two ways, aerobically, with the aid of oxygen, or anaerobically, without oxygen.
In this lab, you will try to determine whether yeast are capable of metabolizing a variety of sugars. Although aerobic fermentation of sugar is much more efficient, in this experiment we will have yeast ferment sugars anaerobically. When the yeast respire aerobically, oxygen gas is consumed at the same rate that CO2 is producedthere would be no change in the gas pressure in the test tube. When yeast ferments the sugars anaerobically, however, CO2 production will cause a change in the pressure of a closed test tube, since no oxygen is being consumed. We can use this pressure change to monitor the respiration rate and metabolic activity of the organism. A gas pressure sensor will be used to monitor the fermentation of sugar.
The fermentation of glucose can be described by the following equation: C6Hl2O6 2 CH3CH2OH +2 CO2 + energy
1. Yeast cannot utilize all of the sugars equally well. While glucose, sucrose, and fructose all can be metabolized by yeast, lactose is not utilized at all.
26
2. Yeast may not have the proper enzymes to either transport lactose across its cell membrane, or it may not have the enzyme needed to convert it from a disaccharide to a monosaccharide.
3. The yeast need to be incubated so that the oxygen in the test tube will be completely consumed. If the yeast respire aerobically, no pressure change occurs, because much oxygen is consumed as CO2 is produced. It also takes a few minutes for the yeast to transport the sugar into the cell, to respire at a constant rate, and to reach the proper temperature.
4. Some yeast live on other organisms. If they are warm blooded, they may be near the optimal temperature for yeast respiration, 37C. Many yeast live in soils. The temperature of soils may easily be measured at different times of the year.
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Chapter 4
28
4.2 EQUIPMENTS REQUIRED 4.2.1 Vertical autoclave This equipment is primarily used for sterilization purpose. It is an enclosed space where steam bath is given to any equipment placed inside it. Water filled in it is heated by electric coils present at bottom. It has a vent at top, from where the steam can be released to maintain the required pressure. For our case, sterilization is required after maintaining the pH and also for each filtration. Pressure around 2.02 kg/cm2 is used. Once the pressure reaches 2.02 kg/cm2, it is maintained for half an hour. The equipments are allowed to cool down before removing from the autoclave.
29
4.2.2 Laminar flow chamber A laminar flow cabinet or laminar flow closet or tissue culture hood is a carefully enclosed bench designed to prevent contamination of semiconductor wafers, biological samples, or any particle sensitive device. Air is drawn through a HEPA filter and blown in a very smooth, laminar flow towards the user. The cabinet is usually made of stainless steel with no gaps or joints where spores might collect. Such hoods exist in both horizontal and vertical configurations, and there are many different types of cabinets with a variety of airflow patterns and acceptable uses
4.2.3 Vaccum Filtration Vacuum filtration is a technique for separating a solid product from a solvent or liquid reaction mixture. The mixture of solid and liquid is poured through a filter paper in a Buchner funnel. The solid is trapped by the filter and the liquid is drawn through the funnel into the flask below, by a vacuum.
4.2.4 UV-Spectrophotometer The UV-Visible spectrophotometer uses two light sources, a deuterium (D2) lamp for ultraviolet light and a tungsten (W) lamp for visible light. After bouncing off a mirror (mirror 1), the light beam passes through a slit and hits a diffraction grating. The grating can be rotated allowing for a specific wavelength to be selected. At any specific orientation of the grating, only monochromatic (single wavelength) successfully passes through a slit. A filter is used to remove
30
unwanted higher orders of diffraction. The light beam hits a second mirror before it gets split by a half mirror (half of the light is reflected, the other half passes through). One of the beams is allowed to pass through a reference cuvette (which contains the solvent only), the other passes through the sample cuvette. The intensities of the light beams are then measured at the end. 4.2.5 Shaking incubator Incubator Shaker is a large capacity shaking incubator which may be stacked up to 3 high, offering a large capacity in a minimum of floor space. The ingenious design of the Infors shakers features a front-loading pullout platform, stainless steel interior panels, panoramic front window and inspection light. The patented drive system provides uniform motion and quiet operation without vibration. 4.2.6 pH- meter A pH meter is an electronic instrument used to measure the pH (acidity or alkalinity) of a liquid. A typical pH meter consists of a special measuring probe (a glass electrode) connected to an electronic meter that measures and displays the pH reading
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Chapter 5
PROCEDURE OF EXPERIMENT
32
5.2 Hydrolysis
The cellulose molecules are composed of long chains are broken down to free the sugar, before it is fermented for alcohol production. Though hydrolysis is of many types, dilute acid hydrolysis is an easy and productive process. Also the amount of alcohol produced in case of acid hydrolysis is more than that of alkaline hydrolysis. Concentrated acid hydrolysis is not used as it is a hazardous and corrosive process and also acid has to be separated out after hydrolysis for the experiment has to be feasible.
7. The bottles were then capped with the help of cotton plugs.
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5.3 pH adjustment
Before addition of any micro-organism to the above prepared samples, pH of these samples has to be adjusted. Otherwise the micro-organism will die in hyper acidic or basic state. A pH of around 5-5.5 is maintained.
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35
Chapter 6
RESULTS
36
16 ml of absolute ethanol was mixed with 24 ml of water and an absorbance of 0.897 nm was found in the UV spectrophotometer. This was assumed to be 100 % ethanol and the standard plot was drawn. SL.NO. Amount of ethanol (in ml) 1 2 3 4 5 6 7 8 9 10 11 0 0.3 0.6 0.9 1.2 1.5 1.8 2.1 2.4 2.7 3.0 Amount of water (in ml) 3.0 2.7 2.4 2.1 1.8 1.5 1.2 0.9 0.6 0.3 0 0 10 20 30 40 50 60 70 80 90 100 0 0.185 0.223 0.291 0.376 0.419 0.611 0.665 0.734 0.8 0.897 Conc. Of ethanol Absorbance
Table 6.1
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The 100 percent of this ethanol is actually the 40 percent concentration of absolute alcohol. Therefore the actual standard plate is given in the next table.
SL.NO.
Amount of water ( in ml) 3.0 2.7 2.4 2.1 1.8 1.5 1.2 0.9 0.6 0.3 0
Conc. Of ethanol
Absorbance
1 2 3 4 5 6 7 8 9 10 11
0 0.3 0.6 0.9 1.2 1.5 1.8 2.1 2.4 2.7 3.0
0 0.185 0.223 0.291 0.376 0.419 0.611 0.665 0.734 0.8 0.897
Table 6.2
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1 0.9 0.8 0.7 Absorbance 0.6 0.5 0.4 0.3 0.2 0.1 0 0 100 200 Ethanol conc. 300 400 500
Figure 6.1
39
Day 1 Sl. No. 1 2 3 4 5 Sample 0.2 0.3 (I) 0.3(II) 0.5(I) 0.5(II)
Table 6.3 Absorbance (210 nm) 0.62 0.54 0.581 0.523 0.568
0.63 0.62 0.61 0.6 0.59 0.58 0.57 0.56 0.55 0.54 0 0.2 0.4 0.6
Figure 6.2
40
Day 2 Sl. No. 1 2 3 4 5 Sample 0.2 0.3 (I) 0.3(II) 0.5(I) 0.5(II)
Table 6.4 Absorbance (210 nm) 0.667 0.605 0.589 0.558 0.529
0.68 0.66 0.64 0.62 0.6 0.58 0.56 0.54 0.52 0.5 0 0.2 0.4 0.6
Figure 6.3
41
Day 3 Sl. No. 1 2 3 4 5 Sample 0.2 0.3 (I) 0.3(II) 0.5(I) 0.5(II)
Table 6.5 Absorbance (210 nm) 0.678 0.667 0.624 0.637 0.563
0.69 0.68 0.67 0.66 0.65 0.64 0.63 0.62 0.61 0.6 0.59 0 0.2 0.4 0.6
Figure 6.4
42
Day 7 Sl. No. 1 2 3 4 5 Sample 0.2 0.3 (I) 0.3(II) 0.5(I) 0.5(II)
Table 6.6 Absorbance (210 nm) 0.712 0.672 0.671 0.667 0.598
0.72 0.71 0.7 0.69 0.68 0.67 0.66 0.65 0.64 0.63 0.62 0 0.2 0.4 0.6
Figure 6.5
43
Day 8 Sl. No. 1 2 3 4 5 Sample 0.2 0.3 (I) 0.3(II) 0.5(I) 0.5(II)
Table 6.7 Absorbance (210 nm) 0.723 0.678 0.691 0.695 0.601
0.73 0.72 0.71 0.7 0.69 0.68 0.67 0.66 0.65 0.64 0 0.2 0.4 0.6
Figure 6.6
44
Day 9 Sl. No. 1 2 3 4 5 Sample 0.2 0.3 (I) 0.3(II) 0.5(I) 0.5(II)
Table 6.8 Absorbance (210 nm) 0.735 0.701 0.742 0.718 0.655
0.74
0.73
0.72
0.71
0.7
0.69
Figure 6.7
45
Day 10 Sl. No. 1 2 3 4 5 Sample 0.2 0.3 (I) 0.3(II) 0.5(I) 0.5(II)
Table 6.9 Absorbance (210 nm) 0.753 0.721 0.742 0.736 0.698
0.755 0.75 0.745 0.74 0.735 0.73 0.725 0.72 0.715 0 0.2 0.4 0.6
Figure 6.8
46
ETHANOL
CONCENTRATION
WITH
INCREASING
Table 6.10 SL. NO. 1 2 3 4 5 6 7 8 9 10 NO. OF DAYS 1 2 3 4 5 6 7 8 9 10 ETHANOL CONC. 276.47 297.43 302.34
Figure 6.9
47
For 0.3 M acid concentration SL. NO. 1 2 3 4 5 6 7 8 9 10 NO. OF DAYS 1 2 3 4 5 6 7 8 9 10 Table 6.11 ETHANOL CONC. 249.94 275.39 287.85
Figure 6.10
48
For 0.5 M acid concentration SL. NO. 1 2 3 4 5 6 7 8 9 10 NO. OF DAYS 1 2 3 4 5 6 7 8 9 10 Table 6.12 The graph of the above table is as follows
340 320 300 Ethanol conc. 280 260 240 220 200 0 2 4 6 No. of days 8 10 12
Figure 6.11
49
Chapter 7
DISCUSSION
50
1. The ethanol concentration in ml/lt of acid hydrolyzed bagasse increases with the increasing number of days. 2. This shows that the sugar is being fermented by the help of micro organism yeast for the production of ethanol. 3. The maximum concentration of ethanol is 328.73 ml/lt of acid hydrolyzed bagasse which was found on the 10th day of fermentation. 4. In absorbance vs molarity curve , after each progressive data the curve tends to come in a straight line. 5. For 0.3 M and 0.5 M we have taken two identical samples for each for better accuracy. 6. For drawing graphs, in case of 0.3 M, 0.5 M samples average is taken. 7. Still some deflection in the graphs can be considered due to precision error, long duration of experiment, and human errors.
51
Chapter 8
REFERENCES
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1. Biosource technology, Volume 83 Issue 1, May 2002. 2. Biosource Technology, Volume 77, Issue 2, April 2001. 3. Cellulosic Ehanol- Wikipedia, The free encyclopedia. 4. Ethanol From Cellulose: A General Review Trends in new crops and new uses. 2002. J. Janick and A. Whipkey (eds.). ASHS Press, Alexandria, VA. 5. Ethanol production via enzymatic hydrolysis of sugar edition 1996 Elba P.S. Bon 6. www.osti.gov/bridge/servlets/purl/755492CufN/webviewable/755492 7. www.ethtec.com.au/downloads/Latest_News/ETHANOL_FROM_BAG ASSE_MEETINGS 8. Renewable and Sustainable Energy Reviews, Volume 13, Issues 6-7, August-September 2009, Pages 1418-1427 9. www.cellulosicroundtable.com/greenfieldethanol.htm 10. www.bioresourcesjournal.com/index.php/BioRes
11. sciencestage.com/d/29329/lime-pretreatment-of-sugarcane-bagasse-
for-bioethanol-production
12. www.academicjournals.org/ajb/PDF/pdf2008/18Jul/Damisa%20et%20
al.
13. aiche.confex.com/aiche/2006/preliminaryprogram/abstract_58322.ht
m
14.
www.rurdev.usda.gov/rbs/pub/sep06/ethanol.htm
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