INVESTIGATION ON CONVERSION OF FLOWER WASTES INTO BIOETHANOL AND PERFORMANCE EVALUATION ON SINGLE CYLINDER IC ENGINE

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1 INVESTIGATION ON CONVERSION OF FLOWER WASTES INTO BIOETHANOL AND PERFORMANCE EVALUATION ON SINGLE CYLINDER IC ENGINE COLLEGE : BAPUJI INSTITUTE OF ENGINEERING AND TECHNOLOGY, DAVANGERE DEPARTMENT : MECHANICAL ENGINEERING GUIDE : Dr. S KUMARAPPA Dr. B M KULKARNI STUDENTS : VEERBHADRAPPA C BALIKAYI Keywords: Flower wastes, Microorganisms, fermentation, bioethanol and Engine Performance. Introduction: The world has been confronted with an energy crisis due to depletion of finite resources of fossil fuel, difficulties in their extraction and processing, leading to an increase of its cost. Also fossil fuels contribute an important role in accumulation of greenhouse gases which can ultimately pollute the environment. The level of greenhouse gasses in the earth s atmosphere has drastically increased. In this scenario, renewable sources might serve as an alternative. Hence it is necessary to look forward for alternative fuels. Wind, water, sun, biomass and geothermal heat can be the renewable sources for the energy industry, where as fuel production and the chemical industry may depend on biomass (bioethanol and biodiesel) as an alternative source in the near future, As a result, production of ethanol by fermentation from renewable carbohydrate materials for use as an alternative liquid fuel has been attracting worldwide interest. Fermentation is defined as the energy yielding process that cells carryout in the absence or near absence of oxygen, thus being close to the natural environment to which microorganisms are adapted. Although fermentation does not provide much usable energy for the cell, it is sufficient for yeast cells. The main function of a fermenter is to provide a controlled environment for the growth of microorganisms or animal cells, to obtain a desired product. Agricultural substrates and crop residues serve as cheap raw materials for use in for production of various bio-products, i.e. organic acids, enzymes, amino acids, and bio-ethanol. Fig. 1- After usage status of flowers Fig. 1 shows the present condition of flowers, indicating that flowers thrown in the holy river, after festival and dumping in a field as a waste, but actually we are not throwing wastes, but losing one of our easily and freely available resources. Flower wastes have many distinct advantages over traditional crops, such as its high fermentable sugars, zero investment and available everywhere on earth. Flower wastes available in nature freely and abundantly hence, can be economical for ethanol production. Recently, the production of ethanol from mahula flowers by free and immobilized Saccharomyces cerevisiae cells in submerged fermentation has been described. The production of ethanol from flower wastes by Fermentation has not been studied. Therefore, the aim of the present investigation was to examine the potential of flower wastes as a source for ethanol production by Saccharomyces cerevisiae cells via Stirrer type Fermentation, as well as to study the comparisons of yield in untreated and treated (Hydrolyzed) samples. Also to study the fuel properties and engine performance. Objectives: To manage disposal of flower waste. To add value to flower waste. Production of bio-ethanol from flower wastes.

2 Utilization of rich source of sugars of the flower wastes, which can be potentially converted into bioethanol. Comparisons of outcome yield from untreated and treated (Hydrolyzed) samples. Evaluation of bioethanol fuel properties. Evaluation of engine performance and emission characteristics. Methodology: Flower wastes were collected from selected local temples, market yards and community halls and dried it for 7 days in sun rays in order to remove the moisture content to some extent. And then by using commercial mixer grinder prepared powder from dried waste flowers. The flower wastes have the following compositions (expressed in g/100 g dry weight basis): moisture, 22-25; starch, ; total sugar (glucose, fructose, sucrose and maltose), 18-20; crude protein, 8-10; undetermined solids, 30 35; ph Inoculation Media and Yeast Cultivation. Saccharomyces cerevisiae (strain CTCRI) was grown in 250 ml Erlenmeyer flasks containing 100 ml sterilized medium SDB (sub rose dextrose broth) ph was adjusted to 5.5 by NaOH. It was cultured for 48 h at 30 0 C in an incubator. This served as the starter culture for ethanol production shown in Fig 2. Fig. 2- Cultured cells Fig. 3- Immobilized cell beads Fig. 4- Autoclave unit Immobilization method The yeast cell suspension was added to sodium alginate solutions in a 1:1 volume ratio and mixed thoroughly. The cell alginate mixture was then cast into beads by dropping from a glass pipette into 2% CaCl 2 solution. These beads had a diameter of approximately 3.0mm and were hardened by keeping in dilute 2% CaCl 2 solution for 24 h and washed with sterile distilled water before being used for the fermentation process shown in Fig 3. Preparation of Acid Hydrolyzed solution In this process 2% concentration H 2 SO 4 were prepared by taking 20 ml of H 2 SO 4 and added in 980 ml of distilled water in a glass beaker, then mixed thoroughly in order to get good acidic nature. Weighed 80gms of waste flowers powder sample and taken in 1Litre Reagent bottle, then was added prepared H 2 SO 4 solution in reagent bottle kept in autoclave unit (Fig. 4) at C and bar for 1 hr. After sterilizing cooled to room temperature and filtered the hydrolyzed sample solution by using filter papers and also ph was adjusted to 5.5 by adding NaOH (1N).

3 Fig. 5- Acid Hydrolyzed Solution Fig. 6- Stirrer type Fermenter unit Fermentation medium Flower wastes were mixer grinded (TTK Prestige Ltd, Bangalore, India) to make powder, then by adding distilled water (Flower powder sample: Distilled water, ratio1:5) again grinded to make a slurry. The slurry was cooked by steam in a auto clave unit at C for 60 min. (NH 4 ) 2 SO 4 was added at the rate of 1 g /1l slurry and the ph was adjusted to 5.5. The cooked mash was used as fermentation medium after cooling down. For fermentation using immobilized cells and free cells, the starter culture was added at the rate of 10% to 1L slurry and fermentation was carried out in stirrer type fermentation unit at room temperature (30±2 0 C) for 96 h. Analytical technique At appropriate time intervals, fermentation sample were removed, and the contents were analyzed. And maintained the constant speed (200 rpm) and constant moisture of about 70% for 96 h. Afterwards sample was taken out and centrifuged in a cooling centrifuge unit and the whole centrifuged sample was distilled to collect the ethanol. Ethanol concentration of the fermentation liquid was determined by Gas Chromatography (carried out by Bangalore Teas House). s of the total carbohydrates (glucose, fructose, sucrose and maltose) in the flowers were determined by Anthrone method (Biogenics, Hubli). The other proximate compositions like starch, crude protein and undetermined solids were estimated as per the standard procedure. The ph is measured by a ph meter using glass electrode. Distillation Fig.7-Distilation unit Fig.8- Fermented sample Fig. 9- Residue Fig. 10- Bioethanol

4 Fig.7 Shows Fractional distillation unit utilizes a packed column before the still head to give an increase surface area for repeated vaporization, condensation cycles. The temperature of packed column closer to round bottom flask is at a higher temperature than the end closer to still head, this results in the vapors with higher water content to condense then revaporize with lower water content. As a water vapor climbs the fractionating column, this cycle continues until the vapor reaches the still head, having an ideal concentration of 75-80% ethanol shown in Fig 10. Results and Conclusions: Analysis of total carbohydrates by Anthrone method Anthrone method is the one used to estimate the concentration of total carbohydrates in a given samples. Here ANTRHONE is a chemical used as a reagent and its melting point is in between C C and its chemical formula C 14 H 10 O. Table 1- Data showing the readings of standard Glucose curve of glucose (µg) Absorbance at 630 nm Fig.7 describes the concentration of glucose (µg) values in X-axis and respective absorbance values at 630nm in Y-axis, observed that absorbance rate increases gradually with respect to increasing the concentration of the glucose. By using this Standard curve and below mentioned formula, we estimated the percentage of carbohydrates in the sample. Y= (OD-0.035)/0.007 Table 2- Total carbohydrates in tested samples Sample Absorbance at 630 nm of Carbohydrate (g/kg) Flower Slurry Fresh Flower Flower powder Fig. 7- Standard Glucose Curve Inference: comparing all the above three samples flower powder having more amount of carbohydrates. So on these results, in our project work study FLOWER POWDER had been used as a feedstock to produce bioethanol. Also compared the yield results of untreated and treated (acid hydrolyzed) Flower powder samples. Estimation of total reducing sugars by DNS method Fig.8 describes the concentration of Maltose (µg) values in X-axis and respective absorbance values at 540nm in Y-axis, observed that absorbance rate increases gradually with respect to increasing the

5 concentration of the Maltose. By using this Standard curve, we estimated the concentration of total reducing sugars in the sample. Table 3- Readings of standard Maltose curve Absorbance of maltose(µg) at 540 nm Fig 8- Standard Maltose curve Table 4- of reducing sugars at 540 nm OD at 540 nm (1:4 dilution) of reducing sugars/0.1 ml of reducing sugars after rectified dilution factor (µg/ml) of reducing sugar (µg/ml) of reducing sugar (mg/ml) Inference: The sample has been detected with mg/ml reducing sugars Percentage of ethanol In this process we used Gas Chromatography (GC) method to estimate the total percentage of ethanol in a treated (Hydrolyzed) and untreated flower powder sample and compared the total ethanol yield from both the samples. Table 5- Shows the percentage of ethanol by GC SL. No. Parameters Description Results in Test method percentage Sample 1 Ethanol by GC Light brown 2.74 GC (Untreated) coloured liquid Sample 2 (treated) Ethanol by GC Light brown coloured liquid 0.51 GC Evaluation of fuel properties Here we evaluated the fuel properties like Flash point, Fire point, Viscosity, Density and Calorific value of bioethanol and compared these results with petrol and diesel Table 6- Comparison of bioethanol properties with petrol and diesel Sl. No Properties Bioethanol Petrol Diesel 1 Flash point 0 C Fire point o C Density Kg/m

6 4 Viscosity mm 2 /s Conclusion: According to the current statistics, there is rise of consumption and thus cost of fossil fuels, from the literature survey concluded that there is increasing demand for the petrol and diesel in world day by day. On the other hand, the utility of flowers cannot be reduced due to its wide applications. Almost all worships, festivals and for any type of functions flowers is a main attractive and necessity material used by all community peoples and thus results in increase in flower waste. However, the huge amount of flower waste contents sugars, as we know that any material having sugars can produce bioethanol so flower waste may be treated as feedstock to produce bioethanol by using fermentation technique. And Saccharomyces cerevisiae is used as a microorganism in this process because it gives better results as compared to Z. mobiles or other bacteria. If we treat this waste as a feedstock, it would also take care of value addition to the disposal of flower waste. Scope for future work: Waste flowers are used as a feedstock to produce bioethanol, results obtained with good yield. If we concentrate on the flowers having rich amount of sugars can improve the total yield. Also by varying the various fermentation parameters like ph, temperature and amount of nutrients good results can be expected.