CHAPTER 1
INTRODUCTION
1.1 Background Study
Galanthamine is a tertiary alkaloid derived from bulbs of Amaryllidaceae family of flowering plants that include daffodils and the common snowdrops. It is a cholinesterase inhibitor and allosteric modulating ligand at nicotinic cholinergic receptors. Nowadays, galanthamine, especially under its brand names Reminyl® and Nivalin®, is commonly used in the treatment of Alzheimer’s disease (AD). Micheal et al (2010) reported that Alzehimer’s disease characterized by Alois Alzheimer in 1907 is a progressive degenerative disorders of the brain. It affect over 20 million individuals worldwide and this number will increase in the future along with the increase number of elderly in the population. According to the chollinergic hypothesis, impairment of the chollinergic function give the critical important in Alzheimer’s Disease especially in brain areas, dealing with memory, learning, behavior and emotional respond.
Ping Jia et al (2008) stated that among all the acetylcholinesterase (AChE) inhibitors in clinic, galanthamine exhibits unique dual mechanism on cholinergic system, not only inhibiting the cholinergic activity but also allosterically modulating nicotinic acetylcholinereceptors (AChER) which will promote the release of acetylcholine. Besides, it is less toxicity compared to tacrine, rivastigmine and donepezil. During the past few years, various galanthmaine derivatives has been synthesized and tested for anticholinesterase activity, structure-activity-relationship (SAR) studies reveal that substitution of nitrogen atom of galanthamine is favourable AChE inhibitory activity because show interaction with peripharal anionic site (PAS).

For this Final Year Project, the researcher synthesis the galanthamine acetate using Novozym435 as a catalyst. Acetic acid react with galanthamine whereby the acetic acid is the most commonly free solvent system that can readily be used. Novozym435 is the enzyme catalyst that able to form peptide bond and speed up the reaction within hours only to give good result.
According to Rao et al (2017) the method on neutralization of galanthamine hydrobromide to galanthamine is by using ammonium hydroxide that will yield about 89% of the product. The galanthamine then will be reacted with other solvent to give new compound under certain condition. The reaction will monitored by thin layer chromatography (TLC).

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Miklosi et al (2006) stated that over the past twenty years, the zebrafish (Danio Rerio) has been emerged as a pre-eminent vertebrates model for studying genetics, development, human disease and the screening of therapeutic drugs. A number of favourable attributes including its small size, rapid development and generation time, optical transparency during early development, tractibility in forward genetic screens and genetic similarity to humans rise in popularity to be used in areas of research.

Bertelli et al (2017) explained that zebrafish is a small fresh water teleost fish that has been used to preclinically drug discovery and toxicological investigations. Considering that the zebrafish embryo is an important tool to access the toxicity profile and cholinergic system, the aim of this study is to analyze the toxicity of galanthamine acetate compound using zebrafish embryo as for the biological assay study.

As zebrafish research is to a large degree predicated on consistent production of large numbers of emryos, information of the reproductive biology and behavior of the animals in the wild is of clear for husbandry.
1.2 Objectives
The purpose of this project are synthesizing galanthamine acetate and determine the toxicity of the compound using zebrafish embyros. The objectives of this project are
To synthesize the galanthamine acetate when mixing galanthamine with acetic acid solution using a catalyst Novozym 435.

To characterize the synthesized compound using various spectroscopic method.

To determine the biological assay of the galanthamine derivatives using zebrafish embryo based on the toxicity test.

CHAPTER 2
LITERATURE REVIEW
2.1 Galanthamine (GL)
Bin Dong et al, (2017) stated that (?)-Galanthamine ( HYPERLINK ;http://www.sciencedirect.com/science/article/pii/S0040402017306385; l ;fig1; Fig.?1) belonging to galanthamine-type Amaryllidaceae alkaloid has unique tetracyclic structure and intriguing biological activities. It was found to be a selective, reversible, and competitive acetylcholinesterase (AChE) inhibitor and it also showed allosteric modulation of the neural nicotinic receptors to increase acetylcholine release.
The molecular formula of galanthamine is C17H21NO3 which has average molecular weight 287.354 g/mol. Besides, it is more crystals than benzene and it melts at temperature range of 115-130C. Galanthamine also can be fairly soluble in dichloromethane, tetrahydrofuran and methanol. (Han ep al., 1991). But, the galanthamine not soluble in chloroform and 2-propanol. The chemical structure of galanthamine can be shown in Figure 1:

Figure 1: The chemical structure of (?)-galanthamine (Bin Dong et al, 2017)
Galanthamine hydrobromide salt was used because it is much cheaper than pure galanthamine. Then, Bin Dong et al (2017) describes In 2001, (?)-Galanthamine hydrobromide salt was approved by FDA in the USA for the treatment of early Alzheimer’s disease. A huge amount of efforts have been put into the total synthesis of galanthamine because of its limited supplies from natural sources, high cost of isolation and purification process.. According to Rao et al, (2010) the hydrobromide salt of galanthamine was neutralized with ammonium hydroxide to produce pure galanthamine in 89% yield.
Trinadhachari et al, (2014) reported that HYPERLINK "http://www.sciencedirect.com/science/article/pii/S0957416613005387" l "b0015" Many other synthetic routes for the preparation of (?)-galanthamine have also been reported in the literature. Besides, Cordina et al (2006) explained that there was other techniques that can be used involving multiple and longer steps to produce pure galanthamine. Then, galanthamine act as an intermediate that will combine with other compound.

HYPERLINK "http://www.sciencedirect.com/science/article/pii/S0040402017308347" l "!" NaoshiYamamotoa et al (2017) stated that HYPERLINK "http://www.sciencedirect.com/science/article/pii/S0040402017308347" l "!" (–)-Galanthamine was synthesized from HYPERLINK "http://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/naltrexone" o "Learn more about Naltrexone" naltrexone in 18 steps with 3% total yield by overcoming many specific side reactions derived from the 4,5-epoxymorphinan skeleton. The key features are cleavage of the D-ring by the Hofmann elimination and the following the one-pot C9–C10 and C9–14 HYPERLINK "http://www.sciencedirect.com/topics/pharmacology-toxicology-and-pharmaceutical-science/bond-cleavage" o "Learn more about Bond cleavage" bond cleavages concomitant with the C9 removal by the OsO4–NaIO4 combination reaction. Then, the treatment with HYPERLINK "http://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/zinc" o "Learn more about Zinc" zinc powder in HYPERLINK "http://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/acetic-acid" o "Learn more about Acetic acid" acetic acid led to not only removal of the 2,2,2-trichloroethoxycarbonyl (Troc) group, but also reductive amination of the resulting imine to give the desired 7-membered ring.

HYPERLINK "http://www.sciencedirect.com/science/article/pii/S0968089613009577" l "!" HiroyukiKimura et al (2014) explained that the synthetic routes to (±)-galanthamine and (±)-norgalanthamine are synthesized step by step. (±)-3-Hydroxy-N-trifluoroacetyl-N-nornarwedine was converted to triflate by a conventional method, and subsequent reduction with formic acid in the presence of palladium acetate and dppf gave narwedine derivative. The trifluoroacetamide group was cleaved by hydrolysis with methanolic sodium hydroxide to afford the amine. Subsequent diastereoselective reduction with l-selectride® gave (±)-norgalanthamine in good yield about 84%. An alternative N-methylation was attempted using methyl iodide or methyl triflate as the methylating agent. But, the former conditions gave no product, while the latter conditions provided only a low yield of (±)-galanthamine about 33%. The overall yield of (±)-galanthamine was 25%.

HYPERLINK "http://www.sciencedirect.com/science/article/pii/S0957416613005387" l "!" Anand et al (2017) claimed that herein (±)-narwedine was chosen as a key starting material for the synthesis of the galanthamine isomers. This approach involves two critical synthetic stages which are the kinetic dynamic resolution of (±)-narwedine and the reduction of the narwedine isomers by complementary stereoselective reduction reactions to give all four isomers of galanthamine with an enantiomeric purity of more than 97%.

2.2 Synthesis of Galanthamine Derivatives
2.2.1 Chemical catalyst
There are many chemical catalyst used to speed up esterification reaction. Baghwat et al (2012) reported that the esters of low molecular weight such as ethyl acetate and butyl acetate usually have pleasant and fruity odor. Also large volumes of esters are used as solvents. According to a recent review industrial acid-base catalyst, out of the more than 120 process identified over 90% are solid catalyzed. The use to conventional liquid acids such as PTSA, H2SO4, HCL, HF, ALCL3, BF3, NZCL2 and SBF5 involves risks in handling, contaminant disposal and regeneration due to their toxic and corrosive nature. It is required to eliminate the ecologically harmful mineral acids to carry out a a large number of acid-catalyzed industrial process. Therefore, the solid catalyst are gaining attention. They have the proven advantages of heterogenous catalysts, like simplified product isolation, mild reaction condition, high selectivity, ease in recovery and reuse of the catalysts as well as reduction in the generation of wasteful byproduct. The example of solid acid catalyst used is sulfated zincornia catalyst, that highly active in acid catalyzed organic reactions but deactivation of activity occurs by the loss of sulfate ions thereby recycling of the catalyst is limited.

Kale et al (2017) explained that Glycerol esterification with acetic acid is usually performed using homogenous catalysts like sulphuric acid and paratoluene sulphonic acid but these catalysts are hazardous, corrosive and not eco-friendly. Therefore it is desired to replace these mineral acids by strong solid acid catalysts such as sulphated mesoporous silica, heteropolyacids or acidic ion exchange resins like Amberlyst-15.

Shanmugam et al (2004) state that the effort to develop green processes has led to the development of solid acid catalysts to an increase in research activities both in academic and industrial sections. These materials can replace the corrosive liquid acids currently used in many industries. A variety of materials have been used as solid acid catalysts such as clays, zeolites, sulfated metal oxides and heteropolyacids. Each of these materials offers unique properties that can influence the catalytic activity. Among these solid acids, heteropoly compounds are unique in the sense, they can be tuned at atomic/molecular level to exhibit a wide variation on properties like acidity and redox behavior. They behave like mineral acids having the protons in protected environments and exhibit Bronsted acidity, which of several orders of magnitude higher in strength than that of conventional mineral acids.

Table 1: Preparation of galanthamine derivatives using chemical catalyst
Structure of galanthamine derivatives Method/ Reaction condition Chemical catalyst used Yield (%) References
Galanthamine was reacted with bis (2-chloroethyl) phosphoramidic dichloride
triethylamine (TEA) in dry tetrahydrofuran (THF)
NA
Rao et al, 2010
Demethylation of galantamine to norgalantamine was accomplished by a non-classical Polonovski
iron(II) sulfate heptahydrate 74%
Atanasova et al,2009
2.2.2 Enzyme catalyst
Lipases are versatile catalysts. In addition to their natural reaction of fat hydrolysis, lipases catalyze a plethora of other reactions such as esterification, amidation, and transesterification of esters as well as organic carbonates. Moreover, lipases accept a wide variety of substrates while maintaining their regioselectivity and stereoselectivity. Lipases are highly stable even under adverse conditions such as organic solvents, high temperatures, and so forth. Applications of lipases include production of food additives, chiral intermediates, and pharmaceutical products. Among these, synthesis of various chiral intermediates in pharmaceutical industry and cocoa butter substitutes is being commercially exploited currently.

Novozyme435 was used for the present study as a biocatalyst because this enzyme has been confirmed to be the most effective to esterify lactic acid in the previous observations. The influence of substrate concentrations on enzymatic esterification of lactic acid was examined in several hydrophobic ethers and ketones that are miscible with lactic acid (exceptionally, lactic acid was not fully dissolved only in disopropyl ether, but the reaction mixtures using this solvent were completely miscible and monophasic by addition of ethanol under the experimental conditions). The molar ratios of lactic acid to ethanol, used as substrates,were 1:1 or 1:2.
Enzyme-catalyzed esterification acquired increasing attention in many applications, due to the significance of the derived products. More specifically, the lipase-catalyzed esterification reactions attracted research interest during the past decade, due to an increased use of organic esters in biotechnology and the chemical industry (Torres and Castro, 2004). For this reason, esterification by lipases was developed a few decades ago (Okumura et al., 1979) and various microbial lipases have been employed in experiments using either primary or secondary alcohols, or both, free-solvent systems, or organic solvents. Zaks and Klibanov (1988) reported that among the important factors which influence the ester yield are the concentrations of enzyme and substrates, their molar ratio, the reaction pH-value and temperature, the mixing rates, and the water content.

2.3 Biological Assay of Galanthamine on Zebrafish
According to Dubinska-Magiera et. al., (2016) skeletal muscles in zebrafish compromise about 60% of adult body mass so study on toxic effect on muscles are of high relevance. The zebrafish contain the skeletal muscles originate from the paraxial mesoderms which undergoes fregmentation into repititive units called somites. There are three compartments to differentiate during early embryonic development which are dermomyotome, myotome and sclerotome. The source of skeletal muscles of the trunk and pelvic fin muscles comes from myotome. Table 2 summarize how the zebrafish model could be applied in accessing the impact of toxicants and bioactive compounds on neuromuscular system development and functions.
Table 2: Toxicants effect on the development and functioning of zebrafish skeletal muscle.

Toxicant Examples Effect References
Drugs GAL(Galanthamine) Motility impairment induced by myopathy Dubinska et al., 2016
Roy et al (2015) claimed that the zebrafish model has become particularly popular in the laboratory setting given its genetic and embryological similarities to higher order vertebrates including humans. For a toxicological perspective, zebrafish are particularly useful as their development is well characterized and all stages of toxicological accessment can be made ex utero. The organs specific to toxins such as the liver can be seen the toxin conversion at the early stages. These zebrafish can give information that cannot be obtained from other models and knowledge of mechanisms as the development of the toxicity is scarce.
Berteli et al (2017) reported that cholinesterase enzyme have a crucial rules in cholinergic neurotransmission once it regulates the amount of neurotransmitters in synaptic cleft. Acetylcholinesterase (AChE) regulates the transmission of nerve impulses through the synapse by hydrolysis of cholinergic neurotransmitter acetylcholine (ACh) and choline acetate. AChE inhibitors represent the first option pharmacotherapy from initial to moderate treatment of Alzheimer’s Disease (AD). One type of drugs that already registered for the treatment of AD is galanthamine. Zebrafish has been used to pre clinical drug discovery and toxicological investigations.
Hill et al (2005) explained that zebrafish have been used majorly in developmental biology and molecular genetics, also in toxicology and drug discovery has been recognized. To evaluate the toxicity of a chemical, it is essential to identify the endpoints of toxicity and their dose-response relationships, elucidate the mechanisms of toxicity, and determine the toxicodynamics of the chemical. Related to detailed toxicological investigations of a single chemical, there also is a need for high-throughput large-scale screening for toxicity of several hundreds of chemicals at a time. Hence, the zebrafish has numerous attributes.

Table 3: Biological Assay of galanthamine on zebrafish
Biological assay of galanthamine on zebrafish embryos Findings
References
Toxicity
The coagulation of fertilized eggs, lack of somite formation, lack of detachment of the tail-bud from the yolk-sac and lack of heartbeat. Pablo et al., (2017)
IC50
Galanthamine has a dual action mechanism on chollinergic systems whereby inhibits AChE and allosterically modulates nACHR activity Yu Pong et al., (2015)
Neurotoxicity
For AChE inhibition, galanthamine modulates nicotic neurotransmission via allosteric potention of pre- and postsynaptic nAChR. Yu Pong et at., (2015)
CHAPTER 3
MATERIALS AND METHODOLOGY
3.1 Materials
All the reagents and solvents used are commercially available from Sigma-Aldrich. Meanwhile, the immobilised lipase Novozym 435 was purchased from Novozym.com. Table 3 shows the list of the chemicals and reagents used in this research:
Table 3 : Materials
Chemicals Enzyme
Galanthamine hydrobromide Novozym 435
Ethanol Acetic acid Hexane Ethyl acetate Acetone Methanol Silica gel Distilled water 3.2 General flow of experiment
41910110490Enzymatic esterification
Enzymatic esterification

32385269240Galanthamine solution was mixed with acetic acid under optimum condition
Galanthamine solution was mixed with acetic acid under optimum condition

13335323215Galanthamine acetate
Galanthamine acetate

-15240225425Galanthamine acetate structure was purified, identified and charcterized using following analysis:
-TLC,GC, GCMS, FTIR and NMR
Galanthamine acetate structure was purified, identified and charcterized using following analysis:
-TLC,GC, GCMS, FTIR and NMR

3810159385Biological assay of galanthamine acetate on zebrafish
Biological assay of galanthamine acetate on zebrafish

3.2.1 Synthesis of galanthamine acetate
The enzymatic synthesis of this reaction started with 0.100 g of galanthamine was dissolved in 5 ml of ethanol solution in a small beaker. Then, 0.017 g of acetic acid and 0.005 g of Novozym 435 were put into solution. Next, the beaker was covered in water bath at 55? for 24 hours. The product the was cooled and filtered from enzyme. The steps followed with the mixture was dried into 80? using rotary evaporator until white powder of pure galanthamine was produced. The solvent was purified using solvent mixture of acetone: methanol 3:1 ratio to produce high purity yield of ester. The sample then was kept in the bottle for characterization using analytical instruments.

The optimum condition of synthesis galanthamine acetate using Novozym 435 were predicted using optimisation function of the Design-Expert software. The optimum condition that had been proposed from this software were the temperature at 73.24? about 15 hours operation time, 2.00 wt% of enzyme amount and used 3.42:1 substrate molar ratio for the galanthamine:acetic acid. The synthesis of galanthamine acetate was done based on the optimum condition in order to get high percentage yield of the product.

Percentage of conversion was measured by titration with 0.1 M NaOH (aq) solution in the presence of phenolphthalein as an indicator. The colourless solution turned pink as the end point was obtained. The ester produced was expressed as equivalent to the acid conversion. The amount of acid reacted was calculated from the data obtained for the control samples (without enzyme) and test samples (with enzyme). The percentage of conversion of each sample was obtained in triplicate.

Percentage conversion: (Vc?Vs/)Vc×100
Where,
Vs : The volume of NaOH (with Novozym 435) in mL
Vc: The volume of NaOH used (without Novozym 435) in mL
3.3 Purification, Identification and Characterization of Galanthamine Acetate
3.3.1 Thin layer chromatography analysis (TLC)
Thin Layer Chromatography (TLC) is an analytical technique to identify the one or more components of the mixture, monitor the progress of the reaction and check the purity of the sample. This method is the most quick, simple and effective ways to analyze the small samples. The stationary phase is the polar absorbance usually finely alumina or silica particles the absorbent is coated on glass side or plastic sheet to create layer of stationary phase where the silica plates was used. Alumina, when anhydrous, is the more active, that is, it will adsorb substances more strongly. To separate the more polar substrates such as alcohols, carboxylic acids, and amines, the less active adsorbent, Silica Gel,was used. Then, almost all mixtures of solvent can be used as the mobile phase and used ethyl acetate: hexane (3:7). A capillary tube was used to spot the samples 1 cm from the bottom of TLC plates. Next, the plates were slightly dried after a few minutes developed. When the solvents were half eluted 1 cm from the top, the plate was removed.
3.3.2 Column chromatography
Column chromatography is the methods for separation and purification of the organic compounds based on its polarity using hydrobromide salts, HBR and the stationary phase was silica gel. The principle of column chromatography based on differential of adsorption of substance by the adsorbent. The component of mixture was separated, and they form in bands. The component that was weaker absorbed will move faster to the column and get separated first compared to the component that is highly absorbed that will move slower to the column and get separated at last. The galanthamine was subjected to column chromatography eluted with ethyl acetate: hexane (3:7) on column chromatography.
3.3.3 Gas Chromatography
Gas chromatography is an analytical separation techniques used to analyze the volatile substances in the gas phase. In gas chromatography, the components of a sample are dissolved in a solvent and vaporized in order to separate the analytes by distributing the sample between two phases: a stationary phase and a mobile phase. The mobile phase is a chemically inert gas that serves to carry the molecules of the analyte through the heated column. The solvent used for this sample is hexane:ethyl acetate (7:3). The analysis was analyzed by injecting 0.5 ?L of the sample into Shimadzu Gas Cromatography using nitrogrn gas as the inert gas. The total time required for this analysis to be done is 25 minutes.
3.3.4 Gas Chromatography Mass Spectroscopy
Gas Chromatography-Mass Spectroscopy is a combination of both the process of GC and MS. Its purpose is to separate the chemical elements of a certain compound and identify the molecular level component. In the process, the mixture will be heated in order to separate the elements. Once it vaporizes, it passes the column through an inert gas, most likely to be helium, and proceeds to the mass spectroscopy process. Once the vaporized compound proceeds to the mass spectroscopy process, it will be then separated and its components will be identified through the mass of the analyte molecule. The sample that is send to GCMS lab can be in the form of liquid and also in the form of solid. For solid sample, it is diluted with suitable solvent in this case methanol is used as a solvent.
3.3.4 Fourier transform infrared (FTiR)
Fourier Transform Infrared (FTiR) is analytical technique that identify chemical bond in a molecule by producing an infrared absorption spectrum. This technique widely used to identify organic and inorganic materials. It measures the absorption of infrared radiation by the sample material against wavelength. The infrared absorption bands determine the molecular components and structures. Once a material is irradiated with infrared radiation, absorbed IR radiation usually excites molecules into a higher vibrational state. The wavelength of light absorbed by a particular molecule is a function of the energy difference between the at-rest and excited vibrational states. The characteristic of its molecular structure for a given sample depending on the wavelength that is absorbed.
.

3.3.5 Nuclear magnetic resonance (NMR)
This method is the most common method used to determine different component in the sample. The NMR system exposes the sample to a magnetic field and measures the resultant resonant frequency and absorption energy.  This allows for the recording of characteristic NMR spectra that can be used to identify known compounds in the sample. The instrumentation involved are sample holder, permanent magnet, magnetic coils, sweep generator, radio frequency transmitter, radio frequency receiver and read out system. The solvent normally used in which hydrogen replaced by deuterium are carbon tetrachloride, carbon disulphide, deuteriochloroform, hexa deuteriobenzene and deuterium oxide.
A chemical shift is the exact field strength in ppm of a nuclei comes into resonance relative to a reference standard (TMS Electron clouds “shield” nuclei from the external magnetic field causing then to absorb at slightly higher energy Shielding: influence of neighboring functional groups on the electronic structure around a nuclei and consequently the chemical shift of their resonance.
3.4 The biological assay of galanthamine acetate on zebrafish.
When the galanthamine acetate had been characterized, this research continued by studying its toxicity using zebrafish embryos.There are three parameters that had been study which are survival rate and LC50 measurement, body curvature or scoliosis determination and heart rate measurement of zebrafish embryo trated with sample until 96 hours.
For survival rate and LC50 measurement, zebrafish embryo is treated with sample galanthamine acetate (0.05g) and incubated for 24 hours per exposure. Then, the treated embryo is observed under an inverted or streomicroscope after 24 hours per exposure. The acute toxicity endpoint after 24 hour exposure to test solutions is analyzed. Next, the Dead or Coagulated column.which are”1″ is dead or coagulated and “0” is not considered dead or coagulated is observed.

The parameters for the body curvature and heart rate measurement are the zebrafish embryo is treated with sample galanthamine acetate (0.05 g) and incubated for 96 hours per exposure. The treated embryo is observed under an inverted or streomicroscope after 96 hours per exposure. The researcher read the given Acute toxicity endpoint after 96 hours per exposure test solutions. In body curvature measurement. Next, the body curvature was observed with column consists of “1” is severe scoliosis and “0.5” is light scoliosis also “0” is absence of scoliosis. In heart rate measurement, the. researcher observe presence of heart beat and measure the heart rate on no heart beat column consists of “1” is absence of heartbeat and “0” is presence of heartbeat.

The procedure for danio assay acute toxicity kit firstly the embryo is checked under a streomicroscope to ensure it is alive and are at stage of pharyngula (24 hpf). Then the embryo is transferred into 96-well plate using a transfer pipette with one healthy embryo (24 hpf) added to each well. The most of the fluid is carefully removed from around each embryo using micropipette (Figure 2) and the fluid is replaced with 100 ?L of Danio-sprintM Embryo Media containing 0.1% DMSO (untreated embryo). this step is handled carefully and quickly, the embryo do not allow to dry out. This is embryo plate.
The sample is prepared in another 96- well plate. About 150 ?L of embryo media in row B1-12 until H1-H12. Then, 300 ?L of sample with concentration 0.1:9.9 in row A1-A12. After that, a 2-fold serial dilution is made by transferring 150 ?L of the sample from row A until row G. This sample dilution is conducted using a single channel micropipette or multiple channel micropipette (12 channels). this is dilution plate (Figure 3).

Then, 100 ?L of each sample dilution from dilution plate into embryo plate. The total volume in each well of embryo plate is 200 ?L. The treated emrbyo plate is incubated at 28±2?. The embryo is observed under an inverted or stereomicroscope as alive or dead.

The kit contents which are live zebrafish embryo (wild type), Danio-SprintM Embro Media containing 0.1% DMSO, Danio-GripM Mounting Solution, 96-well plate with cover, disposable petri dish, disposable transfer pipettes, micropipette tips and statistical acute toxicity template.

Figure 2: Kit contents

Figure 3: Dilution plate
Dead or coagulated embryos look milky white, opaque and appear dark under microscope, with cellular degeneration visible and its normal structure is unrecognizable totally and partially. The dead embryo is dicarded with the help of the disposable transfer pipettes together with the part of the old medium. This embryos is fragile hence avoid direct contact with the living embryos.

Figure 4: Normal healthy embryo Figure 5 : Dead or coagulated embryo
CHAPTER 4
RESULT ; DISCUSSION
4.1 Synthesis of galanthamine acetate
This research overall about reacting galanthamine with acetic acid to produce galanthamine acetate using Novozym435 as a catalyst.Acetic acid act as solvent free system since the solvent is the most common solvent to be found. The reaction to synthesis galanthamine acetate is as follows:
2165985899795?
?
34709101720850Galanthamine
Galanthamine
7181851638935Acetic acid
Acetic acid

28136851949452737485213360
303276029845Novozym435
Novozym435
173672558420Ethanol
Ethanol

Galanthamine acetate
Figure 6 : Esterificaiton reaction to produce galathamine acetate
4.2 Purification, Identification and Characterization of galanthamine acetate
4.2.1 Thin Layer Chromatography (TLC)
The first thin layer chromatography need was used to determine the presence of galanthamine during the neutralization process of galanthamine hydrobromide to form galanthamine by removing HBr. Ammonium hydroxide is the solution used to react with the HBr. Different compound can be seen on the silica plate depending on their polarity. Next, the second TLC used to determine galanthamine acetate when reacting the pure gaalnthamine with the acetic acid to form galanthamine acetate. Galanthamine acetate is the more polar compared to the other compound, so it was elute slower with lower retention time. Then, acetic acid is less polar than galanthamine acetate hence it will elute faster and give higher retention time. The solvent used as a mobile phase for this sample is hexane:ethyl acetate (3:7) successfully showed peak before and after purification. The silica plate that contain sample was put in the chamber with mobile phase inside it and let it absorb the solvent from the base line until the front line. Then, the silica plate was immersed into KMnO4 and let it dry upon air.

Based on the figure below, the retention time was calculated based on
Rf =distance from based line travelled by solute/distance from base lined travelled by solvent
Rf = a/b
The retention time was calculated for each of the compound which are galanthamine, acetic acid and galanthamine acetate. Before purification, there were two peak that was observed on the silica plate. The highest retention time is 0.72 for the acetic acid and the other one is for galanthamine acetate compound with retention time about 0.36. This mean that the galanthamine not fully react with acetic acid, hence the column chromatography was needed to get pure galanthamine acetate.

4.2.2 Column Chromatography
Column chromatography was used to purify the galanthamine acetate compound. The ratio of the solvent used was ethyl acetate:methanol(3:1). the column was packed and let it 24 hours before pouring the product. The total volume of solvent is 500 ml where 375 ml methanol and 125 ml ethyl acetate. The amount of vial needed can be more than 40 depending on the polarity of the compound.The vials from 1 until 27 started with less polar compound that was eluted first followed by the desired compound, galanthamine acetate which is more polar. The product of galanthamine acetate was collected from vial 25 until 36. next, the researcher used tlc plate to check on the presence of the galanthamine acetate either it is successfully purified or not fully purified.
After purification, there was only one peak on the silica plate at retention time 0.40. It means that column chromatography is the crucial step that need to be done so that the researcher will get pure target product.

4.2.3 Gas Chromatography Mass Spectroscopy (GCMS)
Gas Chromatography Mass Spectroscopy was used to determine the molecular weight of the desired compound. The expected molecular weight of galanthamine acetate is 347.65 g/mol. The GC-2010 was used as an instrument to run the sample. The column oven temperature was 50°C, the injection temperature was 250°C, injection mode was split, the flow control mode was linear velocity, the pressure was 53.5 kPa,
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Ping Jia, Rong Sheng, Jing Zhang, Liang Fang, Qiajun He, Bo Yang, Yongzhou Hu, (2009) Design,synthesis and evaluation of galanthamine derivatives as acetylcholinesterase inhibitors. Europian Journal Medicinal Chemistry, 44 pp. 772-784.

Rao, V.K., Rao, A.J., Reddy, S.S., Raju, C.N., Rao, P.V., Gsosh, S.K.. (2010). Synthesis, spectral characterizaiton and biological evaluation of phosphorylated derivaitves of galanthamine. Europian Journal of Medicinal Chemistry, 45 pp. 203-209.
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Stergiou, P-Y., Foukis, A., Filippou, M., Koukouritaki, M., Parapouli, M., Theodorou, L.G., Hatziloukas, E., Afendra, A., Pandey, A., Papamichael, E.M. (2013). Advances in lipase catalyzed esterification reactions. Biotechnology Advance, 31 pp. 1846-1859.

Trinadachari, G.N., Kamat, A.G., Babu K.R., Sanasi P.D.,Prabahar K.J. (2014). Stereoselective syntheses of galanthamine and its stereoisomers by complimentary Luche and L-selectride reductions. Tetrahedron:Assymmetry, 25 pp. 117-124.

Anand, P., Singh, B. (2013). A review on cholinesterase inhibitors for Alzehimer disease, 36 pp. 375-399.

Bertelli, P.R., Biegelmeyer, R., Rico, E.P., Klein-Junior, L.C., Toson, N.S.B., Minetto, L., Bordignon, S.A.L., Gasper, A.L., Moura, S., Oliveira, D.L., Henriques, A.T. (2017) Toxicological profile and acetylcholinesterase inhibitory potential of Palicourea deflexa, a source of ?-carboline alkaloids, Part C 201 pp. 44-50.

Rao, V.K., Rao A.J., Reddy, S.S., Raju, C.N., Rao, P.V., Gsosh, P.V. Synthesis, spectral characterization and biological evaluation of phosporylated derivatives of galanthamine, 45 pp. 203-209.
Lawrence, C. (2007) The husbandry of zebrafish (Danio Rerio) : A review, 269 pp. 1-20.

Shangde, S., Bingxue, H. (2017) A novel method for the synthesis of glyceryl monocaffeate by the enzymatic transesterification and kinetic analysis, 214 pp. 192-198

Chapter 1
Introduction

1.1 Introduction
Mehsana city is one of the important city in the north Gujarat. Number of people come to Mehsana for the job aspect and education aspect from the neighbor’s city like patan and palanpur. So that parking is very important factor in Mehsana city. So that One of the problem created by road traffic is parking. Not only do vehicle required street space to move about, but also do they require space to park where to park where the occupants can be loaded and unloaded. Parking system is very important for the transportation system in India. So it can be design by two major method which are on-street parking and off-street parking. In now a day in India vehicle culture are fast growing so that it create a lack of parking spaces in cities. So it can make a one of the biggest problem in the city. The most of parking and traffic problem in city which making a main CBD area like a shopping mall, bazar. Now a day major cities accepted the smart future parking system like a multy story parking, underground parking, roof parking so it could be helpful to control the parking problem.Parking control has become the chief means available to cities all over the world do limit congestion. It is the enforcement of laws and regulations. The size of average parking space is 14m2. This result in a great demand for parking space.in the CBD and other area where the other acitivites are concentrated. Parking should be control by below method which are:-
There are two type of parking system
1 On-street Parking
2 Off street parking
On street parking
On street parking means the vehicles are parked on the sides of the street itself. This will be usually controlled by government agencies itself. Common types of on-street parking are as listed below. This classification is based on the angle in which the vehicles are parked with respect to the road alignment. As per IRC the standard dimensions of a car is taken as 5× 2.5 meters and that for a truck is 3.75× 7.5 meters.
1. Parallel parking: The vehicles are parked along the length of the road. Here there is no backward movement involved while parking or unparking the vehicle. Hence, it is the most safest parking from the accident perspective. However, it consumes the maximum curb length and therefore only a minimum number of vehicles can be parked for a given kerb length. This method of parking produces least obstruction to the on-going traffic on the road since least road width is used. The length available to park N number of vehicles, L = N 5.9
2. 30? parking: In thirty degree parking, the vehicles are parked at 30? with respect to the road alignment. In this case, more vehicles can be parked compared to parallel parking.
3. 45? parking: As the angle of parking increases, more number of vehicles can be parked. Hence compared to parallel parking and thirty degree parking, more number of vehicles can be accommodated in, length of parking space available for parking N number of vehicles in a given kerb is L = 3.54 N+1.77
4. 60? parking: The vehicles are parked at 60? to the direction of road. More number of vehicles can be accommodated, length available for parking N vehicles =2.89N+2.16.
5. Right angle parking: In right angle parking or 90? parking, the vehicles are parked perpendicular to the direction of the road. Although it consumes maximum width kerb length required is very little. In this type of parking, the vehicles need complex maneuvering and this may cause severe accidents. This arrangement causes obstruction to the road traffic particularly if the road width is less. However, it can accommodate maximum number of vehicles for a given kerb length. Length available for parking N number of vehicles is L = 2.5N.
Off street parking
In many urban centers, some areas are exclusively allotted for parking which will be at some distance away from the main stream of traffic. Such a parking is referred to as off-street. Off-street parking means parking your vehicle anywhere but on the streets. These are usually parking facilities like garages and lots. Off-street parking can be both indoors and outdoors. Off-street parking also includes private lots, garages and driveways. The users of on-street parking are casual users who use the space for a short period of time. Off street parking users differ from short to long-term, i.e. monthly tenants and regular users. In many urban centers, some areas are exclusively allotted for parking which will be at some distance away from the mainstream of traffic. Such a parking is referred to as off-street parking. They may be operated by either public agencies or private firms.

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CHAPTER 1
INTRODUCTION
1.1 INTRODUCTION
Brain tumors do not discriminate. Beginning in the brain they tend to stay there in people of all age groups but are statistically more frequent in older adults and children. Metastatic brain tumors i.e. which begin as cancer else where in the body and reach to the brain are more common in adults.

As per the American Brain Tumor Association 80,000 primary brain tumor cases are predicted to be diagnosed this year in America alone.

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Gliomas are the most common type of tumor and surgery is the most common type of treatment but radiations and chemotherapy may also be used to slow down the growth of the tumors that cannot be physically removed. Magnetic Resonance images imaging may be used to provide detailed images of brain.

Tumors such as meningiomas, gliomas and glioblastomas can be easily segmented using the method of image processing proposed in here. Tumors can be anywhere in the brain in any shape and size. Gray scale values of MRI may vary depending on field of view, voxel resolution, gradient strength, type of MRI machine (1.5 Tesla, 3 Tesla, 7 Tesla) which varies for different hospitals.

Goal of our project is to identify a tumor from a given MRI scan of the brain with its location and extension using digital image processing which is done by identifying abnormal high intensity areas when compared to normal tissues.

1.2 Magnetic Resonance Imaging
Magnetic Resonance Imaging is based on the theory that protons and neutrons of the nucleus have an angular momentum which is called spin. When the number of subatomic particles is even the spins are cancelled whereas nuclei with odd number of subatomic particles will have a spin.

The MRI scanner uses powerful magnets to polarize and excite hydrogen proton in human tissue which produces a signal that can be encoded and detected, resulting in the images of the body being targeted. The part of the body that is to be examined is subjected to radio frequencies emitted by MRI machine which binds only to the hydrogen. Due to the radio frequency pulse, protons in the targeted area absorb the energy needed to make them spin in a specified direction.

MRI uses three electromagnetic fields which is static field (strong static electromagnetic field which polarizes the hydrogen nuclei), gradient field (weaker and time varying field) and weak radio frequency field used for manipulation of hydrogen nuclei to produce measurable signals which are received via radio frequency antennae.

1.3 Challenges
Brain is the key part of the central nervous system. Abnormal and uncontrolled cell division in the brain leads to brain tumor. We have used axial view of the brain from the MRI scan as MRI scan is less harmful as compared to CT scan. Several techniques such as imaging, biopsy, MRI, CT scan of the brain can be done based on the symptoms described by the patient. In biopsy a specimen of the brain tissue which is considered to be an tumor is taken by the pathologist and looks under the microscope to check for the presence of abnormalities in the tissue cells. Though doctors will only be able to know that the patient has tumor or not using the biopsy. To get the exact location of the tumor for being operated, MRI images are taken.

Traditional methods in hospitals is to segment the image manually and this depends on how well the doctor can perceive the image to get the required region extracted out which is made difficult with the minute variations and resemblances and original parts in the image.

The shortage of radiologists and large number of MRIs to be analyzed makes the task labor intensive and also increases the cost. It also depends on the experience and expertise of the doctor examining the images. Estimates indicate that between 10% and 30% are missed by radiologists during the screening process.

1.4 RELATED WORK
Various approaches have been carried out in the field of brain tumor segmentation. Sindhushree. K.S, et. al1 have developed a brain tumor segmentation method on 2D MRI data. Also detected tumors are represented in 3 dimensional view.

High pass filtering, histogram equalization, thresholding, morphological operators and segmentation using connected component labeling was carried out to detect tumor. The 2-dimensional extracted data was reconstructed into 3-Dimensional volumetric data and the volume of the tumor was also calculated.

M.C. Jobin Christ and R.M.S. Parvathi 2 proposed a methodology that integrates K Means clustering with marker controlled watershed segmentation algorithm. The proposed approach is a two stage process. First K Means clustering is used to get a primary segmentation of the input image and secondly marker controlled watershed algorithm is applied to the primary segmentation to get the final segmentation.

P.Vasuda, S.Satheesh 3, proposed a technique to detect tumors from MRI images using fuzzy clustering technique. This algorithm uses fuzzy C-means but the major drawback of this algorithm is the computational time required. Classifiers are also known as supervised methods since they require training data that are manually segmented and then used as references for automatically segmenting new data. The use of the same training data for classifying a large number of images, may lead to biased result. Supervised segmentation method requires considerable amount of training and testing data which comparatively complicates the process.

Vinay Parameshwarappa and Nandish S. et al, 2014 in his paper “Segmented morphological approach to detect tumor in brain images” 4, they proposed an algorithm for segmented morphological approach.

Sentilkumaran N and Thimmiaraja et al, 2014, Compare the image enhancement techniques in his paper “Histogram equalization for image enhancement using MRI brain images” 5, they presented the study of image
enhancement techniques and comparison of histogram equalization basic method like Brightness preserving adaptive histogram equalization (AHE), Local histogram equalization (LHE), global histogram equalization (GHE), Dynamic histogram equalization using different quality objective measures in MRI images.

CHAPTER 2
PROPOSED METHODOLOGY
PROPOSED METHODOLOGY
The part of the image the tumor normally has more intensity then the other portion and we can assume the area, shape and radius of the tumor in the image. We have used these basic conditions to detect tumor in the selected MRI images and the code goes through the following steps:
Preprocessing
Gray Scale Image
High Pass Filter
Enhanced Image
Post Processing
Threshold Segmentation
Watershed Segmentation
Morphological Operators

The proposed block diagram is shown below:
2762250118406
Figure 1: Proposed Block Diagram
PREPROCESSING
In preprocessing some basic image enhancement and noise reduction techniques are implemented. Apart from that different ways to detect edges and doing segmentations have also been used. The purpose of these steps is to improve the image and the image quality to get more surety and ease in detecting the tumor. The basic steps in preprocessing are the following:-
Image is converted to gray scale image in first step.

Noise is removed if any
The obtained image is then passed through a high pass filter to detect edges
Then the obtained image is added to original image to enhance it.

222631043878500The images below show the different preprocessing steps:
123190045339000416623546926500
Figure 2: MRI Images after being converted to gray scale, passing from high pass filter and finally the enhanced image.

2.3 POST PROCESSING
It is the phase of the implementation where main tumor detection will take place. It consist of three steps namely, threshold segmentation, watershed
segmentation and morphological operator.

Threshold Segmentation
Image thresholding is a simple yet effective way of partionioning an image into a foreground and background. This image analysis technique is a type of image segmentation that isolates objects by converting grayscale images into binary images. It is most effective in images with high levels of contrast. Thus in the MRI image segmentation is done on the basis of a threshold due to which whole image is converted into a binary image.

The images below shows the original image (Figure 3) and the image after applying threshold segmentation (Figure 4) using the code snippet:
T = graythresh(c); bw = im2bw(c,T+0.3); imshow(bw);
Figure 3: Original Image

Figure 4: After applying threshold segmentation
2.3.2 Watershed segmentation
The term watershed refers to a ridge that divides areas drained by different river systems. The catchment basin is the geographical area draining into a river or reservoir.

213868046863000
Figure 5: Image displaying watershed line and catchment line (Courtesy: mathworks.com)
Watershed Segmentation is the best method to segment an image or to separate a tumor but it suffers from over and under segmentation as every region makes its own catchment area, due to which we have used it as a check to our output. We have not used watershed segmentation on our input, rather it is only used on our output to check if the result is correct or not.

.2494279152442
(Figure 6: Displaying Watershed Segmentation)

2.3.2 MORPHOLOGICAL OPERATOR
Morphology is the study of shapes and structures from a scientific perspective. Morphological filters are formed from the basic morphology operations. A structuring element is mainly required for any morphological operation. Morphological operations operate on two images, structuring element and the input image. Structuring elements are small images that are used to probe an input image for properties of interest. Origin of a structuring element is defined by the centre pixel of the structuring element. In morphology, the structuring element defined will pass over a section of the input image where this section is defined by the neighbourhood window of the structuring element and the structuring element either fits or not fits the input image. Wherever the fit takes place, corresponding image that represents the input image’s structure is got suppression of the geometric features of the input
image that doesn’t fit the structuring element’s neighbourhood takes place. Two main morphology operations are erosion and dilation where erosion results in the thinning of the objects in the image considered and dilation results in thickening of the objects in the image. Dilation uses the highest value of all the pixels in the neighbourhood of the input image defined by the structuring element and erosion uses the lowest value of all the pixels in the neighbourhood of the input image. The basic purpose of the operations is to show only that part of the image which has the tumor that is the part of the image having more intensity.

The basic commands used in this step are:
Strel
Imerode
Imdilate
For a better understanding the code snippets are given below with their outputs.

SE = strel(‘disk’,0);
bw1 = imerode(bw,SE); imshow(bw1);
SE = strel(‘disk’,0); bw1 = imdilate(bw1,SE); imshow(bw1);

Figure 7: Applying snippet (A) Figure 8: Applying Snippet (B)
Figure 9 : (Imerode With strel(‘disk’,6);)

Figure 10: (Imdilate With strel(‘disk’,6);)
CHAPTER 3
SOFTWARE REQUIREMENT SPECIFICATION
3.1 PURPOSE
The purpose of this project is to create an application that allows the user to select the MRI image with the help of the user friendly interface and get the output which shows the tumor and its area if present in the MRI.

3.2 SCOPE OF THE PROJECT
The application developed in this project has the potential to be used as a means to analyze MRI images for tumor after performing rigorous testing.

3.3 TECHNOLOGY USED
Matlab 7.12.0 is used for developing the application and the user friendly graphical user interface. MATLAB is a high-performance language for technical computing. It integrates computation, visualization, and programming in an easy-to-use environment where problems and solutions are expressed in familiar mathematical notation. MATLAB is an interactive system whose basic data element is an array that does not require dimensioning. This allows you to solve many technical computing problems, especially those with matrix and vector formulations.

3.4 SYSTEM REQUIREMENT
The application was developed using Matlab 2011. Any device that can support Matlab 2011 or its superior versions along with necessary memory space and disk storage space will be able to execute our application.

DESCRIPTION OF MAIN FUNCTIONALITY
This application allows user to select a MRI image and by clicking on the output button the interface displays the tumor region in the MRI if present.

The output is displayed within seconds with preprocessing and post processing taking place in backend.

Figure 11: Graphical User Interface for the application
CHAPTER 4
SCREENSHOTS ; OUTPUT
4.1 The Console
100965044704000
Figure 12: The console window for the application
114490545466000Loading/Selecting the MRI image
Figure 13: The MRI image is loaded into the console detect tumor
Figure 14: Clicking on detect tumor generates the images for different functions
Figure 15: The tumor is shown in the output image.

Other Results: Different types of MRI images taken different angles are passed through this application and the tumor region is successfully found as shown in the results below.

Figure 16: Colored MRI image and its output

Figure 17: Different location and size of tumor
Figure 18: Different location and size of tumor

Figure 19: Different location and size of tumor
Figure 17,18,19 and 20 clearly shows that the application is able to find the tumors located at different places of brain and of different sizes.

Below are few mapped resultant tumor image onto the original grayscale image
20
lab code is able to distinguish between edema and tumor and does not return edema as tumor.

For all the MRI images the output has always been for correct irrespective of the size and location of the tumor.

CONCLUSION
Although there are many applications that implement brain tumor segmentation but this concept helps us to overcome many shortcomings of the popular methods such as cost and time as the methods pertaining to convolution neural network, fuzzy logics etc requires lot of complex operations which turns out to be costly and time consuming.

To improve the productivity and accuracy image processing can be combined with other soft computing techniques to provide a better and more accurate results with numerical values.

5.3 FUTURE WORK
With more research and rigorous testing this project can be used in future to identify brain tumors in low and high grade of MRI images. The code can also be embodied in MRI machines which also displays the result for tumor presence.

REFERENCES
Sindhushree. K. S, Mrs. Manjula. T. R, K. Ramesha, Detection And 3d Reconstruction Of Brain Tumor From Brain Mri Images, International Journal of Engineering Research ; Technology (IJERT), vol. 2, no. 8, pp 528-534, 2013.

M.C. Jobin Christ, R.M.S.Parvathi, “Segmentation of Medical Image using Clustering and Watershed Algorithms”, American Journal of Applied Sciences, vol. 8, pp 1349-1352, 2011.

P.Vasuda, S.Satheesh, “Improved Fuzzy C-Means Algorithm for MR Brain Image Segmentation,International Journal on Computer Science and Engineering (IJCSE), vol. 02, no.05, pp 1713-1715, 2010.

Vinay Parmeshwarappa, Nandish S, “A segmented morphological
approach to detect tumor in brain images”, IJARCSSE, ISSN: 2277 128X , volume 4, issue 1, January 2014.

Senthilkumaran N, Thimmiaraja J,”Histogram equalization for image enhancement using MRI brain images”, IEEE CPS,WCCCT.2014.45.

Pratibha Sharma, Manoj Diwakar, Sangam Choudhary, “Application of Edge Detection for Brain Tumor Detection”, International Journal of Computer Applications, vol.58, no.16, pp 21-25, 2012.

Anam Mustaqeem, Ali Javed, Tehseen Fatima, “An Efficient Brain Tumor Detection Algorithm Using Watershed ; Thresholding Based Segmentation”, I.J. Image, Graphics and Signal Processing, vol. 10,no. 5, pp 34-39, 2012.

Chang Wen Chen, Jiebo Luo, Kevin J. Parker,”Image Segmentation via Adaptive K-Mean Clustering and Knowledge-Based Morphological Operations with BiomedicalApplications”, IEEE Trans. Image Process.,vol.7,no.12, pp1673-1683, 1998.

P.Dhanalakshmi , T.Kanimozhi, “Automatic Segmentation of Brain Tumor using K-Means Clustering and its Area Calculation”, International Journal of Advanced Electrical and Electronics Engineering ,vol. 2,no. 2,pp 130- 134, 2013.

G.Evelin Sujji, Y.V.S Lakshmi, G.Wiselin Jiji,”MRI Brain Image Segmentation based on Thresholding”, International Journal of Advanced Computer Research, vol.3 no.1, pp 97-101, 2013.

Swe zin Oo, Aung Soe Khaing, “Brain tumor detection and segmentation using watershed segmentation and morphological operation”, IJRET, Volume 03,Issue 03, March 2014.

Rajesh Kumar Rai, Trimbak R. Sontakke, “Implementation of Image Denoising using Thresholding Techniques”, International Journal of Computer Technology and Electronics Engineering (IJCTEE),vol.1,no. 2, pp 6-10, 2013.

Mohammed Y. Kamil, “Brain Tumor Area Calculation in CT-scan image using Morphological Operations”, IOSR Journal of Computer Engineering (IOSR-JCE) ISSN: 2278-8727, Volume 17, Issue 2, Ver.-V, pp 125-128, Mar
– Apr. 2015.

Poonam, Jyotika Pruthi, “Review of image processing techniques for automatic detection of tumor in human brain”, IJCSMC, Vol.2, Issue.11, pp.117-122, November 2013.

A.Sindhu1 , S.Meera2 “A Survey on Detecting Brain Tumor in mri Images Using Image Processing Techniques “International Journal of Innovative Research in Computer and Communication Engineering Vol. 3, Issue 1, January 2015.

Team Details:
Project Member name : signature:

Utsav rawat (1501051147)
Manish bhati (160105900)
Kundan singh (1501051198)
Rashesh Dwivedi (1501051158)
Guide Name and Signature:
Mr. Nishant Singh Rathore
Assistant Professor
IT Department
DIT University

Signature(guide): ______________

Chapter 1
Introduction
1.0Background to the Study
Wastes are spent materials. They are any materials that are discarded after its primary use, as it has been deemed unusable. Historically, the amount of wastes generated by mankind was insignificant mainly due to the low population densities, coupled with the fact that there was very little exploitation of natural resources.(Chukwuemeka, 2011) Common wastes produced during the early ages were mainly ashes and human and biodegradable wastes and these were reused as manure and some, discarded. Biodegradable waste typically originates from plant or animal sources, which are broken down by bacteria, fungi or by the activities of other micro-organisms. Non-biodegradable waste cannot be changed to a harmless natural state by the action of bacteria, and may therefore constitute hazard to the environment over a long period of time. With the advent of industrial revolution (1760-1840), waste management became a critical issue. This was due to the population explosion coupled with the massive migration of rural dwellers to urban centres during the 18th century. Consequent upon this, was a massive generation of wastes that posed threat to human survival and the environment.
Today, the generation and disposal of waste is an intrinsic part of any developing or industrial society. Waste, both from domestic and commercial sources has grown significantly in Nigeria over the past decade. Every time a householder shops at the Store and open market, the individual contributes to the mountain of waste. (Jimoh, 2005) According to Chukwukelo Chukwogo, Nigeria generates 24 million tons of waste annually (The Sun, August 2017). The population of urban dwellers in Nigeria has more than doubled in the last 15 years (Jimoh, 2005). This population explosion has aggravated the problems associated with waste disposal.
Studies have shown that the volume of waste does not actually constitute the problem but the ability or inability of governments and the public to keep up with the task of waste management. The quality of life of the citizenry is affected by poor standard of living, which affects aesthetic abilities and the state of health of the people. A corollary to the aforementioned is that improper waste management procedures can constitute hazards to the society. The existing waste management system, where the garbage is collected from the streets, houses and other establishments on quotidian basis, has been largely ineffective.

Currently, most operations of municipal waste collectors focused on emptying containers according to predefined schedules (Fixed route collection process). This is sufficiently inefficient. One of the main attractions of IoT is its ability to efficiently control devices. In the waste industry, the goal is to maximize productivity. One way to achieve this feat is to minimize cost through the adaptation of effective but proficient resource management tools, while at the same time automate the existing process.
The IOT could be a pathfinder in modernizing waste collection methods with an impeccable database. It can change the way waste collectors carry on with their operations, know more information about their bins and bring change in the method of waste collection. Consumers on the other hand, will be able to pay reduced premium and other related costs. In short, the process of providing insurance, servicing the policies and settling the claims will be much more efficient and transparent in terms of processes. (Patil et al). A proper waste management system is necessary to avoid spreading some deadly diseases. Managing the smart bins by monitoring the status of it and accordingly taking the decision. This waste is further picked up by the municipal corporations to finally dump it in dumping areas and landfills. But due to lack of resources, ineffective groundwork, some waste is not collected which poses serious health hazard to the surrounding environment (Khan, Aliasgar, Naik et al, 2017). In the proposed system, multiple waste bins will be used and these bins will be provided with low-cost embedded devices which will help in tracking the level of the garbage bins and a unique ID will be provided for each of the waste bins so that it is easy to identify which garbage bin is full (Khan,et al, 2017). The Smart dust bins are connected to the internet to get the real time information of the smart dustbins. These dustbins are interfaced with arduino uno based system with ultrasonic sensors.
When the level reaches the threshold limit, the device will transmit the level of the trash, along with the unique ID provided and the location of the bin, to the concerned authority with the help of Internet and an immediate action can be made to clean the dustbins. An ultrasonic sensor will detect the level of waste in the bin using height, and send the appropriate level to the system. We would introduce a proximity sensor to detect presence of people who want to dispose waste. A PIR sensor is a motion detector sensor which detects the heat emitted naturally by human. Whenever a person is in the field of vision nearer to the garbage bin, the sensor is triggered and the servo motor automatically opens the smart bin for the disposal of waste. In order to prevent decaying smell around the bin, a harmless chemical (like baking soda) sprinkler will be used as soon as the smell detector senses the decaying smell. As a filled dustbin will be made to stay shut even after sensing human presence, the location of the nearest serviceable bin will be displayed with the help of an LCD.

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Problem Statement:
The greatest problem regarding waste management in developing countries begins at the very beginning of the process. Due to lack of proper systems for disposal and collections, wastes and garbage end up on the roads and surrounding. With the existing methods of collecting and disposal it is near impossible to manage such amount of waste in the future as around 30% of waste end up on the roads and public places due to ineffective disposing and collecting methods. There is no systematic methodology for treating the collected garbage thus most of them end up in landfilling water channels, making the environment unhealthy. The prime impediment of implementing smart waste management system based on IoT in a developing country is the social and economic infrastructure of the country itself. The initial stage of this system comprises of proper disposal and collection, which is the biggest challenge. In addition, to motivate and influence people to follow proper waste disposal methods is also important (Rakib ,Golam, Labib et al, 2017). Therefore, the concise statement of the problem is to optimize cost by:
Maximizing the volume of the waste bin
Minimizing number of trips taken by the waste vehicle to the various locations
Minimize fuel consumption
Minimize time taken to get to the various points of garbage collection.

1.2 Aims and Objectives
The purpose of the project is to investigate the possibility of successfully managing the monitoring of waste in the university of Lagos and also to design and build a prototype for the detection of the level of trash inside the dustbin. A harmless chemical sprinkler will also be used to avoid the decaying smell around the bin.
The main objective of our proposed system are as follows;
To provide real time information on the level of waste in the dustbin and send it to the appropriate authorities.
To detect the presence of people that want to dispose trash
To manage awful smell that may arise from decaying waste in the bins, prior to the time of collection.

To reduce manpower required to handle the garbage collection.

1.3 Significance of the project
Waste management using Internet of things can be applied in optimization of route planning and scheduling in waste collection. This can lead to significant reduction of cost. This technology has the potential to eliminate all forms of environmental contamination and human threatening diseases.
The advantages of this mode of monitoring wastes include the following;
Our smart operating system enables two way communication between the dustbin deployed in the city and service operator. Therefore the focus is only on collection of route based fill level of the containers.
Historical information on collections helps adapt the deployment of containers to the actual needs of the city, therefore reducing the number of containers that clutter up the road and increasing public parking spaces.

It keeps the surroundings clean and green, free from bad odour of wastes, emphasizes on healthy environment and keep cities more beautiful.
Reducing manpower required to handle the garbage collection.
1.4 Scope and limitations
The scope of this study is to build an efficient system that monitors and tracks the level of waste in the garbage bins and informs the appropriate authority about the level of garbage collected in the garbage bins via a web page. It would make use of a smell detector to detect offensive smell caused by decaying waste and take care of this with the aid of harmless chemical sprinklers. It would also detect the presence of people that want to dispose waste and open the lid for them. For this the system uses ultrasonic sensors placed over the bins to detect the garbage level and compare it with the garbage bins depth. The system makes use of AVR family microcontroller, LCD screen, Wi-Fi modem for sending data and a buzzer. The system is powered by a 12V transformer. The LCD screen is used to display the status of the level of garbage collected in the bins, and a web page is built to show the status of the bins to the authority monitoring it. The web page gives a graphical view of the garbage bins and highlights the garbage collected in color in order to show the level of garbage collected. The LCD screen shows the status of the garbage level. The system puts on the buzzer when the level of garbage collected crosses the set limit. Thus this system helps to keep the city clean by informing about the garbage levels of the bins by providing graphical image of the bins via IOT Gecko web development platform. Sensing of refuse dumped around the neighborhood of the waste bins, separation of liquid and solid waste and defining path optimization strategies for garbage collection goes beyond the scope of this work.

Main Equipments Used In The Smart Waste Management System.

Hardware Specifications
AVR family microcontroller
Wi-Fi Modem
LED’s
LCD Display
12V transformer
Ultrasonic sensors
Resistors
Capacitors
Diodes
PIR Sensor
Software Specifications
Arduino uno compiler
IOTGeckoMC Programming Language: Embedded C
Garbage Container
A waste container is a container for temporarily storing waste, and is usually made out of metal or plastic. The curbside dustbins usually consist of three types: trash cans (receptacles made of metal or plastic), dumpsters (large receptacles similar toskips) and wheelie bins (light, usually plastic bins that are mobile). All of these are emptied by collectors, who will load the contents into a garbage truck and drive it to a landfill, incinerator or consuming crush facility to be disposed of.

Ultrasonic Sensor
A special sonic transducer is used for the ultrasonic proximity sensors, which allows for alternate transmission and reception of sound waves. The sonic waves emitted by the transducer are effected by an object and received back in the transducer. After having emitted the sound waves, the ultrasonic sensor will switch to receive mode. The time elapsed between emitting and receiving is proportional to the distance of the object from the sensor. Ultrasonic sensors generate high-frequency sound waves and evaluate the echo which is received back by the sensor, measuring the time interval between sending the signal and receiving the echo to determine the distance to an object.

Arduino Board
Arduino is a software company, project, and user community that designs and manufactures computer open-source hardware, open-source software, and microcontroller-based kits for building digital devices and interactive objects that can sense and control physical devices 3. The project is based on microcontroller board designs, produced by several vendors, using various microcontrollers. These systems provide sets of digital and analog I/O pins that can interface to various expansion boards (termed shields) and other circuits. The boards feature serial communication interfaces, including Universal Serial Bus (USB) on some models, for loading programs from personal computers. For programming the microcontrollers, the Arduino project provides an integrated development environment(IDE) based on a programming language named Processing, which also supports the languages C and C++.

The first Arduino was introduced in 2005, aiming to provide a low cost, easy way for novices and professionals to create devices that interact with their environment using sensors and actuators.

SOFTWARE OF ARDUINO :The Arduino project provides the Arduino integrated development environment (IDE), which is a cross-platform application written in the programming language Java. It originated from the IDE for the languages Processing and Wiring. It is designed to introduce programming to artists and other newcomers unfamiliar with software development. It includes a code editor with features such as syntax highlighting, brace matching, and automatic indentation, and provides simple one-click mechanism to compile and load programs to an Arduino board. A program written with the IDE for Arduino is called a “sketch” 4. The Arduino IDE supports the languages C and C++ using special rules to organize code.

BREAD BOARD
A breadboard is a construction base for prototyping of electronics. Originally it was literally a bread board, a polished piece of wood used for slicing bread. In the 1970s the solderless breadboard (AKA plugboard, a terminal array board) became available and nowadays the term “breadboard” is commonly used to refer to these. “Breadboard” is also a synonym for “prototype”. Because the solderless breadboard does not require soldering, it is reusable. This makes it easy to use for creating temporary prototypes and experimenting with circuit design. For this reason, solderless breadboards are also extremely popular with students and in technological education. Older breadboard types did not have this property. A stripboard (veroboard) and similar prototyping printed circuit boards, which are used to build semipermanent soldered prototypes or one-offs, cannot easily be reused. A variety of electronic systems may be prototyped by using breadboards, from small analog and digital circuits to complete central processing units (CPUs).

A modern solderless breadboard consists of a perforated block of plastic with numerous tin plated phosphor bronze or nickel silver alloy spring clips under the perforations. The clips are often called tie points or contact points. The number of tie points is often given in the specification of the breadboard. The spacing between the clips (lead pitch) is typically 0.1 in (2.54 mm). Integrated circuits (ICs) in dual in-line packages (DIPs) can be inserted to straddle the centerline of the block. Interconnecting wires and the leads of discrete components (such as capacitors, resistors, and inductors) can be inserted into the remaining free holes to complete the circuit. Where ICs are not used, discrete components and connecting wires may use any of the holes.

JUMP WIRES
Jump wires (also called jumper wires) for solderless breadboarding can be obtained in ready-to-use jump wire sets or can be manually manufactured. The latter can become tedious work for larger circuits. Ready to- use jump wires come in different qualities, some even with tiny plugs attached to the wire ends. Jump wire material for ready-made or homemade wires should usually be 22 AWG (0.33 mm2) solid copper, tin-plated wire – assuming no tiny plugs are to be attached to the wire ends. The wire ends should be stripped 3?16 to 5?16 in (4.8 to 7.9 mm). Shorter stripped wires might result in bad contact with the board’s spring clips (insulation being caught in the springs). Longer stripped wires increase the likelihood of short-circuits on the board. Needle-nose pliers and tweezers are helpful when inserting or removing wires, particularly on crowded boards.

REFRENCES
1. Jimoh I.A (2005), A new Approach to Municipal Waste Management in Nigeria, International Conference on Energy, Environment and Disasters- INCEED, Charlotte N.C, USA.- July 24-30.

2. http://sunnewsonline.com/nigeria-generates-24m-tonnes-of-waste-annually/3. Patil et al., International Journal of Advanced Research in Computer Science and Software Engineering 7(4), April- 2017, pp. 407-410
3. “Arduino – Introduction”. arduino.cc.4. “Programming Arduino Getting Started with Sketches”. McGraw-Hill. Nov 8, 2011.

Retrieved 2013-03-28.5. Anton A. Huurdeman, The Worldwide History of Telecommunications, John Wiley & Sons, 31 July 2003, page 529
6. “GSM Global system for Mobile Communications”. 4G Americas. Retrieved 2014-03-22.7. Khan Shauzab, Aliasgar Lightwala, Naik Nadeem and Khan Shanawaz , Smart waste Management system using IoT. Anjuman I Islam Kalsekar technical campus, April 2017
8. B.M Rakib Hasan, A.M.M. Golam Yeazdani, Labib Md Istiaque, Rafee Mizan Khan Chowdhury, Smart Waste Management System Using Iot. Brac University, August 2017.

9. Okafor,Chukwuemeka Ikechukwu, Problems And Prospects Of Waste Management In Enugu State. A Case Study Of Enugu State Waste Management Authority (ESWAMA). University of Nigeria, Nsuka, April 2011

Chapter 2
Literature Review
2.0Preamble
For the sake of clarity, we have organized this chapter in three sections. Section 2.1 contains a review of works on the concept of waste, the environmental and economical impact of improperly managed waste. Section 2.2 gives a review of works on the concept of waste management practices and the historical background of waste management in Nigeria. Section 2.3 reviews works on the internet of things concept and section 2.4 delves into the environmental impact of solid waste management.

The Concept of Waste:
In a publication by Akinwale (2005), Nigel Bell defines wastes as rubbish or materials that are not needed and are economically unusable without further processing. Here, Nigel’s emphasis is that to ascertain something as a waste, it has to be economically unusable. However, Nigel’s position can be questioned because recent practices have shown that what one party considers as unneeded materials, and of course economically unusable, may be of utmost need and of economic importance to another party. For example, after drinking the liquid contents of a bottle of champagne, the empty bottle is considered as a waste by the person who drank the liquid content and perhaps is thrown away. But, another person may pick it up from the point of disposal and either reuse or recycle the empty bottle for containing another liquid substance or some other item of economic importance. The bone of contention here is that it is not clear to say at what point an item constitutes a waste.

Defra in Ogwueleka (2009) succinctly posits that there is no definitive list of what is and is not waste. It goes further to state that whether or not a substance is discarded as waste- and when discarded materials ceases to be waste- are matters that must be determined by the facts of the case and the interpretation of the law. Defra is of the opinion that whether or not a substance is discarded as waste, rests on one hand, with the producer or holder of such substance to decide whether it is being discarded as waste and, on the other hand, with regulations or laws stipulating it as such. Contributing to the subject matter, the Basel convention cast its vote to the school of thought that believes that wastes are “substances or objects which are disposed of or are intended to be disposed of or are required to be disposed of by the provisions of national law”. While the United Nations Statistics Division (UNSD) stated that “Wastes are materials that are not prime products (that is, products produced for the market) for which the generator has no further use in terms of his/her own purposes of production, transformation or consumption, and of which he/she wants to dispose”.
Section 32 of the Lagos State Environmental Sanitation Edict has defined waste as any substance which constitutes a scrap material, an effluent or other unwanted surplus substance arising from the application of any process. It further states that waste is usually classified according to:
(a) Its source,
(b) Its harmful effect on humans and the environment, and
(c) The control which are appropriate to deal with it.
With regards to the source classification, it either comes out of the shop (market) or office (commercial waste) or, out of the factory ( industrial waste), or out of the home (household or domestic waste). (Okafor, April 2011).
Moreover, it is clearly understood that what could be regarded as a waste can as well be valuable to another person and trading opportunities may arise if the cost of transportation of such item does not exceed the worth as perceived by the intending owner. (Thomas H. Christensen, p.4)
In addition to the aforementioned factors, what actually becomes waste depends on which items are being purchased and consumed. Or in other words culture, climate, religious and ethnic background as well as economical abilities affect what becomes waste. Hence, waste quantities and composition vary widely, both geographically (regionally, locally) and over time. (Thomas H. Christensen, p.4)
To be brief, we shall adopt our conclusion from the consensus of the Waste Framework Directive of the European Union (75/442/EC) that “once a substance or object has become waste, it will remain waste until it has been fully recovered and no longer poses a potential threat to the environment or to human health”. Therefore, anything which is discarded or otherwise dealt with as if it were waste shall be presumed to be waste unless it is proved otherwise. Waste, as a concept, does not exist in abstraction but has impacts as well as costs on nature and man. The Wikipedia free Encyclopedia 2010 observes three different costs of waste. These costs include:
Environmental Costs
Waste can attract rodents and insects which cause gastrointestinal parasites, yellow fever, worms, the plague and other bad health conditions. Exposure to hazardous wastes, particularly when they are burned, can cause various other diseases including cancers. Waste can contaminate water, soil, and air which causes more problems for man, other species, and the ecosystem. Waste treatment and disposal produces significant greenhouse gas (GHG) emissions, notably methane, which is contributing significantly to global climate change.

Social costs
Waste management is a significant environmental justice issue. Many of the environmental burdens cited above are more often borne by marginalized groups, such as racial minorities, women, and residents of developing nations. NIMBY (not-in-my-back-yard) is a popular term used to describe the opposition of residents to a proposal for a new development close to them. However, the need for expansion and siting of waste treatment and disposal facilities is increasing worldwide. There is now a growing market in the trans-boundary movement of waste. A significant amount of waste is moved from developed to developing nations. (Okafor, 2011)
Economic costs
The economic costs of managing waste are high, and are often paid for by municipal governments. Money can often be saved with more efficiently designed collection routes, modifying vehicles, and with public education. Environmental policies such as pay as you throw can reduce the cost of management and reduce waste quantities. Waste recovery (that is, refuse, recycling, reuse) can curve economic costs because it avoids extracting raw materials and often cuts transportation costs. The location of waste treatment and disposal facilities often has an impact on property values due to noise, dust, pollution, unsightliness, and negative stigma. The informal waste sector consists mostly of waste pickers who scavenge for metals, glass, plastic, textiles, and other materials and then trade them for a profit. This sector can significantly alter or reduce waste in a particular system, but other negative economic effects come with the disease, poverty, exploitation, and abuse of its workers. We shall now take a look at some of the wastes types that are of interest to the study.

2.2 Types of waste
There are many types of waste but a few shall be mentioned here for the purpose of the study, these are:
1. Municipal wastes
2. Industrial wastes
3. Agricultural wastes
4. Construction and demolition waste
5. Commercial and institutional waste
And Hazardous wastes which can stem from any of the above sources. Therefore it will not be taken as a part of the classification of wastes by source, rather as a cross-cutting character for all these wastes (UNESCAP, 1993). Moreover, the amount of waste generated in either developed or developing countries depends on the population, degree of urbanization and industrialization, and intensity of agricultural activities in such nation. Sources of waste according to their types are described below in details:
2.2.1 Municipal Solid Waste
Municipal solid waste (MSW), also called urban solid waste, is a waste type that includes predominantly household waste (domestic waste) with sometimes the addition of commercial wastes collected by a municipality within a given area. They are in either solid or semisolid form and generally exclude industrial hazardous wastes.

According to World Bank statistic (1999) it was estimated that high income countries generate MSW more than low or middle income countries. For example High income countries (such as Australia, Japan, Hong Kong, China, Republic of Korea, and Singapore) produce between 1.1 and 5.0 kg/capita/ day; middle-income countries (such as Indonesia, Malaysia and Thailand) generate between 0.52 and 1.0 kg/capita/day, whilst low-income countries (such as Bangladesh, India, Viet Nam and Myanmar) have generation rates of between 0.45 and 0.89 kg/capita/day.
The figure 1 below elaborate more on MSW generation by the high, middle and low-income countries of the region.

Figure 1: Municipal Solid Waste Generation in Different Groups of Countries in the Region adopted from (World Bank 1999)
In addition, Asian and Pacific Regions are said to be currently producing about 1.5 million tons of MSW each day and these are expected to be double by 2025 (World Bank 1999). Similarly in Africa, Thousands of tons are generated on daily bases which are approximately 0.5 kilograms per person per day and in some cases reaching as high as 0.8 kilograms per person per day. While this may seem modest compared to the1–2 kg per person per day generated in developed countries, most waste in Africa is not collected by municipal collection systems because of poor management, fiscal irresponsibility, equipment failure, or inadequate waste management budgets (EGSSAA 2009). Sources of Municipal solid waste are elaborated in Table 1. Table1: General sources of municipal wastes(adapted from peavy, Rowey, and Tchobanoglous, 1985)
2.2.2 Industrial Solid Waste
Industrial solid wastes are residues being generated by human activities which could be described as the activities carried out in the industry. These unwanted residues may occur as relatively pure substances or as complex mixtures of vary-ing composition and in varying physiochemical states which significant proportion of it are regarded as hazardous or potentially toxic, thus requiring special handling, treatment and disposal (UNESCAP 1993).
The major sources of industrial solid wastes are the thermal power plants produc-ing coal ash, the integrated Iron and Steel mills producing blast furnace slag and steel melting slag, non-ferrous industries like aluminum, zinc and copper produc-ing red mud and tailings, sugar industries generating press mud, pulp and paper industries producing lime and fertilizer and allied industries producing gypsum (The Ministry of Urban Development Government of India, ref. 20 May 2012).

2.2.3 Agricultural waste
Agricultural wastes are residues produced from different agricultural activities. For example, horticultural and forestry wastes, comprise crop residues, animal manure, diseased carcasses, unwanted agrochemicals and ’empty’ containers (UNESCAP, 1993). The composition and amount of agricultural wastes in a particular country depends on the agricultural system being practiced and similarly, the continued increase in productivity will significantly lead to a proportional increase in Agricultural waste.

2.2.4 Construction and Demolition Waste
C&D wastes is the type of waste generated when there is an activity such as building and demolition of roads, bridges, and fly over, subway, remodeling or remover of construction and to name of few. It comprises of inert and non-biodegradable material such as concrete, plaster, metal, wood, plastics and so on (The Ministry of Urban Development Government of India, ref. 20 May 2012).
In the past, C&D waste has not been given serious attention as a waste that has value rather than using it for Landfill .However in the recent years more concentration has been placed and information were discovered that many material in C&D wastes are contaminated either as part of their original design or through their use and therefore should be managed accordingly. Moreover, it has been noted that a large proportion of C&D waste (about 90 %) can be easily recycled and thus can conserve landfill capacity. (Thomas and Lizzi 2011, p.104).2.2.5 Commercial and Institutional Waste
Commercial and institutional waste is generated from anything from paper and packaging of obsolete equipment in different sectors like, retail (stores), hotels, restaurants, health care (except health risk waste), banks, insurance companies, education, retirement homes, public services and transport.(Thomas H. Christensen p.112). Commercial and institutional waste generated a significant portion of municipal waste (23%), even in small region and environs which are usually collected by private sectors and municipalities have been slower to target this waste stream for recovery. Nowadays many cities and regions have realized the economic importance of commercial and industrial waste recycling system; therefore they are actively participating in the practice.

2.2.5 Hazardous waste
Hazardous wastes are unwanted material that are extremely dangerous and harmful to human health, animal, plants or the environment and it require critical control system by necessary body.
Hazardous wastes can be liquids, solids, or contained gases. They can be the by-products of manufacturing processes, discarded used materials, or discarded un-used commercial products, such as cleaning fluids (solvents) or pesticides. In regulatory terms, a hazardous waste is a waste that exhibits one of the four characteristics of a hazardous waste – ignitability, corrosivity, reactivity, or toxicity. However, material can be hazardous even though it is not labeled as harzardous or have the characteristic of being hazardous (Oluwaleye, 2012). Hazardous wastes may pollute soil, air, surface water, or underground water. Pollution of soil may affect people who live on it, plants that put roots into it, and animals that move over it. Toxic substances that do not break down or bind tightly to the soil may be taken up by growing plants; the toxic substances may later appear in animals that eat crops grown there and possibly in people who do so. Air may become contaminated by direct emission of hazardous wastes. The air above hazardous waste may become dangerously contaminated by escaping gas, as can occur in houses built on mine tailings or old dump sites. River and lake pollution, if it is toxic enough, may kill animal and plant life immediately, or it may injure slowly. For example, fluoride concentrates in teeth and bone, and too much fluoride in water may cause dental and bone problems. Compounds such as dichlorodiphenyltrichloroethane (DDT), PCBs, and dioxins are more soluble in fats than in water and therefore tend to build up in the fats within plants and animals. These substances may be present in very low concentrations in water but accumulate to higher concentrations within algae and insects, and build up to even higher levels in fish. Birds or people that feed on these fish are then exposed to very high levels of hazardous substance. In birds, these substances can interfere with egg production and bone formation. Even pollution that is not toxic can kill. Phosphates and nitrates, usually harmless, can fertilize the algae that grow in lakes or rivers. When algae grow, in the presence of sunlight, they produce oxygen. But
if algae grow too much or too fast, they consume great amounts of oxygen, both when the sun is not shining and when the algae die and begin to decay. Lack of oxygen eventually suffocates other life; some living things may be poisoned by toxins contained in the algae. This process of algal overgrowth, called eutrophication, can kill life in lakes and rivers. In some cases, particular algae can also poison the drinking water of people and livestock. Irrespective of the category or type of wastes involved, the need for an effective and efficient management of wastes in the society becomes inevitable. According to Mowoe (1990:177) the management of waste is a matter of national and international concern. The volume of waste does not actually constitute the problem but the ability or inability of governments, individuals and waste disposal firms to keep up with the task of managing waste and the environment. There is no doubt that a dirty environment affects the standard of living, aesthetic sensibilities, health of the people and thus the quality of their lives. The corollary is that improper disposal or storage of this waste can constitute hazards to the society through the pollution of air, land and especially water as already noted above. What actually is waste management? Let’s take a look at what constitutes waste management and what does not. (Okafor, April 2011).

2.3 Concept Of Waste Management And Historical Background Of Waste Management In Nigeria
Waste management is the process of managing waste materials (normally those produced as a result of human activities). In order to define waste management, we need to include several different processes such as collection, transport, processing, recycling, disposing, and monitoring of waste. Along this line, the Wikipedia Web Encyclopedia 2010 defines waste management as the collection, transport, processing, recycling or disposal and monitoring of waste materials. According to Atsegbua L.A et al (2003:104), waste management does not just end at collection, transporting processing, recycling or disposal and monitoring of wastes materials but refers to the collection, keeping, treatment and disposal of wastes in such a way as to render them harmless to human and animal life, and the environment generally. In other words, the primary aim of managing wastes is for the safety of human, animal, and the environment. We could as well add here that any other aim that goes in contrary to the aims mentioned above does not constitutes waste management. Attah (2009) added that waste management could also be said to be the organized and systematic dumping and channeling of waste into landfills or through pathways to ensure that they are disposed of with attention to acceptable public health and environmental safeguard and that a proper waste management will result in the abatement or total elimination of pollution. Attah?s contribution raises some critical concerns with regards to the situation in Nigeria (generally). Perhaps one would be tempted to ask questions like;
Is dumping of wastes organized and systematic?
Is attention given to acceptable public health and environmental safeguard when it comes to waste disposal?
Have the LAWMA activities resulted in the abatement or total elimination of pollution in Lagos state?
Judging by sight, we can argue unequivocally that waste management in Lagos state does not conform to Attah’s definition of what waste management is. Whatever the perspective, the term waste management, in all its ramifications encompasses all steps taken in controlling the production, storage, collection, transportation, processing and disposal or utilization of wastes, in a sanitary manner (Taiwo: 2010).
2.3.1 Methods of Waste disposal
Practically, there are just two methods- those that are environmentally friendly e.g. sanitary or engineered landfill and composting, and those that are not, including incineration, stream dumping and open burning. Other existing methods include: petrification, bailing, land burial and pyrolysis, just to mention a few. Also of all the listed methods above only the sanitary landfill, stream dumping and land burial are permanent waste disposal systems while the others are just waste treatment i.e. meant to reduce the waste volume.
Landfill method:
Disposing of waste in a landfill involves burying the waste and this remains a common practice in most countries. Landfills were often established in abandoned or unused quarries, mining voids or burrow pits. A properly designed and well-managed landfill can be a hygienic and relatively inexpensive method of disposing of waste materials. Older, poorly designed or poorly managed landfills can create a number of adverse environmental impacts such as wind-blown litter, attraction of vermin, and generation of liquid leachate. Another common byproduct of landfills is gas (mostly composed of methane and carbon dioxide), which is produced as organic waste breaks down anaerobically. This gas can create odour problems, kill surface vegetation, and is a greenhouse gas.
Design characteristics of a modern landfill include methods to contain leachate such as clay or plastic lining material. Deposited waste is normally compacted to increase its density and stability, and covered to prevent attracting vermin (such as mice or rats). Many landfills also have landfill gas extraction systems installed to extract the landfill gas. Gas is pumped out of the landfill using perforated pipes and flared off or burnt in a gas engine to generate electricity.
Incineration method
Incineration is a disposal method that involves combustion of waste material. Incineration and other high temperature waste treatment systems are sometimes described as “thermal treatment”. Incinerators convert waste materials into heat, gas, steam and ash. Incineration is carried out both on a small scale by individuals and on a large scale by industry. It is used to dispose of solid, liquid and gaseous waste. It is recognized as a practical method of disposing of certain hazardous waste materials (such as biological medical waste). Incineration is a controversial method of waste disposal, due to issues such as emission of gaseous pollutants. Incineration is common in countries such as Japan where land is scarcer, as these facilities generally do not require as much area as landfills. Waste-to-energy (WTE) or energy-form- waste (EFW) is broad terms for facilities that burn waste in a furnace or boiler to generate heat, steam and/or electricity. Combustion in an incinerator is not always perfect and there have been concerns about micro-pollutants in gaseous emissions from incinerator stacks. Particular concern has focused on some very persistent organics such as dioxins, furans, PAHs, which may be created within the incinerator and afterwards in the incinerator plume which may have serious environmental consequences in the area immediately around the incinerator. On the other hand this method or the more benign anaerobic digestion produces heat that can be used as energy.
Plasma gasification method
Plasma is a highly ionized or electrically charged gas. An example in nature is lightning, capable of producing temperatures exceeding 12,6000F (6,9800C). A gasifier vessel utilizes proprietary plasma torches operating at +10,0000F (5,5400C) (the surface temperature of the sun) in order to create a gasification zone of up to 3,0000F (1,6500C) to convert solid or liquid wastes into a syngas. When municipal solid waste is subjected to this intense heat within the vessel, the waste’s molecular bonds break down into elemental components. The process results in elemental destruction of waste and hazardous materials. Plasma gasification offers states new opportunities for waste disposal, and more importantly for renewable power generation in an environmental sustainable manner (Alliance Federated Energy).
Recycling Method
The popular meaning of “recycling” in most developed countries refers to the widespread collection and reuse of everyday waste materials such as empty beverage containers. These are collected and sorted into common types so that the raw materials from which the items are made can be reprocessed into new products. Material for recycling may be collected separately from general waste using dedicated bins and collection vehicles, or sorted directly from mixed waste streams.
The most common consumer products recycled include aluminum beverage cans, steel food and aerosol cans, HDPE and PET bottles, glass bottles and jars, paperboard cartons, newspapers, magazines, and corrugated fiberboard boxes. PVC, LDPE, PP, and PS are also recyclable, although these are not commonly collected. These items are usually composed of a single type of material, making them relatively easy to recycle into new products. The recycling of complex products (such as computers and electronic equipment) is more difficult, due to the additional dismantling and separation required. Critics dispute the net economic and environmental benefits or recycling over its costs, and suggest that proponents of recycling often make matters worse and suffer from confirmation bias. Specifically, critics argue that the costs and energy used in collection and transportation detract from (and outweigh) the costs and energy saved in the production process; also that the jobs produced by the recycling industry can be a poor trade for the jobs lost in logging, mining, and other industries associated with virgin production; and that materials such as paper pulp can only be recycled a few times before material degradation prevents further recycling. Proponents of recycling dispute each of these claims, and the validity of arguments from both sides has led to enduring controversy.
2.3.2 Waste management in Nigeria
In Nigeria, especially in major urban centres, solid waste management is a critical problem. In fact, Nigerian Government has taken different steps in the past to combat the problem without success. You don?t need to look far before you see mountain of refuse in most of urban centres. Earlier on, the step taken was based on waste disposal on some designated landfills (that were not sanitary because they were not constructed with underlain (LDPE) to prevent leachate problem). This system i.e. one-fits-all does not work again because of increase in population and urbanization that effect the land use pattern. Then, when and where the municipal government cannot cope with waste collection and disposal successfully, people resolve into waste dumping into storm water, during the rainfall, open dumping and stream dumping. Based on observation, as Taiwo (2010) noted, waste management problem in most of Nigerian communities is multidimensional in nature. It is associated with lack of community participation in solid waste management. Most of policies concern this issue are made without considering the community people who are the waste generators. For instance, in a study conducted at Orita-Aperin communities in the year 2004, it was found out that attitudes and belief of community people affected their waste management practices. Furthermore, Taiwo stated that in Nigeria context, waste disposal is normally seen by the municipal government as a venture without any financial gain. That is the issues of environmental protection and healthful living are not very important to some health planners. In addition to this, the question of whose responsibility is to take care of waste generated in a community has not been clearly answered. Unless in some civilized areas, many people do not realized that they are liable to the disposal of wastes generated by them as they dump them by the road side for government workers to pick up!
Atsegbua (2003:104) observes that the problem associated with waste management in Nigeria does not appear to be a problem of absence of legislative framework for waste management but other factors have been identified as being responsible for penetrating the crises experienced in the management of waste in Nigeria. He highlighted these other factors as:
Lack of Adequate Funding and Excessive Population: waste management is by nature both capital and economic intensive. This requires huge capital outlay. Many state governments spend a good percentage of their funds on domestic waste management. For examples Lagos state Government spend between 2-25% of its funds on waste management. But what this amount could accomplish is dwarfed by the population it caters for. Lagos state, for instance, has a projected population of 12-18 million persons. It is estimated that the average individual in such mega cities as Lagos generates an average of 0.115kg of waste daily. It is that the funds available or at least earmarked for domestic waste management is grossly inadequate, to fund the public agencies and other private sector participants (PSP) involved in collection and disposal of domestic waste; to fund the procurement of equipment and materials required for effective domestic waste disposal.
Lack of Trained/Professional Waste Managers: There are just a few sanitation and environment Engineers in Nigeria. In fact most private sector operators in waste management are mainly party stalwarts; know little or nothing about waste management.
Lack of Effective Monitoring and Control: The Waste regime in the UK provides a quintessence of a system that makes for effective monitoring of domestic waste prior to disposal and the steps to be taken on disposal. The regime distinguishes between controlled and special waste. Under section 30 of the EPA, 1990, waste authorities in charge of waste administration have three basic functions: regulation, collection and disposal. Waste disposal authorities are to award waste disposal contracts through competitive tendering and are to make contracts with waste disposal contractors who may be private sector companies or companies set up by the local authority which must be at arm?s length from the waste authority. The waste regulation authority is responsible for issuing a waste management license. Under the regime, controlled waste may not be deposited, treated, kept or disposed of without a license. The licensing method issued as a means of controlling waste. Section 33(1)(a) of the EPA provides that it is an offence of “treat, keep or dispose of controlled waste in a manner likely to cause pollution of the environment or harm to human health. Pollution of the environment is defined in section 29 to mean the release or escape of the waste into any medium so as to cause harm to man or any other living organisms supported by the health of living organisms or other interference with the ecological systems of which they form part and in the case of man includes offence to any of his sense or harm to his property”. Thus the offensive smell of a waste tip would be covered, as presumably would its unattractive appearance. The offence can be committed whether or not the offender has a license. So the offence focuses on environmental protection, not with enforcing the licensing regime. The penalties are quite prohibitive. Again the duty of care principle under the EPA, designed to satisfy the European ideology on the environment that the polluter pays is an important form of liability on producers of domestic waste. The producer is responsible for the proper disposal of the waste. This means that the producer must ensure it is transferred to a responsible carrier. The producer cannot escape liability simply by passing the waste onto anyone else who could include the fly-tipper. This unbroken chain of waste transmission ensures that indiscriminate dumping and disposal is eliminated. The waste management regime in Nigeria is far from what is described above, so that the house-holder-producer of domestic waste is not deterred by any form of sanctions, because mostly, waste management agencies or contractors hardly exist in many places in Nigeria nor is monitoring and monitoring authorities effective.
Peculiarity of the Nigerians’ Attitude: The “government-does-everything” philosophy of many Nigerians contributes to the domestic waste management problems in Nigeria. A careless attitude permeates the thinking especially, those living in cities and towns. Self-help methods of domestic waste disposal are available and could be explored by individuals and institutions. Domestic incineration, landfill system is practicable, but most Nigerians would take to the easy way of depositing waste along the highway and corners of street for “government” to pick up. Some have founded this attitude on illiteracy but this would be a fallacy. Traditionally, as is still apparent in some of our villages, where a good number of individuals are still illiterate, residents are very conscious of the importance of having a clean environment and this is evidenced by the sanitation arrangements in force in these societies.
2.4Background of IOT
The Internet of Things (IOT) has not been around for very long. However, there have been visions of machines communicating with one another since the early 1800s. Machines have been providing direct communications since the telegraph ( the first landline ) was developed in the 1830s and 1840s. Described as “wireless telegraphy ,” the first radio voice transmission took place on June 3, 1900, providing another necessary component for developing the Internet of Things. The development of computers began in 1950s.
Internet of Things are therefore, sensors and actuators embedded in physical objects which are then linked through wired and wireless networks, often using the same Internet Protocol (IP) that connects the internet.
When thinking of the IOT device, think of a nearly endless supply of opportunities to interconnect our devices and equipments.
2.5Environmental Impact of Waste Management
Waste disposal is key especially when implemented the right way. Garbages, when not properly disposed can have adverse effects on the environment
1. Chemicals contaminating soil: When waste end up at the landfill, chemicals in the trash can leech out in the soil, contaminating it. This could destroy plants as well as weaken animals and humans who come in contact with such environment.
2. Air Pollution: Garbage can create air pollution due to gasses and chemicals being released from such waste. Let’s not forget, most of what we make us of these days are made with chemicals. The chemicals don’t just disappear when they are thrown in the trash can. They will continue to exist and cause environment havoc for a long time.

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