Heavy Metal Analysis on Babylonia Areolata

Heavy Metal Analysis on Babylonia Areolata Heavy Metal Analysis on Babylonia areolata CONTENTS PAGES 1.0 INTRODUCTION 1.1 Background of study 1.2 Problem statements 1.3 Objectives of study 1.4 Hypothesis 1.5 Scope of study 1.6 Significant of study 2.0 LITERATURE REVIEW 3.0 MATERIALS AND METHODS 3.1 Study area 3.2 Research instrument 3.3 Sampling and sampling material 3.4 Research procedure EXPECTED RESULTS 5.0 GANTT CHART 6.0 REFERENCES CHAPTER 1 INTRODUCTION BACKGROUND OF STUDY Mollusks can be found almost everywhere on land and its habitat was including coral reef, estuaries, freshwater lakes and also rivers. Its habitat ranging from deserts to rainforests (Lydeard Lindberg, 2003). Mollusks also had been identified as an important fisheries and mariculture food such as clams, scallops, abalone and conch instead of commercial pearl production (Landman et al., 2001). Babylonia areolata, ivory shell was categorized as a gastropod in family Buccinidae (Hualkasin et al., 2008). B. areolata can easily be recognized by its colour-pattern because it was the only member of the genus with three broadly separated rows of dark spots (Altena et al., 1981). This three widely separated rows of spots are visible on the body-whorl, enabling easy identification of this species (Altena et al.,1981). It can be found 10-20m deep in sandy bottom (Habe, 1997). B. areolata has a buccinoid shell with the aperture approximately half of the total height. There are no notch can be seen from the upper part as the outer lip of the aperture not clearly thickened inside. On the last whorl in the inner lip, a notch consisting of a strong callus can be seen for the umbilicus. The umbilicus is wide open and the initial whorls whitish follows by the reddish-brown spots on a white background for the following whorls (Altena et al., 1981). The spotted babylon, B. areolata which also known as the Hoy Wan in Thailand supports a commercial fishery (Chaitanawisuti Kritsanapuntu, 1999). This species can be found in the Gulf of Thailand by three different shell colors which are brown, cream and white. The different shell colors indicate different values. The brown shell has the highest price while the white shell has the lowest price (Hualkasin et al., 2008). High demand for the brown shell B. areolata was from China, Taiwan, Hong kong and Japan. In Thailand, the distribution of B.areolata was different at the upper and lower Gulf of Thailand. Phetchaburi and Rayong which located at the upper gulf of Thailand only have brown shells B. areolata while in Songkhla and Pattani, all three colors are found there (Hualkasin et al., 2008). B. areolata is a well-known and nutritious food which have been a popular mollusk model used for the study of heavy-metal toxicity and biologic poisoning toxins transmission (Chen Chou, 1998). As the B. areolata has a potential market in Thailand as well as in Malaysia it will be used in this experiment to study its heavy metal contents and concentration. The place chosen for the sample drawing of B. areolata is in Bachok, Kelantan. Table 1: Taxonomy of B. areolata . PROBLEM STATEMENT Since B. areolata are widely used for the food purpose, there should some study on the composition or content of the B. areolata itself in order to ensure it was safe for the human consumption. The B. areolata lives in place like beach and may consume or eat on heavy metal that comes from nearby industry which may become harmful when human consume it. This experiment then can prove or can ensure the safety of B. aerolata for the consumption if the result of the experiments shows it flesh was having small quantity of the heavy metal which was deemed harmless to human. Heavy metal poisoning in human was resulted from the toxic accumulation of heavy metals in soft tissues. There was some level of heavy metal that permitted in the human body which would not give any health problems to the human health. In this study, the level of the expected heavy metal in the flesh of B. areolata will be discover to investigate whether their level was below the acceptable amount. The concentration of the heavy metal and exposure time make the metal’s toxic effects establish in an organ as many toxicants tend to be bio accumulate. When they occur at certain levels, even essential elements that are critical for life, may lead to loss of organ function or death (Goyer, 1996). This determination also important as B. areolata has the potential to become one of the ingredients for the feed preparation. If B. areolata was safe for human consumption, it also can become the ingredients for the feed production. OBJECTIVE OF STUDY To determine the concentration of heavy metal (Arsenic, Cadmium, Copper, Lead, Mercury and Zinc) in the Babylonia areolata. HYPOTHESIS H0 = B. areolata contains heavy metal concentration. Ha = B. areolata do not contains heavy metal concentration. SCOPE OF STUDY The scopes of this research are as follow: Only B. areolata which was draw from Bachok, Kelantan, Malaysia. Only one parameter of were being used in this study, that heavy metal concentration. To determine the component and concentration of the heavy metal in the B. areolata. SIGNIFICANT OF STUDY The study was important to get more knowledge about the contamination of the seafood which is B. areolata by the heavy metals. The importance of the study is to identify the concentration of the heavy metal in the B. areolata compared to the amount that permissible for the human consumption. As the contents of the heavy metal in the B. areolata was safe for human consumption, so the species can be the potential ingredients for the feed preparation. CHAPTER 2 LITERATURE REVIEW Distribution and background of Babylonia areolata B. areolata can be easily identify as it has three widely separated rows of spots which visible on its body whorl. The size of the shells can be up to 93 mm high and 52 mm broad. This species inhabits in sandy or muddy bottoms in shallow water. The distribution of this species is from Ceylon and the Nicobar Islands through the Gulf of Thailand, along the Vietnamese and Chinese coasts to Taiwan (Altena Gittenberger, 1981). B. areolata is well-known for domestic and international consumption which make it one of the economically important aquatic species. The price of this species was quite high which around 300- 350 Baht/kg in Thailand (Sutthinon et al., 2007). B. areolata will be a target species of commercial fisheries in Thailand. Nowadays, the aquaculture activity have been increase and improve to ensure the increment and sustainability of the stock as the number and the size of mollusk catch from the Thai coast is small (Hualkasin et al., 2008). In Thailand, the expanding domestic market of seafood and increasing demand of B. areolata results to the many interest to the commercial culture of this species. This also results due to the catastrophic decline in its populations in the gulf of Thailand. At present, the culture of B. areolata in large-scale production using the flow-through seawater system in concrete or canvas ponds shows a success for the species to survive from juveniles to marketable size (Chaitanawisuti et al., 2011). B. areolata with brown shell has the highest price while the white shell has the lowest price. High demand from countries such as China, Taiwan, Hong Kong, and Japan is for the brown shells (Hualkasin et al., 2008) This species has been cultured and consumed widespread throughout Asia which can be introduce as a new commercial gastropod. In recent years, market demand for this species has increased which results to the decrease in the wild population and shortage of seed stock to support its aquaculture industry. In order to support the increasing demand, the B. areolata should be produced in high quantity and quality by artificial method (Abol-munafi et al., 2010). 2.2 Category and hazards of heavy metal Heavy metal analysis on the source food was very important studies as it will ensure the safety of the food for the human consumption. Seafood was one of the food sources that have a high risk on the contamination with the heavy metal. Seafood may live in the sea where the components of heavy metals may be dissolved in it from the many sources such as the industrial activity nearby it. Heavy metal was divide into two categorized which is biological essential and non-biological essential metals (Kie, 2013) Biological essential metal can be refers to the metal that are needed to fulfill wide and variety range of human body functions. On the other hand, non-biological essential metals are considered to be toxic, nevertheless, modern medicine utilized them in a diverse range of applications including diagnostics, imaging and therapeutic applications (Dyson, 2011). Some examples of non-biological essential metal are lead, mercury, cadmium, chromium and tin. All heavy metals can be toxic when they exceed the threshold concentrations (Kie, 2013). 2.3 Heavy metal analysis in fish Heavy metals can harm human health by contaminating or accumulating in aquatic life and enter the food chain if the contamination and exposure is significant (Fernandes et al. 2007). In addition, this heavy metal are categorizes by the United States Environmental Protection Agency (USEPA) based on their potential for human exposure and health risk (Birungi et al. 2007). Fish is consumed by large population especially those who live nearby the river as a source of protein and the accumulating of heavy metal in fish will be an important issues (Roshasliney et al., 2010) Fish which has low saturated fat and sufficient omega fatty acid are also important in supporting good health to human. Growing rates, metabolism, feeding pattern and ecological need will influence the level of heavy metal contamination in fish (Yilmaz et al. 2005; Yilmaz et al. 2010). Besides, the exposure of fish to heavy metals also depends on the differences in life history patterns among species (including trophic levels and geographical distribution of life stages (Allen-Gil Martynov 1995). 2.4 Heavy metal analysis in bivalve and gastropod Mollusks are able to accumulate heavy metals which will give a hazard to the consumers (Qiang et al., 2001). According to Qiang et al. (2001), the heavy metal concentrations in the local seafood being monitored for many years to ensure the safety of the seafood. The study conducted by Qiang et al. (2001) was to access the current status of heavy metal pollution in shellfish available in major markets in Pearl River Delta and to compare with the tolerable limits. The costal environments of Pearl River become polluted with the liquid waste since the industrial development and the rapid population growth occur there (Qiang et al., 2001). MATERIAL AND METHODS 3.1 STUDY AREA Study area for this experiment is in Bachok, Kelantan. 3.2 RESEARCH INSTRUMENT Instrument that will be used for the analyzing of the heavy metal contents was Atomic Absorption Spectrometer (AAS). 3.3 SAMPLING AND SAMPLING MATERIAL Sample that will be collect is B. areolata 3.4 RESEARCH PROCEDURE Sample of B. areolata is collected from the sampling area which is in Bachok, Kelantan. 25 g of wet tissue of B. areolata (5 g dry weight) will be weighed out into crucible, and samples will be dried at 135 Â °C for 2 hour to obtain a dry weight. The dry sample then transfer to a cool muffle furnace and the temperature will slowly rise to 450Â °C-500Â °C. The ash will be left overnight. The samples then removed from the muffle furnace and let to cool to room temperature. Next, 2 ml of nitric acid (HNO3) will be added cautiously and swirled. The sample will be evaporated carefully just to dryness on warm hot plate or steam bath. It then transfers to cooled furnace, and the temperature slowly rises to 450 Â °C- 500 Â °C and hold at this temperature for 1 hour. The crucible will be removed and cool. To obtain clean, practically carbon-free ash the nitric acid (HNO3) treatment will be repeated if necessary. 10 ml of 1N hydrochloric acid (HCl) will be added and ash will dissolve by heating cautiously on a hot plate. The ash then transfers to a volumetric flask and hydrochloric acid (HCl) add as necessary. The ash then cool and dilute to a volume. As the sample ready, the sample will be test with Atomic Absorption Spectrometer (AAS) for the heavy metal analysis. Atomic Absorption Spectrometer general procedures consist of: 1. The hollow cathode lamp or electrode discharge lamp and D2-lamp were lighted if such background correction is used. The lamp current was set to the value specified by the manufacturer. 2. The monochromator was positioned at wavelength 213.9 nm. 3. The intensity of the hollow cathode lamp and the D2-lamp was carefully balanced if such background correction is used. 4. The burner head was aligned to assure that the center of the light beam passes over the burner slot. 5. The flame was lighted and the flow of fuel and oxidant was regulated to produce an oxidizing flame (lean blue). 6. Calibration blank was aspirate and a zero point was established. 7. Standard solutions were aspirated and a calibration curve was constructed. 8. Distilled water was aspirated after each standard or sample. EXPECTED RESULTS The B. areolata was live in deep sandy bottom which may be in sea or beach. For this experiment, the sample was draw near the industrial place. The expected heavy metals that will be analyzed from this experiment are Arsenic (As), Cadmium (Cd), Copper (Cu) and Zinc (Zn), Lead (Pb) and Mercury (Hg) (Qiang et al., 2001). These heavy metals was hazardous if accumulated in human tissue with exceeding value that recommended by the expertise. Arsenic can be found naturally on earth in small concentration. It occurs in soils and minerals and it may enter air, water and land through wind-blown dust and water run-off. This could be the reason for the contamination in B. areolata. The cadmium can be contaminated in the B. areolata by the exposure with the anticorrosion agent used by the ship for the coat of the ship itself. As the cadmium was mostly used as the anticorrosion agent, this would be the reasons of the cadmium absorption that release from the ship coat by B. areolata. Next, for the copper, it could be released into the environment by both natural sources and human activities. Examples of natural sources are wind-blown dust, decaying vegetation, forest fires and sea spray. Apart from that, it is applied in the industries and in agriculture which eventually will contaminate the sea water. For lead, although native lead is rare in nature but the contamination could be occur due to the lead cycle results from human production such as from the car exhaust. For mercury it can found naturally in the environment such as in metal form and mercury salts. This could be the source of the contamination. Lastly for the zinc, zinc occurs naturally in air, water and soil, but zinc concentrations are rising unnaturally, due to addition of zinc through human activities. Some soils are heavily contaminated with zinc, and these are to be found in areas where zinc has to be mined or refined, or were sewage sludge from industrial areas has been used as fertilizer. This also could be the source of the heavy metal that contaminated in the B. areolata. GANTT CHART