In recent twelvemonth, Malaysia is confronting a rapid development with urbanisation and industrialisation. The population is turning in a fast gait due to this development and therefore increasing the production of waste burden to the aquatic environment. Assorted anthropogenetic activities have produced considerable measures of solid and liquid wastes. Although Malaysia has effluent intervention works installations, nevertheless, the effluent intervention works is non efficient plenty to handle all the pollutants and therefore resulted in important sum of pollutants being discharged into aquatic systems.
Large-scale of Linear Alkylbenzenes ( LABs ) production started being introduced into the environment in the early 1960s when additive alkylbenzenesulfonates ( LASs ) were used to replace alkylbenzenesulfonates ( ABSs ) in detergents ( Eganhouse. et al. , 1983 ) . LABs with C10-C14 normal alkyl concatenation are the natural stuff for the industrial production of LASs which have been in extended usage for the fabrication of domestic man-made detergents ( Tsutsumi. et al. , 2002 ) .
LABs are industrially synthesized during Friedel-Crafts alkylation reaction and later undergo sulfonation reaction to bring forth LASs. During sulfonation procedure, little but important sums of residuary unsulfonated LABs are carried with LASs into detergents and subsequent discharge into the aquatic environments ( Eganhouse. et al. , 1983 and Takada. et al. , 1987 ) . The construction of LAB and LAS are shown in the Figure 1.
Figure 1: The construction of Linear Alkylbenzene ( LAB ) and additive alkylbenzenesulfonate ( LAS )
Takada and Ishiwatari have proposed a ratio of internal to external isomers ( I/E ratio: a ratio of the amount of 6-C 12 and 5-C 12 comparative to 4-C 12, 3-C 12 and 2-C 12 ) as an index of the grade of LAB debasement. Apart from that, they have besides demonstrated how the concentration ratio of internal to external isomers used to gauge the grade of LAB debasement ( Takada. et al. , 1990 ) . Therefore, I/E ratio is utile in measuring the destiny of LABs and sewage-derived atoms in aquatic environments.
In my proposed survey, deposit samples will be collected from selected oyster aquaculture farms of Peninsular Malaysia. The selected oyster aquaculture farms are Batu Lintang ( Kedah ) , Pulau Betong ( Pulau Pinang ) , Muar ( Johor ) , Pasir Panjang ( Negeri Sembilan ) , Kuala Setiu ( Terengganu ) and Merchang ( Terengganu ) . There are more than 100 living oyster species found worldwide. My proposed survey will concentrate on the species of the tropical oyster Crassostrea belcheri.
In general, an oyster farm is a installation where oyster are raised and harvested for nutrient. Oysters are molluscs and classified as pelecypods. Harmonizing to Fisheries and Aquaculture Department, Food and Agriculture Organization of the United Nations, molluscs production constitutes about 11 per centum of the entire piscaries production of Malaysia. In Sabah, experimental civilizations of Crassostrea belcheri are in advancement. The commercial oyster farming in Sabah in done by co-ops. Every twelvemonth, the production of oyster estimation is about 18 dozenss per hectare by utilizing rack-and-raft method. However, oyster civilization operations are still in limited graduated table in Peninsular Malaysia which is practiced on part-time by fishermen.
2.0 Problem Statement
The discharge of ill treated or untreated sewerage wastewaters is one of the major menaces to aquatic environments. Recently, several surveies have proven the bioaccumulation of LABs in some species of aquatic beings. High concentrations of LABs in oyster aquaculture farms non merely potentially degraded aquatic systems but besides hurts to human wellness as worlds are at the top of nutrient concatenation.
3.0 Aim of the Research Project ( Aims )
To find the concentration of Linear Alkylbenzenes ( LABs ) in deposit samples collected from selected oyster aquaculture farms of Peninsular Malaysia.
To find the internal isomer relation to external isomer ratio ( I/E ratio ) of LABs in gathered deposit samples in order to measure the current position of sewerage pollution in oyster aquaculture farms.
4.0 Significance of the Study
At an earlier clip, Linear Alkylbenzenes ( LABs ) were thought to be components of marine affair, nevertheless many research workers have proved that LABs are recognized as omnipresent pollutants. In Malaysia, sewerage and effluent is treated before discharged into the aquatic environments but still carries important sums of sewerage tonss into aquatic environments.
My proposed survey is about the distribution of Linear Alkylbenzenes ( LABs ) in deposit samples collected from selected oyster aquaculture farms of Peninsular Malaysia.
An appraisal the current position of sewerage pollution can be conducted in selected oyster aquaculture farms. The proposed survey will supply information on the application of LABs as a molecular marker to measure the sewerage pollution in selected oyster aquaculture farms.
5.0 Review of Research Literature
Alkylbenzenes were measured in 34 green mussels collected from India, Indonesia, Malaysia, Thailand, Cambodia, Vietnam, and the Philippines together with bluish mussels collected from Tokyo Bay, Japan ( Tsutsumi. et al. , 2002 ) . They discovered that the I/E ratios ( a ratio of internal to external isomers of LABs ) in the South and South East Asiatic states ( 1-3 ) were much lower than those in Tokyo Bay ( 3-8 ) . Apart from that, Alkylbenzenes with branched alkyl ironss and tetrapropylene-based alkylbenzenes ( TABs ) were detected in mussels from Indonesia and Philippines. Based on this find, they concluded that sewerage intervention systems in South and South East Asiatic states are deficient. They besides stated that LABs are a powerful tool to measure the measure and quality of sewerage impacts on coastal environments.
Takada and Ishiwatari have conducted laboratory incubations of Linear Alkylbenzenes ( LABs ) to obtain experimental grounds of systematic microbic change of their isomeric composing ( Takada. et al. , 1989 ) . They stated that the grade of LAB debasement in was quantitatively related to the alteration in their isomeric composing. The consequences indicated that the grade of LAB debasement in Tokyo estuarine and bay deposits is about 45 % . By usage of the I/E debasement diagram, they estimated that 55 % of LABs come ining the aquatic environment of Tokyo remain in the deposits without biodegradation. They proposed that I/E debasement diagram as an index of the grade of LAB debasement.
Linear Alkylbenzenes ( LABs ) were measured in deposits from three countries of Port Phillip Bay in Australia ( Murray. et al. , 1987 ) . They found that the concentrations of LABs in sediment sample from Port Phillip Bay ( 0-19 Aµg g-1 ) were by and large lower than those reported for other coastal deposits from the USA and Japan. The compositional profiles showed that both physical and biological procedures had acted to take LABs during deposition. Apart from that, an unusual alkyl benzine compounds were detected at one side. These compounds were likely synthesized mundanely by the bacteriums of a nearby sewerage farm. They concluded that microbic oxidization and disintegration in H2O were the two procedures moving to take LABs during deposition.
Takada and Ishiwatari have done on the distribution of Linear Alkylbenzenes ( LABs ) and Linear Alkylbenzenesulfonates ( LASs ) in river deposits, suspended river atoms, domestic wastes, and waste wastewaters around the Tokyo metropolis country ( Takada. et al. , 1987 ) . The consequences indicated that LABs are carried into aquatic environments as a consequence of the usage of man-made detergents around the Tokyo metropolitan country. Based on their research, they demonstrated that the isomeric composing of LABs alterations consistently during biodegradation. They besides concluded that the ratio of LASs to LABs decreases with progressive exposure of LAS-type detergent to the aquatic environment.
Linear Alkylbenzenes ( LABs ) were evaluated as waste tracers in the Marine environment by finding their concentration and composing in suspended particulate affair and deposits in the locality of Joint Water Pollution Control Plant ( JWPCP ) effluent outfall system in California ( Eganhouse. et al. , 1983 ) . They found that LABs appear to be preserved in deposits for clip periods of 10-20 old ages. In add-on to residence clip, a complex gathering of wetting agent related branched alkylbenzenes was besides found in the waste affected deposits. The consequence demonstrated that LABs are potentially utile as molecular tracers of domestic wastes and under appropriate conditions, as geochronological tools.
6.0 Thesis Outline
6.1 Sampling Location
In my proposed survey, the selected oyster aquaculture farms in Peninsular Malaysia are Batu Lintang ( Kedah ) , Pulau Betong ( Pulau Pinang ) , Muar ( Johor ) , Pasir Panjang ( Negeri Sembilan ) , Kuala Setiu ( Terengganu ) and Merchang ( Terengganu ) . The sampling locations are shown in the Figure 2.
Figure 2: Map demoing the sampling locations of the selected oyster aquaculture farms in Peninsular Malaysia
Linear Alkylbenzenes ( LABs ) in deposit samples will be analyzed utilizing Gas Chromatography Mass Spectrometry ( GC-MS ) . In general, GC-MS can be used to find molecular weights and elemental composings of unknown organic compounds in complex mixtures.
The Figure 3 shows the proposed research methodological analysis for deposit samples analysis is in conformity with the Laboratory Analytical Procedures ( Zakaria et al. , 2002 ) .
Sediment samples collected from selected oyster aquaculture farms
Soxhlet Extraction with methylene chloride ( DCM )
Silica Gel Column Chromatography
5 % H2O deactivated
65 % ( DCM ) in Hexane
40 % ( DCM ) in Hexane
25 % ( DCM ) in Hexane
Fully activated Silica Gel Column Chromatography
25 % ( DCM ) in Hexane
Alkylbenzenes ( LABs & A ; TABs )
& A ;
Figure 3: Laboratory analytical processs for deposit samples
Sediment samples collected from six selected oyster aquaculture farms in Peninsular Malaysia, which are of Batu Lintang, Pulau Betong, Muar, Pasir Panjang, Kuala Setiu and Merchang.
The undermentioned statements describe the general information on trying locations:
Batu Lintang is situated in Kedah, Malaysia. Its geographical co-ordinates are 5A° 38 ‘ 0 ” North, 100A° 24 ‘ 0 ” East. Its original name is Kampong Pengkalan Batu Lintang.
Pulau Betong is a little island in Penang, Malaysia. Its geographical co-ordinates are 5A° 18.868 North, 100A° 11.018 East. It is it is about 7A estates ( 28,000 M2 ) in country and is in private owned.
Muar is situated in Johor, Malaysia. Its geographical co-ordinates are 2A° 2 ‘ 0 ” North, 102A° 34 ‘ 0 ” East.
Pasir Panjang is situated in Negeri Sembilan, Malaysia. Its geographical co-ordinates are 2A° 26 ‘ 0 ” North, 101A° 56 ‘ 0 ” East.
Kuala Setiu is situated in Terengganu, Malaysia. Its geographical co-ordinates are 5.543956, 102.894974.
Merchang is situated in Terengganu, Malaysia. Its geographical co-ordinates are 5.031379, 103.293958.
Sediment samples were collected from six selected oyster aquaculture farms in Peninsular Malaysia, which are Batu Lintang ( Kedah ) , Pulau Betong ( Pulau Pinang ) , Muar ( Johor ) , Pasir Panjang ( Negeri Sembilan ) , Kuala Setiu ( Terengganu ) and Merchang ( Terengganu ) . The deposit samples were collected in the sum of sufficient measures for the intent of chemical analysis in research lab. These samples were analyzed for the possibility of Linear Alkylbenzenes ( LABs ) adsorbed to oyster aquaculture farm deposits.
At each trying location, the samples were collected by utilizing Eckman Dredge Sediment Sampler and placed in the pre-cleaned chromium steel steel container. After that, the chromium steel steel container incorporating sediment samples were wrapped with aluminum foil. The deposit samples were stored in the zip-log bag every bit shortly as the chromium steel steel containers were labeled. Following, the zip-log bag with sediment samples was stored in a ice chest box with ice or dry ice at 4°C. The deposits samples were later transported to research lab. The homogenize samples were stored in deep-freeze at -18°C for farther chemical analysis.
Sediment samples collected from six selected oyster aquaculture farms in Peninsular Malaysia were usage to find the concentration of Linear alkylbenzenes ( LABs ) in oyster aquaculture farm deposits. Clean setup is an indispensable demand for research lab work. Laboratory should be cleaned prior to LABs analysis. Harmonizing to Analytical Laboratory Procedure ( Figure 2 ) , 20g of homogenized moisture deposits were undergo freeze-dry procedure to extinguish the H2O content in the wet deposits.
After the drying procedure, the deposits were placed into the Whatman Cellulose Thimbles. These sediment samples were extracted in a procedure called Soxhlet Extraction by utilizing Soxhlet Extractor with 250 milliliters methylene chloride ( DCM ) dissolvers and thermoregulators control for about 9 hours at temperature around 40A°C. The infusions incorporating mark compounds resulted from Soxhlet Extraction procedure were collected and undergo Cu intervention in order to extinguish the S content in deposit samples that produced by micro-organism that may interfere with the chromatography reading. After Cu intervention procedure, the infusions volume can be reduced by rotary vaporization procedure at temperature 30A°C until about waterlessness.
Next, the infusions were purified and fractionated by utilizing two-step silicon oxide gel column chromatography. The first measure silica gel column chromatography is use to take unwanted polar compounds from organic hydrocarbon. Normal hexane ( HPLC class ) was transferred into 0.9cm internal diameter glass column. After that, 5 % H2O deactivated silica gel were use to pack the glass column until 9cm tallness for the first measure column chromatography utilizing Pasture pipette. Then, 1cm tallness of adust anhydrous Na sulfate ( Na2SO4 ) is added into the glass column by utilizing Pasture pipette. The infusions were eluted into the glass column by utilizing Pasture pipette and the infusions in the flask were rinsed with 2mL dissolver ( DCM: Hexane with ratio 1:3 ) in the sequence of 0,4mL, 0.3ml, 0.3mL, 0.5mL and 0.5mL severally.
The infusions were transferred into glass column by grazing land pipette each clip. The staying of 18 milliliters DCM: Hexane ( 1:3 ) was so transferred straight into first measure glass column. For each interval, the flask with dissolver was rinsed in the sonicator to guarantee all infusions were reassign into column. The eluted samples ( incorporate hydrocarbon such as
n-alkanes, LABs, PCBs, PAHs and pesticides ) from first measure silica gel column chromatography were collected into a pear form flask. The samples will be
roto-evaporated as about to dryness as possible for the usage of 2nd measure silicon oxide gel column chromatography.
For the 2nd measure silicon oxide gel column chromatography, normal hexane ( HPLC class ) was transferred into 4.5mm internal diameter glass column and 100 % activated silica gel was used to pack the glass column until 18cm tallness. The eluted sample was loaded from first stairss column chromatography to 2nd measure column by utilizing grazing land pipette and followed by 4mL of normal hexane dissolver ( HPLC class ) . The pear form flask incorporating the eluted samples rinsed by normal hexane and load the staying dissolver into 2nd measure glass column by grazing land pipette. For each burden interval, flask with dissolver was rinsed in the sonicator to guarantee all infusions were reassign into column and n-alkane fraction will be collected in a little pear form flask as first fraction. Following, load 4mL of normal hexane dissolver ( HPLC class ) once more as the same methods shown above into glass column and 2nd fraction ( LABs ) was obtained.
For this survey, merely the 2nd fraction is being analyzed. LABs are the organic compound to be analyzed and the 2nd fraction was transferred to nitrogen blowdown procedure and the LABs fraction was evaporated to dryness. Last, the samples later injected into Gas Chromatography-Mass Spectrometer ( GC-MS ) for LABs analysis.
6.3 Restriction of the Study
There are several restrictions in set abouting concluding twelvemonth undertaking. It is understood that the restrictions are a portion of undertaking.These restrictions will indirectly detain the patterned advance of undertaking or act upon the research consequences. Time restraint is one of the restrictions in finishing concluding twelvemonth undertaking. Time becomes an indispensable undertaking restraint when pupil have to do clip agreements for go toing talks and carry oning the research lab works.
Cost is one of the undertaking constraints common to every pupil. Each pupil will be given a fiscal support of RM250 for concluding twelvemonth undertaking. At the beginning of the undertaking readying, I was worried about what resources are needed and how to obtain these resources. I was fortunate plenty in holding several Masterss pupils assisting and helping me in experimental cost direction.