Fyp for Final Submission 2014

Determination of Physicochemical Parameters and the Correlation with Organotin

Compounds Concentration in Core Sediment

from Kong-Kong Laut, Johore ,Malaysia.

SELVAGAHNESH PRASAD

2013

Department of Environmental Sciences

Faculty of Environmental Studies

Universiti Putra Malaysia

Abstract

A quantitative investigation on the levels of tributyltin(TBT), dibutyltin(DBT), and

monobutyltin (MBT) species in core sediment samples was carried out. The sampling

area is located at the Kong-kong Laut estuary in the south east of Johore state,

Peninsula Malaysia, Malaysia. Tributyltin (TBT), Dibutyltin (DBT), and

(monobutyltin) MBT have been analysed in core sediment samples by Gas

Chromatography Mass Spectrometry. The trend of concentration was comparatively

studied with physicochemical parameters namely pH, Total Organic Carbon (TOC)

and Sediment Particle Size Distribution for the presence of possible correlation.The

correlation between sites of sampling and concentration are vital information to infer

on the reintroduction of pollutants occurances and fate and toxicity effect to the

environmental.On the other hand,the correlation between organotin concentration and

physicochemical parameters are very much essential in understanding the chemistry

of organotin on rates of pollution ,degradation of organotin compounds particularlt

butyltin species ,remediation methods to be applied (eg.bioremediation of pollutants)

and environmental mitigation measures and purposes.

.

2

Acknowledgements

I would certainly not have been able to finish my thesis without the supervision of

my supervisor, committee members, assistance from friends, and encouragement from

my family .I would like to explicit my deepest gratefulness to my advisor, Dr. Ferdi-

us@Ferdaus Mohamat Yusuff, for her excellent guidance, compassionate, persever-

ance, and providing me with an vivid atmosphere for doing research and who let me

experience the research of organotin analysis of core sediments in the field and prac-

tical issues beyond the textbooks, patiently improved my writing by rectifying them

and financially and monetarily supported my research.

I would also like to thank Dr. Lutfi Wan Johari, Prof. Kannan Narayanan, and Prof.

Mohamad Pauzi Zakaria for administrating my study for the past numerous occasions

and helping me to cultivate my circumstantial in analytical chemistry, geochemistry,

and ecology. Special thanks goes to Mr.Kit Wui Sien, who was enthusiastic to contrib-

ute in my sample collection committee with helping me much in transportation and

mobility purposes.

I would take this chance to thank Mr.Azwan Mejan, who as a wholesome friend,

was continually alacritous to help and give his best recommendations. It would have

been a lonely laboratory without him. Countless thanks to Mr Samsudin, Mr Gaffar,

Miss Thana, Mr Zairi, Miss Lela and other workers in the laboratory of the Environ-

mental Studies Faculty for helping me putting into place all the materials, apparatus,

reagents and necessary laboratory conditions for my study. My research would not

have been achievable without their assistance.

Finally, I would also like to thank my parents ,my dad, Mr Marimuthu Ayavoo and

my mom, Miss Tannavali Ramasamy, and my elder sister ,Miss Yogalekshumi Mar-

imuthu. They were always accompanying me and boosting me with their best wishes

and always there encouraging me up and stood by me throughout the good times and

bad.

3

Table of Contents

Abstract..................................................................................................................................1

Acknowledgements................................................................................................................2

Table of Contents...................................................................................................................3

List of Figures........................................................................................................................1

List of Tables.........................................................................................................................1

1 Introduction......................................................................................................................2

Background and Context...................................................................................................2

Scope and Objectives.........................................................................................................5

2 Literature Review.............................................................................................................6

Organotin...........................................................................................................................6

2.2.1 Organotin Species.................................................................................................6

2.1.3 Organotin toxicity.................................................................................................7

2.1.4 Transport and Pathway of Organotin Compound and negative effect on Aquatic

Ecosystem...........................................................................................................................8

2.0 Physicochemical Parameters.......................................................................................9

2.1.1 pH........................................................................................................................9

2.1.2 Total Organic Carbon (TOC)...............................................................................9

2.1.3 Sediment Particle Size.......................................................................................10

3.0 Materials and Methods...................................................................................................11

3.1 Method of Selection Criterion of Sampling Site.......................................................11

3.2 Instruments, Apparatus and Glassware, Materials and Reagents Used.....................12

3.2.1 Instruments.........................................................................................................12

3.2.2 Materials and reagent..........................................................................................13

3.2.3 Apparatus and glassware.....................................................................................14

3.3 Sample Collection, Preservation and Storage............................................................14

3.4 Sample Analysis.........................................................................................................15

3.4.1 Gas Chromatography /Mass Spectrometry Analysis..........................................15

3.4.2 Total Organic Carbon Analysis...........................................................................15

3.4.3 pH Analysis.........................................................................................................15

3.4.4 Sediment Particle Size Distribution Analysis.....................................................16

3.5 Data Collection and Statistical Analysis....................................................................16

3.5.1 Data Collection...................................................................................................16

3.5.2 Statistical Analysis..............................................................................................16

4.0 Results and discussion...................................................................................................18

4

4.1 Physicochemical Parameters and Depth....................................................................18

4.1.1 Depth and Organotin Concentration...................................................................18

Figure 6.Organotin Compounds Concentration against Depth of Kong-kong 3.........20

4.1.2 pH and depth.......................................................................................................20

4.1.3 Total Organic Carbon (TOC) and depth.............................................................22

4.1.4 Particle Size Distribution and depth...................................................................24

4.2 Physicochemical parameters and sites.......................................................................26

2.1.4 4.2.1 Organotin compound concentration and sites...........................................26

4.2.2 Particle Size Distribution and Sites....................................................................27

4.2.3 Total Organic Carbon (TOC) and sites...............................................................28

4.2.4 Mean pH and sites..............................................................................................29

5.0 Conclusion.....................................................................................................................31

Reference.............................................................................................................................32

Appendix 1...........................................................................................................................36

Appendix 2...........................................................................................................................37

Appendix 1...........................................................................................................................43

Appendix 2...........................................................................................................................44

5

List of Figures

Figure 1. Important Organotin Compounds................................................................................6

Figure 2 .Pathway of Organotin in the Aquatic Ecosystem........................................................8

Figure 3.Map of Sampling Sites and its Coordinates...............................................................11

Figure 4.Organotin Compound Concentration against Depth in Kong-kong 1........................19

Figure 5.Organotin Compound Concentration against Depth in Kong-kong 2........................19

Figure 6.Organotin Compounds Concentration against Depth of Kong-kong 3......................20

Figure 7.Mean pH against Depth for Kong-kong 1..................................................................20



Figure 10.Total Organic Carbon against Depth for Kong-kong 1............................................22



Figure 13.Particle Size Distribution against Depth for Kong-kong 1......................................24



Figure 16. Organotin Compound Concentration against Sites.................................................26

Figure 17.Particle Size Distribution against Sites....................................................................27

Figure 18.Total Organic Carbon against Sites..........................................................................28

Figure 19.Mean pH against Sites..............................................................................................29

1

List of Tables

Table 1.Historical Concentration of Organotin Compounds in Malaysia................................12

Table 2 .Instruments used in Analysis......................................................................................13

Table 3.Materials and reagent used in Analysis........................................................................14

Table 4.Apparatus and Glassware used in Analysis.................................................................14

1

1 Introduction

Background and Context

Tin is found in many organic and inorganic compounds. It reacts with chlorine,

sulphur, and oxygen to form inorganic tin compounds, which are found in the earth’s

crust in small amounts (Agency for Toxic Substances and Disease Registry

(ATSDR),2011.). Organotin compounds (OTCs) belong to the group of organometal-

lic compounds (Fent, 1996) with at least one covalent Sn-C bond (Batt, 2006; El Has-

sani et al., 2005; Murata et al., 2008). OTCs can be mono-, di-, tri-, or tetra-substi-

tuted (El Hassani et al., 2005) The general formula of an organotin compound is (El-

Hassani et al., 2005): RmSnX4-m where m = 1, 2, 3, or 4, R is an alkyl or aryl sub-

stituent, and X is a halogen: -OH,-SH, or –OR.

Currently , there are existance of over eighthundred organotin compounds which

are synthetic or arise naturally in the environment . The butyltins species (BTs) of

organotin derivatives, such as tributyltin (TBT) and its degradation products namely

the dibutyltin, DBT and monobutyltin, MBT, and the phenyltins species (PhTs) such

as, triphenyltin (TPhT) and its degradation products such as the monophenyltin,

MPhT and diphenyltin, DPhT, are fairly distributed in the marine environment.

Taking into consideration of the extensive distribution and intense toxicity effect of

these compounds on marine organisms, their monitoring and fate analysis in the

environment are significant (M. Hoch et al., 2001).

Incursion and accumulation of considerable amounts of these compounds in the

bottom sediments of the water bodies was the result of several years of using

organotin-based antifouling paints, where in particular tributyltin for inhibiting the

growth of marine organisms on ships’ hulls, and triphenyltin-based pesticides. Estuary

regions in the coastal marine area, and harbour and shipyard basins were

exceptionally affected.

.

2

Lately, there has been a considerable reduction in new loads of OTs into the

marine environment as a result of to the endorsements of the International Convention

on the Control of Harmful Anti-Fouling Systems on Ships (IMO, London, 2001),

framed under the patronages of the International Marine Organisation (IMO), coming

into force.Since 1st January 2003, the use of organotin-based anti-fouling paints on

newly built or painted ships (shorter than 25 metres) has been forbidden (IMO,

London, 2001). However, habours and shipyards are still considered the parts

susceptible to these pollutants because of intensive ship congestion and activities for

instance removal of old paint from hulls and ship scrapping.

Organotin compounds and mainly other highly toxic pollutant have high capacity to

be preserved in the base sediment which acts as the reservoir ; they impose a possible

ecotoxicological risk for live organisms long after the anthropogenic compounds have

dispersed into the environment. OTs compound adsorb onto very small sediment

fractions (particle size 50.063 mm). The portions of these are easily available to

benthic organisms, in particular decomposers and detritus feeders (J. Bolalek et

al.,1999).

The temperature,salinity,qualitative and quantitative composition of organic

matter ,and particle size distribution are namely the physicochemical

parameters ,which alters in timely manner and causes the release of specific forms of

organotin from sediments (D. Schwesig et al.,2005). As a result of the release,they are

widely bioavailable, hence , harmful and damaging to the aquatic orgamnisms living

in the water bodies.

Bottom sediments or core sediment vary in reference to compositions of chemical,

size of particle, source, rate of sedimentation and geographical dispersion and

distribution (J. Pempkowiak et al.,1997).Many synthetic and anthropogenic

sunstances content in core sediments depends on their size of particle,sediment

surface,magnetic properties and density which are cummulatively the physical

properties and chemical properties (i.e., ion exchange capacity, adsorptive properties,

organic matter content and content of inorganic compounds, such as carbonates and

3

oxides as well as on the environmental conditions for instance , pH, salinity and water

temperature) (M.Hoch et al.,2004).

Continental or surface-dweling sediments, particularly from river mouths and

ports, typically comprise of substantial proportions of mineral clays and organic

matter (D. Schwesig et al.,2005). The environmental conditions of river mouths and

ports for instance the depleted salinity is condusive for the buildup of organic

compounds comprising organotin to occur. Moreover, such sites are exceptionally

subjected to the contribution of new fractions of pollutants. The build up of these

sediment become the sink for hydrophobic pollutants. Nevertheless, under certain

environmental acclimatizes they may also undergo transformation as the source of

reintroduced contamination.

4

Scope and Objectives

1. The general objective of this study is to investigate the concentration of organotin

concentration in core sediment at straights of Johore, particularly Kong-kong Laut and

the physico-chemical parameters of the core sediment.

Objectives

1. To determine the physico-chemical parameters in core sediment samples from

Kong-kong Laut,Johore.

2. To identify the relationship of physico-chemical parameters with organotin com-

pounds concentration in core sediments.

3.To compare the concentration of organotin compounds concentration with historical

datas(before the ban of Organotin and after).

5

2 Literature Review

Organotin

Compounds of organotins(OTs) are a cluster of organometallics that have a distinct struc-

ture designated by the subsequent molecular formula of RnSnX(4-n), in which Sn represent the

atom of tin , R represents the alkyl group, such as methyl, butyl , octyl or phenyl and X which

corresponds to O-, OH-, Cl-, F,SH- etc., and n ranges from approximately 1 to 4.

2.2.1 Organotin Species

The amount and type of organic substituents bonded to the tin atom determine the

properties and applications of specific organotin compounds. Group X has almost no

effect on an organotin compounds properties, while the alkyl chain length has a signi-

ficant effect on a compound’s toxicity (Thoonen et al., 2004). Based on literature data,

in the case of diorganotin compounds, R2SnX2, it is the organic groups R that determ-

ine the activity potential, while group X controls the delivery of active ions

R2Sn2+.Table 1 presents information about selected organotin compounds.

6

Figure 1. Important Organotin Compounds.

2.1.2 Organotin Properties and Characteristics

Organotins bind strongly to solid phase particles, e.g., in sediments (forming oxides and

organic compounds), which results in a risk of contaminating the aquatic environment. OTCs

can also easily bond to proteins (Veltman et al., 2006) and display high affinity for cell mem -

branes (Cima et al., 1996). In order to present a full assessment of the impact of organotin

compounds on the aquatic ecosystem, it is necessary to determine thedegree of accumulation

and harmful effects of these compounds on various organisms in the food chain (Antizar-

Ladislao, 2008;Kannan et al., 1999).Organotin compounds have good lipophilic properties,

which facilitate their penetration through cell membranes.

On the other hand, these compounds also have good hydrophilic properties, which facilitate

their acceptance by well-hydrated cells. The lipophilic and hydrophilic properties of organot-

ins affect these compounds toxicity which in turn may result in cell damage or cell death

(Cima et al., 1996; El Hassani et al., 2005; Gray et al.,1987; Omae, 2003).

2.1.3 Organotin toxicity.

Although the toxicity of organotin compounds in enzymatic systems has not been thor-

oughly investigated (Kimbrough,1976), it is known that due to their solubility, OTCs can eas-

ily penetrate into tissues and the nervous system (Bowen, 1988). Based on the results of nu-

merous studies, tributyltin (TBT), which is characterized by embryotoxicity and genotoxicity

(El Hassani et al., 2005; Jha et al., 2000; Marin et al., 2000), and triphenyltin (TPhT) have

been recognized as the most toxic organotins.

The low toxicity compounds are the derivatives of tri-n-octyltin (Antizar-Ladislao, 2008;

Heroult et al., 2008; Hoch, 2001; Omae, 2003; Ramalho et al., 2010; Riepe et al., 1997). The

harmful effect of TBT was observed in samples of plants and microorganisms; it also showed

a negative effect on higher organisms inhabiting the marine environment (Kannan et al.,

1998; Ramalho et al., 2010; Słaba et al., 2010; Zhenget al., 2005). It was established, inter

alia, that endocrine disordersn occur even at low TBT concentrations (Yang et al., 2010). Be -

7

cause of the toxic properties of OTCs, mainly with regard to butyltin compounds, the occur-

rence of these chemicals in the environment is becoming more alarming. For this reason, in

many scientific circles investigations on the toxicity and ecotoxicological effects of tin com-

pounds have been undertaken (Arambarri et al., 2003; Batt, 2006). The results of these studies

are of widespread interest in society due to the significant increase in pro-environmental

awareness.

2.1.4 Transport and Pathway of Organotin Compound and negative effect on

Aquatic Ecosystem.

Figure 2 .Pathway of Organotin in the Aquatic Ecosystem.

The negative effect of organotin compounds on the aquatic ecosystem is the reason behind at-

tempts to detect and determine the level of these compounds in various parts of the environment.At

present, it is assumed that the trisubstituted tin compounds pose a significant threat to proper function-

ing of the environment, while the decomposition products, or the di- and mono-substituted derivatives,

are less harmful (Belfroid et al., 2000; Guruge et al., 1996, 1997; Harino et al., 1997; Wasik,2012). In

recent years, it was noted that organotin compounds of aquatic origin have begun to enter land organ-

isms, causing negative effects in many of those from the highest level in the trophic chain (Wasik,

2012).

8

2.0 Physicochemical Parameters

Sediment physical properties such as the sediment surface proper, size of particles, ,density

and magnetic properties of unconsolidated marine sediments are vital variables of understand-

ing change in geological event of deposition environment effect of mechanical and chemical

diagenesis with burial depth after deposition.(Hamilton and Bachman.,1982).The concentra-

tion of Organotin compound depends on chemical properties for example the exchange of

capacity of ion, organic matter substance, adsorptive properties and content of inorganic com-

pounds, as well as on the environmental circumstances and conditions (i.e., salinity, pH, and

water temperature) (Cima et al., 1996; El Hassani et al., 2005; Gray et al.,1987; Omae, 2003).

2.1.1 pH

pH= -log [H+]

The pH is the most imperative parameter influencing the sorption capacity of adsorption

medium (Sravani et. al., 2012). With the decreasing of pH (up to pH4)(M.Hoch et

al.,2002),adsorption of organotin compounds onto organic matter and clay mineral increases.

Referring to the adsorptivity of clay minerals, for instance, TBT is the most effectively adsorp

at pH=6 for montmoryllonite, and at pH=7 for kaolinite. At such pH of these values , the in-

tensity of the cations of organotin compounds is the highest. This happens when the pH is

closed to the pKa – for TBT pKa=6.3 and for TPhT pKa=5.2 (C.G Arnold,1998). The formed

cations can effortlessly adsorbed onto the negatively charged superficial of minerals

(K.Medrzycka et al., 2006),while the consequential bonds have electrostatic character. Sedi-

ments rich in organic matter have high adsorptivity which is most effective at pH 6–7, when a

cation is the dominant TBT form. Then the formation of complex bonds between TBT+ ion

and depronated ligands of organic matter are formed (K.Medrzycka et al., 2006).

2.1.2 Total Organic Carbon (TOC)

Organic carbon is the most substantial factor that controls the concentration of organotins

in sediments (M.Hoch et al.,2002).The partition coefficient of sediment and water or known

as kd for organotin undertakes very low values in the case of pure and low organic carbon

content minerals,e.g. kd for pure kaolinite is 51L kg-1,while the supplement of 5% of organic

matter result in the abrupt kd escalation up to 2700L kg-1.(Alonso-Azcarate et al .,2003).

9

2.1.3 Sediment Particle Size

The finest size fractions (<0.063mm) exhibit principally good sorptive properties, for in-

stance silty-clays (M.Hoch et al.,2004).This is for the reason that the physicochemical factors

(high proportion of clay minerals)(J.Pempkowiak et al.,1997).Reference to the surface proper

of sediment particles such as Silty-clay sediments are considered by the surface suitable with

high sorptive properties at the level of number of metres squared per gram, while for gravels,

it does not surpass centimetres squared per gram (J.Pempkowiak et al.,1997).Percentage of

inflatable clay minerals in sediments are characterised by clay minerals which show a high

sorption capacity to organic contaminations of cationic or polar character (M.Hoch et

al.,2002). It depends on their surface area and reactivity (negatively charged surface). Sorp-

tion coefficient (Kd) for TBT (at pH=6) and surface proper values for the mineral particles can

be arranged in the following sequence: montmorillonite (Kd=89 L kg_1; surface area ca 32m2

g_1)> kaolinite (Kd=51 L kg_1; surface area=10m2 g_1)> quartz (Kd=25L kg_1; surface

area=0.31m2 g_1)(M.hoch et al.,2004). Particularly strong sorption of TBT in pure montmoril-

lonite is caused by its big surface area. (Hoch and Schwesig.,2004) observed the increase in

Kd value from 25 to 67 L kg_1 for pure quartz sand after adding 10% of montmoryllonite, and

to Kd=94L kg_1 after adding 20% of montmorillonite. In addition of organic matter, higher

sorption of TBT can be expected onto kaolinite as different minerals adsorb organic matter

with fluctuating strength. The arrangement in which organic matter adsorbs on minerals is as

respects of : kaolinite>montmorilonitequartz (Schwesig et al.,2004).

10

3.0 Materials and Methods

3.1 Method of Selection Criterion of Sampling Site.

Figure 3.Map of Sampling Sites and its Coordinates.

The sampling site of Kong-kong Laut ,Pasir Gudang,Johor which is situated to the south-

east of southern peninsula state of Johore, Malaysia was chosen as the sampling site for the

study due to its geographically highest in concentration of organotin compound in whole Pen-

insula Malaysia(Harino et al .,2008). Coastal and estuaries of Johor Straits and its surrounding

vacinity had the highest concentration among other sites in Peninsula Malaysia. (Harino et

al .,2009).Table 2 below shows the concentration of organitin compounds in Peninsula Malay-

sia.[Red Circle: highest concentration of Organotin]

11

Table 1.Historical Concentration of Organotin Compounds in Malaysia.

3.2 Instruments, Apparatus and Glassware, Materials and Reagents Used

3.2.1 Instruments

List of instruments used during

analysis Instruments

Model

Core sampler (1metre) Self-made silicone core sampler

Gas chromatography/mass spectrometry Hewlett-Packard Model 6890 GC with

Quadra pole mass spectrometry

(HP5973MSD)

152H Hydrometer -

Thermometer -

12

Stop watch Casio 98D

Incubator vertical rotary shaker -

pH meter ORION 2 STAR

Centrifuge Rotofix 32 (Hettich Zentrifugen)

Electronic balance Shimadzu AY 220

Dessicator -

Oven

TOC Analyser Shimadzu TOC VCN-S

Table 2 .Instruments used in Analysis.

3.2.2 Materials and reagent

Materials and reagent Materials and Reagents Grade

Monobutyltin trichloride Reagent grade

Dibutyltin dichloride Reagent grade

Tributyltin chloride Reagent grade

Monophenyltin trichloride Reagent grade

Diphenyltin dichloride Reagent grade

Triphenyltin chloride Reagent grade

MBT trichloride-d5 Reagent grade

DBT dichloride-d10 Reagent grade

TBT chloride-d27 Reagent grade

Tetrabytyltin (TeBT)-d20 Reagent grade

Sodium Tetraethylborate >98%

Sodium acetate Reagent grade

Hydrochloric acid Metal analysis

Ethyl acetate Pesticide residue analysis

Sodium sulphate anhydrous Pesticide residue analysis

Copper(grain,20-60 mesh) Reagent grade

Potassium Hydroxide Reagent grade

Sep-Pak plus Florisil Content (50mg)

n-Hexane Pesticide residue analysis

Acetone Pesticide residue analysis

Methanol Pesticide residue analysis

13

Ethanol Pesticide residue analysis

Diethyl ether Pesticide residue analysis

Sodium hexametaphosphate 250g

Distilled water -

Milli-Q water -

CaCO3 standards (TOC analysis) 5g

Table 3.Materials and reagent used in Analysis.

3.2.3 Apparatus and glassware

List of apparatus and glassware

used during analysis

Volume Apparatus and Glassware

Beakers 500mL x 5

Measuring cylinder 1000mL x 8

Conical flasks 100mL x 5

Volumetric flasks 100mL, 500mL, 1000mL

Pipettes 1mL, 10mL

Centrifuge tubes 50mL

Mortar and pestle 1 unit

Sedimentation cylinder 1000mL x 2

Sieve (500,250,150,100,70μm) 1 unit (each)

Spatula -

Test tubes -

Zip lock bags -

Vacuum filter set -

Table 4.Apparatus and Glassware used in Analysis.

3.3 Sample Collection, Preservation and Storage.

After the sample is collected using a core sampler, the sediment sample is refrigerated at -

20oC and free from light from the time of collection until extraction. A 3-day maximum ex-

tract storage time is recommended. The sediment are kept at -20oC until analysis procedure

applied.

14

3.4 Sample Analysis

3.4.1 Gas Chromatography /Mass Spectrometry Analysis

Organotin compounds were extracted from the sediment followed the application sugges-

ted by (L.Edbon et al,.1998):Freezed-dried 0.5g of sediment in a centrifugal tube,was added

with 100ml of TPrT (50 mgL-1) as an internal standard. After treatment for about 10 minutes,

2 g of sodium chloride (NaCl), 15mL of toluene containing 0.1% tropolone, and 10mL of 1

mol L-1 of methanolic HCl were added. Then the capped tubes were mixed for sixty minutes,

and 10mL of Milli-Q water was added. The tubes were mechanically shaken well for 10 min

and then centrifuged at 2000 rpm for 4 minutes. Next, toluene part was concentrated to 5mL

for further analysis. NaBEt4 was pertained as a derivatization agent for organotin compounds

in sediments as stated : 5mL of 1 molL-1 acetate buffer of pH 5 was added to the acetone part.

Fifteen millilitres of milli-Q water and 1mL of 5% NaBEt4 were added. The tubes were

shaken mechanically for 10 min for extraction and ethylation step, and then centrifuged. Pas-

teur pipette was used to collect the layer of hexane from the centrifugal tube. Water traces in

the sample were removed by smearing 3 g of anhydrous Na2SO4. Finally, the hexane extract

was evaporated to 100 mL applying stream of nitrogen gas. 1 mL was injected into the gas

chromatography instrument for organotin compound concentration analysis.

3.4.2 Total Organic Carbon Analysis

Total organic carbon or TOC was measured in percentage at Faculty of Agrobio ,UPM oper-

ating a Total Organic Carbon Analyzer (Shimadzu TOC-VCSH), in which was well-appointed

with a Solid Sample Module(Shimadzu SSM-5000A) by following the manufacturer’s

method (Shimadzu, 2001). The difference in combustion and oxidation and the carbonate

acidification reaction (analysis of inorganic carbon) was used to calculate and compute TOC.

3.4.3 pH Analysis.

The pH reading was determined by applying (METHOD 9045D, USEPA). Weighted 20 g

of dried sediment was placed into 50-mL beaker, then mixed with 20 mL Milli-Q water,

covered, and the suspension is stirred vigorously for 5 minutes. Additional dilutions are per-

missible if occupied with hygroscopic sediments and soil or other tough matrices. Then, the

sediment suspension was let to stand for about an hour to allow most of the suspended clay to

settle out from the suspension or filtered or centrifuged off from the aqueous phase for pH

measurement. Next, the pH meter electrodes are adjusted using the clamps of the electrode

holder in order to, upon lowering the electrodes into the beaker, the glass electrode will be

immersed just deep enough into the clear supernatant solution to ascertain a good electrical

15

contact through the fibre-capillary hole or ground-glass joint. The electrodes are inserted into

the sample solution in this manner. For permutation of electrodes, immerse just below the sus-

pension. The pH reading is determined.

3.4.4 Sediment Particle Size Distribution Analysis.

The Hydrometer method as recommended by the USDA Soil Survey and Canadian Soil

Survey Committee was used in determining the particle size distribution in the core sediment.

Qualitatively, this method regulates the quantities of 3 distinct sizes of primary sediment

particles, in which is determined via their settling rates in the aqueous solutions using well

calibrated standard hydrometer. Proportion of sediments are characterized by class sizes with

the range from 2000 - 50 um; Silt ranging from50-2.0 um and clay < 2.0 um. The principal Of

particle settling rates are based primarily on sedimentation confined by the Stroke’s Law and

measured using a hydrometer. In this investigation, the standard ASTM 152H-Type hydro-

meter is established on a standard temperature of 20 oC and a particle density of 2.65gcm-3

and units are articulated as grams of soil per litre. For specific samples the method may need

the pre-treatment removal of soluble salts, carbonates, organic matter and iron oxides with

successive dispersion using sodium hexametaphosphate (Day 1965). Determining the hydro-

meter reading of a blank solution will cater the correction for temperature and solution viscos-

ity. The method employs a detection limit of 2.0% sand, silt and clay (dry basis) and is gener-

ally reproducible to within ± 8%.

3.5 Data Collection and Statistical Analysis

3.5.1 Data Collection

All analytical data are collected, tabulated and generated into suitable graphical representation

using Microsoft Excel 2013.Data are rearranged, classified according to intervals and suitable

bin values and stored with backup of hardcopy.

3.5.2 Statistical Analysis.

Correlation analysis is the most widely used and reported statistical analysis method

.It is vital in determining a relationship exist between two distinct variables(Goldrath

N et al.,1988). For instance, depth and physicochemical parameters (eg. total organic

carbon, pH, particle size distribution and orgatotin compounds concentration) and also

depth within sites and sites by concentration of organotin compounds in this investig-

16

ation.The analysis of variance or commonly abbreviated as ANOVA is a statistical

tool used to perceive variances between means of experimental group. ANOVA is

justified in experimental designs with a dependent variable which is a continuous

parametric numerical outcome measure, and multiple experimental groups within one

or more independent or categorical factor. In ANOVA terminology, independent vari-

ables are called factors, and groups within each factor are referred to as levels(Wack-

erly DD et al.,2002).ANOVA is actually a very convincing technique and there are

many accounts which will not be mentioned in this writing, the interested reader can

pursue these in Armitage and Berry (chapters 7 and 8, 1994).

17

4.0 Results and discussion.

4.1 Physicochemical Parameters and Depth

4.1.1 Depth and Organotin Concentration

Concentrations of TBT in the investigated area of Kong-kong 1(Figure 2.) ranged between

790.0 to 7.0 μg/L with corresponding to each depth intervals. It is clear that TBT is the pre-

dominant species of OTC with a ratio to total OT species ranged to 73.82%. The maximum

value of 790.86 ug/L. Overall OT concentration and increasing depth was found to correlate

negatively with r=-0.464(p=0.05) and the significance is strongest for DBT species against

depth. A strong significant correlation with r = 0.745 (p = 0.01) was found between the con-

centration of TBT and DBT reflecting the degradation of TBT as a main source of DBT in the

Kong-kong 1 site. At site Kong-kong 2(Figure 3.), the concentration of both TBT and DBT

correlates positively strong with r=0.804(p=0.01) which emulating degradation of TBT as a

dominant source of DBT in the Kong-kong 2 site. However, total mean concentration of MBT

of site 2 is relatively higher than site 1, and concentration of TBT is relatively is relatively

smaller in site 2 than site 1, this scenario can be contributed by high degradability factor at

site 2 or minimal current reintroduction of organotin compounds at site 2 than site 1 which is

concentrated with anthropogenic activities. Reference to the depth, DBT has negatively strong

correlation against depth, r=-0.685(p=0.05),this might due to certain physicochemical factors

affecting the adsorption of butyltin compounds. At Kong-kong 3(Figure 3.), there is a strong

positive correlation between MBT and depth with r=0.733(p=0.05) which resonates the occur-

rences of MBT settling at the site, which is the furthest site from site 1, which is highly pol -

luted. There is no statistically significant evidence of TBT concentration correlating with

depth at site 3, which suggest the site is far away from the pollutant sources or current reintro-

duction. However, there is a strong negative correlation between DBT and MBT of r=-

0.812(p=0.05) which reflect degradation of DBT as a main source of MBT in the Kong-kong

3 site. Thus, there is statistical evidence that organotin compounds concentration have

significant negative correlation towards depth. Hence, this condition sets a benchmark for

further investigation on other parameters compared to depth.

18

Figure 4.Organ-

otin Compound Concentration against Depth in Kong-kong 1.

Figure 5.Organotin Compound Concentration against Depth in Kong-kong 2.

19

0-3cm 7-9cm 13-15cm

19-21cm

25-27cm

30-33cm

36-39cm

42-45cm

0

200

400

600

800

1000

1200

1400

Organotin Compounds Concentration against Depth of Kong-kong 1

MBT DBT TBTDepth /cm

OT

conc

entr

ation

ug/

L

0-3cm

6-9cm

12-15cm

18-21cm

24-27cm

30-33cm

36-39cm

42-45cm

48-51cm

54-57cm

0

50

100

150

200

250

300

350

400


MBT DBT TBT

Depth /cm

OT

conc

entr

ation

ug/

L

Figure 6.Organotin Compounds Concentration against Depth of Kong-kong 3

4.1.2 pH and depth

At site 1(Figure 5), pH of core sediments by depth intervals has a strong positive correla-

tion with r=0.929(p=0.01).Thus, as depth increases, pH also increases almost linearly, in

which the basicity of the sediment increases. This condition may due to the increase of bac-

terial activity with depth which elevates the pH (Wang and Alva, 2000; Weston et al., 2010).

Statistically there is no significant correlation between pH and depth in sample from site

2(Figure 6) and 3(Figure 7).

Figure 7.Mean pH against Depth for Kong-kong 1

20

0-3cm 6-9cm 12-15cm 18-21cm 24-27cm 30-33cm0

20

40

60

80

100

120

140

160

180


MBT DBT TBT

Depth /cm

OT

conc

entr

ation

ug/

L

Figure 8.Mean pH against Depth for Kong-kong 2.

Figure 9.Mean pH against Depth for Kong-kong 3.

21

4.1.3 Total Organic Carbon (TOC) and depth.

Statistically no significance difference in correlation between TOC and depth in sample

from site 1(Figure 8).However, there is a strong negative correlation between TOC and depth

of site 2(Figure 9) with r=-0.733(p=0.05).The reduction in TOC with increasing depth display

that site 2 is relatively further to industrial and anthropogenic influence which affects the

TOC usually with positive correlation. There is a strong negative correlation between TOC

and depth of site 3(Figure 10) with r=-0.868 (p=0.05).The higher reduction ratio of TOC cor-

responding to its depth at site 3, reflects site 3 is even relatively further than site 2 to indus-

trial and anthropogenic influence which affects the TOC usually with positive correlation.

This is true in the case of sampling site selection in which site 1 with busy anthropogenic

activities, site 2 and 3 is further from site 1 relatively.

Figure 10.Total Organic Carbon against Depth for Kong-kong 1.

22

Figure

11.Total Organic Carbon against Depth for Kong-kong 2.

Figure 12.Total Organic Carbon against Depth for Kong-kong 3.

23

4.1.4 Particle Size Distribution and depth

There is no significant correlation between particle size distribution of site 1(Figure11) and

depth. However percentage of gravel and silt correlate with r=-0.850(p=0.01). At site 2(Fig-

ure12), gravel and depth have strong correlation with r=0.687(p=0.05).However sand and silt

do not display any significant correlation towards depth. There is no significant correlation

between particle size distribution of site 3(Figure13) and depth. However percentage of sand

and silt correlate strongly with r=-0.986(p=0.01).To windup, there is no evidence statistically

significant correlating depth to particle size distribution.

Figure 13.Particle Size Distribution against Depth for Kong-kong 1.

24


25


4.2 Physicochemical parameters and sites.

2.1.4 4.2.1 Organotin compound concentration and sites.

Figure 16. Organotin Compound Concentration against Sites.

Butyltin species were detected throughout the sediment core in all sites’ intervals. It

has been reported that The Australian sediment quality guidelines for TBT are 5 ug/L

and 70 ug/L for low and high threshold values (S.Diez et al,.2002). The concentra-

tions of TBT in sediment samples from all 3 stations in Kong-kong Laut were found

to be higher than the highest threshold value, suggesting these sediments may pose a

threat to a benthic biota(S.Diez et al,.2002). Moreover, high levels of TBT in Kong-

kong Laut sediments reflects its widespread contamination and could be an indicative

of the continuing usage of TBT based antifouling paints on ship hulls. Via graphical

representation in (Figure 14), qualitatively Kong-kong 1 has high TBT/DBT ratio may

be attributed to recent input of TBT and/or to low degradation of TBT to DBT into

this station. It is known that TBT degradation rates in sediment are slower than in wa-

ter column, particularly in anaerobic conditions. The half-life of TBT in sediments is

in the range of years rather than days or weeks in the water column (F.Cima et al,.2003)

26

Kong-kong 1 Kong-kong 2 Kong-kong 30

200

400

600

800

1000

1200

Organotin Compound Concentration against Sampling Sites

Monobutyltin Dibutyltin Tributyltin

Sampling Site

Conc

entr

ation

ug/

L

.OT compound concentration at site 1 is the highest compared to site 2 and 3 respectively. Overall, the

concentration mean of Dibutyltin is the highest compared to Tributyltin and Monobutyltin ,which is

38.99 ug/L for MBT,85.90ug/L and 61.46ug/L for DBT and TBT respectively, it can be inferred that the

highest concentration of DBT is from the source of degradation of TBT. There is a significant negative

correlation between DBT concentration and site with r=-0.523(p=0.01).Thus, it can be concluded that

further the site from main polluted site (Kong-kong 1/site 1), there is a linear decrease in organotin

concentration.

4.2.2 Particle Size Distribution and Sites.

Figure 17.Particle Size Distribution

against Sites.

The current investigation confirms that sediments in kong-kong laut, Johore are

generally consist of silty-clay (>80%) USDA and UK-ADAS textural

triangle.Statistically there is a correlation significantly between sand and sites, which

is r=-0.437(p=0.05),hence when sand and organotin tin concentration were tested for

correlation, there is a significantly strong correlation between them with

r=0.690(p=0.01). A study to assess the degradation kinetics of butyltin and phenyltin

in sandy soil collected in INRA (Pierroton Experimental Unit, near Bordeaux, France)

noted that the order of persistence in the soil was as follows:

MBT>DBT>MPhT>TBT>DPhT>TPhT. Therefore, it can be deduced that sand has high

adsorptivity towards OT compounds, particularly butyl species. Therefore, TBT, DBT

and MBT accumulated in sediments of the Kong-kong laut was affected by grain size,

significantly, sand even though with the least percentage of its constituent at each site.

27

Kong-kong 1 Kong-kong 2 Kong-kong 30%

10%20%30%40%50%60%70%80%90%

100%

Particle Size Distribution against Site

Gravel Sand SiltSand Silt Clay

4.2.3 Total Organic Carbon (TOC) and sites.

Figure 18.Total Organic Carbon against Sites.

The highest TOC content percentage (39.97%) was recorded for kong-kong 2 ,this

may due to it being near to small residential area and the nearby area composes of

small food industry region,hence total organic carbon may due to influx of organic

material to the site (Walkley.A et al.,1947).TOC percentage for site kong-kong 1 and

kong-kong 3 are 22.70% and 19.86% respectively whereby kong-kong 1 have

substantial amount of anthropogenic activities and site 3 with least anthropogenic

activities .Correlation between TOC and organotin compound concentration is

significant between MBT species and TOC which correlates negatively with r=-

0.383(p=0.01). To relate between MBT,TOC and particle size distribution(sand),in

brief MBT correlates negatively with TOC ,but MBT correlates positively with

particle size distribution (sand),hence it can be inferred that MBT is more strongly

adsorbed to mineral phase (sand) than organic matter (TOC) and to further strengthen

this point,the study area(south Johor) investigated has high constituent of

montmorillonite mineral which makes up the sand majorly (Khairuddin et

al.,1992) ,in which montmorillonite has the highest adsorptivity towards butyltin

species (refer 2.2.3 Sediment Particle Size). In addition, the adsorption properties of

OTs cannot be solely determined or correlated with only considering TOC because

OTs adsorption properties also depends on properties of minerals in sediment (eg.

montmorillonite, kaolinite and quartz) which has different sorption coefficient kd.To

wrap up, the total organic carbon has least affects on the organotin concentration at

28

Kong-kong 1 Kong-kong 2 Kong-kong 305

10152025303540

TOC against Site

Kong-kong laut than the type of mineral content which makes the component of the

particles of the sediments.

4.2.4 Mean pH and sites.

Figure 19.Mean pH against Sites.

The mean pH value of all three site is in the range of ±0.24 and tend to display low

variation between sites. From the statistical correlation test done, all species of OT

concentration have strong negative correlation with pH with MBT having r=-

0.929(p=0.01), r=-0.881(p=0.01), and r=-0.833(p=0.05) for TBT and DBT

respectively. Therefore, in lower pH the concentration of OT is higher than in higher

pH. At low pH where pH<pKa, the organotin compound is present primarily as cation

and the main adsorption process is expected to be a cationic exchange mechanism. At

pH higher than pKa, the neutral organotin species are prevailing and adsorption is

mainly controlled by hydrophobic interactions. In general, the highest adsorption of

TBT is detected between pH 6 and 7, which reveals the area of maximum overlap

between the total negative surface charge and the concentration of TBT cations in

solution (Fent,1996; Weidenhaupt, 1997; Hoch, 2001; Hoch, 2004).Thus, the optimal

pH value at Kong-kong Laut is suitable for adsorption and cationic exchange

mechanism on sediments.Thus ,the monitoring of the concentration of organotin in

29

Kong-kong laut is very vital to protect the marine ecosystem and human health

toxicity effect(Howells et al .,1997).

30

5.0 Conclusion

The recent study exposed that the concentration of organotin compounds in Kong-

kong Laut is still higher than the recommended threshold level and considered highly

polluted. Primarily this is due to increased recreational boating activities, perhaps due

to anthropogenic inputs from marinas activity, which is still moderately active till to-

day. Furthermore, the concentration mean of OTs(tributyltin,dibutyltin dan

monibutyltin) had elevated from 232 ug/L [2006 data], (Harino et al.,2008) to

398ug/L [2013 data] at recent study, hence after seven years there is an increase of

72%,hence if divided equally, it is of 9% increment annually of organotin concentra-

tion in Johore. Therefore, the banned organotin compound containing paints are still

being utilized till now, thus ,it is recommended that the enforcement and environmen-

tal departments should investigate the usage of anti-fouling paints as soon as possible

to avoid future contamination and severe ecotoxicological consequences . Factor of

depth correlates negatively with organotin concentration due to the various factor in-

fluencing the penetration of these compounds into the sediments which acts as the

sink, this might due to adsorptivity which was discussed earlier correlates positively

with influence of type of minerals which it constitute in sediment particle,the sand

portion of the Kong-kong Laut sediment which consist of montmorillonite(mineral

that makes up the sand proportion of sediment particle distribution) ,in which highly

adsorps butyltin compounds than kaolinite and quartz (Khairuddin et al.,1992).In ad-

dition, total organic carbon (TOC) influences the least significance in concentration of

organotin compounds in Kong-kong Laut due to the greater effects of sediment min-

erals which suppresses the effect of TOC. In relation to pH and depth, pH increases

with increase in depth due to increase in bacterial activity (Wang and Alva et

al .,2000) consequently increase in pH causes decrease in organotin concentration

due to adsorptivity of organotin species decreases with increasing pH (Fent,1996;

Weidenhaupt, 1997; Hoch, 2001; Hoch, 2004) . In conclusion,the in depth and further

understanding of the physicochemical parametres effects towards organotin concen-

tration particularly the butyltin species, should be studied for greater understanding of

the nature and fate of organotin compounds.

31

Reference

Adehnan, D., Hinga, K. R. & Pilson, M. E. Q. (1990). Bio- geochemistry of butyltins in an enclosed

marine ecosystem. Environ. Sci. Technol., 24, 1027-32.

Alzieu, C. & Portman, J. E. (1984). The effect of tributyltin on the culture of Crassostrea gigas and

other species. 15th Annual Shellfish Conference, Shellfish Association, London, pp. 87-100

Anon (1994). TBT on the way out. Mar. Pollut. Bull., 28, 519. Clark, E. A., Sterritt, R. M. & Lester,

J. N. (1988). The fate of tributyltin in the aquatic environment. Environ. Sci.Technol., 22, 600-k

Harris, J. R. W., Hamlin, C. C. & Stebbing, A. R. D. (1991). A simulation study of the effectiveness

of legislation and improved dockyard practice in reducing TBT concentrations in the Tamar estuary.

Mar. Environ. Res., 32, 279-92.

Hawes, E. L., (1992). A legacy in sediment: Exposing Casco Bay’s dirty history. Habitat, 9, 241.

K. Fent, “Ecotoxicology of Organotin Compounds,” Re- vised Marine Toxicology, Vol. 26, 1996,

pp. 117-121.

G. W. Bryan and P. E. Gibbs, “Impacts of Low Concen- trations of Tributyltin (TBT) on Marine

Organisms,” In: M. C. Newman and A. W. McIntosh, Eds., Metal toxicology, Concepts and applica-

tions, Academic Press, Lon- don, 1991.

M. H. Salazar and S. M. Salazar, “Assessing Site-Specific Effects of TBT Contamination with Mus-

sel Growth Rates,” Marine Environment Research, Vol. 32, No. 1-4, 1991, pp. 131-150.

doi:10.1016/0141-1136(91)90038-A

C. Alzieu, J. Sanjuan, J. P. Deltreil and B. Borel, “Tin Applied,” Organometallic Chemistry, Vol. 17,

1986, pp. 81-105.

T. Horiguchi, H. Shiraishi, M. Shimizu and M. Morita, “Imposex and Organotin Compounds in

Thais Clavigera and T. Bronni in Japan,” Journal of Marine Biolology, Vol. 74, 1994, pp. 651-669.

Harino,H.,Arai,T.,Ohji,M.Ismail,A.B,Wattayakoon,G.andMiyazaki,N.2007,Occurrence of

Antifouling Biocides in Southeast Asia-Malaysia,Thailand and Vietnam –The ASEAN

international

Conference ‘Conservation of Coastal Environment’ pp91-111

Harino, H., Fukushima, M., Mori, Y. and Nakadoi, T. (1990) Distribution of organotins in the har-

bour area of Osaka City, Japan. Hazard Assessment and Control of Environment Contaminants in Wa-

ters, 158±164.

33

Harino, H., Fukushima, M. and Tanaka, M. (1992) Simulta- neous determination of butyltin and

phenyltin compounds in the aquatic environment by gas chromatography. Analy- tica Chimica Acta

264, 91±96.

Harino, H., Fukushima, M., Kurokawa, Y. and Kawai, S. (1997) Susceptibility of bacterial popula-

tions to organotin compounds and microbial degradation of organotin compounds in environmental

water. Environmental Pollution (this issue).

Hattori, Y., Kobayashi, A., Nonaka, K., Sugimae, A. and Nakamoto, M. (1988) Degradation of trib-

utyltin and dibutyltin compounds in environmental water. Water Science and Technology 20, 71±76.

Huggett, R. J., Unger, M. A., Seligman, P. F. and Valkirs, A. O. (1992) The marine biocide trib-

utyltin, assessing and managing the environmental risks. Environmental Science and Technology 26,

232±237.

Harino, H., Fukushima, M., Kawai, S., 2000. Accumulation of butyltin and phenyltin compounds in

various fish species. Archives of Environmental Contamination and Toxicology 39, 13–19.

Hong, H.K., Takahashi, S., Min, B.Y., Tanabe, S., 2001. Butyltin residues in blue mussels (Mytilus

edulis) and arkshells (Scapharca broughtonii) collected from Korean coastal waters. Environmental

Pollution 117, 475–486.

Horiguchi, T., Shiraishi, H., Shimizu, M., Morita, M., 1994. Imposex and organotin compounds in

Thais clavigera and T. bronni in Japan. Journal of the Marine Biological Association of the United

Kingdom 74, 651–669.

Hung, T., Lee, T., Liao, T., 1998. Determination of butyltins and phenyltins in oysters and fishes

from Taiwan coastal waters. Environmental Pollution 102, 197–203.

Hwang, H.M., Oh, J.R., Kahng, S., Lee, K.W., 1999. Tributyltin compounds in mussels, oysters and

sediments of Chinhae Bay, Korea. Marine Environmental Research 47, 61–70.

Kan-Atireklap, S., Tanabe, S., Sanguansin, J., Tabucanon, M., Hungspreugs, M., 1997a. Contamina-

tion by butyltin compounds and organochlorines residues in green mussel (Perna viridis, L.) from Thai-

land coastal waters. Environmental Pollution 97, 79– 89.

Arnold, C.G., Weidenhaupt, A., David, M.M., Muller, S.R., Haderlein, S.B., Schwarzenbach, R.P.,

1997. Aqueous speciaton and 1-octanol–water partitioning of tributyl- and triphenyltin: Effect of pH

and ion composition. Environ. Sci. Technol. 31, 2596–2602.

Arnold, C.G., Berg, M., Muller, S.R., Dommann, U., Schwar- zenbach, R.P., 1998a. Determination

of organotin compounds in water, sediments, and sewage sludge using perdeuterated internal standards,

accelerated solvent extraction, and large- volume-injection GC/MS. Anal. Chem. 70, 3094–3101.

34

Arnold, G.C., Ciaoni, A., Muller, S.R., Amirnahman, A., Schwarzenbach, R.P., 1998b. Association

of triorganotin compounds with dissolved humic acids. Environ. Sci. Technol. 32, 2976–2983.

Batley, G.E., 1996. The distribution and fate of tributyltin in the marine environment. In: De Mora,

S.J. (Ed.), Tributyl- tin: a case study of an environmental contaminant. Cam- bridge Environmental

Chemistry Series, Cambridge University Press, Cambridge.

Gomez-Ariza, J.L., Beltran, R., Morales, E., Giraldez, I., Ruizbenitez, M., 1995. Acid-extraction

treatment of sediment samples for organotin speciation; occurrence of butyltin and phenyltin com-

pounds on the Cadiz coast, south-west Spain. Appl. Organomet. Chem. 9, 51–64.

Ko, M.M.C., Bradley, G.C., Neller, A.H., Broom, M.J., 1995. Tributyltin contamination of marine

sediments of Hong Kong. Marine Pollution Bulletin 31, 249–253.

35

Hellawell JM (1986) Biological indicators of freshwater pollution and environmental management. In:

Melanby, K. (Ed.), Pollution Monitoring Series. Amsterdam, Elsevier; 546pp

Ishak A, Hassan ZNC, Edros NH, Zamberi MH, Rahman MNA (2008) The effect of local on urban

heat island trend; a case study in urban areas of Ipoh and Kuantan. http://www.met.gov.my/images/

pdf/research/researchpaper_201101.pdf

Juahir H, Zain SM, Aris AZ, Yusoff MK, Mokhtar M (2010) Spatial assessment of Langat river water

quality using chemometrics. Journal of Environmental Monitoring 12: 287–295

Mazlum N, Ozer A, Mazlum S (1999) Interpretation of water quality data by principal components

analysis. Turkish Jour Engg Environ Sci 23: 19-26

Metcalfe JL (1989) Biological water quality assessment of running waters based on macroinvertebrate

communities: History and present status in Europe. Environ Pollut 60: 101–139

Nedeau EJ, Merritt RW, Kaufman MG (2003) The effect of an industrial effluent on an urban stream

benthic community: water quality vs. habitat quality. Environ Pollut 123: 1–13

Pinel-Alloul B, Methot G, Lapierre L, Willsie A (1996) Macro-benthic community as a biological in-

dicator of ecological and toxicological factors in Lake Saint-Francois (Quebec). Environ Pollut 9:

65–87

Rizwan AM, Dennis LYC, Liu C (2008) A review on the Generation, Determination and Mitigation of

Urban Heat Island. Journal of Environmental Science 20: 120-128

Wright JF, Furse MT, Armitage PD, Moss D (1993) New procedures for identifying running-water

sites subject to environmental stress and for evaluating sites for conservation, based on the macroin -

vertebrate fauna. Arch. Hydrobiologia 127: 319–326

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Appendix 1

37

Appendix 2

40

Appendix 1

You may have one or more appendices containing detail, bulky or reference material that is relevant

though supplementary to the main text: perhaps additional specifications, tables or diagrams that would

distract the reader if placed in the main part of the dissertation. Make sure that you place appropriate

cross-references in the main text to direct the reader to the relevant appendices.

46

Appendix 2

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