LEVELS OF Zn, Pb, As AND Hg IN SELECTED MARINE FISHES FROM STRAITS OF MALACCA,

BY

RINA SHARLINDA HJ. ZABRI

A thesis submitted in fulfilment of the requirement for the degree of Master of Science

Kulliyyah of Science International Islamic University Malaysia

JUNE 2010 ABSTRACT

A study of the accumulation of four trace elements (Zn, Pb, As and Hg) in tissues of selected marine fish species namely Japanese Threadfin Bream, Dorab Wolf-herring, White Pomfret, Tiger-toothed Croaker and Emperor Red Snapper from Straits of Malacca was conducted from January 2008 to February 2009. Fish samples were obtained from 16 sampling sites namely; Pontian, Kukup, Muar, Linggi, Pasir Penambang, Tanjung Karang, Hutan Melintang, , , Batu Maung, Teluk Bahang, Tanjung Dawai, Kuala Kedah, Langkawi, Kuala Perlis and Kuala Sg. Baru. Sampling sites were comprised of Lembaga Kemajuan Ikan Malaysia (LKIM) fish landing ports and local jetties along the states situated on the west coast of Peninsular Malaysia. Two type of fish tissues were studied; namely muscle and gill. They were dissected and dried before the digestion process took place using HNO3, HF, HCl and H2SO4 in both open and closed digestion system. Correspondingly, determination of heavy metals was carried out by using Inductively Coupled Plasma Spectrometry (ICP-MS). Results for heavy metal accumulation were divided into zonation of the states consisting of the north zone (Perlis, Kedah and Pulau Pinang), middle zone ( and Selangor) and south zone (Negeri Sembilan, Melaka and Selangor). The fish samples were found to contain Zn within the range of 2.327 to 5.890 μg/g (dry wt.) in the muscles and 8.722 to 13.960 μg/g (dry wt.) in the gills. The highest Pb levels in the muscles were at 0.263 μg/g (dry wt.) while the lowest was at 0.014 μg/g (dry wt.). In the gills, Pb accumulated from 0.032 μg/g (dry wt.) to 0.193 μg/g (dry wt.) respectively. For As, concentrations in the muscles ranged from 0.004 μg/g (dry wt.) to 0.025 μg/g (dry wt.) while in the gills As accumulated from 0.013 μg/g (dry wt.) to 0.062 μg/g. Interestingly, levels of Hg were between 0.009 μg/g (dry wt.) and 0.026 μg/g (dry wt.) in the muscles and were from 0.005 μg/g (dry wt.) to 0.055 μg/g (dry wt.) in the gills; with both the highest accumulation recorded in White Pomfret and the lowest concentration recorded in Tiger-toothed Croaker. Comparison of heavy metal concentration in the fish from different locations showed significant differences with p<0.05 suggesting the fish is influence by the heavy metal concentration in the surrounding area. Positive correlations were acquired from heavy metal concentrations in the fish with fish size and length, thus proved bigger and larger fish were possible to contain greater amount of metals. A comparative study was also carried out between the existing data on the metals in the fish tissues and species of marine and coastal fish from different regions in the world to observe their trend and status in regional and global contexts. On the whole, the findings from this study revealed that all the metal concentrations in the tissues were lower than the maximum permissible limit as recommended by the Malaysian Food Regulations (1985) and World Health Organization (1990). Results from Provisional Tolerable Weekly Intake (PTWI) indicate that concentration of Zn, Pb and Hg are still within the permissible limits. However, As concentration may exceed the permissible limits in a human’s body if consumed according to the weekly intake recorded; thus pose a threat to Malaysian public.

ii ﻣﻠﺨﺺ اﻟﺒﺤﺚ

وآﺸﻔﺖ دراﺳﺔ ﻟﺘﺮاآﻢ اﻟﻤﻌﺎدن اﻟﺜﻘﻴﻠﺔ اﻷرﺑﻊ (اﻟﺰﻧﻚ ، اﻟﺮﺻﺎص ، واﻟﺰﺋﺒﻖ و) ﻓﻲ أﻧﺴﺠﺔ ﻣﻦ أﺳﻤﺎك اﻟﻘﺎع اﻟﻤﺨﺘﺎرة ﻣﻦ اﻷﺳﻤﺎك اﻟﺒﺤﺮﻳﺔ اﻟﻴﺎﺑﺎﻧﻴﺔ هﻤﺎ Threadfin ﺑﺮﻳﻢ، Dorabاﻟﺬﺋﺐ اﻟﺮﻧﺠﺔ، واﻷﺑﻴﺾ ﺑﻮﻣﻔﺮﻳﺖ واﻟﻨﻤﻮر اﻟﻨﺎﻋﺐ ﻣﺜﻠﻢ واﻻﻣﺒﺮاﻃﻮر ﺳﻤﻚ اﻟﻨﻬﺎش اﻷﺣﻤﺮ ﻣﻦ ﻣﻀﻴﻖ ﻣﻠﻘﺎ أﺟﺮي ﺧﻼل اﻟﻔﺘﺮة ﻣﻦ ﻳﻨﺎﻳﺮ 2008 اﻟﻰ ﻓﺒﺮاﻳﺮ 2009. وﺗﻢ اﻟﺤﺼﻮل ﻋﻠﻰ ﻋﻴﻨﺎت ﻣﻦ اﻷﺳﻤﺎك ﻣﻦ 15 ﻣﻮاﻗﻊ أﺧﺬ اﻟﻌﻴﻨﺎت وهﻤﺎ؛ Pontian، آﻮآﺐ، ﻣﻮار، Linggi، ﺑﺎﺳﻴﺮ Penambang، ﺗﺎﻧﺠﻮﻧﻎ آﺎراﻧﻎ، ﻏﺎﺑﺔ Pantai ،Melintang رﻳﻤﺲ، Terong، ﺑﺎﺗﻮ ﻣﻮﻧﻎ ، ﺗﻴﻠﻮك ﺑﺎهﺎﻧﻎ ، ﺗﺎﻧﺠﻮﻧﻎ دواي، آﻮاﻻ ﻟﻜﻴﺪا ، ﻻﻧﻐﻜﺎوي، آﻮاﻻ ﺑﻴﺮﻟﻴﺲ وآﻮاﻻ ﻟﺴﺎن ﺟﺮﻣﺎن. ﺑﺎرو. هﺬﻩ اﻟﻤﻮاﻗﻊ آﺎﻧﺖ ﻣﺒﺎﻏﺎ Kemajuan اﻟﺴﻤﻜﻴﺔ ﻣﺎﻟﻴﺰﻳﺎ (LKIM) ﻣﻮاﻧﺊ إﻧﺰال اﻷﺳﻤﺎك اﻟﺮﺋﻴﺴﻴﺔ واﻷﺳﻮاق اﻟﻤﺤﻠﻴﺔ ﻋﻠﻰ اﻣﺘﺪاد اﻟﺪول اﻟﺘﻲ ﺗﻘﻊ ﻋﻠﻰ اﻟﺴﺎﺣﻞ اﻟﻐﺮﺑﻲ ﻣﻦ ﺷﺒﻪ ﺟﺰﻳﺮة ﻣﺎﻟﻴﺰﻳﺎ. وهﻤﺎ ﻧﻮع ﻣﻦ اﻧﺴﺠﺔ اﻻﺳﻤﺎك ﺗﻤﺖ دراﺳﺘﻬﺎ ، وﺗﺤﺪﻳﺪا ﻓﻲ اﻟﻌﻀﻼت واﻟﺨﻴﺸﻮﻣﻴﺔ. آﺎﻧﻮا ﺗﺸﺮﻳﺢ وﺗﺠﻔﻴﻔﻬﺎ ﻗﺒﻞ ﻋﻤﻠﻴﺔ

اﻟﻬﻀﻢ وﻗﻌﺖ ﺑﺎﺳﺘﺨﺪام HNO3، ﻓﻠﻮرﻳﺪ اﻟﻬﻴﺪروﺟﻴﻦ وآﻠﻮرﻳﺪ اﻟﻬﻴﺪروﺟﻴﻦ و H2SO4 ﻓﻲ آﻼ اﻟﻤﻔﺘﻮﺣﺔ واﻟﻤﻐﻠﻘﺔ ﻧﻈﺎم اﻟﻬﻀﻢ. ﻓﻲ اﻟﻤﻘﺎﺑﻞ، ﻟﻢ ﻳﻨﻔﺬ اﻟﻤﺼﻴﺮ ﻣﻦ اﻟﻤﻌﺎدن اﻟﺜﻘﻴﻠﺔ ﻋﻦ ﻃﺮﻳﻖ اﺳﺘﺨﺪام اﻟﺒﻼزﻣﺎ اﻟﻤﺘﻘﺎرﻧﺔ ﺑﺎﻟﺤﺚ اﻟﻄﻴﻒ (ﺑﺮﻧﺎﻣﺞ اﻟﻤﻘﺎرﻧﺎت اﻟﺪوﻟﻴﺔ ﻟﻤﺮض اﻟﺘﺼﻠﺐ اﻟﻌﺼﺒﻲ اﻟﻤﺘﻌﺪد). ﻧﺘﻴﺠﺔ ﻟﺘﺮاآﻢ اﻟﻤﻌﺎدن اﻟﺜﻘﻴﻠﺔ آﺎﻧﺖ ﻣﻘﺴﻤﺔ اﻟﻰ ﺗﻘﺴﻴﻢ ﻣﻨﺎﻃﻖ ﻣﻦ اﻟﺪول اﻟﺘﻲ ﺗﺘﺄﻟﻒ ﻣﻦ ﻣﻨﻄﻘﺔ اﻟﺸﻤﺎل (ﺑﻴﺮﻟﻴﺲ وآﻴﺪاﻩ وﺑﻮﻻو ﺑﻴﻨﺎﻧﻎ) ، واﻟﻤﻨﻄﻘﺔ اﻟﻮﺳﻄﻰ (ﺑﻴﺮاك وﺳﻴﻼﻧﻐﻮر) وﻣﻨﻄﻘﺔ اﻟﺠﻨﻮب (ﻧﻴﻐﻴﺮي ﺳﻴﻤﺒﻴﻼن ، ﻣﻠﻘﺎ وﺳﻴﻼﻧﻐﻮر). ﻓﻲ ﻋﻴﻨﺎت ﻣﻦ اﻷﺳﻤﺎك ووﺟﺪ أﻧﻬﺎ ﺗﺤﺘﻮي ﻋﻠﻰ اﻟﺰﻧﻚ ﺿﻤﻦ ﻣﺠﻤﻮﻋﺔ ﻣﻦ 2.327 إﻟﻰ 5.890 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) ﻓﻲ اﻟﻌﻀﻼت و8.722 إﻟﻰ 13.960 ﻣﻴﻜﺮوﻏﺮام / ﻏﺮام (ﺑﺎﻟﻮزن اﻟﺠﺎف) ﻓﻲ اﻟﺨﻴﺎﺷﻴﻢ. أﻋﻠﻰ ﻣﺴﺘﻮﻳﺎت اﻟﺮﺻﺎص ﻓﻲ اﻟﻌﻀﻼت وﺻﻠﺖ 0.263 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) ﻓﻲ ﺣﻴﻦ آﺎن أدﻧﻰ ﻣﺴﺘﻮى ﻓﻲ 0.014 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف). ﻓﻲ اﻟﺨﻴﺎﺷﻴﻢ ، اﻟﺮﺻﺎص اﻟﻤﺘﺮاآﻢ ﻣﻦ 0.032 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) إﻟﻰ 0.193 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) ﻋﻠﻰ اﻟﺘﻮاﻟﻲ. وآﻤﺎ ل، ﺗﺮآﻴﺰات ﻓﻲ ﻋﻀﻼت ﺗﺮاوﺣﺖ ﺑﻴﻦ 0.004 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) إﻟﻰ 0.025 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) ﻓﻲ ﺣﻴﻦ أن اﻟﺨﻴﺎﺷﻴﻢ واﻟﻤﺘﺮاآﻤﺔ ﻣﻦ 0.013 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) إﻟﻰ 0.062 ﻣﻴﻜﺮوﻏﺮام .g / وﻣﻦ اﻟﻤﺜﻴﺮ ﻟﻼهﺘﻤﺎم ، وﻣﺴﺘﻮﻳﺎت اﻟﺰﺋﺒﻖ آﺎﻧﺖ أﻋﻤﺎرهﻢ ﺗﺘﺮاوح ﺑﻴﻦ 0.009 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) و 0.026 ﻣﻴﻜﺮوﻏﺮام / ﻟﺘﺮ (ﺑﺎﻟﻮزن اﻟﺠﺎف) ﻓﻲ اﻟﻌﻀﻼت ، وآﺎﻧﻮا ﻣﻦ 0.005 إﻟﻰ 0.055 ﻓﻲ اﻟﺨﻴﺎﺷﻴﻢ ، ﻣﻊ آﻞ ﻣﻦ أﻋﻠﻰ اﻟﻤﻌﺪﻻت اﻟﻤﺴﺠﻠﺔ ﻓﻲ ﺗﺮاآﻢ اﻻﺑﻴﺾ وﺑﻮﻣﻔﺮﻳﺖ ﺳﺠﻠﺖ أدﻧﻰ ﺗﺮآﻴﺰ ﻓﻲ اﻟﻨﻤﻮر اﻟﻨﺎﻋﺐ ﻣﺴﻨﻨﺔ. ﻣﻘﺎرﻧﺔ ﺗﺮآﻴﺰ اﻟﻤﻌﺎدن اﻟﺜﻘﻴﻠﺔ ﻓﻲ ﻣﻮاﻗﻊ ﻣﺨﺘﻠﻔﺔ ﻣﻦ اﻷﺳﻤﺎك ﻟﻢ ﻳﻈﻬﺮ اﺧﺘﻼف آﺒﻴﺮ ﻣﻊ ع > 0.05 ﻣﻤﺎ ﻳﺪل ﻋﻠﻰ اﻷﺳﻤﺎك آﻤﺨﻠﻮق اﻟﻤﺤﻤﻮل اﻟﺬي هﻮ أﻗﻞ ﻣﻦ اﻟﺘﺄﺛﻴﺮ ﻋﻠﻰ ﺗﺮآﻴﺰ اﻟﻤﻌﺎدن اﻟﺜﻘﻴﻠﺔ ﻓﻲ اﻟﻤﻨﻄﻘﺔ اﻟﻤﺤﻴﻄﺔ. ﻋﻼﻗﺎت إﻳﺠﺎﺑﻴﺔ ، ﺗﻢ اﻟﺤﺼﻮل ﻋﻠﻴﻬﺎ ﻣﻦ ﺗﺮآﻴﺰات اﻟﻤﻌﺎدن اﻟﺜﻘﻴﻠﺔ ﻓﻲ اﻷﺳﻤﺎك ﻣﻊ ﺣﺠﻢ اﻟﺴﻤﻚ واﻟﻄﻮل ، وﺑﺬﻟﻚ ﺛﺒﺖ أآﺒﺮ وأآﺒﺮ ﺳﻤﻜﺔ ﻣﻦ اﻟﻤﻤﻜﻦ أن ﺗﺤﺘﻮي ﻋﻠﻰ أآﺒﺮ آﻤﻴﺔ ﻣﻦ اﻟﻤﻌﺎدن اﻟﺜﻘﻴﻠﺔ. دراﺳﺔ ﻣﻘﺎرﻧﺔ

iii أﺟﺮﻳﺖ أﻳﻀﺎ ﺑﻴﻦ اﻟﺒﻴﺎﻧﺎت اﻟﻤﻮﺟﻮدة ﻋﻠﻰ اﻟﻤﻌﺎدن ﻓﻲ أﻧﺴﺠﺔ اﻟﺴﻤﻚ وأﻧﻮاع اﻷﺳﻤﺎك اﻟﺒﺤﺮﻳﺔ واﻟﺴﺎﺣﻠﻴﺔ ﻣﻦ ﻣﺨﺘﻠﻒ اﻟﻤﻨﺎﻃﻖ ﻓﻲ اﻟﻌﺎﻟﻢ ، ﻟﻤﺮاﻗﺒﺔ اﻟﻮﺿﻊ واﺗﺠﺎهﻬﺎ ﻓﻲ اﻟﺴﻴﺎﻗﺎت اﻹﻗﻠﻴﻤﻴﺔ واﻟﻌﺎﻟﻤﻴﺔ. ﻋﻠﻰ اﻟﻌﻤﻮم ، ﻓﺈن ﻧﺘﺎﺋﺞ هﺬﻩ اﻟﺪراﺳﺔ آﺸﻔﺖ أن ﺟﻤﻴﻊ ﺗﺮآﻴﺰات اﻟﻤﻌﺎدن اﻟﺜﻘﻴﻠﺔ ﻓﻲ اﻷﻧﺴﺠﺔ آﺎﻧﺖ أﻗﻞ ﻣﻦ اﻟﺤﺪ اﻷﻗﺼﻰ اﻟﻤﺴﻤﻮح ﺑﻪ ﻋﻠﻰ اﻟﻨﺤﻮ اﻟﻤﻮﺻﻰ ﺑﻪ ﻣﻦ ﻗﺒﻞ وزارة اﻟﺼﺤﺔ ﻣﺎﻟﻴﺰﻳﺎ (1998) وﻣﻨﻈﻤﺔ اﻷﻏﺬﻳﺔ واﻟﺰراﻋﺔ (2002). وﻣﻊ ذﻟﻚ ، وﺗﺤﺪﻳﺪ اﻟﻤﺘﺤﺼﻞ اﻷﺳﺒﻮﻋﻲ اﻟﻤﻘﺒﻮل ﻣﺒﺪﺋﻴﺎ (PTWI) وآﻤﺎ ﻳﺘﻀﺢ أن اﻟﺘﺮآﻴﺰ ﻗﺪ ﺗﺘﺠﺎوز اﻟﺤﺪود اﻟﻤﺴﻤﻮح ﺑﻬﺎ ﻓﻲ ﺟﺴﻢ اﻹﻧﺴﺎن إذا ﺗﺴﺘﻬﻠﻚ وﻓﻘﺎ ﻟﻜﻤﻴﺔ أﺳﺒﻮﻋﻴﺔ ﻣﺴﺠﻠﺔ ﺑﺬﻟﻚ ﺗﺸﻜﻞ ﺧﻄﺮا ﻋﻠﻰ اﻟﺠﻤﻬﻮر اﻟﻤﺎﻟﻴﺰي.

iv APPROVAL PAGE

I certify that I have supervised and read this study and that in my opinion; it conforms to acceptable standards of scholarly presentation and is fully adequate, in scope and quality, as a thesis for the degree of Master of Science.

...... Kamaruzzaman Bin Yunus Supervisor

………………………..……... Ahmed Jalal Khan Chowdhury Co-Supervisor

I certify that I have read this study and that in my opinion, it conforms to acceptable standards of scholarly presentation and is fully adequate, in scope and quality, as a thesis for the degree of Master of Science.

…………………………… Shahbudin Saad Examiner

This thesis was submitted to the Department of Biotechnology and is accepted as a fulfilment of the requirement for the degree of Master of Science.

...... ……………… Tg. Haziyamin Tg. Abd. Hamid Head, Department of Biotechnology

This thesis was submitted to the Kulliyyah of Science and is accepted as a fulfilment of the requirement for the degree of Master of Science.

……………………………… Kamaruzzaman B. Yunus Dean, Kulliyyah of Science

v DECLARATION

I hereby declare that this dissertation is the result of my own investigations, except where otherwise stated. I also declare that it has not been previously or concurrently submitted as a whole for any other degrees at IIUM or other institutions.

Rina Sharlinda binti Hj. Zabri

Signature ……………………………… Date ………………...

vi

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Copyright © 2010 by Rina Sharlinda binti Hj. Zabri. All rights reserved.

LEVELS OF Zn, Pb, As AND Hg IN SELECTED MARINE FISHES FROM STRAITS OF MALACCA, MALAYSIA

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1. Any material contained in or derived from this unpublished research may only be used by others in their writing due acknowledgement.

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Affirmed by Rina Sharlinda binti Hj. Zabri

………………………………… ……………… Signature Date

vii ACKNOWLEDGEMENTS

Bismillahirrahmanirrahim. In the name of Allah, all Praise is due to Him, the Sustainer of the Heavens and Earth and all that is within it and may His blessings be upon the Prophet Muhammad SAW, peace be upon him. I would like to express my sincere appreciation to my supervisor, Prof. Dr. Kamaruzzaman bin Yunus for his inspiring guidance, opinion, tolerance, encouragement and support to all my endeavours in the field of research and in the preparation of this thesis. Without his unstinted cooperation, my efforts would not have been as successful as it has been. His moral support was inspirational to me. May Allah bless you. I would also like to register my gratitude to Assoc. Prof. Dr. Jalal Khan Chowdury and Asst. Prof. Dr. Shahbuddin bin Saad, Kulliyyah of Science, for their supervision and assistance. Without their persistence trust in me, I would not have finished my research and thesis on time. I am also grateful to Ministry of Science, Technology and Innovation (MOSTI) for granting me a scholarship through National Science Fellowship (NSF) for financial support. I am also indebted to Kulliyyah of Science members and administrative staff for their cooperation during my study. Appreciation also goes out to officers from Lembaga Kemajuan Ikan Malaysia (LKIM) and Persatuan Nelayan for their assistance in collecting fish samples for my study. I am also fortunate to have the officers and staffs of Institute Oceanography and Maritime Studies (INOS), Universiti Malaysia Terengganu (UMT) whom had assisted and cooperated with me even in odd hours in attending to my samples for detection in their respective labs. Special thanks also go to many people who helped me in sharing and solving my problems during my laboratory work. I am highly indebted to Joseph Bidai for training me on the use of Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and for his valuable help especially during method development when running the ICP-MS program. My most sincere gratitude also goes to Mohd Nahar and Wadi for their consideration, kindness and patience in supplying all the equipments and apparatus needed for my research analysis and training me on the use of various instruments. They were all friends whom were ever willing to share their knowledge and experience in research. To conclude, I gratefully thank all my colleagues (in no chronological order); Azfar, Zuhairi, Nadia, Shuhada and Siti for their assistance directly and indirectly with my project on whole. Great deals of work were much owed to their cooperation and support. Special dedication goes to my loving parents, Hj. Zabri and Hjh. Fatimah Rohana for putting up with my schedules and and be encouraging at any point of my study period. Your prayers and support had always been a pillar of strength for me. I would also love to convey my gratitude to my siblings; Shahrulniza and Roza Maria along with their families for their undying support for my studies all this while. Lastly, special thanks also expressed to Mohd Rozani whom had been very helpful and cooperative with me in completing my studies. Thank you very much.

viii TABLE OF CONTENTS Abstract ...... ii Abstract in Arabic ...... iii Approval Page ...... v Declaration ...... vi Copyright Page...... vii Acknowledgements ...... viii List of Tables ...... xii List of Figures ...... xiv List of Symbols/ Abbreviation ...... xvii

CHAPTER ONE: INTRODUCTION ...... 1 1.1 Background of the Study ...... 1 1.2 Objectives of Study ...... 4

CHAPTER TWO: LITERATURE REVIEW ...... 6 2.1 Fish Consumption in Malaysia ...... 6 2.2 Pollution in the Marine Environment ...... 8 2.3 Straits of Malacca ...... 11 2.4 Changes of Physico-chemical Parameters of Water due to Pollution ...... 14 2.4.1 Physical Parameters ...... 14 2.4.1.1 Temperature ...... 14 2.4.1.2 Turbidity and Colour ...... 15 2.4.1.3 Depth and Flow ...... 15 2.4.1.4 Light ...... 15 2.4.2 Chemical Parameters...... 15 2.4.2.1 pH ...... 15 2.4.2.2 Dissolved Oxygen ...... 15 2.4.2.3 CO2 ...... 16 2.4.2.4 Alkalinity ...... 16 2.4.2.5 Salinity ...... 17 2.4.3 Dissolved Solids ...... 17 2.4.3.1 Nitrates and Phosphates ...... 17 2.4.3.2 Heavy Metals ...... 17 2.5 Source of Heavy Metal Pollution ...... 18 2.5.1 Transport of Heavy Metals in the Environment...... 18 2.5.1.1 Atmospheric Transport ...... 20 2.5.1.2 Aquatic Transport ...... 20 2.5.1.3 Biological Transport ...... 20 2.5.2 Transformation of Heavy Metals in Aquatic Environment ...... 21 2.6 Bioaccumulation of Heavy Metals in Marine Species ...... 22 2.6.1 Bioaccumulation ...... 24 2.6.2 Bioconcentration ...... 24 2.6.3 Biomagnification ...... 25 2.7 Bioaccumulation of Heavy Metal in Fish Tissues ...... 26 2.8 Fish as a Biomonitor ...... 27 2.9 Metals ...... 29 2.9.1 Zinc (Zn) ...... 31 2.9.2 Lead (Pb) ...... 33

ix 2.9.3 Arsenic (As) ...... 36 2.9.4 Mercury (Hg) ...... 39

CHAPTER THREE: MATERIALS AND METHODS ...... 44 3.1 Study Area Description ...... 44 3.2 Sampling Sites ...... 46 3.2.1 Classification of Sampling Sites ...... 47 3.2.1.1 South Zone ...... 47 3.2.1.2 Middle Zone ...... 48 3.2.1.3 North Zone ...... 48 3.3 Fish Species Selected ...... 51 3.3.1 Japanese Threadfin Bream ...... 51 3.3.2 Dorab Wolf-herring ...... 52 3.3.3 Emperor Red Snapper ...... 53 3.3.4 Tiger-toothed Croaker ...... 54 3.3.5 White Pomfret ...... 55 3.4 Sampling Methodology ...... 56 3.4.1 Preparation before Sampling ...... 56 3.4.2 During Sampling ...... 56 3.5 Laboratory Analysis ...... 58 3.5.1 List of Chemicals used ...... 58 3.5.2 Instruments and Glasswares used ...... 59 3.5.3 Apparatus Preparation ...... 59 3.5.4 Sample Preparation ...... 60 3.5.4.1 Dissection Process ...... 60 3.5.4.2 Drying of Samples ...... 63 3.5.4.3 Determination of Constant Weight ...... 63 3.6. Analysis of Heavy Metals ...... 63 3.6.1 Open Digestion Method ...... 63 3.6.2 Closed Digestion Method ...... 64 3.6.2.1 Before Analysis ...... 64 3.6.2.2 During Analysis ...... 64 3.7 Detection with ICP-MS ...... 65 3.7.1 The Origin, Realization and Performance of ICP-MS System ...... 65 3.7.2 Standard Stock Solutions for ICP-MS ...... 69 3.7.3 Preparation of Calibration Standards ...... 69 3.7.4 Blank Solutions ...... 70 3.8 Quality Assurance ...... 71 3.9 Statistical Analysis ...... 71

CHAPTER FOUR: RESULTS ...... 73 4.1 General Findings ...... 73 4.2 Heavy Metals Study ...... 73 4.3 Accuracy and Precision ...... 74 4.3.1 Recovery Test ...... 74 4.4 Calibration Curve of all Studied Elements ...... 75 4.5 Heavy Metals Concentrations in Fish and Zones ...... 78 4.5.1 Zinc (Zn) ...... 78 4.5.1.1 Japanese Threadfin Bream ...... 78

x 4.5.1.2 Dorab Wolf-herring ...... 79 4.5.1.3 Emperor Red Snapper ...... 80 4.5.1.4 Tiger-toothed Croaker ...... 81 4.5.1.5 White Pomfret ...... 82 4.5.2 Lead (Pb) ...... 83 4.5.2.1 Japanese Threadfin Bream ...... 83 4.5.2.2 Dorab Wolf-herring ...... 84 4.5.2.3 Emperor Red Snapper ...... 85 4.5.2.4 Tiger-toothed Croaker ...... 86 4.5.2.5 White Pomfret ...... 87 4.5.3 Arsenic (As) ...... 88 4.5.3.1 Japanese Threadfin Bream ...... 88 4.5.3.2 Dorab Wolf-herring ...... 89 4.5.3.3 Emperor Red Snapper ...... 90 4.5.3.4 Tiger-toothed Croaker ...... 91 4.5.3.5 White Pomfret ...... 92 4.5.4 Mercury (Hg) ...... 93 4.5.4.1 Japanese Threadfin Bream ...... 93 4.5.4.2 Dorab Wolf-herring ...... 94 4.5.4.3 Emperor Red Snapper ...... 95 4.5.4.4 Tiger-toothed Croaker ...... 96 4.5.4.5 White Pomfret ...... 97

CHAPTER FIVE: DISCUSSION ...... 98 5.1 Heavy Metal Accumulation in Fish ...... 98 5.1.1 Comparison on Heavy Metal Level among Tissues ...... 98 5.1.1.1 Zinc (Zn) ...... 98 5.1.1.2 Lead (Pb) ...... 101 5.1.1.3 Arsenic (As) ...... 104 5.1.1.4 Mercury (Hg) ...... 107 5.1.2 Comparison of Heavy Metal Level among Species ...... 111 5.1.2.1 Zinc (Zn) ...... 111 5.1.2.2 Lead (Pb) ...... 113 5.1.2.3 Arsenic (As) ...... 115 5.1.2.4 Mercury (Hg) ...... 118 5.1.3 Comparison of Heavy Metal Level among Locations ...... 120 5.2 Correlation of Heavy Metal Accumulation ...... 124 5.2.1 With Fish Weight ...... 124 5.2.2 With Fish Length ...... 126 5.3 Fish as a Biomonitoring Agent ...... 129 5.4 Comparison of Permissible Limits ...... 131 5.5 Assessment of Exposure to Heavy Metals due to Consumption ...... 132

CONCLUSION ...... 136

BIBLIOGRAPHY ...... 138

APPENDICES ...... 165

xi LIST OF TABLES

Table No. Page No.

2.1 Total of marine fish landing according to states in 2006 7

2.2 The Interim Marine Water Quality Standards 9

2.3 Marine Water Quality 2004 10

2.4 Distribution of major sources of water pollution in Malaysia, 1991 11

2.5 Risk-based consumption limits based on methylmercury concentrations in fish tissue 43

3.1 Chemicals and reagents used 58

4.1 Recovery test results of analysis of the standard reference material, Dogfish muscle, (DORM-2) 74

4.2 Descriptions of fish species used for analysis 77

5.1 Comparison of Zn concentration in fish tissues studied and other documented studies 101

5.2 Comparison of Pb concentration in fish tissues studied and other documented studies 104

5.3 Comparison of As concentration in fish tissues studied and other documented studies 107

5.4 Comparison of Hg concentration in fish tissues studied and other documented studies 111

5.5 Comparison of Zn concentration in fish species studied and other documented studies 113

5.6 Comparison of Pb concentration in fish species studied and other documented studies 115

5.7 Comparison of As concentration in fish species studied and other documented studies 117

5.8 Comparison of Hg concentration in fish species studied and other documented studies 120

xii

5.9 Mean concentration of heavy metal distribution among location 121

5.10 Maximum permitted levels in fish enforced by Malaysia, Singapore and Australia 132

5.11 Estimated Weekly Intake (EWI) of Zn, Pb, As and Hg from studied fish 134

xiii LIST OF FIGURES

Figure No. Page No.

2.1 Marine Water Quality Status 2002-2004 9

2.2 General model describing the fate of heavy metals in living organisms 22

2.3 Diagram of heavy metal uptake in marine organisms 23

2.4 Movement of trace metals in hydrological cycle 31

3.1 3D surface and cross sectional plot of the Straits of Malacca 45

3.2 Map of sampling sites along the west coast of Peninsular Malaysia 50

3.21 Yellow-threadfin bream 51

3.22 Dorab Wolf-herring 52

3.23 Emperor Red Snapper 53

3.24 Tiger-toothed Croaker 54

3.25 White Pomfret 55

3.26 Local fishermen gathering their catch 57

3.27 Identifying fish species needed for analysis 57

3.28 Measurement of standard length and total length of the fish 61

3.29 Diagram of fish tissues selected for analysis 61

3.30 Fish samples; Disected and Dried in the oven 62

3.31 Inductively-coupled Plasma Spectrometry (ICP-MS) 68

3.32 The ICP Torch showing the fate of the sample 68

3.33 The interface region of an ICP-MS 68

3.24 Schematic of quadruple mass filter 69

4.1 Zn calibration curve 75

xiv 4.2 Pb calibration curve 75

4.3 As calibration curve 76

4.4 Hg calibration curve 76

4.5 Zn concentration in Japanese Threadfin Bream 78

4.6 Zn concentration in Dorab Wolf-herring 79

4.7 Zn concentration in Emperor Red Snapper 80

4.8 Zn concentration in Tiger-toothed Croaker 81

4.9 Zn concentration in White Pomfret 82

4.10 Pb concentration in Japanese Threadfin Bream 83

4.11 Pb concentration in Dorab Wolf-herring 84

4.12 Pb concentration in Emperor Red Snapper 85

4.13 Pb concentration in Tiger-toothed Croaker 86

4.14 Pb concentration in White Pomfret 87

4.15 As concentration in Japanese Threadfin Bream 88

4.16 As concentration in Dorab Wolf-herring 89

4.17 As concentration in Emperor Red Snapper 90

4.18 As concentration in Tiger-toothed Croaker 91

4.19 As concentration in White Pomfret 92

4.20 Hg concentration in Japanese Threadfin Bream 93

4.21 Hg concentration in Dorab Wolf-herring 94

4.22 Hg concentration in Emperor Red Snapper 95

4.23 Hg concentration in Tiger-toothed Croaker 96

4.24 Hg concentration in White Pomfret 97

5.1 Correlation of Zn concentration with fish weight 124

5.2 Correlation of Pb concentration with fish weight 124

xv

5.3 Correlation of As concentration with fish weight 125

5.4 Correlation of Hg concentration with fish weight 125

5.5 Correlation of Zn concentration with fish length 126

5.6 Correlation of Pb concentration with fish length 126

5.7 Correlation of As concentration with fish length 127

5.8 Correlation of Hg concentration with fish length 127

5.9 Formula for Estimation of Weekly Heavy Metal Intake 133

xvi

LIST OF SYMBOLS / ABBREVIATION

/ Per : Ratio ˚ Degree < Less than = Equal > Greater than ± Plus Minus C Celsius cm Centimeter dw Dry weight g Gram kg Kilogram L Liter mg Miligram mg/g Miligram per gram mg/kg Miligram per kilogram mg/L Miligram per liter ml Mililiter mm Milimeter mol Molar ppb Parts per billion ppm Parts per million wt. Weight μ Micro μg/g Microgram per gram μg/L Microgram per liter μm Micrometer

xvii CHAPTER ONE

INTRODUCTION

1.1 BACKGROUND OF THE STUDY

Metals are non-biodegradable and are considered as major environmental pollutants causing cytotoxic, mutagenic and carcinogenic effects in animals (More et al., 2003).

Aquatic organisms have the ability to accumulate heavy metals from various sources including sediments, soil erosion and runoff, air depositions of dust and aerosol, and discharges of waste water (Labonne et al., 2001; Goodwin et al., 2003). Therefore, accumulation of heavy metals in aquatic organisms can pose a long lasting effect on biogeochemical cycling in the ecosphere.

Fish are often at the top of aquatic food chain and may concentrate large amounts of some metals from the water (Mansour and Sidky, 2002). Metal bioaccumulation is largely attributed to differences in uptake and depuration period for various metals in different fish species (Tawari-Fufeyin and Ekaye, 2007).

Multiple factors including season, physical and chemical properties of water (Kargin,

1996) can play a significant role in metal accumulation in different fish tissues.

Fish is an important source of food for humans and is a key component in many natural food webs. Fish is also one of the sources of biologically valuable protein, fats and fat-soluble vitamins (Belitz and Grosch, 1987). The high quality protein of fish is better for health than that in meat and poultry. Fish consists of 15-

24% protein; 1-3% carbohydrate; 0.1-22% lipid; 0.8-2% inorganic substances and 66-

84% water (Suzuki, 1981). Each of these is important for human health, growth and intelligence. In human nutrition, fish plays an important role as it provides an

1 important source of trace minerals and calcium. Fish also provides calories, nutrients such as fat, vitamins (B complex and D), elements such as, phosphorus, sodium as well as trace elements (Mn, Mg, I, Zn, etc.) (Abdullah and Idrus, 1977).

Hadzley (1997) stated that distribution of fish in the sea is related to certain physical and chemical parameters of the water. In conjunction to that, since physical parameters in Malaysian waters have not changed much over the years, it may be assumed that the distribution of species has also not changed in the whole area, thus the availability and distribution of food resources and seabed conditions are the main factors that affected the distribution of fish and its behaviour (Mohsin et al., 1987,

1988).

Despite its nutritional value, fish is a commodity of potential health concern as it can be contaminated with a range of environmentally persistent chemicals. At the top of the aquatic food chain, fish is known to be able to accumulate large amounts of toxic contaminants from their living environment. One group of the contaminants accumulated by aquatic organisms is heavy metals such as mercury, arsenic, cadmium and lead. They are cumulative poisons which are not detoxified by metabolic activities.

The knowledge of levels and distribution of heavy metals, particularly within the body of a marine animal, is of interest in understanding the role they play in the biochemical and physiological mechanisms of the organisms. Heavy metal enters the marine environment naturally through the earth crust. Nowadays, heavy metal increased in concentration because of human activities which has led to a steady state background for a period of time. Accumulation of heavy metal in fish from different sources (atmospheric deposition, agricultural practices, urban-industrial activities,

2 etc.) is of a great environmental concern because of metal persistence and toxicity.

Furthermore, after the catastrophe in Minamata, Japan, caused by fish consumption containing methyl mercury, the study of the effects of heavy metals present in fresh fish as part of human diet is of particular interest.

In Malaysia, increased concentrations of metals have been observed in both marine and freshwater fish. Studies on heavy metal pollution in fish started early as reported by Babji et al., (1979) in West Malaysia. The importance of the study was justified for the fact that heavy metal concentration in the water column correlates positively with concentrations in fish tissue (Svobodova et al., 1996; Kamaruzzaman et al., 2007). Law and Singh (1988) also stated that monitoring heavy metal content in organisms can give meaningful information on the pollution status of a water body than merely monitoring the metals in water and sediments. The level of heavy metal bioaccumulation in fish tissues is influenced by biotic and abiotic factors, such as fish biological habitat, chemical form of metal in the water, water temperature and pH value, dissolved oxygen concentration, water transparency, as well as by fish age, gender, body mass, and physiologic conditions (Has-Schon et al., 2006).

Over the last 15 years, research on heavy metal content in fish tissues in the water column has been conducted in Malaysia (Din and Jamaliah, 1995; Ismail et al.,

1995; Shazili et al., 1995, Naransa et al., 2001; Tukimat et al., 2002, 2006; Agusa et al., 2005; Kamaruzzaman et al., 2007; Ikram et al., 2008; Irwandi and Farida, 2009;

Hajeb, et al., 2009 and Ahmad et al., 2009). Results of a few studies stated also showed the distribution trend of heavy metals in the fishes has increased eventually.

Ikram et al., (2008) had studied a Threadfin species from Straits of Malacca and concluded that metabolic activities of the matured and young fishes contributed for the

3 accumulation of heavy metals in its parts. Tukimat et al., (2002, 2006) on the other hand had conducted research of heavy metal concentration in local seafood from

Kemaman, Terengganu and Tanjung Karang, Selangor both representing South China

Sea and Straits of Malacca. They concluded that demersal fish species that consume mostly detritus, benthic algae and small fishes are most likely to accumulate more heavy metal in their body parts respectively. Irwandi and Farida (2009) whom studied mineral and heavy metal contents in marine fin fish from Pulau Langkawi, Kedah had concluded that heavy metal concentration in the fish muscles could be attributed to natural anthropogenic metal sources affecting their habitats

However, given the wide range of marine fish species available, not many researchers had investigated the heavy metal concentration for certain classes of fish such as pelagic or demersal respectively and include several heavy metals in their studies at the same time. To enhance the scarcity of data, a research on the accumulation of heavy metals in any commercial fish species in our country is urgently needed. Therefore, this study was designed to determine the concentrations of Zinc, Lead, Arsenic and Mercury in the muscle and gill tissues of five selected fish species from Straits of Malacca, Malaysia.

1.2 OBJECTIVES OF STUDY

In order to enhance information on the trace elements’ accumulation in fish from

Straits of Malacca, several objectives were listed as guidelines for this study:

1. To determine the concentrations of zinc, lead, arsenic and mercury in the

muscle and gill of five selected marine fish.

4 2. To determine the distribution of zinc, lead, arsenic and mercury accumulation

in the fishes from different zones along the straits.

3. To ascertain whether concentrations of zinc, lead, arsenic and mercury in the

analyzed fishes exceed the national and international standards for food and

human health.

5 CHAPTER TWO

LITERATURE REVIEW

2.1 FISH CONSUMPTION IN MALAYSIA

Malaysia is among the countries with highest fish consumption in the world. Average fish consumption has increased from 49 kg per capita in 2000 to 53 kg in 2005

(Ahmad et al., 2005). This figure is expected to increase to 56 kg in 2010. According to Fisheries Department figures, Malaysia produces about 1.5 million metric tonne

(mt) of fishery products annually of which about 85% are marine captured fish.

Fisheries resources in Malaysian waters can be divided into two types; coastal water fisheries and deep water fisheries. Coastal fisheries resources were caught in an area within 30 nautical miles while deep water fisheries resources catching area exceeds 30 nautical miles from the coastal shores (Annual Fisheries Statistics, 2006)

Most of the marine catches are pelagic fish with Indian mackerel, round scad, squid, tuna and bream being among the major species caught. Demersal species on the other hand, were reported to dominate in the catches of most of the fish resource surveys conducted off the Malaysian waters since 1970’s but the abundance was reported to decrease (Mansor et al., 1999). The decline in the abundance of demersal fish resources is always an issue in fishing industry (Hadzley, 1997). This declining trend is thought to prevail due to either overexploitation of the demersal resources using highly efficient harvesting gears or factors relating to availability of food in the area (Chang et al., 1975; Hadzley, 1997). Currently Malaysia imports about 30% of its total fish consumption requirements. Sardines, Indian mackerel and Pomfrets are

6 among the major imported species to date. Table 2.1 states the total marine fish landing according to states.

Table 2.1 Total of marine fish landing according to states in 2006 (Annual Fisheries Statistics, 2006)

Coastal Water Deep water Total States Quantity (mt) Perlis 109, 177 53, 861 163, 038 Kedah 63, 478 3, 644 67, 122 P.Pinang 31, 313 1, 799 33, 112 Perak 180, 228 28, 925 209, 153 Selangor 146, 388 0 146, 388 Negeri 374 0 374 Sembilan Melaka 1, 829 0 1, 829 Johor Barat 19, 026 0 19, 026 Pantai Barat 551, 813 88, 229 640, 042

As an important diet of Malaysians and most people of the Association of

South East Asean Nation (ASEAN) countries, fish provides approximately 49% of the total animal protein consumed and 12 % of the total protein intake in the Malaysian diet. In a human health perspective, an ASEAN per capita seafood consumption rate of 40 kg/person/year (i.e., 110g/person/day) was adopted for calculation of water quality criteria for protection of human health from contaminated seafood consumption. This is based on the agreement of the ASEAN Marine Water Quality

Criteria-Working Group (Chongprasith et al., 1999). The need to include seafood, particularly fish, in the human diet has been emphasized with regard to its lower levels of saturated fat, cholesterol, and caloric intake compared with meat, poultry, and dairy products (Velez and Montoro, 1998).

7