JAMSHORO Ph.D Thesis Persistent, Bio-accumulative and Toxic Contaminants in Coastal Marine Environment of Pakistan Nuzhat Khan Submitted to University of Sindh Jamshoro towards fulfillment of the requirement to award of the degree of Doctor of Philosophy in Analytical Chemistry National Centre of Excellence in Analytical Chemistry University of Sindh, Jamshoro, Pakistan 2012 Persistent, Bio-accumulative and Toxic Contaminants in Coastal Marine Environment of Pakistan A thesis submitted in fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY IN ANALYTICAL CHEMISTRY By Nuzhat Khan National Centre of Excellence in Analytical Chemistry University of Sindh, Jamshoro, Pakistan 2012 In the name of Allah Subhanutallah He is Allah, other than whom there is no deity, the Sovereign, the Pure, the Perfection, the Bestower of Faith, the Overseer, the Exalted in Might, the Compeller, the Superior. Exalted is Allah above whatever they associate with Him. sūrat l-ḥashr 59 (59:23). My humblest thanks to Allah, He who is my acquaintance and my greatest strength. Thanks for the courage and determination to pursue my dreams. DEDICATION This work is dedicated to My mother A pillar of strength, my best friend, my buddy, my inspiration, my protector, my elation, wind beneath my wings without her in it, my life would be zero. My mother remains the love of my life. She is always there for me even before I am there myself, When she is around I am never alone. I thank her for believing in me and always making me see good, even at the most unpleasant times, and helping me make my weakness my greatest strengths. She always encouraged me to peruse my dreams. To my mother I owe the person I am. Thanks for not just believing, but knowing that I could do this! Love You Maa ACKNOWLEDGEMENTS This PhD was researched while I was enrolled as a candidate, as well as employed in a full-time as a Senior Research Officer at National Institute of Oceanography, Karachi. It included many hours of sea-time collecting data and samples over Pakistan coastal area. The intensive field work and samples pre-treatment was carried out with the financial assistance provided by Pesticide Action Network of N. America (PANNA) and WWF-Pakistan Grant WWF-50033101 is greatly appreciated. The opportunity to carry out my research work at National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia would have not been possible without the fellowship, granted by Partnership for Observation of the Global Oceans POPGO, Intergovernmental Oceanographic Commission (IOC) and the Scientific Committee on Oceanic Research (SCOR). Special appreciation to Prof. Dr M. Iqbal Bhanger, Prof. Dr. Shahid Amjad and Dr. Jochen Müller, it would not have been possible to complete this doctoral thesis without your help and support. Prof Dr.Des Connell, Chairman, School of Public Health, Griffith University to accept me as visiting researcher at National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia, where I have conducted most part of my PhD analytical work. Olaf Päpke facilitate me to obtain very first report on dioxins from Pakistan and to the ERGO laboratories, where these analysis were performed. Mary Hodge provided access to the laboratory at Queensland Health Scientific Services, where I used methodology and techniques, which allowed the analysis of OCPs and PAHs samples that would have been otherwise possible. My sincere thank to Director General, National Institute of Oceanography, Pakistan for the continued institutional support and encouragement. Thanks to Prof. Dr. Shafi M. Nizamani, University of Sindh for his editorial advice and support. I would like to acknowledge my team members Miss Saira Ishaq (RO) for providing me assistance in identification and biometric measurements and for all the lab and field work. Mr. Danish (RO) for the geological study, I would also like to thank NIO supporting Field and laboratory staff. Page i ACKNOWLEDGEMENTS A very sincere and extra special thanks to my dear friend Ms Tanya, EnTOX, Australia, I thank her also for the support, company and positive attitude always - a truly sincere person. I am so lucky for her friendship. I always remember my stay at Brisbane just because of you Tanya. My thanks are extended to the many people who helped along the way from National Research Centre for Environmental Toxicology (EnTox), Brisbane Australia and Centre of Excellence in Analytical Chemistry University of Sindh Jamshoro. Finally my very especial thank to my lovely family, my Sisters, my Neisse, my Nephews and my Khala for their love and support. They are the one who always there for me. Page ii ABSTRACT The present work is the first of its kind to describe in detail the fate and distribution pattern of Persistence Bio-accumulative and Toxic (PBTs) in the coastal area of Pakistan bordering North Arabian Sea. The PBTs included OCPs, PAHs and Dioxins and Dioxin like PCBs. Despite their widespread use and injurious effect of PBTs, little information is available on contamination levels of these pollutants in the coastal marine environment of Pakistan. Mangrove swamps, intertidal mudflats and Indus Deltaic creek represent main feature of the coastal marine environment. The coastal areas are of significance as there are spawning, breeding and nursery grounds of commercially important fishery resources. These living resources are under continuous threat of untreated industrial effluents and sewages discharges into coastal areas via several sources. To evaluate the contamination levels of PBTs, 61 sampling sites were selected along coastal Pakistan, 09 sites were identified along the Balochistan coast, while 47 sites were identified along the coastal Sindh, and 5 sites were located in the Indus River. In the current study, the analytical method routinely used in Environmental Toxicological Lab and Queensland Health Lab was adopted for identification quantification of organochlorine Pesticides, PCBs and PAHs. Analysis was carried out by using Varian 3400 Gas Chromatograph equipped with a Finnigan A 200S auto sampler and Finnigan SSQ710 Single Stage Quadrapole Mass Spectrometer. Sediments were sent to the ERGO Laboratory in Hamburg for the determination of 2,3,7,8-substituted PCDD/PCDF and dioxin-like PCB. Identification of PCDD/Fs was carried out by using retention times of the 13C-labelled standard and isotope ratios. The results on the contamination levels of PBTs revealed that residual concentration of Organochlorine Pesticides were considerably higher (17.5 ng g-1 dry wt.) in semi-enclosed area (Creeks and Karachi Harbour) in the effluent discharge areas, this was attributed to low tidal flushing. However, in the areas where port and harbour Page iii AN ABSTRACT OF THE THESIS activities and untreated effluents from industrial and domestic sources are discharged, are reflective of highest PAHs contamination levels (2610.812 ng g-1 dry wt.) in the vicinity of Kemari jetty, Karachi Harbour area. Dioxin and Dioxin-like chemicals were detectable in all samples collected from Pakistan coastal environment. However, OCPs contamination levels in the sediment collected from Balochistan coastal environment were found below detection limit at almost all the sampling sites. Spatial distribution pattern were significantly different among the localities sampled (p<0.004). Distribution pattern of most of the PBTs were well correlated with total sediment organic carbon contents (p>0.767 & R2=0.66). A larger variation of ∑OCPs contamination levels (>0.002-17.5 ng g-1 dry wt. with a mean concentration of > 4.5 ng g-1 dry wt.) was observed in the samples collected from various localities of coastal area of Pakistan. The DDT’s metabolites DDE were found in soft tissue of the marine biota (fishes, crab, shrimps and molluscs) collected from Coastal area. The residue of DDT mainly its metabolites DDE and DDD were detected in most of the samples in relatively higher concentrations, compared with the concentration obtained for other OCPs. The high proportion of pp′-DDE at most of the site (41–95%) and ratio of ∑DDT and DDT (0.04 –0.24) suggests old inputs of DDTs in the environment, it is restriction have been made on the use of these chemicals. Pakistan has also switched over to natural pest control or using safer formulas. The concentrations of ∑16PAHs varied amongst the localities, highest concentrations (2610.81 ng g-1 dry wt.) were detected in sediment samples collected in vicinity of Karachi harbour. Relatively lower levels (>400ng/g) observed in Korangi creek area south-east of Karachi. Higher concentrations of higher molecular weight PAHs such as (Benz(b+k) Fluoranthenen (>442.5 ng g-1 dry wt.), Ind(123cd)pyr (>270 ng g-1 dry wt.) and B(ghi) pryl (242.1 ng g-1 dry wt.) were detected near the discharge points of Lyari and Malir River. The Phen/Anth and Flth/Pyr concentration ratios indicated that mixture of pyrolytic and petrogenic PAHs sources at most of the site along the coast. First time water borne PAHs contaminations were estimated using Triolein- passive sampler Semi-Permeable Membrane Devices (SPMDs) as an alternative Page iv AN ABSTRACT OF THE THESIS monitoring tool for coastal waters of Pakistan. The estimated water concentration was 2 found to be highest in the harbour area (CwSPMDs 4.6ng/l) that is well correlated (R =0.5) with the evaluated contamination levels (CwSed. 35.67ng/l) using the levels observed in the sediment. According to the sediment quality standards of the USEPA and Canadian Council of Ministers of the Environment, observed levels of OCPs, PAHs and Dioxin contamination levels were generally lower than the threshold known to harm wildlife by OCPs. However, PAHs levels demonstrate moderate to low risk. Overall PCDD contributed to about 50 % of the TEQ in the samples with concentrations above 2 pg TEQ g-1 dwt and TCDD together with 1,2,3,7,8-PeCDD and 3,3',4,4',5-Penta-CB were the key contributors to the TEQ.