Characterization of Dioxin-like and Estrogenic Compounds in Environmental Samples from Onsan Bay and its Inland Region, Korea JS Khim 1,2 , KT Lee 1 , DL Villeneuve 2 , K Kannan 2 , JP Giesy 2 , CH Koh 1 1 Seoul National University, Seoul, Korea 2 Michigan State University, East Lansing, MI, USA
Industrial Complexes in Onsan Bay Area
Petrochemical Sediment sampling Korea I.C. locations in Onsan W1 W0 Onsan Bay Yongyeon Woihang River Bay Study Area Heavy I.C. W2 ON1 10 m Samples were collected Onsan Bay ON2 Onsan City ON3 from 22 locations O5 O4 in April 1999 O6 OS3 O8 O7 O10 OS1 OS2 O9 Daejeong Onsan Nonferrous Metal I.C. Stream D1 D0 D2 D3 D4 D5 D6 0 1 km I.C. : Industrial Complex
Objectives � To quantify chemicals of PCBs, OC pesticides, PAHs, Alkylphenols in Onsan Bay sediments � To examine cell line responses of dioxin-like and estrogenic compounds associated with sediment samples � To identify chemicals showing cell line responses
Methods � Sediment extracts were obtained from Soxhlet extraction and Florisil fractionation (3 fractions) � Cell line bioassay were performed using H4IIE-luc cells for dioxin-like activities and MVLN cells for estrogenic activities � Mass balance analyses were applied to evaluate chemicals showing cell line activities
Extraction & Fractionation Sequence Frozen Whole Freeze Drying Sediment Homogenization Soxhlet Extraction Cu treatment Raw Extract Bioassay Florisil fractionation Non-polar Mid-polar Polar (Hexane) (20% DCM in Hexane) (50% DCM in MeOH) PCBs, PAHs, DDD, DDT, NP, OP, HCB, DDE HCHs, CHLs BPA GC/MS & GC/ECD GC/MS & GC/ECD HPLC & GC/MS Bioassay Bioassay Bioassay
In vitro Bioassay Procedure H4IIE-luc or MVLN cells • cells were trypsinized • plated at 75 K cells/well (250 μ l/well) • dosing test samples (2.5 μ l) • After 72 h. exposure, culture media aspirated • cells were rinsed with PBS • luclite reagent added into the wells • after 10 minutes, luciferase activity was determined with luminometer TCDD SC Test E2 blank Sample
Results and Discussions Concentration Levels in Sediments 1. Concentrations of PCBs, OC pesticides, PAHs, Alkyl phenols in sediments 2. Comparison of concentrations with other Korean sediments 3. Potential biological effects of sediment concentrations Cell Bioassay Activities 4. Dioxin-like and estrogenic activity in vitro
1. Concentrations of PCBs, OC pesticides, PAHs, Alkylphenols in sediment � Concentrations of PCBs in sediments were generally less than method detection limit of 10 ng/g, dry wt � Among different OC pesticides analyzed, DDT concentrations (sum of p,p’ -DDD, DDE, DDT) were the greatest, ranging from <0.01 to 7.6 ng/g, dry wt � PAHs were detected in all the sediment samples. Highest concentrations of 300 to 1000 ng/g, dry wt. were found in 5 sampling locations in landward region. Outer bay region showed less than 30 ng/g, dry wt of concentration � High concentrations of NP, OP, and BPA were found only at Daejeong stream with mean concentration of 218 ng/g, dry wt. (sum of NP, OP, BPA). Other locations showed very low concentrations of 1.0 to 10 ng/g, dry wt
Unit 325 ng/g dry wt Petrochemical I.C. 50 ng/g dry wt Mean Concentrations of W1 W0 Woihang River (W) PCBs: W2 OC pesticides: ON1 10 m Onsan North Part (ON) ON2 PAHs: ON3 Onsan Bay (O) O5 O4 Alkyl phenols: O6 OS3 O8 O7 O10 OS1 OS2 O9 Onsan South Part (OS) Onsan Nonferrous Metal I.C. D1 D0 D2 218 ng/g dry wt D3 D4 D5 D6 PCBs Pesticides PAHs Alkylphenols Daejeong Stream(D)
2. Comparison of concentrations with other Korean sediments � Sedimentary PCBs and PAHs in Korea have been reported to range from a few ng/g to several µg/g � Concentrations of PCBs and PAHs in Onsan Bay were approximately 2 to 4- fold less than those reported in other industrialized coastal areas such as Kyeonggi, Masan, Ulsan, and Yeongil Bays � Concentrations of AP and BPA in sediment from Onsan Bay were similar to those in Ulsan Bay but far less than those in other bay areas
3. Potential biological effects of sediment concentrations � Concentrations of total TCDD equivalents (TEQs) ranged from 0.003 - 8.86 pg/g, dry wt � The TEQs estimated for selected PAHs and PCBs in Onsan sediment were at the lower end of the sediment quality guideline (SQG) of 0.014-210 pg/g, dry wt, reported for dioxin equivalents � None of the locations exceeded the Effect Range Lows (ERLs) for 16 PAHs and PCBs � When Threshold Effect Concentration (TEC) for total PCBs were compared, only one location (D4) exceeded the limit of 35 ng/g, dry wt
4. Dioxin-like and estrogenic activity in vitro � Screening of raw extracts (whole sediment extract) for dioxin-like activity with the H4IIE-luc cell bioassay revealed 17 of 22 samples had significant dioxin-like activity � Further fractionation of sediment extracts indicated that mid polar and more polar fractions were responsible for the significant dioxin-like activity � Estrogenic activities were low in the most samples. Only one raw extract sample (D3) yielded a significant estrogenic activity in the MVLN bioassay � The low estrogenic activity was consistent with low concentrations of measured estrogenic compounds in the Onsan sediments
H4IIE-luc cell line activity: Outer Bay Region 100 Outer bay region RE (O: Onsan Bay) 80 F1 %-TCDD-max F2 60 F3 40 20 0 ON1 OS1 O4 O5 O6 O10 -20 Response magnitude presented as percentage of the maximum response observed for a 2000 pM 2,3,7,8 tetrachlorodibenzo- p -dioxin standard (%- TCDD-max.) indicates cells exhibited an altered or “stressed” morphology
H4IIE-luc cell line activity: Landward Area 100 Landward Area (W: Woihang River 80 %-TCDD-max D: Daejeong stream) 60 40 20 0 W0 W1 W2 D0 D1 D2 D3 D4 D6 -20 Response magnitude presented as percentage of the maximum response observed for a 2000 pM 2,3,7,8 tetrachlorodibenzo- p -dioxin standard (%- TCDD-max.) indicates cells exhibited an altered or “stressed” morphology
Conclusion � The relative abundance of organic residues in sediment was in the order of PAHs ≈ APs > PCBs > OC pesticides � Concentration levels of organic compounds were high in landward region and those in outer bay were relatively low � Mid polar (PAHs) and more polar fractions of sediment extracts were responsible for the significant dioxin-like activity � Estrogenic activities were low in the most sediment
Acknowledgement • National Institute of Environmental Research (NIER), Ministry of Environment, Korea (Sediment Organic Compounds Bioassay Study; SORGBIOS ’98-2000) • Dr. MD Pons, INSERM Montpiellier France • Colleagues from Benthos Lab at Seoul National University, Korea and Aquatic Toxicology Laboratory at Michigan State University, USA
Recommend
More recommend
