1. Analysis of Low Brominated DD/F Takeshi Nakano (Hyogo Pref. Inst. of Pub. Health and Env.Sci.) PCDD/DF/PCB in environment

2. ２, ３, ４, ７, ８－ PeCDF ３, ４, ５, ３ ’, ４ ’ － Co- PCB ２, ３, ７, ８－ TCDD １, ２, ３, ５, ６, ７－ HxCN ７５ congeners １３５ congeners ２０９ congeners

3. Trend of PCB Use in Japan Total 58,000t Amount of PCB use (t)

4. PCB product use in Japan Ar-1242 1248 1254 1260 1969-1971 KC-300 400 500 600

5. KC300 KC400 KC500 KC600 Japan Homologue pattern reflects product use

6. PCB Homologue Distribution and Source Estimation

7. Source estimation KC ３０ ０ KC ４０ ０ KC ５０ ０ KC ６０ ０ Sediment A Sediment B Ship paint N.C.P. Insulator capacitor

8. To evaluate environmental risk

9. Daily variation of PCDD/Fs and PCBs in ambient air

13. Daily variation of Dioxin Concentration

14. Time trend of PCDD/F levels in ambient air

15. Time trend of PCDDs/PCDFs （ TEQ ） Matsumura et al, symposium of J.Env.Chem, 2003.6 Dioxin Emission Reduction Plan

16. 0.00 0.10 0.20 0.30 0.40 0.50 0 10002000300040005000600070008000 Emission inventry (g-TEQ/year ） PCDDs/PCDFs （ pg-TEQ/m3) 1997 1998 1999 2000 2001 Emission inventry vs Level in air Matsumura et al, symposium of J.Env.Chem, 2003.6

17. Time trend of Co-PCBs （ TEQ ）

19. Time trend of PCB levels in ambient air (log scale)

20. 0.01 0.1 1 10 100 1000 19701975198019851990199520002005 Time trend of PCBs concentration in ambient air (pg/m 3 ) Time trend of PCBs concentration in ambient air (ng/m 3 ) (pg/m 3 ) (ng/m 3 ) Ambient air

21. Low volume air sampler

22. Time trend of PCB levels in breast milk

23. Time trend of breast milk levels Dioxins PCB pg TEQ / g-fat µg g-fat 60 40 20 0 Year (1973-2000) Konishi et al. (2003) Konishi et al (2003) Organohalogen compds pg TEQ/g-fat µg / g-fat Year (1973-2000) 60 40 20 0 0.3 0.2 0.1 0

24. Development of congener specific analysis for PCBs using HT8-PCB

25. Congener number number of isomerof peak mono 3 3 di 12 10 tri 24 23 tetra 42 39 penta 46 38 hexa 42 40 hepta 24 23 octa 12 12 nona 3 3 deca 1 1 total 209 192

26. Matsumura et al, J. Env. Chem. (2002) GC/MS-SIM chromatogram of TrCB (HT8-PCB) PCB product 13 C-TrCB

27. Matsumura et al, J. Env. Chem. (2002) GC/MS-SIM chromatogram of PeCB (HT8-PCB) 13 C-PeCB PCB product

28. Congener profiles of PCBs in environmental and human samples

29. Congener profiles of PCBs(Hexa-) in environmental and human samples

30. Congener profiles of PCBs(Hepta-) in environmental and human samples

31. Congener profiles of PCBs in human samples (pooled blood)

32. ＃ 28 ＃ 31 ＃ 74 ＃ 80 ＃ 99 ＃ 118 ＃ 153 ＃ 138 ＃ 163/ ＃ 164 ＃ 187 ＃ 180 ＃ 170 Tri-CBs Tetra-CBs Penta-CBs Hexa-CBs Hepta-CBs Chromatogram of PCB in human blood （ 3~7Cl ） Hirai et al, (2003) Organohalogen compounds #187 #180 #153 #138 #74 #99 #118 #28 245-2345 245-245 245-234 245-34 245-2356 245-4 245-24 24-4

33. (n=24) CongenersMean ± 2S.D. Existence Ratio* (BZ#)(ng /g-lipid)(%) #153 28.0 ± 22.3 #18014.9 ± 16.611.8 #13810.4 ± 9.28.3 #1878.3 ± 8.56.6 #1186.9 ± 5.95.5 #163,1646.4 ± 6.25.1 #994.7 ± 4.23.8 #744.6 ± 2.63.6 #1464.1 ± 3.73.3 #1703.8 ± 4.13.0 #1562.8 ± 3.02.3 Sum75.6 * Mean is compare to total PCBs level Levels of the predominant PCB congeners in human blood Hirai et al, (2003) Organohalogen compounds 245-2345 2345-234 245-245 245-234 245-4 245-24 245-34 245-2356 245-235 2345-34 22.3 34.1 42.4 49.0 54.5 59.6 63.4 67.0 70.3 73.3 75.6 cumulative (%)

34. human milk/adipose tissue #74 (2,4,4',5-) #99 (2,2’,4,4',5-) #118 (2,3’,4,4',5-) #153 (2,2',4,4',5,5'-) #138 (2,2',3,4,4',5'-) #187 (2,2',3,4’,5,5’,6-) PCB congeners predominant in human samples

35. Analysis of Low Brominated DD/F ANALYSIS of MBDD/MBDF to T 3 BDD/T 3 BDF on a SP2331 - COLUMN

36. Introduction In the last two decades the increased use of brominated organic compounds especially brominated flame retardants (BFR) resulted in the presence of brominated compounds indoor, in the waste stream and in the environment.

37. Introduction Some of the BFR, in particular the PeBDE, are associated with the presence and the formation potential of brominated dibenzo-p-dioxins and furans (PBDD/PBDF). Therefore the congener specific analysis of PBDD/PBDF received more attention in recent time.

38. Introduction In this respect also the analysis of low brominated DD/F homologues might be interesting in some processes and samples: Brominated aromatic compounds are subjected to debromination reaction under various conditions are dehalogenated more rapidly compared with chlorinated aromatic compounds.

39. Introduction Under UV or exposure to sunlight, brominated aromatic compounds can undergo rapid debromination. photo degradation Debromination reactions are also observed during thermal degradation of polybrominated aromatics. thermal degradation

40. Introduction Therefore it might be interesting to have selected samples analysed for low brominated DD/DF. For this study we synthesised a low brominated DD and DF mixture and optimised a temperature program on a SP2331 column.

41. Introduction We have been reported complete assignment of M 1 CDD/F to T 3 CDD/F on SP-2331 (Dioxin 2000) DB5MS (Dioxin 2001) DB-Dioxin (Dioxin 2002) In this study, as the first step of complete assignment of M 1 BDD/F-T 3 BDD/F (74congeners)

42. Standards. (synthesised) The PBDD standards were synthesized by pyrolysis of bromophenols in Pyrex ampoules. The PBDD was synthesized by condensation of 2-monobromophenol, 2,4- and 2,6- dibromophenol(DBP) 2,4,6-tribromophenol (TBP) at 350 º C (30 min). Materials and Methods

43. Standards PBDD by condensation 2-BP + 2,4- DBP 2-M 1 BDD 2-BP + 2,6- DBP 1-M 1 BDD 2,4-DBP + 2,4- DBP 2,7/2,8-D 2 BDD 2,4-DBP + 2,6- DBP 1,7/1,8-D 2 BDD 2,6-DBP + 2,6- DBP 1,6/1,9-D 2 BDD 2-BP + 2,4,6-TBP 1,3-D 2 BDD 2,4- DBP + 2,4,6-TBP 1,3,7/1,3,8-T 3 BDD 2,6- DBP + 2,4,6-TBP 1,3,6/1,3,9-T 3 BDD 2,4,6- TBP + 2,4,6-TBP 1,3,6,8/1,3,7,9-T 4 BDD

44. Standards. (synthesised) For the PBDF mixture all three 2-/3-/4- bromophenol isomers, phenol, 2,4-DBP were pyrolysed at 370 º C (15 minutes) in the presence of trace amounts of CuBr 2 Materials and Methods

45. Standards PBDF 4-BP + phenol 2-M 1 BDF 3-BP + phenol 1-/3-M 1 BDF 2-BP + phenol 4-M 1 BDF 2,4-DBP + phenol 2,4-D 2 BDF 4-BP + 4-BP 2,8-D 2 BDF 3-BP + 3-BP 1,7-/3,7-/1,9-D 2 BDF 2-BP + 2-BP 4,6-D 2 BDF ------------------------------------------------------- 2,4-DBP + 4-BP 2,4,8-T 3 BDF ------------------------------------------------------- 2,4-DBP + 2,4-DBP 2,4,6,8-T 4 BDF

46. Standards. (commercially available ) We obtained commercially available standards 2,7/2,8-D 2 BDD, 2,3,7-T 3 BDD 1,2,3,4-, 1,3,6,8-, 1,3,7,9-, 1,3,7,8-, 2,3,7,8-T 4 BDD 2,7-D 2 BDF, 2,8-D 2 BDF, 2,3,8-T 3 BDF Materials and Methods EBC-2500 8-Br-2,3-CDF, 8-Br-2,3,4-CDF, 7-Br-2,3-CDD, 2-Br-3,7,8-CDD, EDF-2046: 13 C-2,3,7,8-TeBrDF, 13 C-2,3,7,8-TeBrDD (Te ～ HxBrDD/DF)

47. GC/MS Analysis The analysis was carried out using HP 5890 II gas chromatograph connected to JMS-700 mass spectrometer (JEOL Ltd. Japan) operating at a resolution >10,000.

48. Temperature program Temperature program used for congener specific separation of the MBDD/F-T 3 BDD/F on SP2331 column (60m, 0.32mm i.d., 0.20um,): 120°C, 1 min. isothermal; 20°C/min. to 200°C, 2°C /min. to 260°C, 30min. isothermal. Carrier gas flow rate: He 1.2mL/min.

49. Table.1 Monitoring ion M 1 BrDD 261.9629263.9610 D 2 BrDD 339.8735341.8715 T 3 BrDD 419.7820421.7800 T 4 BrDD 497.6925499.6904 M 1 BrDF 245.9680247.9661 D 2 BrDF 323.8785325.8765 T 3 BrDF 403.7870405.7850 T 4 BrDF 481.6975483.6955

50. Results and Discussion The use of the 2,4-DBP (resulting in 2- and 3- (7- and 8-) PBDD substitution) and 2,6-DBP (resulting in 1- and 4- (6- and 9-) PBDD substitution) produced a wide range of congeners within the D 2 BDD.

51. 138 139 237 129 136 137 2 1 13 19 27/28 M 1 BrDD D 2 BrDD T 3 BrDD GC/MS-SIM chromatogram of M 1 BrDD ～ T 3 BrDD SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min.

52. Reliable assignment (blue color) Confirmation by authentic standards Progressing assignment （ red color ） Estimation from -formation ratio (major/minor) -regularity of elution order -comparison of RRT for PXDD/F (Cl and Br)

53. Brominated DD Mixture (Synthesis) Major component DiBrDD 13-, 27/28-, 16-/19-, 17/18- TrBrDD 137-, 138-, 136-, 139- TeBrDD 1368-, 1379-

54. 2-MBP 2,6-DBP 2,4-DBP 2,4,6-TBP 1,3-DBDD 2,8-DBDD 2,7-DBDD 1,3,6-D 1,3,7-D 1,3,8-D 1,3,9-D

55. 138 139 237 129 136 137 2 1 13 19 12 27/28/23 18 16 14/17 M 1 BrDD D 2 BrDD T 3 BrDD GC/MS-SIM chromatogram of M 1 BrDD ～ T 3 BrDD SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min.

56. Results and Discussion The use of all three monobromophenols produced a wide range of congeners within the D 2 BDF. Due to the higher pyrolysis temperature and the addition of CuBr 2 the isomerisation/bromination reactions had a significant impact and hence a broad range of T 3 BDF were formed.

57. 46 13 27 28 346 238 137 13 2 4 M 1 BrDF D 2 BrDF T 3 BrDF GC/MS-SIM chromatogram of M 1 BrDF ～ T 3 BrDF SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min.

58. Reliable assignment (blue color) Confirmation by authentic standards Progressing assignment （ red color ） Estimation from -formation ratio (major/minor) -regularity of elution order -comparison of RRT for PXDD/F (Cl and Br)

59. Brominated DF Mixture (Synthesis) Major component M 1 BDF 2-, 1-, 3-, 4- (4:2:2:1) D 2 BDF 24-, 28-, 18-, 27-, 17-, 19-, 26-, 37- 16-, 26-, 46- T 3 BDF 248-, 247-, 246-, 249-(168-) T 4 BDF 2468-

60. 2,8-DBDF 2-MBDF 1-MBDF3-MBDF 4-MBDF 2,4-DBDF 2,4-DBP 2-MBP 3-MBP 4-MBP

61. 46 17 13 18 16 37 24 27 36 28 26 247 248 246 346 238 138/136 137 168 13 2 4 M 1 BrDF D 2 BrDF T 3 BrDF 347/236 237/149 GC/MS-SIM chromatogram of M 1 BrDF ～ T 3 BrDF SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min.

62. PCDF PBDF MCDF DCDF TCDF MBDF DBDF TBDF

63. On the high polar SP2331-column, all 4 M 1 BDF were separated and also for D 2 BDF and T 3 BDF a reasonable selectivity was achieved. However on the less polar DB5 column even the 4 brominated M 1 BDF were not completely separated while the 4 chlorinated M 1 CDF isomers could be separated also on this low polar column. This demonstrates that the selectivity for brominated congeners is smaller compared with the chlorinated congeners.

64. Separation of low chlorinated DD/F (Dioxin 2000, 2001, 2002) congener ULTRA-2 DB-5MS SP-2331 DB-DIOXIN MCDF 4 4 4 4 4 D2CDF16 8 10 15 16 T3CDF28 8 10 23 21 congener ULTRA-2 DB-5MS SP-2331 DB-DIOXIN MCDD 2 2 2 2 2 D2CDD10 5 4 8 9 T3CDD14 6 5 12 12

65. Separation of low brominated DD/F on BPX35 & SP-2331 congener BPX35 SP-2331 MBDF 4 4 4 D2BDF16 8 13 T3BDF28 16 15 congener BPX35 SP-2331 MBDD 2 2 2 D2BDD10 5 7 T3BDD14 6 9

66. Comparison of Separation for PXDD/F on SP-2331 (Cl/ Br) congenerClBr MXDF 4 4 4 D2XDF161513 T3XDF282315 congenerClBr MXDD 2 2 2 D2XDD10 8 7 T3XDD1412 9 the selectivity for brominated congeners is smaller compared with the chlorinated congeners.

67. Comparison of Separation D 2 CDF and D 2 BDF 1,9-D 2 CDF 1,9-D 2 BDF

68. Low Brominated DD/F in Stack Gas Samples

69. 162024 Retention Time (min) 0 649323 162024 Retention Time (min) 0 568627 D 2 BrDF STD mix Stack Gas 27 28 #118 #105 #114 28 27 GC/MS-SIM chromatogram(D 2 BrDF) SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min. PeCB(323.8834,325.8805) DiBrDF(323.8785, 325.8765) #126

70. 14161820 Retention Time (min) 0 171522 14161820 Retention Time (min) 0 86858 BrClDF Sample-B Sample-A GC/MS-SIM chromatogram(BrClDF) SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min. BrClDF ＞ BrCl 2 DF, BrClDD ＞ BrCl 2 DD

71. Thank you for your attention

72. Low Chlorinated DD/F in Environmental Samples

73. 環境大気 Ambient air

74. 環境大気の異性体分布 (MCDF ～ TrCDF) Ambient air Incineration (flyash) Isomer distribution of ambient air(MCDF-TrCDF)

75. 環境大気の異性体分布 (MCDD ～ TrCDD) Isomer distribution of ambient air(MCDD-TrCDD) Incineration (flyash) Ambient air

76. 塩素漂白パターンの異性体分布 (MCDF ～ TrCDF) 28- 23- 27- 37- 2- 238- 128- 237- 1278-TCDF 2378-TCDF 238-TrCDF 237-TrCDF 128-TrCDF 37-DiCDF 27-DiCDF 23-DiCDF 28-DiCDF Isomer distribution of bleaching process(MCDF-TrCDF)

77. 塩素漂白パターンの異性体分布 (MCDF ～ TrCDF) 環境大気の異性体分布 (MCDF ～ TrCDF) 28- 27- 238- 128- 237- 2- 124- 24- Isomer distribution of ambient air(MCDF-TrCDF) Isomer distribution of bleaching process(MCDF-TrCDF) Ambient air

78. 農薬 CNP 中の異性体分布 (MCDF ～ TrCDF) 環境大気の異性体分布 (MCDF ～ TrCDF) 248- 246- 124- 24- Isomer distribution of ambient air(MCDF-TrCDF) Isomer distribution of pesticide CNP (MCDF-TrCDF) pesticide CNP Ambient air

79. 4-MCP 3,4-DCP 2,3,8-TrCDF 1,2,8-TrCDF

80. Synthesis of 1,4,X-T 3 CDF isomers from chlorophenol Authentic standard (1,4,7- : NMR) 2,5-DCP 4-MCP 3-MCP 2-MCP O-O- m-m- p-p- 1,4,6-TrCDF 1,4,8-TrCDF 1,4,9-TrCDF 1,4,7-TrCDF

81. NMR 1,4,7-TrCDF Authentic standard H9H9 H9H9 H6H6 H6H6 H8H8 H8H8 H 2 ; H 3

82. PCB levels and Homologue profiles in Environmental samples

83. PCB levels in air, water, sediment and biota (2002, MOE Japan) Air Sediment Water Biota 0.1-10ng/m 3 0.1-10ng/L 0.05-800ng/g-dry 0.1-800ng/g-wet

84. Homologue profiles of PCBs in human samples

85. Air Water Bird 0 5 10 15 20 25 30 35 40 45 50 abundance(%) Human blood Breast milk

86. Congener profiles of TrCB,TeCB in human and combustion sample (A)

87. PCB congeners predominant in human samples (TrCB, TeCB) PCB product Combustion Breast milk Combustion PCB product Breast milk

88. Congener profiles of TrCB,TeCB in combustion sample (B)

89. (stack gas) TrCB TeCB #68 #51 #47 24-26 24-24 24-35

90. Congener profiles of PCB in human sample

91. 12 DL-PCB(TEF)+di-ortho-PCB Predominant PCB in product Contribution of DL-PCBs in breast milk

92. Congener profiles of PCB in food sample

93. PCB congener profiles in food sample (Konishi et al ) 2001 1982 ng/day

94. Congener profiles of PCB in human sample

95. 1973 2000 PCB congener composition in human breast milk #187 #180 #153 #138 #74 #99 #118 #187 #180 #153 #138 #74 #99 #118

96. Salmon Breast milk PCB congener composition in human breast milk PCB congener composition in salmon #187 #180 #153#138 #66 #101 #118 #187 #180 #153 #138 #74 #99 #118

97. PCB product human #74 #66 245-4 24-34 #99 #101 245-24 245-25 #146 #149 245-235 245-236

98. Isomer distribution (P 7 CB, Port Island in June,2001) KC300-600 Sea Water Mussel Intensity #174 #180 #170

99. Isomer distribution (O 8 CB, Port Island in June, 2001) KC300-600 Sea Water Mussel Intensity #202#201 #196 #203#194#195 #199