1. ANALYSIS OF LOW BROMINATED DD/F ANALYSIS OF MBDD/MBDF TO T 3 BDD/T 3 BDF ON A SP2331 - COLUMN Takeshi Nakano, Chisato Matsumura, Roland Weber 1 Hyogo Prefectural Inst. of Pub. Health and Env. Sci. 1 Ulmenstrasse 3, 73035 Goeppingen, Germany.

2. 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.

3. 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.

4. 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 subjected to debromination reaction under various conditions are dehalogenated more rapidly compared with chlorinated aromatic compounds. are dehalogenated more rapidly compared with chlorinated aromatic compounds.

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

6. 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.

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

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

9. 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

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

11. 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

12. Standards. (commercially available ) We obtained commercially available standards 2,7/2,8-D 2 BDD, 2,7/2,8-D 2 BDD, 2,3,7-T 3 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 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, 2,7-D 2 BDF, 2,8-D 2 BDF, 2,8-D 2 BDF, 2,3,8-T 3 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)

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

14. 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,): on SP2331 column (60m, 0.32mm i.d., 0.20um,): 120°C, 1 min. isothermal; 120°C, 1 min. isothermal; 20°C/min. to 200°C, 20°C/min. to 200°C, 2°C /min. to 260°C, 30min. isothermal. 2°C /min. to 260°C, 30min. isothermal. Carrier gas flow rate: He 1.2mL/min.

15. 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

16. 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.

17. 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.

18. 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)

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

20. 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

21. 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.

22. 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. the isomerisation/bromination reactions had a significant impact and hence a broad range of T 3 BDF were formed.

23. 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.

24. 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)

25. Brominated DF Mixture (Symthesised ) 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-

26. 2,8-DBDF 2-MBDF 1-MBDF3-MBDF 4-MBDF 2,4-DBDF 2,4-DBP 2-MBP 3-MBP 4-MBP

27. 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.

28. PCDF PBDF MCDF DCDF TCDF MBDF DBDF TBDF

29. 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.

30. 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

31. 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

32. 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.

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

34. Low Brominated DD/F in Stack Gas Samples

35. 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

36. 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

37. Thank you for your attention

38. Low Chlorinated DD/F in Environmental Samples

39. 環境大気 Ambient air

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

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

42. 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)

43. 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

44. 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

45. 4-MCP 3,4-DCP 2,3,8-TrCDF 1,2,8-TrCDF

46. 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

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

48. ● BPX-35 (35%phenylpolysilphenylene-siloxane) 120 ℃ (2 min) – 20 ℃ /min-180 ℃ – 2 ℃ /min- 220 ℃ – 10 ℃ /min- 300 ℃ (10min). OSi C H 3 6 CN OSi C H 3 6 CN C H 3 6 yx ● SP-2331 (95%cyanopropyl 5%phenyl-siloxane) 120 ℃ (1 min.), 20 ℃ /min. to 200 ℃, 2 ℃ /min. to 260 ℃ (30min)

49. Brominated DF Mixture (Symthesised ) 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-

50. 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.

51. 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)

52. 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.

53. BPX-35 ; 30m x 0.32mm id. 0.25um ; 120°C (2 min.), 20°C/min. to 180°C, 2°C /min. to 220°C, 10°C /min. to 300°C (10min).; He 1.0mL/min. 1011 Retention Time (min) 0 2726152 Intensity M1BDF / Average 131415 Retention Time (min) 503 14887 Intensity BrClDF / Average 16171819 Retention Time (min) 0 792528 Intensity D2BDF / Average 13 2 4 13 46 24 27 28 26 17 18

54. BPX-35 ; 30m x 0.32mm id. 0.25um ; 120°C (2 min.), 20°C/min. to 180°C, 2°C /min. to 220°C, 10°C /min. to 300°C (10min).; He 1.0mL/min. 26272829 Retention Time (min) 0 312320 Intensity T3BDF / Average 313233 Retention Time (min) 0 87023 Intensity T4BDF / Average

55. Brominated DD Mixture (Symthesised ) Major component DiBrDD 13-, 27/28-, TrBrDD 137-, 138-, 136-, 139- TeBrDD 1368-, 1379-

56. 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.

57. BPX-35 ; 30m x 0.32mm id. 0.25um ; 120°C (2 min.), 20°C/min. to 180°C, 2°C /min. to 220°C, 10°C /min. to 300°C (10min).; He 1.0mL/min. 10.410.8 Retention Time (min) 0 4457267 Intensity M1BDD / Average 141516 Retention Time (min) 524 9893 Intensity BrClDD / Average 171819 Retention Time (min) 0 2389477 Intensity D2BDD / Average 2 1 13 27/28

58. BPX-35 ; 30m x 0.32mm id. 0.25um ; 120°C (2 min.), 20°C/min. to 180°C, 2°C /min. to 220°C, 10°C /min. to 300°C (10min).; He 1.0mL/min. 272829 Retention Time (min) 0 1307231 Intensity T3BDD / Average 31.031.532.032.5 Retention Time (min) 0 191064 Intensity T4BDD / Average 1368 1379

59. 分析方法 使用した機器は JEOL JMS ‐ 700 質量分析計、 HRGC/HRMS-SIM 分析条件は、ｲｵﾝ源温度： 280 ℃. ｲｵﾝ 化ｴﾈﾙｷﾞｰ： 38eV 標品 EBC-2500 (3-Br-2,7,8-CDF), 8-Br-2,3-CDF, 8-Br- 2,3,4-CDF, 7-Br-2,3-CDD, 2-Br-3,7,8-CDD, EDF-2046 : 13C-2,3,7,8-TeBrDF, 13C-2,3,7,8- TeBrDD (Te ～ HxBrDD/DF) 。 ロックマスは、 PFP(PerFluoroPhenanthrene) の 454.9729 を用いた。

60. Monitoring Ion PB DD/DF MBrDF: 245.9680, 247.9661, MBrDD: 261.9629, 263.9610, DiBrDF: 323.8785, 325.8765, DiBrDD: 339.8735, 341.8715, TrBrDF : 403.7870, 405.7850, TrBrDD: 419.7820, 421.7800, TeBrDF ： 481.6975, 483.6955, TeBrDD ： 497.6925, 499.6904, 13 C-TeBrDD ： 509.7327, 511.7306, 13 C-TeBrDF ： 493.7378, 495.7357 PX DD/DF BrClDF ： 279.9291, 281.9269, BrClDD ： 295.9240, 297.9218, BrCl 2 DF ： 313.8901, 315.8877, BrCl 2 DD ： 329.8850, 331.8827, BrCl 3 DF ： 349.8487, 351.8460, BrCl 3 DD ： 365.8436, 367.8410

61. 標品混合物の合成 標品混合物はパイレックスアンプル中、臭素化フェノー ルの縮合によって合成した。 ●PBrDF 混合標品は、フェノール、 2-/3-/4- ブロモフェ ノール、 2,4- ジブロモフェノール (DBP) を微量の CuBr 2 と ともに加熱して合成（ 370 ℃、 15 分）。 Authentic standards （市販） 2,7-D 2 BDF, 2,8-D 2 BDF, 2,3,8-T3BDF, ●PBrDD 混合標品は、 2- ブロモフェノール、 2,4-DBP 、 2,4,6-TBP を 300 ℃、 1 時間、熱分解して合成した。 Authentic standards （市販） 2,7-/2,8-DiBrDD, 2,3,7-TrBrDD, 1,2,3,4- TeBrDD, 1,3,6,8-TeBrDD, 1,3,7,9-TeBrDD, 1,3,7,8-TeBrDD, 2,3,7,8-TeBrDD

62. 使用した 2-/3-/4- ブロモフェノールの量を反映し、 2->1-,3->4-(4:2:2:1) であった。 DiBrDF の場合、 24- 以外に 28-, 18-, 27-, 17-, 19-, 26-, 37-, 16-, 36-, 46- が含まれ、 TrBrDF は、 248-, 168-, 247-, 246- 、 TeBrDF の場合、 2468- が主要な異性体である。 低臭素化 DD/DF 異性体について、市販標品の RT 、標品混 合物中の異性体予測生成比、異性体の溶出順序の規則性か ら、推定した。

63. はじめに 低塩素化 DD/DF の異性体分布は、生成機構や分解過 程の解明、毒性評価、起源推定などに非常に重要な情 報を提供するが、これまであまり関心が払われなかっ た。我々は、 SP-2331 [1] 、 DB5MS [2] DB-Dioxin カラム で 1 ～ 3 塩素化 DD/DF の全 74 異性体について、同定結果 を報告してきた。 極性の中間的な BPX35 (35%phenylpolysilphenylene-siloxane) および強極性の SP-2331 (95%cyanopropyl 5%phenyl-siloxane) カラムによる低臭素化ダイオキシン類の異性体分析に ついて報告する。