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

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

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

PCB product use in Japan Ar KC

KC300 KC400 KC500 KC600 Japan Homologue pattern reflects product use

PCB Homologue Distribution and Source Estimation

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

To evaluate environmental risk

Daily variation of PCDD/Fs and PCBs in ambient air

Daily variation of Dioxin Concentration

Time trend of PCDD/F levels in ambient air

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

Emission inventry (g-TEQ/year ） PCDDs/PCDFs （ pg-TEQ/m3) Emission inventry vs Level in air Matsumura et al, symposium of J.Env.Chem,

Time trend of Co-PCBs （ TEQ ）

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

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

Low volume air sampler

Time trend of PCB levels in breast milk

Time trend of breast milk levels Dioxins PCB pg TEQ / g-fat µg g-fat Year ( ) Konishi et al. (2003) Konishi et al (2003) Organohalogen compds pg TEQ/g-fat µg / g-fat Year ( )

Development of congener specific analysis for PCBs using HT8-PCB

Congener number number of isomerof peak mono 3 3 di tri tetra penta hexa hepta octa nona 3 3 deca 1 1 total

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

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

Congener profiles of PCBs in environmental and human samples

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

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

Congener profiles of PCBs in human samples (pooled blood)

＃ 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 #

(n=24) CongenersMean ± 2S.D. Existence Ratio* (BZ#)(ng /g-lipid)(%) # ± 22.3 # ± # ± # ± # ± #163, ± #994.7 ± #744.6 ± # ± # ± # ± Sum75.6 * Mean is compare to total PCBs level Levels of the predominant PCB congeners in human blood Hirai et al, (2003) Organohalogen compounds cumulative (%)

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

Analysis of Low Brominated DD/F ANALYSIS of MBDD/MBDF to T 3 BDD/T 3 BDF on a SP COLUMN

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.

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.

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.

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

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.

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)

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

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

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

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 ,4-DBP + 4-BP 2,4,8-T 3 BDF ,4-DBP + 2,4-DBP 2,4,6,8-T 4 BDF

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 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)

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.

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.

Table.1 Monitoring ion M 1 BrDD D 2 BrDD T 3 BrDD T 4 BrDD M 1 BrDF D 2 BrDF T 3 BrDF T 4 BrDF

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.

/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.

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)

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

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

/28/ /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.

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.

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.

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)

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-, , 26-, 46- T 3 BDF 248-, 247-, 246-, 249-(168-) T 4 BDF 2468-

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

/ M 1 BrDF D 2 BrDF T 3 BrDF 347/ /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.

PCDF PBDF MCDF DCDF TCDF MBDF DBDF TBDF

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.

Separation of low chlorinated DD/F (Dioxin 2000, 2001, 2002) congener ULTRA-2 DB-5MS SP-2331 DB-DIOXIN MCDF D2CDF T3CDF congener ULTRA-2 DB-5MS SP-2331 DB-DIOXIN MCDD D2CDD T3CDD

Separation of low brominated DD/F on BPX35 & SP-2331 congener BPX35 SP-2331 MBDF D2BDF T3BDF congener BPX35 SP-2331 MBDD D2BDD T3BDD14 6 9

Comparison of Separation for PXDD/F on SP-2331 (Cl/ Br) congenerClBr MXDF D2XDF T3XDF congenerClBr MXDD D2XDD T3XDD the selectivity for brominated congeners is smaller compared with the chlorinated congeners.

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

Low Brominated DD/F in Stack Gas Samples

Retention Time (min) Retention Time (min) D 2 BrDF STD mix Stack Gas #118 #105 # 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( , ) DiBrDF( , ) #126

Retention Time (min) Retention Time (min) 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

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Low Chlorinated DD/F in Environmental Samples

環境大気 Ambient air

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

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

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

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) Isomer distribution of ambient air(MCDF-TrCDF) Isomer distribution of bleaching process(MCDF-TrCDF) Ambient air

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

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

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

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

PCB levels and Homologue profiles in Environmental samples

PCB levels in air, water, sediment and biota (2002, MOE Japan) Air Sediment Water Biota ng/m ng/L ng/g-dry ng/g-wet

Homologue profiles of PCBs in human samples

Air Water Bird abundance(%) Human blood Breast milk

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

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

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

(stack gas) TrCB TeCB #68 #51 #

Congener profiles of PCB in human sample

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

Congener profiles of PCB in food sample

PCB congener profiles in food sample (Konishi et al ) ng/day

Congener profiles of PCB in human sample

PCB congener composition in human breast milk #187 #180 #153 #138 #74 #99 #118 #187 #180 #153 #138 #74 #99 #118

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

PCB product human #74 # #99 # #146 #

Isomer distribution (P 7 CB, Port Island in June,2001) KC Sea Water Mussel Intensity #174 #180 #170

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