Transcript 低臭素化ダイオキシン類の 異性体分析 兵庫県立健康環境科学研究センター 中野 武

低臭素化ダイオキシン類の
異性体分析
兵庫県立健康環境科学研究センター
中野 武
現状
PCDD/DF分析法は、毒性係数の示されている17異
性体、4～8塩素化DD/DFのみを対象
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
低塩素化ダイオキシン類
の異性体分析
●
●
●
SP-2331
DB-DIOXIN
DB5-MS
Column;
SP-2331 and DB-DIOXIN
Polarity: DB-DIOXIN <
DB-Dioxin
44% methyl, 28% phenyl,
20% cyanopropyl
8% polyoxyethylene
SP-2331
90% biscyanopropyl
10% phenylcyanopropyl
SP-2331
Standard:
a) Authentic standards:
13, 27, 23, 36, 28, 26, 34, 46-DiCDFs
Authentic standards of
136,138,146,147,149,236,238,246,267,346 -TrCDFs
b) Synthesis of PCDD by pyrolysis phenols
in presence of KOH at temperature between
250C-300C
c) Synthesis of PCDF by pyrolysis of
respective Phenols in pyrex ampoule
(370-400C, 15-30min)
d) Synthesis of PCDF by pyrolysis of PCB in
pyrex ampoule (400-450C, 15min)
PCDF formation from chlorophenols
Phenol 2-MCP
3-MCP
4-MCP
23-DCP
24-DCP
25-DCP
Phenol
4-F
1/3-F
2-F
34-F
24-F
14-F
2-MCP
46-F
16/36-F
26-F
346-F
246-F
146-F
168/247-F
147/149-F
248-F
148-F
3-MCP
4-MCP
17/37/19-F
18/27-F 167/347-F
28-F
267-F
Synthesis PCDF
Synthesis of single isomers (in one
homolog):
2-chloroPhenol : 4,6-DiCDF
Synthesis of mixtures
3,4-dichloroPhenol
(result in 1,2- and 2,3-fragment)
3-chloroPhenol
(result in 1- and 3-fragment)
OH
Cl
Cl
Cl
4-MCP
O
1,2,8-TrCDF
1,2,8-
Cl
+
OH
Cl
Cl
Cl
3,4-DCP
Cl
O
2,3,8-TrCDF
2,3,8-
Cl
Cl
Cl
2-MCP
Cl
OH
3-MCP
Cl
OH
O
Cl
O
1,4,6-
Cl
1,4,9-
Cl
OH
1,4,6-TrCDF
Cl
1,4,9-TrCDF
Cl
Cl
Cl
Cl
OH
2,5-DCP
Cl
Cl
1,4,7-
O
1,4,8-
O
Cl
1,4,8-TrCDF
Cl
Cl
1,4,7-TrCDF
4-MCP
Authentic standard
(1,4,7- : NMR)
Synthesis of 1,4,X-T3CDF isomers from chlorophenol
Authentic standard
H9
Cl1
H8
H2
Cl7
H3
H6
Cl4
H2 ; H3
H6
H9
H8
NMR 1,4,7-TrCDF
Synthesis PCDD
Synthesis of single isomers
1-MCDD, 2-MCDD,
12-DCDD, 13-DCDD, 23-DCDD,
123-TCDD, 124-TCDD, 236-TCDD, 237-T3CDD
Synthesis PCDD
Most cases simple PCDD mixtures with two
isomers (smiles rearangement):
2,3+2,3,4 result in 1,2,6- and 1,2,9-T3CDD
Assignment in mixture of two isomers:
With the same ring fragment substitution
the more polar compound elutes later.
Chromatogram on Columns;
Comparison Retention Behavior
Chromatograms
- DB-Dioxin
- SP2331 column
9
10
11
12
13
14
129
15
17
16
18
19
20
21
346
347/236
19
267
137
12
24
19
237/149
18
248
46
37
27
36 23
28
26
19
34
18
18
246
247
17
128
16
17
126
15
139/127
14
16
167
TrCDF
15
146
13
14
178
148
123
12
13
124/147
11
12
138
136
168
134
10
11
137
9
10
16
14
9
17
13
4
3
1
2
Ambient air
SP-2331
MCDF
DiCDF
20
21
22
20
21
22
22
DB-DIOXIN
3
MCDF
4
24
1
2
34
23
SP-2331
D2CDF
46
19
19
28
9
10
11
12
13
14
14
29.5
30.0
30.5
31.0
31.5 32.0
18
19
20
21
22
21
22
DiCDF
D2CDF
12
14
347/236/234
CDF
TT3CDF
17
46
46
29.0
16
37
27
36 23
28
26
19
34
28.5
18
28.0
16
27.5
17
27.0
13
13
26.5
15
24
16
17
18
13
37
13
26
12
27
36
D2CDF
38
346
267
237/149
246
347/236
20
248
247
128
346
17
123
15
19
137
18
126
17
139/127
137
137
16
167
129
15
146
14
178
148
13
124/147
12
129
129
346
267
238
37
11
TrCDF
T3CDF
149
36
10
138
136
168
134
35
126/128/246
237
34
9
247
139
167/248
33
148/127
168
134/147
124
32
138
136
137
137
178/146/123
3
39
9
10
11
12
13
14
16
18
19
GC/MS-SIM chromatogram of PCDF
20
21
22
DB-DIOXIN
16
SP-2331
CDD
DD22CDD
2
19
19
12
18
17
14
23
3
126
123/178
9
129
34.4
34.8
35.2
35.6
11
12
36.0
36.4
36.8
37.2
13
14
15
137
37.6
38.0
10
11
12
13
22
DiCDD
D
2CDD
19
T3CDD
TrCDD
9
34.0
10
146
127
128
139
147
237
124
136
137
138
137
21
16
17
18
19
20
21
22
14
15
16
17
18
19
38.4 38.8
GC/MS-SIM chromatogram of PCDD
20
129
129
T
CDD
T 3CDD
20
127
128
146
126
13
32.0
19
139/237
31.6
18
123147
178
31.2
17
124
30.8
16
19
30.4
15
138
30.0
14
136
29.6
13
137
29.2
12
14/17
18
16
12
28.8
11
129
28.4
10
27/23/28
9
13
13
27/28
1
13
28.0
MCDD
21
22
Assignment for MCDD/F to T3CDD/F
MCDD
2
1
DCDD
13
27
28
23
14
17
18
16
12
19
23.76
24.69
28.51
29.57
29.57
29.73
30.05
30.26
30.39
30.85
31.26
31.77
TrCDD
137
138
136
124
139
147
237
123
178
146
127
128
126
129
34.18
34.24
34.61
35.07
35.40
35.60
35.63
36.19
36.24
36.65
36.86
37.00
37.31
38.21
MCDF
1
3
2
4
DCDF
13
17
14
18
24
37
16
12
27
22.57
23.09
23.56
24.51
26.43
27.71
28.48
28.65
28.96
29.08
29.28
29.34
29.49
DCDF
23
28
36
26
34
46
19
TrCDF
137
138
136
168
134
147
29.87
29.96
30.16
30.69
31.09
31.31
31.46
31.77
32.58
32.87
33.44
33.52
33.52
TrCDF
124
127
148
123
146
178
247
139
248
167
126
128
246
237
33.80
34.43
34.43
34.63
34.65
34.65
34.75
35.08
35.22
35.23
35.48
35.48
35.58
35.94
TrCDF
238
347
236
234
149
267
346
129
36.33
36.88
36.88
36.88
36.98
37.37
37.73
38.49
Separation of low chlorinated DD
The No. of separation peaks
isomer ULTRA-2 DB-5MS SP-2331 DB-DIOXIN
MCDD 2
D2CDD 10
T3CDD 14
2
5
6
2
4
5
2
8
12
2
9
12
Separation of low chlorinated DF
The No. of separation peaks
isomer ULTRA-2 DB-5MS SP-2331 DB-DIOXIN
MCDF 4
D2CDF 16
T3CDF 28
4
8
8
4
10
10
4
15
23
4
16
21
R.T. Behavior : Substitution Fragments
SP-2331 and DB-DIOXIN
PCDF
Elution order on DB-DIOXIN
2,4-, 3,7- isomers : earlier
1,2-, 1,6-, 1,9- isomers : later
compare with SP-2331
23
DB-Dioxin
24
24
34
SP-2331
GC/MS-Chromatogram of D2CDF
27.5
28.0
28.5
46
26
12
27
19
28
14
27.0
19
16
17
18
37
37
13
26.5
36
12
SP-2331
D2CDF
16
29.0
29.5
30.0
30.5
31.0
31.5 32.0
33
34
35
139
149
36
37
129
149
178/146/123
T3CDF
267
238
346
SP-2331
126/128/246
SP-2331
237
247
139
167/248
148/127
168
134/147
124
32
138
136
137
GC/MS-Chromatogram of T3CDF
DB-Dioxin
347/236/234
129
38
39
R.T. Behavior : Substitution Fragments
SP-2331 and DB-DIOXIN
PCDD
Elution order on DB-DIOXIN
1,4- isomers : earlier
compare with SP-2331
GC/MS-Chromatogram of D2CDD
29.2
17
12
18
29.6
14
23
28.8
19
DB-Dioxin
27/28
13
28.4
SP-2331
14
D2 CDD
28.0
16
DB-Dioxin
30.0
30.4
30.8
31.2
31.6
32.0
GC/MS-Chromatogram of T3CDD
129
DB-Dioxin
34.0
34.4
34.8
35.2
126
146
35.6
146
127
128
139
147
237
124
136
137
138
147
123/178
T3CDD
36.0
36.4
36.8
37.2
37.6
38.0
38.4 38.8
低臭素化ダイオキシン類
の異性体分析
CN
CN
C3 H
C3 H
6
6
O Si
O Si
C3 H
6
CN
●
●
x
y
SP-2331 (95%cyanopropyl 5%phenyl-siloxane)
BPX-35 (35%phenylpolysilphenylene-siloxane)
CH3
O Si
CH3
CH3
CH3
Si
CH3
O Si
x
y
Standards.
For the PBDF mixture all three monobromophenol isomers,
Phenol and 2, 4-DBP were pyrolysed at 370ºC (15 minutes) in
the presence of trace amounts of CuBr2.
Authentic standards
2,7-D2BDF, 2,8-D2BDF, 2,3,8-T3BDF,
合成した臭素化DF標品混合物は、
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。
2
M 1BDF
1587256
M1BrDF
1
3
4
0
11.5
D2BDF
497896
12.0
12.5
13.0
13.5 14.0 14.5
Retention Time
D2BrDF
15.0
15.5
16.0 16.5
28
27
18
17
13
24
16
36
37
26
46
0
16
17
18
T3BrDF
22
248
25
347/236
137
24
238
246
138/136
23
237/149
T3BDF
102569
19
20
21
Retention Time
247
346
168
0
24
26
28
30
Retention Time
32
34
M1BrDF～T3BrDFのGC/MS-SIMクロマトグラム
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.
Intensity
M1BDF / Average
2726152
2
1 3
4
0
10
11
Retention Time (min)
Intensity
BrClDF / Average
14887
503
13
14
15
Retention Time (min)
28
Intensity
D2BDF / Average
792528
13
27
18
17
24
26
46
0
16
17
18
19
Retention Time (min)
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.
T3BDF / Average
Intensity
312320
0
26
27
28
29
Retention Time (min)
T4BDF / Average
Intensity
87023
0
31
32
33
Retention Time (min)
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.
Standards. The PBDD standards were synthesized by pyrolysis
of bromophenols in Pyrex ampoules.
The PBDD was synthesized by condensation of 2-monobromo
phenol, 2, 4- and 2, 6- dibromophenol (DBP) and 2, 4, 6tribromophenol (TBP) at 350ºC (30 min).
Authentic standards:
2,7/2,8-D2BDD, 2,3,7-T3BDD.
合成した臭素化DD標品混合物は、
DiBrDDは、13-、27/28- 、
TrBrDDは、137-, 138- 、 136-, 139-
TeBrDDは、1368-, 1379- が含まれていた。
M 1BDD
3095833
1
M1BrDD
2
0
11
12
D2BDD
760425
13
14
Retention Time
13
14/17
15
D2BrDD
18
27/28/23
16
16
19
12
0
16
17
18
19
20
21
Retention Time
22
23
24
T3BrDD
T3BDD
398216
136
138
137
124
129
237
0
25
26
27
28
29
Retention Time
30
31
32
M1BrDD～T3BrDDのGC/MS-SIMクロマトグラム
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.
1
M1BDD / Average
4457267
Intensity
2
0
10.4
10.8
Retention Time (min)
BrClDD / Average
Intensity
9893
524
14
15
16
Retention Time (min)
D2BDD / Average
27/28
Intensity
2389477
13
0
17
18
19
Retention Time (min)
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.
Intensity
T3BDD / Average
1307231
0
27
28
29
Retention Time (min)
1368
1379
Intensity
T4BDD / Average
191064
0
31.0
31.5
32.0
32.5
Retention Time (min)
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.
排ガス試料
PCB濃度が高い場合、
DiBrDF(323.8785, 325.8765) は
PeCB(323.8834,325.8805)の妨害を受ける。
臭素系DD/DF は
BrDF>>BrClDF＞BrCl2DF、
BrDD>>BrClDD＞BrCl2DD
PCB濃度が高い場合、DiBrDF(323.8785, 325.8765) はPeCB(323.8834,325.8805)の妨害を受ける。
28
STD mix
27
649323
D2BrDF
0
16
20
24
Retention Time (min)
#118
Sample
568627
#114
#105
D2BrDF
28
27
0
16
20
24
Retention Time (min)
GC/MS-SIMクロマトグラム(D2BrDF)
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.
臭素系DD/DF はBrDF>>BrClDF＞BrCl2DF、BrDD>>BrClDD＞BrCl2DD
Sample-A
171522
BrClDF
0
14
16
18
20
18
20
Retention Time (min)
Sample-B
86858
BrClDF
0
14
16
Retention Time (min)
GC/MS-SIMクロマトグラム(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.