Transcript 人血清中クロルデン類の 光学異性体分析

POPs類における
光学異性体分析の検討
○松村千里、鈴木元治、鶴川正寛、
奥野俊博、中野 武
（兵庫県立健康環境科学研究センター）
光学異性体
• クロルデン異性体には光学異性体が存在。生体内
で一方の異性のみが選択的に代謝されることによ
り光学異性対比ER（(+)／(－)）が変化することが
知られている。これらの光学異性体の把握は、生
体中での代謝情報やリスク評価に役立つことから、
重要なファクターになると考えられる。
Example of chlordane component
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
O
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
H
H
trans-chlordane
Cl
chiral
oxychlordane
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
O
Cl
Cl
Cl
trans-nonachlor
cis-nonachlor
achiral
Cl
Cl
Cl
heptachlor
Cl
Cl
Cl
component-U82
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
trans-chlordane
Cl
Cl
Cl
component-MC4
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
cis-chlordane
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
component-MC5
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
trans-nonachlo
Cl
cis-nonachlor
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
component-MC7
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
component-MC6
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
heptachlor-exoepoxide
Cl
Cl
Cl
H
Cl
Cl
C*
C*
H
H
Cl
Cl
H
H
Cl
Cl
H
Cl
H
Cl
Cl
oxychlordene
Cl
Cl
Cl
Cl
Cl
O
O
Cl
H
H
H
Cl
H
Cl
H
H
H
H
H
H
Cl
H
Cl
H
Cl
Cl
Cl
H
Cl
Cl
H
Cl
H
Cl
alpha-HCH
O
Cl
Cl
o,p’-DDD
Cl
Cl
Cl
O
Cl
Cl
H
Cl
Cl
Cl
Cl
Cl
Cl
H
Cl
H
Cl
H
H
O
Cl
Cl
C*
Cl
Cl
Cl
Cl
Cl
Cl
O
Cl
Cl
oxychlordane
Cl
C*
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
o,p’-DDT
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
H
Atropisomer of PCB 1
#45 (2,2’,3,6-TeCB)
#84(2,2’,3,3’,6-PeCB)
#95(2,2’,3,5’,6-PeCB)
#91(2,2’,3,4’,6-PeCB)
#88(2,2’,3,4,6-PeCB)
Atropisomer of PCB 2
#132(2,2’,3,3’,4,6’-HxCB)
#135(2,2’,3,3’,5,6’-HxCB)
#136(2,2’,3,3’,6,6’-HxCB)
#149(2,2’,3,4’,5’,6-HxCB)
#131(2,2’,3,3’,4,6-HxCB)
#139(2,2’,3,4,4’,6-HxCB)
#144(2,2’,3,4,5’,6-HxCB)
Atropisomer of PCB 3
#171(2,2’,3,3’,4,4’,6-HpCB)
#175(2,2’,3,3’,4,5’,6-HpCB)
#183(2,2’,3,4,4’,5’,6-HpCB)
#176(2,2’,3,3’,4,6,6’-HpCB)
#174(2,2’,3,3’,4,5,6’-HpCB)
#196(2,2’,3,3’,4,4’,5,6’-OcCB)
#197(2,2’,3,3’,4,4’,6,6’-OcCB)
P309～P312
光学異性体
• 今回は、ＧＣ／ＭＳ分析用キャピラリーカラムとして
液相にβ－シクロデキストリンを用いたカラム（ＢＧ
Ｂ－１７２、20% tert-butyldimethylsilylated betacyclodextrin in
methylphenylcyanopropylpolysiloxane ）を使用する
ことにより、クロルデン類およびAtropisomer
（PCB）の光学異性体分析を行った。人血清、クロ
ルデン製品や環境試料の光学異性体パターンを
比較したのでその結果を報告する。
HRＧＣ／HRＭＳ and Column
GC Hewlettpackard HP-5890II
MS JEOL MS-700
Column： BGB-172 (BGB Analytik)
30mX0.25mmID,0.25μmFilmThickness
（20%-tert-butyldimethylsilylatedbeta-cyclodextrin in 15%-phenyl-85%methyl-polysiloxane ）
Table1 GC/MS conditions
GC:HP6890Ⅱ
Column:BGB-172 (BGB Analytik) 30mX0.25mmID,
0.25μmFilmThickness
Carrier gas:Helium(1.0ml/min constant flow)
Oven temp.:120℃(0min)-4℃/min-200℃(0min)-1℃/min230℃(25min), Inj.temp.:230℃
MS:JEOL JMS-800D
Ionization mode:EI, Ion source temp.:250℃, Interface
temp.:245℃, Ionization current:500μA, Ionization
energy:38eV
G#: 1 Repeat: 1.01[sec] m/z: 218.9 - 337.9 (1.54M)
Ch.#
Compound name
m/z
1
HCH
218.9116
2
HCH
220.9087
3
13C6-HCH
224.9317
4
Mass Lock
230.9856
5
Lock Check
230.9856
6
DDT
235.0081
7
DDT
237.0053
8
D8ring-DDD
243.0575
9
13Cring-DDT
247.0484
10
heptachlor
271.8102
11
heptachlor
273.8072
12
13C-Heptachlor
276.8269
13
PCB4
289.9224
14
PCB4
291.9195
15
Chlordene
302.8883
16
13C-PCB4
303.9597
17
Chlordene
304.8853
18
13C-Chlordene
312.9218
19
Oxy-chlordene
318.8832
20
Oxy-chlordene
320.8803
21
PCB5
325.8805
22
PCB5
327.8775
23
13C-PCB5
337.9207
GC/MS
channel list 1
(2 injections)
GC/MS
channel list 2
(2 injections)
G#: 1 Repeat: 1.01[sec] m/z: 352.8 - 441.8 (1.25M)
Ch.#
Compound name
m/z
1
heptachlor-O
352.8442
2
heptachlor-O
354.8413
3
PCB6
359.8415
4
PCB6
361.8386
5
13C-Heptachlor362.8778
6
13C-PCB6
371.8818
7
CHL
372.826
8
CHL
374.8231
9
Mass Lock
380.976
10
Lock Check
380.976
11
13C-CHL
384.8566
12
Oxy-CHL
386.8052
13
Oxy-CHL
388.8023
14
PCB7
393.8025
15
PCB7
395.7996
16
13C-Oxy-CHL
398.8358
17
13C-PCB7
405.8428
18
nonachlor
406.787
19
nonachlor
408.7841
20
13C-Nonachlor
418.8176
21
PCB8
427.7636
22
PCB8
429.7606
23
13C-PCB8
441.7999
Technical chlordane import (tons)
2500
2000
1500
1000
500
technical chlordanes (including heptachlor)
1986
1984
1982
1979
1980
1972
1970
1968
1966
1964
1962
1960
1958
0
chlorine number
Basic
component
of technical
chlordane in
Japan
chlorine
component
ratio (%)
α- chlordene
0.7
γ- chlordene
3
β- chlordene
3.6
Cl7
heptachlor
4.8
Cl8
MC-4
0.3
trans- chlordane
18
Cl6
MC-5
5.8
Cl7
cis-chlordane
16
MC-6
1.5
trans-nonachlor
14
cis-nonachlor
4.5
Cl8
Cl9
Cl6
Cl9
chlordane
Cl
Cl
Cl
Cl
Cl
Technical
Chlordane
trans-
Cl
Cl
Cl
trans-
cis- cis-
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
MC-5
Cl
racemic
trans-
MC-4 MC-4
trans-
MC-5
racemic
Cl
Atmosphere
cis- cisMC-5
Cl
Cl
Cl
Cl
MC-5
Cl
Cl
Cl
Cl
Cl
MC-4 MC-4
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
transtranscis- cis-
River Water
Cl
Cl
Cl
Cl
Cl
MC-5
MC-5
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
racemic
MC-4 MC-4
trans-
Cl
Cl
trans-
River Sediment
racemic
MC-5
cis- cisMC-5
MC-4 MC-4
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
trans-
Technical
Chlordane
trans-
Cl
Cl
cis- cis-
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
MC-5
racemic
MC-5
MC-5
Cl
MC-4 MC-4
trans-
nonracemic
MC-5
Cl
Cl
Cl
cistrans-
Cl
Serum
cis-
MC-4 MC-4
Cl
chlordane
Technical
Chlordane
Air
racemic
Water
Sediment
nonracemic
Serum
trans-
cis-
MC-5
heptachlor-epoxide and oxychlordane
Cl
Cl
Cl
Cl
Cl
oxychlordane
O
Cl
Cl
Cl
H
nonracemic
exo-
exoCl
heptachlor-epoxide
Cl
Cl
Cl
Cl
O
Cl
endo- endo-
Cl
H
H
heptachlorOxychlordane
epoxide
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
O
Cl
Cl
O
Cl
Air
Cl
H
Cl
H
H
Water
nonracemic
Serum
Bivalve
(corbicula)
beta-
Technical
Chlordane
Atmosphere
alpha-
gamma-
chl6-2
chlordene
River Water
River Sediment
Serum
Figure2
GC/MS-SIM chromatogram showing elution of chlordene on
BGB-172 HRGC column. (average of m/z 302.8883 and 304.8853)
Table1 The enantiomeric excess (EE is absolute of 100 X [Area-first –
Area-last]/[Area-first + Area-last]) values of chlordane component in
serum sample. (%)
chl6-2*3
beta-chlordene
41
3.8
gamma-chlordene
21
heptachlor-exo-epoxide
50
MC-5
71
trans-chlordane
24
cis-chlordane
48
oxy-chlordane
21
•1 Area-first : area of first eluted enantiomer
•2 Area-last : area of last eluted enantiomer
•3 unknown isomer on chlordane
4,4'-DDD
(2)2,4'-DDD
(1)2,4'-DDD
4,4'-DDD
4,4'-DDD
4,4'-DDT
(2)2,4'-DDT
(1)2,4'-DDT
(1)2,4'-DDD
(2)2,4'-DDD
(1)2,4'-DDD
(2)2,4'-DDD
DDD&DDT（Air）
DDD&DDT（Water1）
DDD&DDT（Water2）
DQ Main View
Page 1
DqData : h:\DiokData\POPsPCBChiralHi2new\07POPsChiralHi-003
Injection : 06EDAirItamiCityHVFeb
PCB7
Intensity
80000
40000
0
34
36
38
40
42
44
46
48
Retention Time (min)
50
52
50
52
54
56
58
56
58
60
#189
#190
#170
#181
#172
(1)#171
#192
(2)#171
#173
#177
#193
#180
#191
#185
(1)#174
(2)#174
#187
(1)#183
(2)#183
#184
#188
#179
(1)/(2)#176
#186
#178
#182
(1)/(2)#175
Calculated Retention Time
Intensity
13C-PCB7
8000
4000
0
34
36
38
40
42
44
46
48
Retention Time (min)
54
60
DQ Main View
Page 1
DqData : h:\DiokData\POPsPCBChiralHi2new\07POPsChiralHi-003
Injection : 070208Fat98Yf
PCB7
Intensity
20000
10000
0
34
36
38
40
42
44
46
48
Retention Time (min)
50
52
50
52
54
56
58
56
58
60
#189
#190
#170
#181
#172
(1)#171
#192
(2)#171
#173
#177
#193
#180
#191
#185
(1)#174
(2)#174
#187
(1)#183
(2)#183
#184
#188
#179
(1)/(2)#176
#186
#178
#182
(1)/(2)#175
Calculated Retention Time
Intensity
13C-PCB7
2000
1000
34
36
38
40
42
44
46
48
Retention Time (min)
54
60
#170
(1)#183
(2)#183
PCB7
(adipose
tissue)
#180
(1)#174
(2)#174
#170
(1)#171
(2)#171
(1)#183
(2)#183
#180
#187
PCB7
(Air)
#187
atropisomer
(2)#174
(1)#174
#185
(2)#183
(1)#183
PCB7
(adipose
tissue)
#187
(2)#174
(1)#174
#185
(2)#183
(1)#183
#187
PCB7
(Air)
E(+)
EF = ---------E(+)+ E(-)
EF (-)
EFs of alpha-HCH in sea waters
POPs in
the
oceans
0.5
The Japan Sea
EF (-)
0.8
0.5
The South China Sea
EF (-)
0.4
0.5
EF (-)
The North Atlantic Ocean
0.5
The Pacific Ocean
S-9
S-8
S-10
S-11
S-7
S-6
S-5
S-4
S-3
S-2
S-1
The correlation between EFs and alphaHCH/ΣHCHs ratios
0.6
POPs in
the oceans
The North Atlantic Ocean
The South China Sea
The Japan Sea
The Pacific Ocean
EF
北大西洋
日本海
南シナ海
太平洋
0.5
0.4
0.0
0.2
0.4
0.6
alpha -HCH/ΣHCH
0.8
1.0
POPs in
the oceans
The EFs of some POPs in the sea waters
The North Atlantic Ocean
The South China Sea
The Japan Sea
The Pacific Ocean
0.8
0.7
EF
0.6
0.5
0.4
0.3
0.2
Heptachlor-exo-epoxide
trans-chlordane
0
1
2
3
4
5
6
7
8
9cis-chlordane
10 11 12
13 α
14-HCH
15 16
17