Enantioselective%20Analysis%20of%20Chlordanes%20in%20Serum

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Enantioselective Analysisof Chlordanes in Serum

• Chisato MATSUMURA,
• Masahiro TSURUKAWA, Hiroaki KITAMOTO, Toshihiro
  OKUNO, Takeshi NAKANO
• (Hyogo prefectural institute of public health and
  environmental sciences, Kobe Japan)

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Technical chlordane

• from 1945 in U.S.
• a ubiquitous and persistent
• killing lawn and garden pests
• termite control
• pest control on crops

3
In Japan

• From 1950
• In 1968, it was banned for agriculture
• Used for termite control
• The Chemical Substances Control Law restricted it
  in 1986.

Termite
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Technical chlordane import (tons)
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Basic component of technical chlordane
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major component of technical chlordane
trans-chlordane chiral
oxychlordane
trans-nonachlor cis-nonachlor achira
l
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Enantiomeric excess (EE)

• Efirst Elast EL - ES
• EE 100 X ------------------- or 100 X
  ------------
• Efirst Elast EL ES
• E peak area or height Efirst
  Elast
• Efirst first eluting enantiomer
• Elast last eluting enantiomer
• EL larger enantiomer
• ES smaller enantiomer ES
  EL
• Retention time

8
Whats known from EE
production
diffusion
use
transport
agriculture
accumulation
house
degradation
For termite control
e.g. Physical process Biological process
Change EE
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Analytical Flow
Blood
Concentrated to 1ml under the nitrogen streem
Centrifuged (3000rpm, 10minuten)
Clean-up with LC-Florisil (1g) / LC-Si (1g)
(Supelco)
2g of upper layer (Serum)
Eluted with 10ml of 15 diethylether/hexane
? CS solution (2.5ng each) 2.5ml of
diethylether 5ml of ethanol 10ml of
n-hexane X2
Concentrated to 0.1ml under the nitrogen streem
Shake (10minuten) and centrifuge (3000rpm,
10minuten)
? ISS solution (0.5ng)
Analyzed with HRGC/MS
n-hexane layer
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Internal standard solution

• CS solution
• 13C10 chlordene, 13C10 heptachlor,
• 13C10 heptachlor-exo-epoxide,
• 13C10 trans-chlordane, 13C10 oxychlordane,
• 13C10 trans- and cis-nonachlor

• ISS solution
• 13C12 2,3,4,5-TeCB (PCB70)
• 13C12 2,2,4,5,5-PeCB (PCB101)

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HRGC/HRMS and Column
GC Hewlettpackard HP-5890II MS JEOL MS-700
Column BGB-172 (BGB Analytik) 30mX0.25mmID,0.25µm
FilmThickness (20-tert-butyldimethylsilylated-bet
a-cyclodextrin in 15-phenyl-85-methyl-polysiloxa
ne )
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HRGC/HRMS conditions
Carrier gasHelium (1.5ml/min constant flow) Oven
temp.120?(0min)-4?/min-180?(0min) -1?/min-230?(15
min), or 120?(0min)-4?/min-200?(0min) -1?/min-2
30?(25min), Ion source 250? Inj.Temp.
230? Ionization Energy 38eV Ionization Current
500µA Transfer line Temp. 245? Injected
amount 1µl
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GC/MS compound list (2 injections)
m/z 352.8 - 441.8 (1.25M) Heptachlorepoxide Chlo
rdane Oxychlordane Nonachlor Hx6CB Hp7CB
Oc8CB 13C10-heptachlorepoxide 13C10-chlordane 13C
10-oxychlordane 13C10-nonachlor 13C12-Hx6CB
13C12-Hp7CB 13C12-Oc8CB
m/z 218.9 - 337.9 (1.54M) HCH
DDTDDD Heptachlor Chlordene Te4CB
Oxychlordene Pe5CB 13C6-HCH 13Cring-DDTDDD
13C10-heptachlor 13C10-chlordene 13C12-Te4CB
13C12-Pe5CB
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GC/MS-SIM chromatogramon BGB-172 HRGC column
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chlordane
trans- trans- cis- cis-
MC-5 MC-5
MC-4 MC-4
Technical Chlordane
Atmosphere
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chlordane
trans- trans- cis- cis-
MC-5 MC-5
MC-4 MC-4
River Water
River Sediment
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chlordane
trans- trans- cis- cis-
MC-5 MC-5
MC-4 MC-4
Technical Chlordane
Serum
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chlordene
beta- gamma- alpha-
chl6-2 chlordene
Technical Chlordane
Atmosphere
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chlordene
beta- gamma- alpha-
chl6-2 chlordene
River Water
River Sediment
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chlordene
Technical Chlordane
beta- gamma- alpha-
chl6-2
chlordene
Serum
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heptachlor-epoxide and oxychlordane
oxychlordane
exo-
exo- endo- endo-
heptachlor-epoxide
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Enantiomeric excess (EE) Enantiomeric excess (EE)
chl6-23 41
beta-chlordene 3.8
gamma-chlordene 21
heptachlor-exo-epoxide 50
MC-5 71
trans-chlordane 24
cis-chlordane 48
oxy-chlordane 21
3 unknown isomer on chlordane 3 unknown isomer on chlordane
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(No Transcript)
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heptachlor
component-U82
trans-chlordane
component-MC4
cis-chlordane
component-MC5
trans-nonachlo cis-nonachlor achiral
component-MC7
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component-MC6
o,p-DDT
oxychlordane
o,p-DDD
heptachlor-exo-epoxide
alpha-HCH
oxychlordene
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GC/MS channel list 1 (2 injections)
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GC/MS channel list 2 (2 injections)
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chlordane
trans- trans- cis- cis- MC-5
MC-5 MC-4 MC-4
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chlordene
beta-
gamma- alpha- chl6-2
chlordene
Technical Chlordane
Atmosphere
River Water
River Sediment
Serum