1. Enantioselective Analysis of Chlordanes in Serum Chisato MATSUMURA, Masahiro TSURUKAWA, Hiroaki KITAMOTO, Toshihiro OKUNO, Takeshi NAKANO (Hyogo prefectural institute of public health and environmental sciences, Kobe Japan)

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

4. Technical chlordane import (tons)

5. Basic component of technical chlordane Our study

6. Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl trans-nonachlor cis-nonachlor achiral trans-chlordane chiral oxychlordane major component of technical chlordane

7. | E first – E last | E L - E S EE = 100 X ------------------- or 100 X ------------ E first + E last E L + E S E : peak area or height E first E last E first : first eluting enantiomer E last : last eluting enantiomer E L : larger enantiomer E S : smaller enantiomer E S E L Retention time Enantiomeric excess (EE)

8. What’s known from EE production house degradation agriculture diffusion Change EE accumulation e.g. Physical process Biological process use For termite control transport

9. ← CS solution (2.5ng each) 2.5ml of diethylether 5ml of ethanol 10ml of n-hexane X2 Analytical Flow Blood Shake （ 10minuten ） and centrifuge (3000rpm, 10minuten) n-hexane layer Concentrated to 1ml under the nitrogen streem Clean-up with LC-Florisil (1g) / LC-Si (1g) (Supelco) Eluted with 10ml of 15% diethylether/hexane Concentrated to 0.1ml under the nitrogen streem Centrifuged (3000rpm, 10minuten) Analyzed with HRGC/MS 2g of upper layer (Serum) ← ISS solution (0.5ng)

10. CS solution 13 C 10 chlordene, 13 C 10 heptachlor, 13 C 10 heptachlor-exo-epoxide, 13 C 10 trans-chlordane, 13 C 10 oxychlordane, 13 C 10 trans- and cis-nonachlor ISS solution 13 C 12 2,3’,4’,5-TeCB (PCB#70) 13 C 12 2,2’,4,5,5’-PeCB (PCB#101) Internal standard solution

11. HR ＧＣ／ HR ＭＳ and Column GC Hewlettpackard HP- 5890II MS JEOL MS-700 Column ： BGB-172 (BGB Analytik) 30mX0.25mmID,0.25μmFilmThickness （ 20%-tert-butyldimethylsilylated-beta- cyclodextrin in 15%-phenyl-85%-methyl- polysiloxane ）

12. HR ＧＣ／ HR ＭＳ ｃｏｎｄｉｔｉ ｏｎｓ Carrier gas:Helium (1.5ml/min constant flow) Oven temp.:120 ℃ (0min)-4 ℃ /min-180 ℃ (0min) -1 ℃ /min-230 ℃ (15min), or 120 ℃ (0min)-4 ℃ /min-200 ℃ (0min) -1 ℃ /min-230 ℃ (25min), Ion source ： 250 ℃ Inj.Temp. ： 230 ℃ Ionization Energy: 38eV Ionization Current: 500μA Transfer line Temp. ： 245 ℃ Injected amount ： 1μl

13. GC/MS compound list (2 injections) m/z: 218.9 - 337.9 (1.54M) HCH DDT&DDD Heptachlor Chlordene Te4CB Oxychlordene Pe5CB 13 C 6 -HCH 13 Cring- DDT&DDD 13 C 10 -heptachlor 13 C 10 -chlordene 13 C 12 -Te4CB 13 C 12 -Pe5CB m/z: 352.8 - 441.8 (1.25M) Heptachlorepoxide Chlordane Oxychlordane Nonachlor Hx6CB Hp7CB Oc8CB 13 C 10 -heptachlorepoxide 13 C 10 -chlordane 13 C 10 -oxychlordane 13 C 10 -nonachlor 13 C 12 -Hx6CB 13 C 12 -Hp7CB 13 C 12 -Oc8CB

14. GC/MS-SIM chromatogram on BGB-172 HRGC column

15. Technical Chlordane Atmosphere chlordane trans- trans- cis- cis- cis- cis- MC-5 MC-5 MC-4 MC-4

16. River Water River Sediment chlordane trans- trans- cis- cis- cis- cis- MC-5 MC-5 MC-4 MC-4

17. Technical Chlordane Serum chlordane trans- trans- cis- cis- cis- cis- MC-5 MC-5 MC-4 MC-4

18. beta- gamma- alpha- chl6-2 chlordene Technical Chlordane Atmosphere chlordene

19. beta- gamma- alpha- chl6-2 chlordene River Water River Sediment chlordene

20. beta- gamma- alpha- chl6-2 chlordene Technical Chlordane Serum chlordene

21. heptachlor-epoxide and oxychlordane heptachlor-epoxide oxychlordane exo- exo- endo-

22. Enantiomeric excess (EE) chl6-2* 3 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

24. 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 Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl trans-nonachlo cis-nonachlor achiral component-MC7 cis-chlordane component-MC5 trans-chlordane component-MC4 heptachlor component-U82

25. component-MC6 o,p’-DDT o,p’-DDD alpha-HCH oxychlordane heptachlor-exo-epoxide oxychlordene

26. GC/MS channel list 1 (2 injections)

27. GC/MS channel list 2 (2 injections)

28. Technical Chlordane Atmosphere River Water River Sediment Serum chlordane trans- trans- cis- cis- MC-5 MC-5 MC-5 MC-5 MC-4 MC-4

29. beta- gamma- alpha- chl6-2 chlordene Technical Chlordane Atmosphere River Water River Sediment Serum chlordene