1. 1 2011/05/23 Development of SPE for urine sample

2. 2 2011/05/23 hydrophobic interaction type SPE hydrophobic interaction Interaction between hydrophobic chemicals Aromatic compounds dissolve in organic solvent effective for hydrophobic chemicals widely used for collection of the chemicals in water not effective for ionic, hydrophilic chemicals not effective for ionic, hydrophilic chemicals modify polymer with multifunctional group for ionic chemicals  ionic group for hydrophilic chemicals  hydrophilic group

3. 3 2011/05/23 highly-functional solid phase extraction(SPE) polymer Hydrophobic interaction Hydrophilic interaction Ion-exchange interaction Complex formation Cation exchange Anion exchange  －  interaction Basel resin polymer New functional group

4. 4 2011/05/23 Chemical structure of SPE (reverse phase ion exchange type) hydrophilic ion exchange hydrophobic polar

5. 5 2011/05/23 Chemical structure of SPE for urine sample RP-SAX RP-WAX RP-OH

6. 6 2011/05/23 Property of SPE 樹脂名単位RP-SAXRP-WAXRP-OH median diameter µm 68 58 specific surface area m 2 /g 550560250 micropore diameter nm 8.89.714 micropore capacity mL/g 0.660.590.67 ion exchange capacity meq/g 0.330.50 － unit capacity (dry) mL/g 2.422.523.40 unit capacity(swelling) mL/g 3.043.234.19 degree of swelling － 1.261.281.24 ion exchange － － N(CH 3 ) 2 C 2 H 4 OH － N(C 2 H 5 ) 2 －

7. 7 2011/05/23 Treatment for urine sample

8. urine sample add. x1, 20mM Phosphate buffer (pH 4.0 ) add. surrogate ： OH-PCB 13C mix (100ppb) 20uL, Solid PhaseExtraction （ RP-WAX.) Concentration elution, 8 mL 0.1% NH 3 in MeOH adjust pH 4.0 by 10% HCOOH aq. add. 5mL, 20mM Phosphate buffer (pH 7.0 ) add. 20 μL β-Glucuronidase / Aryl sulfatase, Enzymegenation Extraction Centrifuge 2600rpm. 10min Concentration add. x 9 CH 3 CN Concentration LC/MS analysis by N 2 gas, 1 mL by CH 2 Cl 2, 2 mL x 2 37 ℃, 1.5 hrs dryness by N 2 gas, GC/MS analysis methylation

9. 92011/05/23 LC/MS Analysis

10. 4-OH-146 3-OH-138 4-OH-153 100 ppb 0.5 ppb 4-OH-187 4-OH-107 4-OH-172 100 ppb 0.5 ppb dominant OH-PCB in human blood sample

12. S/N ratio of 0.5ppb 4-OH-CB-54 on selected ion chromatogram(mass window: 30mDa)

13. Std 2.0 ppb urine blank Spike 2.0 ppb urine sample urine blank 4-OH-146 3-OH-138 4-OH-153 3-OH-138 ND

14. Std. 2.0 ppb Urea blank Spike 2.0 ppb Urea sample Urea blank Std 2.0 ppb urine blank + 2.0 ppb urine sample urine blank 4-OH-107

16. urineurine 4-OH CB-18700 4-OH-CB-1074.7 4.5 4-OH CB-1460.0 0 3’-OH CB-1385.3 5.5 3-OH CB-15369.5 4-OH-CB-17200 4'-OH-CB-16500 OH-PCB concentration in urine sample (ng/mL)

18. PCB & PFCs Ca metabolism

19. blood sampling PFHxA/PFOA anethetize swelling 19  24g

20. kidney brain liver gallbladder

21. 0 1 2 3 4 5 6 7 Plasma Ca conc. (mg/dl) Initial 1 day PFOAPFHxA

22. chiral PCB Enantioselective analysis

23. What is enantiomer? enantiomer is one of two stereoisomers one of two stereoisomersstereoisomers that are mirror images of each other that are not identical, mirror imagesmirror images much as one's left and right hands are "the same" but opposite. hands

24. Enantiomer Fraction (EF) EF = -------------- E(+) E(+)+ E(-) E(+) : (+) enantiomer peak area E(-) : (-) enantiomer peak area technical products EF = 0.5 metabolism EF > 0.5 racemates EF < 0.5 non-racemic residues The EF variation may distinguish POPs transfer of current use and that of past applications. POPs in the oceans (+) (-) (+) (-) (+) (-)

25. To identify the behavior of POPs The results of Enantioselective analysis were shown using Enantiomer Fraction (EF). for tracking and apportioning chemical movement between environmental compartments for investigating microbial degradation processes. Enantiomeric compositions of POPs in seawater were investigated

26. Enantiomer Fraction(EF ） EF= E(+) E(+) + E(-) E(+): amount of (+)enantiomer E(-) : amount of (- ） enantiomer (+) (-) (+) (-) (+) (-) (+) (-) (+) (-) EF= E 1 E 1 + E 2 E 1 : amount of the first enantiomer E 2 : amount of the second enantiomer or

27. Enantiomer Fraction(EF) Changes in physicochemical process, It is possible to distinguish between newly caused pollution and old pollution by monitoring EF the EF value does not change. Changes in metabolic process, the EF value changes.

28. Enantioselective determination of chiral PCBs in Spanish breast milk samples by heart-cut MDGC, Luisa R Bordajandi, Maria Jose Gonzalez (2005 ) : Organohalogen Compounds. 67, 1361-

29. DISTRIBUTION OF CHIRAL PCBs IN SELECTED TISSUES IN THE LABORATORY RAT, Hans-Joachim Lehmler, et al (2004): Organohalogen Compounds. 66, 443- PCB-91PCB-95PCB-149 Chlorofenn.d.0.5 soil0.460.5 Adipose0.73, n=10.66, n=20.50±0.04 (0.62, n=1)(0.53, n=1)(0.48, n=2) Brainn.d. Heartn.d. (0.53,n=1) Kidneyn.d. 0.52±0.04 (0.55,n=2) Livern.d.0.47,n=10.52±0.04 (0.54,n=2) Lungn.d. 0.47,n=2 Skinn.d.0.54,n=10.49±0.04 (0.41,n=1) Spleenn.d.0.51,n=20.54±0.03 (0.56,n=1)(0.54±0.04) WholeBloodn.d. 0.52,n=1 Serumn.d. 0.06,n=2 Enantiomeric fraction of PCB 91, 95 and 149 in tissues and blood from soil extract- treated animals

30. 302011/05/23 GC/MS Analysis

31. 10121416182022242628303234363840 Retention Time (min) 0 200000 400000 600000 800000 1000000 1200000 1400000 Intensity 2PM1214PH104 3PM982M83 2M1114PM1213M1212PM112 4PM93 2M1093M119 3M84 4PM1013M1132PM1142PM1064PM120 4PM86 4PM1124PM107 3M118 4M1093M110 4M97 4PM1274PM106 OH-PCB （ 5Cl ）のクロマトグラム（ STD ）

32. 10121416182022242628303234363840 Retention Time (min) 0 10000 20000 30000 40000 50000 60000 70000 80000 90000 100000 110000 Intensity 2PM121 4PH104 3PM98 2M832M111 4PM121 3M121 2PM1124PM93 2M1093M1193M84 4PM101 3M113 2PM1142PM1064PM1204PM864PM1124PM107 3M1184M1093M1104M97 4PM1274PM106 OH-PCB （ 5Cl ）のクロマトグラム（尿試料）

33. OH-PCB （ 6Cl ）のクロマトグラム（ STD ） 20222426283032343638404244 Retention Time (min) 0 100000 200000 300000 400000 500000 600000 700000 Intensity 4M134 4PM165 4M146 3PM1384PM130 4M163 4PM159 4M162

34. OH-PCB （ 6Cl ）のクロマトグラム（尿試料） 20222426283032343638404244 Retention Time (min) 0 10000 20000 30000 40000 50000 60000 70000 80000 90000 100000 Intensity 4M134 4PM165 4M146 3PM1384PM130 4M163 4PM159 4M162