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Development of SPE for urine sample

Development of SPE for urine sample. 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

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Development of SPE for urine sample

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  1. Development of SPE for urine sample

  2. 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 modify polymer with multifunctional group • for ionic chemicals　ionic group • forhydrophilic chemicals　hydrophilic group

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

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

  5. RP-WAX RP-SAX RP-OH Chemical structure of SPE for urine sample

  6. Property of SPE

  7. Treatment for urine sample

  8. urine sample add. x1, 20mM Phosphate buffer (pH 4.0 ) adjust pH 4.0 by 10% HCOOH aq. add. surrogate：OH-PCB 13C mix (100ppb) 20uL, Solid PhaseExtraction （RP-WAX.) elution, 8 mL 0.1% NH3 in MeOH dryness by N2 gas, Concentration add. 5mL, 20mM Phosphate buffer (pH 7.0 ) add. 20 μL β-Glucuronidase / Aryl sulfatase, 37 ℃, 1.5 hrs Enzymegenation by CH2Cl2, 2 mL x 2 Extraction Centrifuge 2600rpm. 10min Concentration by N2 gas, 1 mL add. x 9 CH3CN Concentration by N2 gas, 1 mL methylation GC/MS analysis LC/MS analysis

  9. LC/MS Analysis

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

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

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

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

  14. OH-PCB concentration in urine sample (ng/mL) urine urine 4-OH CB-187 0 0 4-OH-CB-107 4.7 4.5 4-OH CB-146 0.0 0 3’-OH CB-138 5.3 5.5 3-OH CB-153 6 9.5 4-OH-CB-172 0 0 4'-OH-CB-165 0 0

  15. PCB & PFCs Ca metabolism

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

  17. gallbladder liver brain kidney

  18. Initial 1 day 7 6 5 4 Plasma Ca conc. (mg/dl) 3 2 1 0 PFOA PFHxA

  19. chiral PCB Enantioselective analysis

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

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

  22. Enantiomeric compositions of POPs in seawater were investigated • 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.

  23. EF= EF= E1 E1+ E2 E(+) E(+)+ E(-) (-) (+) (-) (-) (+) (+) (+) (-) (+) (-) Enantiomer Fraction(EF） or E1: amount of the first enantiomer E2: amount of the second enantiomer E(+): amount of (+)enantiomer E(-) : amount of (-）enantiomer

  24. Enantiomer Fraction(EF) Changes in physicochemical process, the EF value doesnot change. Changes in metabolic process, the EF valuechanges. It is possible to distinguish between newly caused pollution and old pollution by monitoring EF

  25. 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-

  26. Enantiomeric fraction of PCB 91, 95 and 149 in tissues and blood from soil extract-treated animals DISTRIBUTION OF CHIRAL PCBs IN SELECTED TISSUES IN THE LABORATORY RAT , Hans-Joachim Lehmler, et al (2004): Organohalogen Compounds. 66, 443-

  27. GC/MS Analysis

  28. 1400000 1200000 1000000 800000 Intensity 600000 400000 200000 0 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 Retention Time (min) 2PM121 4PM121 2PM112 4PM101 2PM114 2PM106 4PM120 4PM112 4PM107 4PM127 4PM106 4PH104 3PM98 4PM93 4PM86 2M111 3M121 2M109 3M119 3M113 3M118 4M109 3M110 2M83 3M84 4M97 OH-PCB（5Cl）のクロマトグラム（STD）

  29. 110000 100000 90000 80000 70000 60000 Intensity 50000 40000 30000 20000 10000 0 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 Retention Time (min) 4PM127 4PM106 2PM121 4PM121 2PM112 4PM101 2PM114 2PM106 4PM120 4PM112 4PM107 4PH104 3PM98 4PM93 4PM86 3M118 4M109 3M110 2M111 3M121 2M109 3M119 3M113 4M97 2M83 3M84 OH-PCB（5Cl）のクロマトグラム（尿試料）

  30. 700000 600000 500000 400000 Intensity 300000 200000 100000 0 20 22 24 26 28 30 32 34 36 38 40 42 44 Retention Time (min) 4PM165 3PM138 4PM130 4PM159 4M134 4M146 4M163 4M162 OH-PCB（6Cl）のクロマトグラム（STD）

  31. 100000 90000 80000 70000 60000 50000 Intensity 40000 30000 20000 10000 0 20 22 24 26 28 30 32 34 36 38 40 42 44 Retention Time (min) 4PM165 3PM138 4PM130 4PM159 4M134 4M146 4M163 4M162 OH-PCB（6Cl）のクロマトグラム（尿試料）

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