外因性内分泌攪乱化学物質調査暫定マニュアル(水質、底質、水生生物)

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1 N- 1) 2) 1) GC/MS HPLC GC/MS 2) 0.01~0.05µg/l 0.5~50µg/kg GC/MS SIM GC/MS SIM GC/MS SIM a) b) -d 10 -d 10 -d 12 -d 12 c) d) 600 C 4 e) % f) a) b) - IX-1 -

2 c) (ODS) 200-1,000mg 5ml 5ml 5ml d) e) 3ml 1g f) 3ml g g) KD h) i) j) 1µm k) / (GC/MS) a) SS 50µg/ml 1µm 5% l 50g 50ml ml 100ml 4 KD 2ml 3) 200ml 4) b) 1µm 5-10ml 500ml 10ml 10ml 10 3ml 20% / 10ml 2ml 5ml 2ml - IX-2 -

3 - IX-3-10~25g 25ml ,000rpm % 400ml 1l 50ml ml 100ml 10 30g 50ml KD 2ml KD 10ml 250ml 5% 5~10ml/min 10ml 10 3ml 20% / 10ml 2ml 5ml 2ml 5) 10~25g 50ml ,000rpm 15 50ml 1l 30ml 5 30ml 5 5ml 5% 500ml 50ml 10 50ml 40 C 10ml 10ml 2ml 5) 5% 5.2

4 10ml 5~10ml 1ml 2 10ml 20% 10ml 2ml Envi-Carb 3ml 0.25g 20% 10ml 50% 5ml 2ml 130 C 10g 1.0 ~1.5 30cm 2g 10ml 2ml 100ml 20% 2ml 0.5ml GC/MS 100µg/ml 1.0µg/ml 6) 0-1.0ml 0.5ml 0.5ml -d 10 -d 10 -d 12 -d µg/ml 6) - IX-4 -

5 a) GC/MS (GC) 0.25mm 30m 0.25µm 50 C 1min -20 C/min-200 C -5 C/min-280 C(5min) min 40-50cm/sec (MS) 280 C 270 C 70eV 300µA (EI) MS DFTPP (decafluorotriphenylphosphin) PFTBA(Perfluorotributylamine) PFK Perfluorokerosene MS (M/Z) IS-1 -d IS-2 -d IS-3 -d IS-4 -d (215) IS (188) IS (186) IS (291) IS (115) IS (127) IS (202) IS (335) IS (144) IS (181) IS-4 - IX-5 -

6 (167) IS-4 (2 ) (167) IS-4 (2 ) (163) IS (212) IS-1 b) 2µl GC/MS c) 2µl GC/MS d) C µg/l S abs ng V conc ml V inj GC/MS µl V spl ml GC/MS 5 (s) 3s (µg/l kg) 10s (µg/l kg) C( µ g / l) = S ( ng) Vconc( ml) 1,000 abs V ( µ l) V 10 1 ±20% 2 10% 2 30% inj spl ( ml) - IX-6 -

7 2,4,5-2 4-D 2,4-2,4,5-T 0.05µg/l 5~10µg/kg ph2 GC/MS SIM 1N - 1:9 ph2 20% / GC/MS a) 2,4- (2,4-D) b) 2,4,5- (2,4,5-T) c) -d 10 d) 2,4- e) f) g) 600 C 4 h) N- -N - -N- i) j) k) l) a) b) c) d) e) f) g) h) 3ml 0.5 1g : i) / (GC/MS) - IX-7 -

8 - IX ) 1l 25g 20% ph2 100ml ml 25 C 2) 10ml 1N 10ml 10ml 70 C 15 3) 100ml 30ml 5 4) 20% ph2 30ml 10 30ml 25 C 1) 2) 3) 2,4-D 2,4,5-T 4) 10~25g 50ml 2,4-20µg/ml 0.1ml 1N - 1:9 20ml ,000rpm 10 1N - 1:9 20ml 2 25 C + 5ml 1N 10ml 70 C ml 30ml 5 20% ph2 20% 50ml 10 20% 50ml 3 20% 25 C 10~25g 2,4-20µg/ml 0.1ml 1N - 1:9 30ml ,000rpm %

9 0.5ml / 1ml 5) d µg/ml 10µl 5) N- -N - -N- 10% / 5ml 20% 5ml 0.5ml ,4-D 2,4,5-T 100mg 100ml 1,000µg/ml 10µg/ml -d 0.5µg/ml a) GC/MS (GC) 0.25mm 30m 0.25µm 50 C 1min -20 C/min-200 C -5 C/min-280 C(5min) min 40-50cm/sec (MS) 280 C 270 C 70eV 300µA (EI) m/z188 -d 10 - IX-9 -

10 m/z ,4-D-Me m/z ,4,5-T-Me b) 0-1.0µg/ml 5 0.5ml / 1ml 1 0.5ml -d µg/ml 10µl 2µl GC/MS c) 2µl GC/MS d) C µg/l S abs ng V conc ml V inj GC/MS µl V spl ml GC/MS 5 (s) 3s (µg/l kg) 10s (µg/l kg) C( µ g / l) = S ( ng) Vconc( ml) 1,000 abs V ( µ l) V 10 1 ±20% 2 10% 2 30% inj spl ( ml) - IX-10 -

11 2- MBC: Methylbenzinidazol-2-yl carbamate MBC 0.1µg/l 2µg/kg 20µg/kg MBC 1.52 /MBC GC/MS 0.1N ph2 GC/MS 0.1N a) 98% b) -d 10 c) d) e) 600 C 4 f) N- -N - -N- g) h) PCB i) j) a) b) c) d) e) KD f) g) / (GC/MS) - IX-11 -

12 - IX l 50g 50ml ml 1) ml 100ml 0.1N 50ml 5 5N 0.2N ph ml 50ml 5 50ml 50ml KD 3-5ml 1) 200ml 2) N- -N - -N- 10% / / 10 25g 50ml ,000rpm 10 50ml 40 C 0. 1N 30ml 50ml 200ml 5 50ml 40 C 50ml 100ml 0.1N 30ml 5 0.1N 20ml 5 5N 0.2N ph6.4 50ml 5 30ml g 50ml ,000rpm 10 50ml 30ml 2 40 C 0.1

13 0.1N 30ml 50ml 0.5ml / 1ml 2) 1 -d µg/ml 0.2ml mg 100ml 1,000µg/ml 1ml 0.2N 50ml 100ml 5 5N 0.2N ph g 20ml MBC 100ml 2 100ml 10µg/ml MBC ml 0.5ml / 1ml 1 -d µg/ml 0.2ml MBC 1.52 a) GC/MS (GC) 5% 0.25mm 30m 0.25µm 50 C 1min -20 C/min-200 C -5 C/min-280 C(5min) min 40-50cm/sec (MS) 280 C 270 C 70eV 300µA (EI) m/z188 -d 10 m/z MBC-Me - IX-13 -

14 b) 2µl GC/MS c) 2µl GC/MS d) C( µ g / l) = S ( ng) Vconc( ml) 1,000 abs V ( µ l) V ( ml) C µg/l S abs ng V conc ml V inj GC/MS µl V spl ml GC/MS 5 (s) 3s (µg/l kg) 10s (µg/l kg) 10 1 ±20% 2 10% 2 30% inj spl 3- -1,2,4- - IX-14 -

15 1µg/l 1µg/kg ph4.0 ODS HPLC- 2% o- HPLC 40% HPLC a) (amitrol) 3- -1,2-,4-98% b) (fluorescamine) (fluram) c) 0.1M (ph9.5)50ml o- 5ml(5ml 270mg o- ) 2-0.2ml 2-50µl d) 60g 16.4g 600ml ph e) f) 700 C 4 g) h) No.5C 5 C i) ODS j) k) AG50W-X8 l) IRC-50 a) b) c) d) e) f) g) - IX-15 -

16 5 C 20ml 50ml 2ml (ph4.0) 1) 0.2% - 4ml 2) 2 20 C 50ml 17g 1N 1ml 5-10ml ODS 2ml 2ml 3, 4) 1) ph ) 0.2% - 4ml 15-30% 0.2% - 3) o- o- 1 R-NH 2 - o- 4) g 50ml 2% 10ml 10 3,000rpm 5 No.5C 2 100ml 0.1M ph9.0 10ml 20ml 50ml o- 1ml 10 O- 10ml ml 10 25g 20ml 5 5 3,000rpm 10 40% 20ml 50ml 20ml 5 / 100ml 10ml 1ml 75 C 30 10ml 5) 10ml 2.8% 12ml n- 30ml 45 5ml 6) 5ml 50ml 2.8% 35ml n- 100ml 0.1M 2ml 1ml 0.2% / 0.1ml 1 0.1M 0.5ml HPLC - IX-16 -

17 6) 10mm 300mm µm 1ml 5ml 7) µm 5ml 1N 2.8% 1N mg 100ml 1,000µg/ml 5µg/ml 5 a) HPLC- ODS 5mm 15cm 5µm 40 C 9% - 10:9 2ml/min 395nm 480nm b) 10ml 1ml 1ml ph ml 0.2% - 0.4ml 2 20 C 10ml 0.5µg/ml 20µl HPLC -20 C 1 o- c) 20µl HPLC d) C( µ g / l) = S ( ng) Vconc( ml) 1,000 abs V ( µ l) V ( ml) C µg/l S abs ng inj spl - IX-17 -

18 V conc ml V inj HPLC µl V spl ml HPLC 5 (s) 3s (µg/l kg) 10s (µg/l kg) 10 1 ±20% 2 10% 2 30% N- 0.03µg/l 2µg/kg 1µg/kg N- CH 3 NH 2 o- OPA OPA 10% 10% HPLC - IX-18 -

19 a) 99% b) c) d) e) o- 2- f) g) 650C 24 h) i) j) k) l) 99.99% a) b) c) d) KD e) f) g) HPLC l l 100g 70g 50 ml ml 100ml 2, 3, 30 C 4) 1µg/ml / 1:9 1ml 5) 1) 200ml 2) 30 C 3) KD 4) 50ml 5% - IX-19 -

20 - IX-20-1g 10ml 10% 20ml 5) HPLC 10 25g 100ml 50ml ,000rpm 10 50ml 10% 500ml l 50ml 5 l 50ml 50ml 5 50ml 2 10ml 30 C 10~12mm 300mm 5% 1g 2cm 10ml 50ml 10% / 20ml 1ml 1µg/ml : 1:9 1ml mg 100ml 1000µg/ml 1: µg/ml µg/ml 10 a) HPLC ODS (1:9)

21 1ml/min 40 C 1 50mM 100 C 0.4ml/min 2 o- / 50 C 0.4ml/min 345nm 450nm b) 20µl HPLC c) 20µl HPLC d) C µg/l S abs ng V conc ml V inj HPLC µl V spl ml HPLC 5 (s) 3s (µg/l kg) 10s (µg/l kg) C( µ g / l) = S ( ng) Vconc( ml) 1,000 abs V ( µ l) V 10 1 ±20% inj spl ( ml) - IX-21 -

22 2 10% 2 30% 1) (1993).. 2) (1983) 1- (NAC). 57, pp , ( 58 5 ) 3) (1988) (BPMC MTMC PHC MIPC X MC NAC methyl 2-benzimidazol carbamate). 62, pp83-99, ( 63 6 ) 4) (1992). 3, pp35-54, ( 4 6 ) 5) (1993) P (MEP) (MPP) (EDDP) EPN (PMP) α- (α-cvp) β- (β-cvp). 4, pp65-94, ( 5 6 ) 6) (1994). 5, pp35-55, ( 6 6 ) 7) (1996) 2,4- p-t-. 7, pp , ( 8 6 ) 8) (1984) 3- -1,2,4-. 58, pp , ( 59 5 ) 9) (1992). 3, pp55-68, ( 4 6 ) 10) (1994) N-. 35, ) (1977). 2, GC/MS LC/MS GC/MS/MS - IX-22 -

23 , 150g, 50ml 1.0l 0.5 1l 0.5l SS:50mg/l 10, 2 GC/MS IS: phenanthrene-d 10, fluoranthene-d 10 chrysene-d 12, perylene-d 12, 25ml 3, 10 25g, 3,000rpm, %, 400ml, 50ml 2, 10, 10ml, 250ml 10ml 20% / 10ml, 50ml 2 50% / 10ml,, 10~25g 10 3,000rpm, 10 5%, 500ml, 50ml 2, 30ml 3, ml 20% / 100ml 1 - IX-23 -

24 ,, 25g 100ml 2, 10ml 1N, 10ml, 10ml 1l, 30ml ph, 20%, 10 20%, 30ml 2, 25 70, 15, 5 ph 2, 10, 0.5ml /, 1ml GC/MS, 1, 5ml 20% /, 5ml IS: 2,4-1N / 1:9, 20ml 2 IS: -d g, 10 3,000rpm, 10, 10ml 1N, 10ml 20% /,, 10ml 20% 5ml 2 ph 2 70, 15, 0.5ml /, 1ml, 10 IS: 2,4-1N / 1:9, 20ml g, IX-24 -

25 NaCl, 150g, 50ml, 10 20ml, 1, 10, 50ml 3 0.1N, 50ml 5N 0.5N, 10 20ml, 50ml 2, 5 ph (6.5),50ml 2 GC/MS, 1 IS: phenantherene-d 10, 0.5mg/ml, 0.2ml 10 25g, rpm, N, 30ml, 50ml 0.1N, ml 5N 0.5N, 50ml 2 ph (6.5) GC/MS IS: -d10,50ml 2 30ml 2, 10 25g rpm, N, 30ml, 50ml 0.1N, 30+20ml 5N 0.5N ph (6.5), 50ml 2 GC/MS IS: -d IX-25 -

26 , No.5C, 17g 1N, 1ml, 2ml ODS, 5-10ml/min, 20ml 2M ph4.0, 2ml 0.2%, 4ml!! ph !! 20, 2!!!! HPLC-, 2ml ODS 9% - (10:9) 395nm, : 480nm 2%, 20ml g O-, 1ml, 2ml, M (ph9.0), 10ml No.5C (ph4.0), 2ml 0.2%, 5ml, 17g 1N, 1ml, 2ml HPLC-, 20ml 40%, 2 4 ODS, 5-10ml/, g, 100ml, 10ml, 5ml, 1ml 70, 30 50ml 50ml 2 n- 10ml, 2.8% 12ml n-, 100ml 50ml, 2.8% 35ml 0.05M, 2ml 0.25% /, 0.1ml 0.1M, 0.5ml HPLC- ODS - IX-26 -

27 , 100g, 100ml, 1l, 10 3, 50ml 2, 50ml IS: 1mg/l, 1ml HPLC - [ 10%, 500ml 10~25g, 10 HPLC - IS: 1mg/l, 1ml 10ml 10% / 10ml, 50ml 2 10%, 500ml 10~25g, 10 10ml 10% / 10ml HPLC - IS: 1mg/l, 1ml 5 - IX-27 -

外因性内分泌攪乱化学物質調査暫定マニュアル(水質、底質、水生生物)

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